Chng 1

Trao i cht v nng lng sinh hc

1.1. Khi nim chung v trao i cht Mi c th sng u tn ti trong mi trng v lin h mt thit vi mi trng . Hin tng c th ly mt s cht t mi trng kin to nn sinh cht ca mnh v thi ra ngoi nhng cht cn b c gi l s trao i cht. S trao i cht gii v sinh khc vi gii hu sinh. gii v sinh, trao i cht lm cho cc cht hu c v v c b phn hu. V d, vi (canxi carbonate) b xi mn v H2CO3 c trong nc tc dng vi vi thnh canxi bicarbonate, m b i ho thnh mt s cht khc l do tc dng vi oxy. th gii sinh vt, mi c th sng lun lun trao i cht vi mi trng, ly thc n vo chuyn ho thnh cc cht s dng cho c th v thi ra ngoi cc cht cn b. Qu trnh c thc hin l do cc bin i ho hc lin tc xy ra trong c th. Ton b cc bin i ho hc c gi l s trao i cht. Qu trnh trao i cht gm nhiu khu chuyn ho trung gian. Mi chuyn ho l mt mt xch ca mt trong hai qu trnh c bn: ng ho v d ho. ng ho v d ho l hai qu trnh i lp, nhng li thng nht vi nhau trong mt c th: chng xy ra ng thi v lin quan mt thit vi nhau. Cc cht c tng hp nn trong qu trnh ng ho l nguyn liu cho qa trnh d ho (v d gluxit l sn phm ca qu trnh quang hp, l nguyn liu cho qa trnh h hp). Nng lng gii phng ra trong qu trnh d ho c s dng mt phn cho qu trnh tng hp. 1.2 Nng lng sinh hc H thng sng cn nng lng chuyn ng, ln ln, tng hp cc phn t sinh hc, s vn chuyn ion v phn t qua mng. Tt c i hi nng lng a vo. Cc c th ly nng lng t mi trng sng v s dng nng lng thc hin cc qu trnh sng c hiu qu. nghin cu nng lng sinh hc i hi phi c hiu bit v nhit ng hc, mt s nh lut, nguyn l m t ngun, trao i nhit, nng lng v vt cht trong h thng nghin cu.

Nhit ng hc cho chng ta xc nh qu trnh ho hc v phn ng c th t xy ra hay khng. Mc d nhit ng hc l khi nim phc tp, nhng n da trn ba nh lut tng i n gin v d hiu. Mt vi nguyn l ca nhit ng hc c bn c a ra trong chng ny bao gm phn tch ngun nhit, sn sinh entropy, hm nng lng t do v mi lin quan gia entropy v thng tin. Chng ny cng cp n ATP v nhng hp cht cao nng khc. Khi nim v nhit ng hc c bn Bt k mt s quan tm no ca nhit ng hc phi phn bit gia h thng v mi trng. H thng l mt phn ca v tr m chng ta quan tm, trong khi mi trng l gm tt c nhng g cn li. C ba trng thi c bn: h thng c lp, h thng ng v h thng m. H thng c lp: Khng c s trao i cht v nng lng vi mi trng. H thng ng: C trao i nng lng, nhng khng c trao i cht vi mi trng. H thng m: C trao i cht v nng lng vi mi trng. C th sng l h thng m in hnh m trao i cht (dinh dng v sn phm thi ra) v nng lng (nhit t trao i cht) vi mi trng. nh lut 1: Nhit, cng v cc dng nng lng khc Trc y trong s pht trin ca nhit ng hc ngi ta cho rng nhit c th bin i thnh nhng dng nng lng khc v hn na tt c cc dng nng lng mt cch c bn c th bin i thnh mt s dng khc. nh lut 1 ni rng: tng nng lng ca mt h thng c lp l khng thay i. Cc nh nhit ng hc m phng thnh mt hm ton hc nghin cu s bin i nhit v s dng cng trong nhng h thng nhit ng hc. Hm ny c gi l nng lng ni nng, thng k hiu l E hoc U. Nng lng ny ch ph thuc vo trng thi hin ti ca mt h thng v v vy c coi l hm trng thi. Nng lng ni nng khng ph thuc vo h thng xy ra nh th no v v vy khng ph thuc vo ng hng. Mt cch m rng suy ngh ny l chng ta c th thay i h thng bng bt c con ng no v cho n khi no h thng tr v trng thi ban u nng lng ni nng s khng thay i.

Nng lng ni nng, E ca bt k h thng no c th thay i nu ngun nng lng vo hoc ra khi h thng dng nhit hoc cng cho bt k qa trnh no bin i mt trng thi (1) sang mt trng thi khc (2) thay i nng lng ni nng l: E = E2 - E1 = q + w (1.1) q l lng nhit c h thng hp th t mi trng w l cng thc hin trn h thng do mi trng Cng c hc c nh ngha l s chuyn ng t ch ny n ch khc, gy ra do s dng lc. C hai phi xy ra cng mi c thc hin. V d: Mt tu ch khch cha y khch nhng khng di chuyn, theo nh ngha nhit ng hc cng khng c thc hin. Trong h thng ho sinh hc v ho hc cng thng lin quan vi p sut v th tch ca h thng. Cng c hc c xc nh w = -PV Trong P l p sut, V l s thay i th tch v = V2-V1 Khi cng c xc nh theo cch ny, k hiu bn phi ca phng trnh (1.1) l dng (i khi w c xc nh l cng c thc hin bi h thng, trong trng hp ny phng trnh l E = q - w). Cng c th c thc hin nhiu dng: c hc, in, t v ho hc. E, q, w phi c cng n v: calorie (cal) v kilocalorie (kcal) c s dng theo truyn thng, nhng theo n v SI: Joule c ngh nn dng. Enthalpy: Hm c nhiu tin li cho h thng sinh hc Nu nh ngha cng c gii hn bi cng c hc, trong trng hp ny E ch l thay i nhit th tch khng i. V vy nu V khng i, khng c cng c thc hin. E = q. V vy E l mt nh lng rt tin li trong qu trnh th tch khng thay i. E khng cn thit bng bin i nhit. V l do ny cc nh ho sinh hc, ho hc xc nh mt hm c bit ph hp cho qu trnh p sut khng i. N c gi l enthalpy, H c nh ngha: H = E + PV (1.2) Nu p sut khng thay i chng ta c: H = E +PV = q + w +PV = q - PV +PV = q (1.3) R rng H tng ng vi bin i nhit trong qu trnh p sut khng i. V cc phn ng ho sinh thng xy ra trong th lng hoc rn hn l th kh nn thay i th tch l nh v enthalpy v nng lng ni nng thng l nh nhau. so snh cc ch s nhit ng hc ca cc phn ng khc nhau iu thun li l xc nh iu kin tiu chun. ho tan trong mt dung dch trng thi tiu chun thng l hot tnh n v (n gin l nng 1M). Enthalpy, nng lng ni nng v nhng nh lng nhit

ng hc khc thng a ra hoc xc nh cho nhng iu kin tiu chun v c k hiu l H0, E0... Enthalpy thay i cc qu trnh ho sinh c th c xc nh bng vic o nhit hp th (hoc to ra) bng mt calorimeter. Mt khc cho bt k qu trnh no A ' B trng thi cn bng, enthalpy trng thi tiu chun thay i qa trnh c th c xc nh t s ph thuc vo nhit hng s cn bng: d (ln Keq) 0 (1.4) H = ---------------d (1/T)

1.1 S thay i enthapy, H0, cho 1 phn ng c th c xc nh dc ca s RlnKeq ngc vi 1/T. minh ha phng php ny nhng gi tr hai bn ca 327K (54,50C) c nu ra. S liu c s dng tnh H0 54,50C

y R l hng s kh = 8.314 J/mol K V d: trong mt nghin cu bin tnh ca protein chymotripsinogen (qa trnh thun nghch, gy bin tnh bi nhit ). Trng thi nguyn thu (N) ' Trng thi bin tnh (D) Keq = [D] / [N]

John F. Brandts o hng s cn bng cho s bin tnh mt lot pH v nhit , s liu pH = 3 Biu ca R (ln Keq) i vi 1/T (s Vant Hoff) c ch ra hnh 1.1. H0 cho qu trnh bin tnh bt k nhit no l gi tr m ca ng dc ca s nhit nh s Gi tr H0 c ngha g i vi bin tnh ca protein? Gi tr dng ca H0 biu din s b gy lin kt hydro cng nh gii phng nhng nhm a nc t bn trong phn t protein ban u trong qa trnh bin tnh, nh vy s nng nng lng ca dung dch protein. ln ca enthalpy ny thay i (533 kJ/m) 54,50C l ln so vi gi tr tng t ca H0 ca protein khc nhau v cho cng loi protein 25 0C (bng 1.1). Nu chng ta xem xt s ch s thay i enthalpy dng ca qu trnh bin tnh th trng thi gp t nhin l rt thun li, nhng ch s khc phi c gii thch.

nh lut th hai v entropy: Mt cch trt t v suy ngh v khng trt t nh lut th hai ca nhit ng hc c m t v th hin trong nhiu cch bao gm nhng im sau: H thng c xu hng tin t trng thi trt t sang trng thi khng trt t (entropy ln). Entropy ca h thng + mi trng l khng i bi qu trnh thun nghch. Entropy ca h thng + mi trng tng do qu trnh khng thun nghch.

Tt c cc qu trnh xy ra trong t nhin hng ti trng thi cn bng, l trng thi nng lng nh nht. Mt s im ca nh lut 2 dn n khi nim entropy, l thc o s mt trt t v s ngu nhin ca h thng, trong trng thi mt trt t l trng thi c entropy cao. Entropy c th c xc nh theo mt vi cch. Nu W l s cch sp xp thnh phn ca mt h thng m khng thay i nng lng ni nng hoc enthalpy ( l s lng ca trng thi knh hin vi c a ra nhit , nh sng v tng vt cht). Entropy c tnh: S = klnW (1.5) k l hng s Boltzmann = 1,38.10-23 J/K nh ngha ny c li cho tnh ton thng k, nhng dng ph bin hn lin quan entropy n s bin i nhit trong mt qu trnh. dq dS thun nghch = -------T (1.6)

dS thun nghch l thay i entropy ca h thng trong mt qu trnh thun nghch, q l nhit c bin i, T l nhit s bin i nhit xy ra. nh lut 3: Ti sao0 tuyt i quan trng nh vy? nh lut 3 ca nhit ng hc ni rng: entropy ca bt k cht no hon ton c trt t, tinh th phi tin n 0. nhit tin n 0 K v T= 0 K entropy chnh xc = 0. Da trn iu ny c kh nng thit lp mt h thng t l entropy tuyt i, s lng S = Cp 0 d lnT (1.7) Cp: kh nng bin i nhit p sut khng i. Kh nng nhit ca mt cht l tng s nhit ca 1 M c th d tr khi nhit ca cht c nng ln 1 . i vi qu trnh p sut khng i n c m t bng ton hc dH Cp = ------(1.8) dt Nu kh nng nhit c th c tnh tt c nhit gia 0 K v nhit no , entropy tuyt i c tnh i vi qu trnh sinh hc thay i entropy c nhiu tin li hn entropy tuyt i. Thay i entropy
T

cho mt qu trnh c th c tnh nu thay i enthalpy v nng lng t do bit. Nng lng t do: Mt gi thuyt nhng l cng c tin li Mt cu hi quan trng i vi nh ho hc, c bit i vi nh ho sinh hc l: Phn ng s xy ra theo hng t phi sang tri? Gibbs, mt trong nhng ngi xy dng nn nhit ng hc nhn thy cu tr li cho cu hi ny nm trong s so snh thay i enthalpy v thay i entropy mt nhit no , nng lng t do Gibbs c nh ngha nh sau: G = H - TS (1.9) Cho bt k qu trnh A ' B nhit v p sut khng i. S thay i nng lng t do c tnh: G = H - T S (1.10) Nu G gn = 0 qu trnh cn bng, khng i theo hng thun hoc ngc li khi G = 0 S = H/T v thay i enthalpy v entropy l cn bng chnh xc. Bt k qu trnh vi G khc 0 thc hin t ng n trng thi cui cng c nng lng t do thp. Nu G m th qu trnh xy ra theo hng t tri sang phi. Nu G > 0 phn ng xy ra theo hng ngc li (k hiu v gi tr ca G cho php xc nh qu trnh s xy ra nhanh nh th no). Nu qu trnh c G m th qu trnh t xy ra, nu G dng th qu trnh khng t xy ra (hay t xy ra theo chiu nghch). Thay i nng lng t do tiu chun Thay i nng lng t do G cho bt k phn ng no ph thuc vo cht tham gia phn ng v sn phm phn ng v cng b nh hng bi iu kin phn ng k c nht , p sut v pH v nng ca cht phn ng v sn phm. Nh gii thch, rt tin li xc nh trng thi tiu chun cho nhng qa trnh nh vy. Nu thay i nng lng t do cho mt phn ng l nhy cm vi iu kin ho tan, iu g c bit c ngha cho s thay i nng lng t do trng thi tiu chun. tr li cho cu hi ny xem xt mt phn ng gia hai cht A v B to nn sn phm C v D A + B C + D (1.11) Thay i nng lng t do cho nng khng trng thi tiu chun l [C ] [D ]

G = G0 + RT ln ---------------- (1.12) [A] x [B ] [C ] [D ] -------------- = Keq [A] x [B ] trng thi cn bng G = 0 Chng ta c G0 = - RT ln Keq (1.13) hoc logarit c s 10 G0 = - 2,3 RT/ log 10 Keq (1.14) N c bin i Keq =10 -G/2,3 RT (1.15) Trong bt c dng no mi lin h cho php xc nh thay i nng lng t do tiu chun cho bt k qu trnh no nu hng s cn bng c bit. Quan trng hn, iu ni rng cn bng thit lp cho mt phn ng trong dung dch l mt hm ca s thay i nng lng t do tiu chun cho qu trnh, ngha l G0 l cch vit khc ca hng s cn bng. V d hng s cn bng xc nh bi Brandts mt s nhit vi s bin tnh ca chymotrypsinogen c th c dng tnh s thay i nng lng t do cho qu trnh bin tnh. V d h s cn bng 54,50C l 0,27, nh vy G0 = - (8,314 J/mol. K (327,5K) ln (0,27) G0 = - (2,72 kJ/mol . ln (0,27) G0 = 3,56 kJ/mol K hiu dng ca G0 ngha l qu trnh bin tnh khng u th. Dng gp l dng bn ca protein 54,50C. Mt khc ln tng i nh ca G0 ngha l dng gp chim u th nh.

1.2 S ph thuc ca G0 vo nhit nhit trong qu trnh bin tnh ca chymotrypsinogen

Hnh 1.2 ch s ph thuc ca G0 vo nhit bin tnh pH = 3. Tnh c H0 v G0 ca s bin tnh chymotrypsin, c th tnh S0 s dng phng trnh (3.10). (G0 - H0) S0 = ---------------(1.16) T 54,50C (327,5 K) S0 = - (3560 - 533,000 J/mol) / 327,5 K S0 = 1,620J/mol.K Hnh 1.3 biu din s ph thuc ca S0 vo nhit bin tnh ca chymotrypsin pH = 3. Gi tr dng S0 ch rng dung dch protein tr nn khng trt t khi protein b bin tnh. So snh gi tr 1,62 kJ/mol.K vi gi tr S0 bng 1.1 ch ra rng gi tr hin ti cho chymotrypsin 54,50C l hon ton ln. ngha vt l ca ch s nhit ng hc cho s bin tnh ca chymotrypsin s r hn trong phn sau. ngha vt l ca c tnh nhit ng hc Nhng ch s nhit ng hc cho ta bit nhng hin tng sinh ho g? Cch tt nht tr li cu hi ny l mt ch s ring r (v d H

hoc G ) khng c nhiu ngha. Mt gi tr dng H0 cho s bin tnh ca mt protein c th phn nh hoc l s gy cc lin kt hydro trong protein hoc s xut hin cc nhm a nc ra bn ngoi. Tuy vy s so snh mt s ch tiu nhit ng hc c th cung cp nhng hiu bit bn trong c ngha v mt qu trnh.

1.3 S ph thuc ca S0 vo nhit trong qu trnh bin tnh ca chymotrypsinogen

nh hng ca nng n thay i nng lng t do thc t Phng trnh (3.12) ch ra rng thay i nng lng t do i vi mt phn ng rt khc nhau so vi gi tr trng thi tiu chun nu nng ca cht phn ng v sn phm khc vi nng hot ng (1M cho dung dch). Xem xt s thu phn ca phosphocreatine: Phosphocreatine + H2O Creatine + Pi Phn ng ny to nhit rt mnh v G0 370C l -42,8 kJ/mol Nng sinh l ca phosphocreatine, creatine v Pi thng l gia 1 mM v 10 mM. Cho rng nng 1 mM v s dng phng trnh (3.12) G cho thu phn phosphocreatine l

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[0,001] [0,001] G = - 42,8 kJ/M + (8.314J/M) (310 K) ln----------------------[0,001] G = - 60,5 kJ/M 370C s khc nhau gia trng thi tiu chun v nng 1 mM cho mt phn ng nh vy l khong -17,7 kJ/mol Tm quan trng ca cc qu trnh kt hp trong c th sng Nhiu phn ng cn thit gi t bo v c th chng li th nhit ng hc, l theo hng G dng. Trong c s tng hp ATP v nhng phn t cao nng khc v to nn gradient ion trong tt c t bo ng vt c v. Nhng qu trnh ny c thc hin theo hng bt buc nhit ng hc. Bng s kt hp vi cc qu trnh c kh nng t xy ra nhiu qu trnh gn lin vi nhau c cp phn sau ca chng ny. Chng rt quan trng trong trao i cht trung gian, phosphoryl ho oxy ho v vn chuyn qua mng. Chng ta c th on nhng cp phn ng kt hp s xy ra t ng bng s thay i tng nng lng t do cho mi phn ng. V d, xem xt phn ng t qu trnh ng phn lin quan n s bin i phosphoenolpyruvic acid n pyruvat. S thu phn PEP gii phng nng lng v c s dng phosphoryl ho ADP thnh ATP, mt qu trnh m v mt nng lng khng t xy ra. 1.2.1 c tnh nng lng ca s trao i cht Nng lng ca cc qu trnh trao i cht (nng lng sinh hc) khc vi nng lng c thc hin trong bn cht khng sng ba c im sau y: c tnh th nht l s chuyn ho nng lng thnh cng v thnh nhng dng khc m khng km theo s chuyn ho s b nng lng ny thnh nhit nng. Xut pht t nguyn tc ny chng ta cn xem h thng sng nh l mt ng c ho ng hc ch khng phi l ng c nhit. c tnh th hai l vic gii phng nng lng trong cc qu trnh oxy ho sinh hc sinh ra t t, tng phn mt, trong mt chui di cc qu trnh k tip nhau cho n khi no tt c cc nguyn t hydro v cacbon u bin thnh cc sn phm cui cng l CO2 v H2O. V d s oxy ho mt phn t gam ng gii phng ra 686 kcal. Nu nng lng ny c

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gii phng ra cng mt lc th s gy ting n v h thng sng khng th s dng ton b nng lng trong mt khong thi gian ngn nh vy.

1.4 Tin trnh gii phng nng lng ho hc trong s trao i cht c chia lm 3 giai on

c tnh th ba l nng lng ho hc c gii phng ra khi phn gii glucid, lipid v nhng hp cht cao phn t khc u c th c tch lu trong nhng hp cht tch tr nng lng c th, c gi l hp cht cao nng.

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Tin trnh ca vic gii phng nng lng ho hc c bn c chia lm 3 giai on (hnh 1.4): Giai on th nht cc hp cht cao phn t (tinh bt, glycogen, proteine, lipid ...) b thu phn thnh cc cht c phn t b (monosaccharide, amino acid, axit bo, glycerine...). Nng lng gii phng ra trong giai on ny khng ng k, ch bng gn 1% d tr nng lng t do ca cc cht ny c gii phng ra di dng nhit. Giai on th hai l qu trnh ng phn, oxy ho cc axit bo v cc amino acid. Nng lng gii phng ra trong giai on ny gn bng 1/3 nng lng t do d tr trong cc cht . Sn phm chnh ca giai on ny l acetyl-CoA, -xetoglutaric acid v oxaloaxetic acid. Giai on th ba l oxy ho tip tc cc cht trn trong chu trnh Krebs. Khong 2/3 nng lng c gii phng ra giai on ny. 1.2.2 Cc hp cht cao nng Nhng hp cht cao nng: Tt c s sng trn tri t ph thuc vo nng lng mt tri, trong nhng dng sng c mt h thng th bc v nng lng. Mt s tip nhn nng lng mt tri trc tip, mt s khc nhn nng lng t nhm trn trong nhng qu trnh tip theo. Nhng sinh vt hp thu nng lng nh sng trc tip c gi l c th t dng. Nhng c th ny d tr nng lng mt tri trong cc phn t hu c khc nhau. Nhng sinh vt s dng nhng phn t , gii phng nng lng d tr trong mt lot cc phn ng oxy ho kh c gi l sinh vt ho dng. Mc d khc nhau c hai loi u c c ch chung v ti sinh mt dng nng lng ho hc, nng lng c th c gii phng trong nhng phn ng to nhit thc hin cc qu trnh sng a dng (cn nng lng). Mt nhm nh cc phn t l cht trung gian chuyn nng lng t cc phn ng gii phng nng lng n cc phn ng cn nng lng ca c th. Nhng phn t ny l coenzyme dng kh, nhng hp cht phosphate c gi l cao nng nu chng gii phng ra nng lng t do c gi tr m ln khi thu phn (G 0 c gi tr m ln hn -25 kJ/M). bng 1.2 sau y l danh sch nhng hp cht cao nng quan trng nht, nhng phn t nh phosphoric anhydric (ATP, ADP), enol phosphate (PEP), acyl phosphate (acetyl phosphate), guanidinophosphate (creatine phosphate). C nhng hp cht thioeste, nh acetyl CoA khng cha phospho nhng gii phng mt nng lng t do ln khi thu phn.

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Tng s nng lng chnh xc gii phng ra khi thu phn ph thuc vo nng , pH, nhit ... nhng gi tr G 0 khi thu phn nhng hp cht ny c gi tr dng ln hn ng k so vi nhng cht trao i khc. Chng c hai c im quan trng: Nhng cht phosphate cao nng (high- energy phosphate compounds) khng phi l cht d tr nng lng lu di, chng l nhng cht chuyn tip nng lng d tr, l cht mang nng lng t im ny sang im khc, t mt h thng ny n mt h thng khc. Nng lng hot ho c cung cp ng k t ATP khi thu phn nhm -phosphat.

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Nng lng lm gy lin kt O-P thng l 200-400 kJ/M, ln hn ng k so vi 30,5 kJ/M khi thu phn ATP. Cc nh ho sinh hc quan tm nhiu n nng lng gii phng thc t. Vai tr trung tm ca ATP trong nng lng sinh hc ATP cha hai pyrophosphoryl (hnh 1.4). Nhng phn t c lin kt anhydric, ADP, GTP, GDP v cc nucleoside triphosphate khc, nucleotide-ng nh UDP-glucose v pyrophosphate v c th hin nng lng t do G0 ln khi thu phn. Nguyn nhn ho hc ca gi tr G0 m ln l do s khng bn vng ca cht phn ng do s cng lin kt gy ra bi s y tnh in. S bn vng ca sn phm phn ng do s ion ho, s cng hng v nhng yu t entropy gy ra do thu phn v s ion ho tip theo. Mc d PEP, cyclic AMP, 13 DPG, phosphocreatine, acetylphosphate v pyrophosphate u c gi tr G0 ln hn, nhng ATP l duy nht nh v gia cc cht phosphate cao nng, (ATP c tng hp khi phn gii cc cht hu c) v cc cht nhn nng lng (khi cc cht ny c phosphoryl ho tham gia cc phn ng tip theo trong trao i cht). Ni mt cch khc ATP l mt xch ni lin hai qa trnh ngc nhau, l ng ho v d ho. Vic hnh thnh tt c cc hp cht cao nng khc cng xy ra do s tiu ph nng lng vn tch lu trong ATP. ADP c th nhn c phosphate v nng lng t cc phosphate cao nng. ATP cho c gc phosphate v nng lng i vi cc phn t c nng lng thp. Nh vy ATP c vai tr d tr nng lng cng nh tiu hao nng lng. Xt v c ch bin i v chuyn ho nng lng trong s phn gii ATP v cc hp cht cao nng tng t ATP ta thy nng lng cn thit thc hin phn ng ho hc c gii phng ra mt im, c th c chuyn n mt im khc, y nng lng c s dng mt cch trc tip. iu ny c ngha l trong c th sng khng nht thit phi tip xc vi nhau bng cch va chm (c trng cho ngoi c th sng) gia cc phn t cho v nhn nng lng.

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1.5 Chui ba gc phosphat ca ATP cha 2 lin kt pyrophosphat, c hai lin kt ny gii phng nhiu nng lng khi thu phn

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Chng 2

Vitamin
Vitamin l nhng cht hu c c trng lng phn t b, c cu to ho hc rt khc nhau v u c hot tnh sinh hc nhm m bo cho cc qu trnh ho sinh v sinh l trong c th tin hnh c bnh thng, v do , c nh hng rt ln i vi s trao i cht. Vitamin khng c tng hp ng vt bc cao, v vy chng phi c tip nhn cng vi thc n. Nhiu vitamin l tin cht ca cofactor (vitamin nhm B) tham gia vo cc phn ng enzyme, trong khi nhng vitamin khc tham gia vo qu trnh nhn v iu khin s sao chp (vitamin A), cc phn ng kh (vitamin C v E), to xng (vitamin D), ng mu (vitamin K) v.v Cc vitamin thuc cc nhm ho hc khc nhau. Thng chng c phn loi da vo ho tan: Nhm vitamin ho tan trong nc: B1, B2, B6, B12, folate, pantothenate, biotin, C. Chng c tch lu ch vi lng t. Lng d tha c thi ra qua nc tiu. Nhm vitamin ho tan trong cht bo: A, D, E, K. Chng c tch lu. Lng tip nhn d tha dn n hin tng tha vitamin (c bit vitamin A v E). 2.1 Cc vitamin ho tan trong nc 2.1.1 Thiamin (vitamin B1) Thiamin c pyrophosphoryl ho thnh coenzyme thiaminpyrophosphate (TPP), y phn ng kh carboxyl ho bng cch oxiho v phn ng chuyn nhm aldehyd hot ho ng mt vai tr quan trng. TPP l nhm prostetic ca pyruvat-dehydrogenase v pyruvatdecarboxylase, ca 2-oxoglutarat-dehydrogenase v ca transcetolase v v vy n tham gia vo qu trnh ng phn, chu trnh citrate, pentosephosphate v Calvin. a. Decarboxylase: xc tc cho phn ng loi nhm carboxyl ca cc pyruvic acid, -cetoglutaric acid. b. Transcetolase: xc tc cho phn ng vn chuyn glycoaldehyd (CH2OH-CO-). V d phn ng chuyn on 2C (C1 v C2) ca xylulose 5-phosphate n ribose 5-phosphate to thnh sedoheptulose 7-phosphate v glyceraldehyd-3-phosphate.

Sinh tng hp Hai thnh phn ca thiamine l pyrimidine v thiazol c tng hp ring v sau c kt hp. Cc con ng tng hp khc nhau sinh vt nhn chun v sinh vt nhn s (nm men, thc vt). Trong E.coli v S.thyphimurium c 1 (5-phosphoribosyl-5aminoimidazol (AIR) l tin cht ca gc pyrimidine, trong khi nm men cht ny c to nn bng cch khc. Trong E.coli pyruvat v Dglyceraldehyd l nhng tin cht ca thiazol. Chng lng kt thnh 1desoxy-D-xylulose, cht ny c gn cc nguyn t C 4, 4, 5, 6, 7. Ngun gc ca C2 v N3 l tyrosine, nguyn t S bt ngun t cysteine. Pyrophosphatester ca thnh phn pyrimidine tham gia vo phn ng gn. Bng mt phn ng pyrophosphoryl ho vitamin c chuyn thnh coenzyme.

B1 c nhiu trong cm go, gan, thn ... mm ng cc v nm men l ngun rt giu vitamin ny. Lng tip nhn hng ngy i vi ngi l 1-1,5 mg. Thiu vitamin ny nh hng n qu trnh trao i carbohydrate dn n bnh ph thng, hay cn gi l bnh beri-beri, ri lon thn kinh v nh hng n chc nng ca tim. B1 ch bn vi nhit trong mi trng acid, cn trong mi trng kim n b phn hu nhanh chng khi un nng. Khi oxy ho B1 chuyn thnh mt hp cht gi l thiocrome pht hunh quang. Tnh cht ny c s dng nh lng vitamin B1. Hm lng B1 trong nguyn liu c th thay i ng k tu thuc iu kin bo qun v ch bin. V d: go xt k hm lng B1 c th b gim n 4 ln so vi ban u. m khi bo qun nguyn liu (thc, go) cng cao, hm lng vitamin B1 b gim cng mnh.
B

2.1.2 Riboflavin (vitamin B2) Trc kia c gi l lactoflavin v ln u tin c tch ra t sa. Bn cnh nicotinamid-nucleotide NAD+ v NADP+ flavin-nucleotide: flavin-adenin-dinucleotide (FAD) v flavin-mononucleotide (FMN) l nhng nhm vn chuyn hydro quan trng. Chng tham gia hn 100 phn ng oxy ho kh. V d, qu trnh kh carboxyl ho bng cch oxiho pyruvic acid, oxy ho acid bo, kh amin ho bng cch oxiho amino acid... Coenzyme ny c to nn t vitamin riboflavin bng phosphoryl ho (FMN) v tip theo adenyl ho (FAD).

Sinh tng hp v bin i Vitamin riboflavin c to nn t GTP v ribulose-5-P trong thc vt, nm men v nhiu vi sinh vt, c t hn trong sn phm ng cc. Phn ng tng hp bt u bng vic m vng imidazol ca GTP v tch ra mt gc pyrophosphate. Bng phn ng kh amin ho, phn ng kh v tch ra gc phosphate cn li lm xut hin 5-amino-6ribitylamino-2,4-pyrimidindion. Phn ng ca hp cht ny vi 3,4dihydroxy-2-butanon-4-P (cht ny xut hin t ribulose-5-P) to nn phn t c hai vng 6,7 dimethyl-8-ribityllumazin. Hp cht ny kt hp vi 5-amino-6-ribitylamino-2,4-pyrimidindion thnh riboflavin. Phosphoryl ho vitamin ny to nn flavinmononucleotide (FMN). Bng phn ng adenyl ho tip theo lm xut hin flavin-adenin-dinucleotide (FAD). S phosphoryl ho riboflavin ng vt c iu khin chnh xc bng hormone tuyn gip trng.

Thiu vitamin B2 nh hng n qu trnh oxy ho kh lm nh hng n qu trnh to nng lng cn thit cho s sinh trng v pht trin ca c th. Thiu vitamin gy ra nhng ri lon v sinh trng v cc bnh v da. Tuy nhin nhng bnh ny tng i him. Nhu cu vitamin B2 hng ngy ca ngi ln khong 1,4 - 2 mg. Sn phm phn gii v sn phm thi ra ngi l riboflavin. Cc bc phn gii khc (c bit vi khun) l hydroxyl ho cc gc methyl v lm ngn chui ribityl. Chc nng ho sinh Chc nng oxy ho kh ca FMN v FAD l do vng isoalloxazin chu trch nhim. mt phn ng kh hon ton hai in t v hai proton c tip nhn. Dng oxy ho hon ton ca riboflavin l nhng tinh th vng da cam, hp th bc sng 370 v 450 nm. Dng kh ca n khng mu v mt tnh hp th 450 nm. Cc coenzyme flavin nucleotide dng kh cng c th b oxy ho tr li khi c cc cht nhn in t nh xanh methylen, 2,6-diclorophenolindophenol FADH2 + xanh methylen FAD + xanh methylen (dng oxy ho mu xanh) (dng kh khng mu) C th s dng cc tnh cht trn theo di cc phn ng do flavin dehydrogenase xc tc. Tinh th vitamin B2 dng kh tng i bn vi nhit hn vitamin B1, tuy nhin vitamin ny khng bn di tc dng ca nh sng. Ngc vi vitamin B1, hm lng vitamin B2 trong go, tht, trng, sa bin i khng nhiu trong qu trnh bo qun v ch bin.
B

2.1.3 Pyridoxin (vitamin B6) Pyridoxin (pyridoxol), pyridoxal v pyridoxamine to nn nhm vitamin B6. C 3 dng ny d dng chuyn ho ln nhau. Dn xut ca vitamin B6 l pyridoxalphosphate (PLP) l coenzyme ca nhiu enzyme xc tc. Pyridoxal- v pyridoxamin-phosphate l coenzyme quan trng cng tc ng mt s ln cc phn ng bin i amino acid, v d phn ng transaminase, decarboxylasev dehydratase. Vitamin B6 cng l thnh phn cu to ca phosphorylase xc tc cho phn ng phn gii glycogen. Ngoi ra vitamin B6 cn c vai tr quan trng trong qu trnh sinh tng hp NAD+, coenzyme A... Do thiu vitamin ny nh hng

n nhiu qu trnh trao i proteine, saccharide, lipid, c bit dn n ri lon v thn kinh v trao i amino acid. Sinh tng hp v bin i Mt s vi khun, nm men v thc vt to nn pyridoxine. ng vt nhai li cng khng cn c vitamin B6 trong thc n v vi sinh vt trong rut ca chng c th tng hp vitamin ny cung cp cho c th ng vt ch. Mt s vi khun khc v ng vt cn phi c cung cp t bn ngoi. Sn phm ngng t ca pyruvate v D-glyceraldehyd l 1-desoxy-Dxylulose cung cp nguyn t C 2, 2, 3, 4 v 4. Erythrose 4-P c bin i thnh 4-hydroxy-L-threonine. Hp cht ny cung cp nguyn t C 5, 5, 6 v N-1. Pyridoxine iu khin t l tng hp bng c ch ngc. Loi hydro ca pyridoxine thnh pyridoxal cho n nay ch c quan st thy trong vi khun.

ng vt pyridoxine, pyridoxal v pyridoxamine sau khi hp th c phosphoryl ho. Kinase ny b c ch bng sn phm ca n. Bng mt phn ng oxy ho pyridoxinephosphate v pyridoxaminephosphate bin i thnh pyridoxalphosphate. Phn ng ny cng b c ch bng sn phm ca n. Lng b sung hng ngy cn thit ngi ln khong 2 mg. S phn gii bt u bng phn ng do phosphatase xc tc. S phn gii tip theo ca vitamin ny thc hin bng mt phn ng oxy ho to thnh carbonic acid-4-pyridoxat, cht khng c hot tnh sinh hc. 2.1.4 Cobalamin (vitamin B12) Cobalamin l mt trong nhng phn t sinh hc khng trng hp ln nht. Nguyn t coban trung tm gn lin hp vi 6 thnh phn. Bn thnh phn trong l pyrrol ca vng corrin, mt thnh phn l

nucleotide bt thng dimethylbenzimidazol v thnh phn th 6 c th l gc hydroxyl (vitamin B12), gc methyl (methylcobalamin), hoc gc 5desoxyadenosyl (desoxyadenosylcobalamin, coenzyme B12). Dng bn th trng: thnh phn th 6 l mt gc cyanid. Sinh tng hp coenzyme v phn ng kh vitamin Coenzyme B12 c tng hp nhiu bc trong vi sinh vt. Bc th nht ging nhau gia hem v chlorophyll. Nhng phn ng tip theo c nhiu bin i.

Sau nhiu bc methyl ho vng porphyrin bin thnh vng corrin. Sau nhiu ln methyl ho tip theo v amid ho Co++ c a vo v c kh thnh Co+, ri kt hp vi adenosine. Tip theo -ribazol c gn vo qua mt cu propionate-aminpropanol v to nn thnh phn th 6 ca Co. Co nm trong phc h lin hp l Co+++. ng vt c v nhu cu B12 ( ngi < 10 g/ngy) c p ng do vi khun rut. hp thu vitamin B12 t rut cn protein c hiu (yu t ni ti, 50 kDa). Nu thiu loi protein ny s dn n hin tng thiu vitamin B12 trm trng (thiu mu c tnh). vn chuyn trong mu B12 lin kt vi nhiu plasmglobulin khc nhau, c gi l transcobalamin. Phn ln B12 c tip nhn dng oxy ho (hydroxy-cobalamin, Co+++). Sau hai bc phn ng xut hin tr li dng desoxyadenosyl (coenzym B12). Vitamin B12 tham gia trong thnh phn cu to ca cc enzyme xc tc cho cc phn ng: - Phn ng chuyn ribonucleotide thnh desoxyribonucleotide - Phn ng chuyn nhm methyl - Sp xp li cc nhm th ca 2 carbon lin nhau trong phn t.
B B

2.1.5 Vitamin H (Biotin) Biotin l mt vitamin cn thit cho cc phn ng carboxyl ho, c gn kt ng ho tr vi enzyme bng nhm -amin ca gc lysine. Sinh tng hp v bin i Qu trnh tng hp xy ra vi khun (c vi khun rut ca ng vt), nm men v thc vt bc cao. Phn t ban u l pimeloyl-CoA. Ngun gc ca phn t ny cha c gii thch y , n xut hin c th do s ngng t ca 3 phn t malonyl-CoA v gii phng ra 2 phn t CO2. Tip theo n ngng t vi L-alanine tch ra pyridoxal-pp, CO2 v HS-CoA. 8-amino-7oxopelargonat (7-KAP) xut hin c bin i thnh 7,8diaminopelargonate bng mt phn ng do transaminase xc tc. phn ng ny S-adenosylmethionine l cht cho nhm amin (thay v chc nng thng xuyn l cht cho nhm methyl). Vi s gn CO2 vo (ph thuc vo ATP) vng imidazol c ng li. Sau xy ra s gn S (t Lcysteine) v ng vng thiophan. Kt qu l to nn (+)-Biotin. S gn kt biotin vo enzyme tng ng cn hot ho bi ATP, c xc tc bng enzyme c hiu biotin-ligase. Ngi ln cn 100-200 g/ngy. Biotin c trong nhiu thc phm v c tng hp bi vi khun ng rut, v vy t khi b bnh thiu

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vitamin ny tr khi n nhiu trng sng c nhiu avidin kt hp rt cht vi biotin, ngn cch vic hp thu n qua rut non.

Phn gii bng vi khun Pseudomonas (c th cc loi vi khun khc) c th lm ngn chui bng -oxy ho v nguyn t C ca vng thiophan c tch ra. Gc ca vng imidazol c thi ra nc tiu. ng vt c v ch c th tch chui bn v to nn hp cht sulfoxid. Tuy nhin thnh phn chnh ca biotin c thi ra dng khng bin i. 2.1.6 Ascorbate (vitamin C) Vitamin C tham gia vo nhiu phn ng bin i cht v thng c chc nng bo v. Sinh tng hp v trao i cht

Thc vt bc cao, to, nm men v phn ln ng vt c th tng hp ascorbate. Mt s ng vt c v (v d ngi) cng nh cn trng, phn ln cc loi c khng c kh nng ny. nhng ng vt c v cn cung cp vitamin C, v thiu enzyme L-Gu-ionolacton-oxidase.

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Sinh tng hp ng vt v phn ln thc vt t D-glucose v i qua UDP-glucose, UDP-glucuronate, D-glucuronate, L-gulonate, Lgulonolacton v 2-dehydro-L-gulonolacton. Do ta thy vitamin C l sn phm oxy ho ca cc ng. N c tng hp t glucose ( ng vt v thc vt), t galactose ( mt s thc vt). Phn t vitamin C c cha eniol, d dng b loi hydro to thnh dehydroascorbic acid (cng c hot tnh vitamin C). Vitamin C mt hot tnh khi vng lacton b thu phn. Trong t nhin, vitamin C c th tn ti dng lin kt vi cc cht khc, gi l ascorbigen l dng d tr vitamin C quan trng. Vitamin C ho tan tt trong nc, dung dch c v acid, c tnh hot quang v c tnh kh mnh. Ascorbic acid c kh nng kh dung dch Fehling, AgNO3. C th s dng tnh cht ny nh lng ascorbic acid. Ngoi ra, cng c th s dng vitamin ny lm cht chng oxy ho trong cng nghip thc phm. Ascorbate i vo cng thc n c vn chuyn n cc c quan khc nhau. mt s c quan s tip nhn l mt qu trnh ch ng, c kch thch bi insulin. Nng ascorbate cao nht tm thy nang trn thn, no thu, mt, gan, d dy v no. Bnh hoi huyt ngi c ngn nga khi tip nhn 10mg ascorbate /ngy. Stress, ngi ht thuc v ngi c thai cn nhiu ascorbate hn. Ngi ta khng nh rng, mt liu lng cao ascorbate tng cng h thng min dch, gim s mt mi v nguy c bnh ung th. Tuy nhin s phn gii ascorbate xy ra mnh m, v vy khi lng cn hng ngy l 50 mg th cn a vo c th l 1 500 mg. Vitamin C c nhiu trong rau qu ti, c bit l cc loi: cam, chanh, da chut... Vitamin C tham gia trong nhiu qu trnh quan trng ca c th sng nh: - Qu trnh hydroxyl ho do oxygenase. V d hydroxyl ho proline, lysine trong phn t collagen. Thiu vitamin C, collagen mi c to thnh khng c hydroxyl ho, khng to c dng xon ba nn c bn km, do da b tn thng v thnh mch mu mng manh d b v, gy ra bnh hoi huyt. Vitamin C tham gia trong qu trnh chuyn ho protocollagen thnh collagen nn c tc dng lm cho vt thng chng thnh so - Vn chuyn hydro trong qu trnh oxy ho - kh thc vt - Lm tng tnh khng ca c th i vi nhng iu kin khng thun li ca mi trng ngoi, cc c t nhim trng. Ngoi ra vitamin C cn tham gia trong nhiu qu trnh tng hp, chuyn ho cc cht khc nhau.

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- Duy tr cn bng gia cc ion Fe2+ /Fe3+ , Cu+ /Cu 2+ Phn gii v s loi thi loi linh trng (primat) mt phn ascorbate tn ti trong c th c thi ra ngoi dng khng bin i qua nc tiu. Nhng sn phm loi thi khc l oxalate v dioxogulonate (xut hin t dehydroascorbate bng mt phn ng khng thun nghch v khng xc tc). Ascorbate c th c oxy ho hon ton thnh CO2. 2.2 Cc vitamin ho tan trong cht bo 2.2.1 Retinol (vitamin A) Vitamin A l tn thng gi cho ru retinol, aldehyd retinal v retinic acid, chng l nhng isoprenoide c c tnh lipid. Chng tn ti cc dng ng phn cis v trans khc nhau. Sinh tng hp v bin i Tin cht tng hp vitamin l provitamin -carotene, s tng hp chng xy ra trong thc vt. ng vt c th tch carotene thnh retinal, bng mt phn ng dioxygenase (xy ra trong rut) v s dng n nh l vitamin. Vitamin A c hp th vo cng vi cht bo v c tch lu dng retinylpalmitat trong gan sau khi kh v ester ho. vn chuyn n nhng c quan khc nhau ester c thu phn v retinol c a vo mu, n c tip nhn bng mt protein kt hp c hiu vi retinol. S bin i gia retinol, retinal v retinic acid c thc hin bng nhng phn ng ph thuc NADP+. Carotene c a vo cng vi thc n ca ngi c ngun gc trc ht l t thc vt, trong khi retinol c ngun gc ng vt (du gan c thu). Nhu cu hng ngy i vi ngi ln l khong 1 mg retinol hoc 6 mg -carotene. Thiu vitamin rt ph bin nhm dn s suy dinh dng, dn n chng qung g, c ch s pht trin ca xng v s phn ho m biu b, hn ch s pht trin v chc nng sinh sn. Chc nng ho sinh Retinic acid l mt cht iu ho sao chp. Sau khi i qua mng t bo phn t ny kt hp vi cht nhn c hiu: all-trans-retinic acid vi RAR-receptor, 9-cis retinic acid vi RXR-receptor. Sau phc h retinic acid -RAR-RXR kt hp vi yu t hormone c to ra t DNA trong nhn. Thuc nhng gen c iu khin theo cch ny l gen m ho cho protein kt hp vi retinol, enzyme PEP-carboxylase v apolipoprotein

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A1. Retinic acid cng tham gia vo s iu khin s pht sinh phi v pht sinh hnh thi ( liu lng nh vitamin A trong thi gian c thai l nguyn nhn gy qui thai), pht trin, phn ho v kh nng sinh sn. S bin i t 11-cis thnh all-cis-retinal nh nh sng l c s cho qu trnh nhn ng vt. Trong vitamin nhm A c 2 dng quan trng l vitamin A1 v A2. Vitamin A2 khc vi A1 ch trong vng c thm mt ni i gia C3 v C4. Di tc dng ca enzyme retinoldehydrogenase nhm alcol ca vitamin A d dng b oxy ho n aldehyd (all-cis-retinal, vn c hot tnh vitamin A). Lin kt i gia C11 v C12 ca retinal c th chuyn thnh dng cis (11-cis-retinal), 11-cis-retinal kt hp vi protein opxin to nn sc t ca mt l rodopxin. y l protein nhn nh sng (photoreceptor protein) c trong t bo hnh que ca mng li mt ngi v ng vt c v. T bo ny hot ng trong nh sng yu, thch nghi vi bng ti. Khi c nh sng, nhm thm ca rodopxin chuyn t dng cis sang dng trans nn mt kh nng kt hp vi opxin. Ngc li trong ti s ti to li dng 11-cis-retinal v rodopxin c tng hp tr li, lm tng nhy cm ca mt trong nh sng yu. Halobacterium s dng s bin i t all-trans thnh 13-cis-retinal nh nh sng lm ng lc cho nhng bm proton ca chng. Retinylphosphate tham gia vo qu trnh sinh tng hp cc glycosaminoglycane trong cc t bo biu b, y n c chc nng tng t nh dolicholphosphate. Bng cch ny n tham gia gin tip vo s ton ven ca mng t bo v ty th, n nh biu b ca da v ca mng nhy v gim nhng ri lon v pht trin.

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Calciferol (vitamin D) ng vt bc cao nhng sn phm hydroxyl ca vitamin D ng mt vai tr trung tm trong trao i calcium.

2.2.2

Thc t khi nim vitamin D l gm mt nhm cc cht h hng, khc nhau v ngun gc. Nu ni cht ch th chng khng phi l vitamin, v da c th tng hp mt trong chng (D3) t mt steroid (7Dehydrocholesterin, provitamin D) khi c nh sng t ngoi.

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Sinh tng hp v s bin i Cc vitamin D khc nhau c to nn t 5,7-steroid (sn phm trung gian tng hp zoo-, phyto- v mycosteroid) bng vic m vng B nh nh sng. Chng ch khc nhau cc mch bn. Hai vitamin D u tin sau y l quan trng nht: - Vitamin D2 (ergocalciferol): to nn t ergosterin, cht ny tn ti trong thc vt v nm. - Vitamin D3 (cholecalciferol): to nn t 7-dehydrocholesterin, cht ny c trong ng vt bc cao. - Vitamin D4: xut hin t 22-dihydroergosterin, c trong thc vt v nm - Vitamin D5: to nn t 7-dehydrositosterin, c trong thc vt. Tng hp ni sinh Trong phn ng u tin khi chiu nh sng t ngoi dn n s ng phn ho ng thi vi vic m vng B ca cu trc sterin, lm xut hin provitamin D. di bc sng thch hp nht cho quang phn ng l 295 nm. Chiu sng vi bc sng ln hn s to lumisterin, vi bc sng ngn hn to tachysterin. nh sng mnh hn xut hin suprasterin I v II. Vitamin D c to nn do mt s ng phn ho ph thuc vo nhit . Vitamin D i vo mu v gn vi protein c hiu (DBP: vitamin D-bindende-protein).

a vo cng vi thc n: Mc d s chiu sng c th cung cp y lng vitamin D, vic b sung thm vn l iu cn thit ( ngi ln 5 g/ngy, tr em 10 g/ngy). Sau khi hp th (c h tr bi acid mt) vitamin D c gn vi chylomikronen hoc DBP v c vn chuyn n gan. trong gan xy ra s hydroxyl ho C25 bng mt enzyme ph thuc vo cytochrome P-450. Cht ny c chuyn n thn, y xut hin hp cht c hot tnh sinh hc 1 25-Dihydroxycalciferin (calcitriol. DHCC) do s hydroxyl ho tip theo C1. Mt bc hydroxyl ho lun phin to nn 24 (R).25dihydroxycalciferin. Nhng phn ng hydroxyl ho c th thy gi non (nhau thai), i thc bo

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S phn gii thc hin bng s hydroxyl ho v/hoc oxy ho ca nhm hydroxyl thnh nhm carboxyl. Nhng hp cht ny phn cc mnh hn, th hin hot tnh sinh hc thp, n c thi ra qua phn hoc nc tiu. Chc nng sinh ho ng vt c v vitamin D (thc t l 1 25-Dihydroxycalciferol) iu khin s trao i calcium v phosphate. N hot ho trc tip s hp thu (ti hp th) Ca++ v phosphate rut v thn v cng tham gia cu to vi PTH v calcitonin trong xng. Mt khc n gii phng Ca++ t xng cho s iu chnh tm thi mc Ca++ trong mu. Hiu qu sao chp c to iu kin bi vitamin D-receptor. y vitamin c vai tr nh mt hormon. trong rut, d dy v xng n kch thch s sao chp ca cc gen m ho cho cc protein gn vi calcium (CaBP, calbindin, osteocalcin...). Nhng protein ny tham gia vo s tip nhn calcium v khong ho xng. Lnh vc y hc: Nhng bnh gn lin vi mc vitamin D khng bnh thng: Ci xng: (Khong ho xng thp, phn ln tr em) sinh ra do s hp th Ca++ gim khi thiu vitamin D, thnh thong cng do tng hp khng calcitriol t calciferol.

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Bnh long xng: xut hin khi tng hp calcitriol b ri lon, do thn khng hot ng hoc mc ostrogen thp v n kch thch bc hydroxyl ho. Tocopherol (vitamin E) Tocopherol (tocos = con chu, pherol = sinh ra) kh ph bin cy xanh, rau x lch, ht ng cc, du thc vt, gan b, lng trng. Nhm tocopherol gm 8 hp cht, bao gm vng 8 - chromanol v mt chui bn isoprenoide Sn phm cui cng ca qu trnh sinh tng hp l -tocopherol (5,7,8-trimethyltocopherol), th hin hot tnh sinh hc cao nht. thc vt v ng vt tocopherol bo v lipid trc s thit hi do oxy ho. Cc tocopherol phn bit vi nhau bng s lng v v tr ca nhm methyl vng v cc lin kt i trong chui bn. Tt c chng u a lipid. Tocopherol c tng hp dng ng nht ch thc vt, trc ht trong lc lp ca l. N c ng vt v thc vt trc ht mng t bo (v d mng ty th, hng huyt cu, v lp th). Lng b sung hng ngy i vi ngi ln l 8-12 mg. Cn lng ln hn khi thc phm c cha nhiu acid bo cha no. Him khi thy thiu vitamin ny, ngoi tr trng hp non v ri lon hp thu lipid. Tuy nhin trong chn nui ch dinh dng khng y gy ra s thoi ho cc c quan v v sinh. Chc nng ho sinh ca tocopherol trc ht l bo v lipid mng trc phn ng peroxid-ho (c bit acid bo c nhiu lin kt cha no). N lm gim tc hi do LDL b oxy ho, dn n bnh sng khp xng. Tocopherol l ni ether ca cc hydroquinon. Chng tc dng l nhng cht nhn gc, y chng chuyn sang trng thi tocopheroxy (semiquinon). y ch l mt gc phn ng yu vi thi gian bn hu l nhiu gi, n lm gy cc phn ng chui. Phn ng to nn l thun nghch, rt c th ascorbate thc hin s kh tr li thnh tocopherol. y n b oxy ho b mt gia lipid v nc thnh monodehydroascorbate. Gia hai vitamin ny c s ng tc dng. -carotene cng c coi l cht kh cho tocopherol. Ngc li semiquinon c oxy ho tip tc thnh quinon, cht ny dn n mt s m vng thun nghch. 2.2.3

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Phylloquinon v menoquinon (vitamin K) Phylloquinon l mt thnh phn ca h thng quang ho ca thc vt quang hp. Menoquinon ng mt vai tr trong h hp ym kh ca vi khun. ng vt hai hp cht (vitamin K1 cng nh vitamin K2) l cofactor s cacboxyl ho ca glutamat. l monomethylnaphtochinon vi mt chui bn isoprenoid. Phyllochinon v menachinon phn bit nhau ch s lng lin kt i v chiu di ca chui bn. Lng tip nhn hng ngy cn thit ngi ln 70-140 g. Lng ny c p ng bng thc n v sn phm ca vi khun ng rut. Trc khi c thi phn ln ch cc chui bn c lm ngn bng oxy ho. Chc nng ho sinh ng vt chng l cofactor to nn -cacboxylglutamate trong protein xy ra endoplamatische reticulum th nh enzyme carboxylase. Protein to nn c phn gii. Qu trnh bin i sau l cn thit cho kh nng kt hp Ca++ v cho s c nh protein bng cc cu Ca++ mng phospholipid (v d qu trnh ng mu). qu trnh bin i protein ny vitamin-K-hydroquinon tch ra mt proton ca glutamate, tip theo n c carboxyl ho. ng thi xut hin mt vitamin K-quinon-2,3-epoxid (nhng chi tit vn cn cha bit). Epoxid c kh bng quinon-epoxid-reductase gn vi dithiol thnh quinon v sau thnh hydroquinon. Dn xut ca cumarine c th km hm bc reductase, v d dicumarol v warfarin c tc dng l cht khng ng.

2.2.4

19

20

CHNG 3

ENZYME V S XC TC SINH HC
sng v pht trin, c th sinh vt phi thng xuyn trao i cht v trao i nng lng vi mi trng bn ngoi. Cc qu trnh ny tin hnh c l nh cc phn ng ha sinh xy ra trong c th theo quy lut c t chc v c lin quan cht ch vi nhau. c im chung ca cc phn ng ny l c th xy ra mt cch c hiu, d dng vi vn tc ln ngay trong iu kin mi trng sinh l v nhit ca c th. S d cc phn ng tin hnh trong c th c nhng c im trn chnh l nh cc cht xc tc c bit c kh nng lm tng vn tc phn ng ln gp bi, l cc cht xc tc sinh hc. Enzyme l cht xc tc sinh hc c bn cht protein lm nhim v xc tc cho cc phn ng ha sinh xy ra trong c th sinh vt. Enzyme tn ti trong tt c cc t bo sng ca ng vt, thc vt, vi sinh vt. Cc phn ng do enzyme xc tc c th xy ra ngay trong c th sng hoc ngoi c th. Hin nay tch c rt nhiu enzyme khc nhau t ngun ng vt, thc vt, c bit l t vi sinh vt, trong c nhiu loi c thu nhn vi thun khit cao. Xc tc enzyme c nhiu u im hn hn xc tc thng thng nh cng lc xc tc ln, tnh c hiu cao, khng c, c th tc dng trong iu kin nh nhng, n gin. Hn na c th sn xut enzyme t cc nguyn liu gi r nhng em li hiu qu kinh t cao, nh vy vic ng dng enzyme rng ri trong cc ngnh cng nghip nh, cng nghip thc phm, nng nghip, y hc v nghin cu khoa hc nhanh chng c nhng bc tin di v ngy cng c nhiu trin vng tt p.

I KHI NIM V S XC TC NI CHUNG

Tc phn ng ha hc c xc nh bi gi tr nng lng hot ha tc l nng lng cc cht tham gia phn ng phi t c trn mc nng lng bnh thng ca chng ct t cc lin kt cn thit v hnh thnh cc lin kt mi. Nng lng hot ha cc phn ng no cng ln th tc phn ng cng chm v ngc li. Vic a mt s cht no vo trong h thng vn c tc dng lm tng tc phn ng ha hc c gi l s xc tc. Trong qu trnh xc tc, cc cht xc tc lm gim nng lng hot ha ca phn ng ha hc. Nng lng cn xy ra s va chm c hiu lc dn n phn ng ha hc c gi l nng lng hot ha.

37

Trong qu trnh xc tc, cc cht xc tc ch tham gia vo cc phn ng trung gian sau chng li c phc hi nhanh chng, cht xc tc khng ng vai tr nh nhng cht tham gia phn ng. Tc cc phn ng thun nghch cng c cc cht xc tc lm tng theo c hai chiu. iu ni ln rng: cht xc tc khng quyt nh chiu i ca phn ng, chng ch thc y cho phn ng t n cn bng mt cch nhanh chng. nh hng ca cht xc tc l lm gim nng lng hot ha. Trong th sau: A l cht phn ng, B l sn phm ca phn ng.

Nng lng t do

Phn ng c enzyme xc tc A

Phn ng khng c xc tc Nng lng Nng lng hot ha hot ha c enzyme xc tc B

6G1 Chiu phn ng

6G2

II ENZYME L CHT XC TC SINH HC

Nhng lun c sau y chng minh bn cht xc tc ca enzyme: a) Cng nh cc cht xc tc ni chung, enzyme lm tng nhanh tc phn ng. Enzyme khng quyt nh chiu hng ca phn ng. b) Enzyme khng ng vai tr l cht tham gia phn ng trong phng trnh phn ng. Trong qu trnh xc tc, lng enzyme khng thay i. c) Cng nh mi cht xc tc khc, enzyme lm gim nng lng hot ha cn thit ca cc phn ng ha hc. Ni cch khc nng lng hot ha phn ng c gim i nhiu khi c s xc tc ca enzyme.

38

III - BN CHT HA HC CA ENZYME 3.1. Enzyme l nhng cht xc tc c bn cht protein.

Trong s pht trin ca ha sinh hc, bc nhy vt t c khi ngi ta thc hin thnh cng vic tch rt cc cht xc tc sinh hc ra khi t bo v nghin cu tnh cht ca chng, lc ngi ta nhn bit rng enzyme c bn cht protein. Nm 1926, Sumner l ngi u tin thu c urease dng kt tinh. Cho n nay c khong hn 150 enzyme c rt ra dng tinh khit. Trong s cc enzyme , mt s c bit trn vn v cu trc bc I nh ribonuclease, trypsin, chymotrypsin, Ngy nay ngi ta xc nhn rng, cc enzyme chnh l nhm protein quan trng. Chng c hnh thnh trong t bo nh cc protein n gin (enzyme mt thnh phn) hoc nh cc protein phc tp (enzyme hai thnh phn). Trong s cc enzyme th a s l enzyme hai thnh phn. Dng hot ng ca enzyme hai thnh phn bao gm phn protein v phn khng c bn cht protein gi l nhm prostetic (nhm ngoi, nhm ghp, ) Enzyme mt thnh phn l cc protein n gin thc hin chc nng xc tc. V d: Ribonuclease A v mt s enzyme thy phn protein v mt s enzyme khc.

3.2. Cc nhm ghp, cc coenzyme

Bn cnh phn protein th enzyme hai thnh phn cn cha phn khng c bn cht protein. Ngi ta gi phn khng phi protein cn thit bt buc i vi hot ng ca enzyme l nhm ghp (nhm ngoi, nhm thm, yu t ph, ) v phn protein vn lin kt vi nhm l apoenzyme, phc hp ca hai thnh phn trn l holoenzyme (enzyme hai thnh phn). Trong trng hp nhm ghp l nhng cht hu c c trng lng phn t b c lin kt vi phn protein th nhm ghp c gi l coenzyme. Theo cch th: Apoenzyme + Coenzyme Holoenzyme

Nu ng ring r th c coenzyme cng nh apoenzyme u khng c kh nng xc tc. Ch c lc no 2 phn ny kt hp vi nhau th hot tnh xc tc ca enzyme mi th hin. Bn cht ha hc ca coenzyme rt khc nhau: - Mt s loi ny chnh l cc vitamin. S lin quan v chc nng gia cc vitamin v cc coenzyme c gii thiu bng sau:

39

Coenzyme NAD, NADP FAD. FMN Coenzyme A Thiaminpyro(P) Pyridoxal(P)

Chc nng - Chuyn H+ v e- Chuyn H+ v e- Vn chuyn gc acyl - Phn gii ho kh v tng hp acid bo - Kh carboxyl ha - Chuyn nhm aldehyd - Chuyn amine ha - Kh carboxyl ha

Vitamin tng ng PP B2 Pantotenic acid(B3)

B

Thiamine(B1) Pyridoxine(B6)

- Cc ion kim loi c vai tr cn thit bt buc cho s hot ng ca enzyme. Cc ion (cation) c chc nng ging vi cc coenzyme. Ngi ta gi cc ion kim loi l cc coenzyme n gin. Cc enzyme cn ion kim loi cho vic thc hin chc nng ca mnh c gi l metalloenzyme. Chc nng ca cc ion kim loi ni chung phn nhiu l chng to ra cc phc hp kiu chelate gia mt nhm nht nh no ca c cht v enzyme. Trc tin cc cation to phc ion kim loi c cht, sau phc hp ny mi phn ng vi enzyme. Cc ion Ca, Cu, Mg, Mn, Mo, Zn, u l nhng ion tham gia trong s hot ng ca cc enzyme.

3.3. Tnh c hiu ca enzyme

Kh nng xc tc vi tnh c hiu cao l mt trong nhng c tnh c bn v quan trng nht ca enzyme. Tnh c hiu ca enzyme th hin ch: enzyme ch xc tc cho mt trong v s nhng chuyn ha c th c c i vi cc cht. C hai loi c hiu c bn l c hiu phn ng v c hiu c cht. * Tnh c hiu phn ng: c th hin ch enzyme ch c kh nng la chn mt dng phn ng trong s cc phn ng v xc tc cho phn ng . iu thy r trong v d sau y:

40

H2O oxidase 2H R1 CH COOH NH2 Aminoacid(1) decarboxylase Transaminase

R1 C COOH

NH3

O Cetoacid(1) R1 CH2 NH2 + CO2

Amine

R2 CH COOH NH2 R2 C COOH O Cetoacid(2) R1 C COOH O Cetoacid(1) Aminoacid(2)

Di tc dng ca oxidase, aminoacid b kh amine ha bng cch oxy ha to ra cetoacid v NH3. Vi s c mt ca oxidase, mt phn ng khc kh carboxyl ha li khng th xy ra. Vic xc tc ny i hi phi c enzyme khc, l decarboxylase. Cng nh vy, phn ng chuyn amine ha i hi phi c transaminase. * Tnh c hiu c cht: Enzyme c th la chn i vi cc cht tham gia phn ng. Khng phi mi c cht c kh nng phn ng u c enzyme tip nhn nh nhau. Mi enzyme ch chuyn xc tc cho mt hoc mt vi c cht nht nh v mc c hiu ca n ty thuc vo tng loi enzyme. C 3 mc c hiu c cht ch yu: a) c hiu tng i: Enzyme ch c th tc dng ln mt kiu lin kt ha hc nht nh m khng ph thuc vo cc nhm ha hc nm hai bn lin kt. V d cc esterase c th tc dng ln hng lot cc ester ca phosphoric acid.

41

b) c hiu nhm: biu hin l enzyme ch c th tc dng ln mt kiu lin kt ha hc nht nh v mt trong hai nhm nm hai bn lin kt cng phi c cu to nht nh. V d carboxylpeptidase c kh nng phn hy lin kt peptide gn nhm COOH t do, ngha l lin kt peptide cui mch polypeptide, .. CH CONH CH COOH .. R1 R2 .. CH COOH .. + H2N CH COOH R1 R2

c) c hiu tuyt i: Enzyme ch tc dng ln mt kiu lin kt nht nh v cc nhm ha hc hai bn lin kt cng phi xc nh. V d: Enzyme trypsine thy phn cc lin kt peptide gia lysine hoc arginine vi bt c aminoacid no. Sn phm l nhng on peptide c lysine hoc arginine cha nhm COOH t do pha tn cng ca peptide. O O

C NH CH C Lysine hay Arginine R V tr thy phn ca trypsine

- Enzyme Trombine cn c tnh c hiu cao hn trypsine: n ch thy phn lin kt peptide pha carboxyl ca gc arginine no c gc glycine ng lin k sau n: O O

C NH CH C Arginine Glycine

V tr thy phn ca trombine * Ngoi cc tnh c hiu trn nhiu enzyme cn biu hin rt cao v tnh c hiu ha hc lp th (c hiu quang hc): Enzyme ch tc dng ln nhng dng ng phn lp th no ca cc cht hu c. V d: Enzyme L-lactatdehydrogenase ch tc dng ln L-lactic acid m khng tc dng ln D-Lactic acid. Mun tc dng ln D-Lactic acid phi c enzyme D-lactatdehydrogenase.

42

IV CC YU T NH HNG TI HOT TNH XC TC CA ENZYME 4.1. Nhit

Trong phm vi l hc, tc ca phn ng tng ln cng vi s tng ca nhit . Nhng khi vt qu phm vi no , cc phn ng c enzyme xc tc b nh hng do s bin tnh ca phn t protein-enzyme. Kt qu ny ph thuc vo nhit ti thch ca enzyme, l nhit m ti tc phn ng enzyme t cc i. Mi enzyme c nhit ti thch khc nhau. S khc nhau ny ty thuc vo ngun gc ca cc enzyme, ty theo tng iu kin hoc tng s khc nhau v tnh nhy cm vi nhit ca phn t protein-enzyme. a s enzyme mt hot tnh xc tc nhit cao(>80oC), tr papain, myokinase c th tn ti 100oC. Hot tnh Nhit ti thch

20

30

40

50

60

70

80

toC

4.2. nh hng ca pH
Mi enzyme u c tr s pH ti thch no i vi hot tnh ca chng. ngoi phm vi ca tr s ny hot tnh ca enzyme u b gim thp. Tr s pH ti thch ca mt s enzyme nh sau: Enzyme Pepsine Amylase(mch nha) Amylase(nc bt) Trypsine Arginase Catalase Peroxidase pH ti thch 1,5 2,5 4,6 5,0 6,8 7,2 7,8 9,5 9,8 6,8 7,0 6,0

43

Nhng nguyn nhn sau y c th dn ti s ph thuc vo pH ca enzyme: a) Nu trong s cc nhm bn tham gia trc tip trong s hot ng ca enzyme cha nhm c kh nng phn ly. b) pH nh hng ti cc nhm phn ly khc ca protein-enzyme vn c tc dng trong vic duy tr cu hnh c hot tnh ca enzyme. c) S thay i pH ca mi trng c th nh hng ti cc nhm phn ly ca c cht hay ca coenzyme vn c kt hp vi enzyme.

4.3. nh hng ca cht hot ha v cht km hm enzyme

Nhng cht no c kh nng lm tng hot tnh xc tc ca enzyme th c gi l cht hot ha enzyme. Cc cht thng l cc ion kim loi nh: K+, Na+, Mg+2, Ca+2, Co+2, Zn+2, Mn+2, V d: Mg+2 lm tng hot tnh phosphatase Ca+2 lm tng hot tnh lypase. S hot ng ca cc enzyme u c th b km hm bi cc tc ng gy bin tnh protein. Ngi ta phn bit cc hnh thc km hm enzyme v phn bit cc cht km hm enzyme nh sau: a) Cht km hm chung: cc cht ny km hm hot tnh xc tc ca tt c cc enzyme. Cc cht ny l cc mui kim loi nng;cht tannin. b) Cht km hm ring: c tc dng km hm mt hay mt nhm enzyme c cu to gn ging nhau. V d: cc cht cha nhm CN km hm enzyme h hp.

4.4. Nng c cht v nng enzyme

Khi mi trng c y c cht th tc phn ng t l thun vi lng enzyme. Khi nng c cht thp, khng li ko tt c lng enzyme vo phn ng th tc phn ng tng t l thun vi nng c cht.Tc phn ng t ti a khi tt c enzyme u kt hp vo c cht.

V C CH XC TC CA ENZYME
Khi t vn nghin cu v c ch xc tc ca enzyme ngi ta xut pht t gi thit cho rng trong cc phn ng c enzyme xc tc, phc tm thi Enzyme C cht c to thnh. Qu trnh ny gm 3 giai on: E+S 1 ES 2 ES* 3 E+P

44

 giai on 1 phn ng xy ra tng i nhanh c cht (S) c lin kt vi enzyme (E) nh cc lin kt yu. Lc ny s lin kt khng gian gia cc phn t c cht v enzyme cha hiu qu i vi s xc tc ca enzyme. giai on tip theo xy ra s bin i ca c cht (S) c lin quan ti vic ph v hay hnh thnh cc lin kt cng ha tr. giai on ny, c cht c hot ha (mt hoc vi phc cht ES chuyn tip c hot ha). y, cu trc bc 3 ca enzyme lun bin i to kh nng tip xc gia cc nhm hot ng ca enzyme vi c cht ang bin i. Enzyme lm bin i phn t c cht lm cho cc lin kt bn trong phn t tr nn lng lo hn, do ch cn mt lng nng lng nh cng lm cho c cht bin thnh cc sn phm (P) khc nhau. Ngi ta chng minh rng trong khi hnh thnh phc cht ES c 2 qu trnh ng thi xy ra nhanh chng l: a) S thay i mt in t gy nn s phn cc ha cc lin kt. b) S bin dng v mt ha hc ca cc lin kt ko cng trong phn t c cht. C hai yu t ny (s bin hnh v s phn cc ha cc lin kt ng ha tr) u lm tng th nng nhit ng hc ca cc lin kt ny, ngha l xc tin vic vt qua hng ro nng lng hot ha ca qu trnh chuyn tip phc Enzyme-C cht.

VI - NG HC CA PHN NG ENZYME 6.1. ng hc ca phn ng enzyme trong trng hp khng c cht km hm. 6.1.1. giai on: E + S ES
E+S K+1 K-1 K+1: hng s vn tc ca phn ng thun. K-1: hng s vn tc ca phn ng nghch. Gi V1 l tc phn ng thun. V-1 l tc phn ng nghch. [E]: nng enzyme. [S]: nng c cht. Ta c: V1 = K+1([E]. [S]) V-1 = K-1[ES] (1) ES

45

Khi phn ng t n cn bng (enzyme phn ng ht vi c cht) th V1 = V-1 ngha l: K+1([E]. [S]) = K-1[ES] K1 [E] [S] t ta c: = [ES] K+1 Nu gi Ks l hng s cn bng cc phn ng bc I, ta c: [E][ . S] K Ks = Ks = 1 (2) [ES] K+1 * Nu Ks c gi tr ln th K-1 s ln v K+1 s nh. T ta thy phc hp ES d phn gii thnh cc cht S v E. Phn ng enzyme tin hnh chm. * Nu Ks c gi tr nh th tc to ES s nhanh ng thi phn ng enzyme cng tin hnh nhanh. Vy nu Ks cng nh th nng ES cng cao. giai on 1 ta tnh c:

Ks =

[E] [S] [ES]

Hay vit di dng khc l: [E] [S] = Ks[ES] (3) Nu gi [Eo] l nng chung ca enzyme trc khi bt u tham gia phn ng. [E]: nng enzyme dng t do (khng to phc hp) [ES]: nng ca phc h ES. th nng enzyme khng to phc hp ( dng t do) l: [E] = [Eo] - [ES] (4) Thay vo phng trnh (3), ta c: ([Eo] - [ES]). [S] = Ks. [ES] Trin khai: [Eo] [S] - [ES] [S] = Ks. [ES] hay l: Ks [ES] + [ES] [S] = [Eo] [S] hay l: [ES](Ks + [S]) = [Eo] [S] T [ES] = [Eo]

[S] K s + [S]

[ES] = [Eo ]

[S] K s + [S]

(5)

Nh vy nng [ES] cng ln th tc phn ng enzyme cng ln (V Ks lc s nh).Tc s t ti ti a khi nng phc h [ES] bng nng enzyme ban u [Eo] ([ES] = [Eo]) ngha l khi tt c enzyme u c kt hp vi c cht th phn ng enzyme l ti a.

46

T c th lp thnh t l:

[ES] V = [Eo ] Vmax [ES] = [Eo ] [S] K s + [S]

V: tc phn ng enzyme ( giai on u) Vmax: tc ti a. T phng trnh (5) ta c: Vy T :

[S] V = Vmax K s + [S]

V = Vmax

[S] K s + [S]

y l phng trnh Michaelis v Menten (1913) dng tnh tc phn ng enzyme t E + S ES. Nu Ks = [S] th V =
1 Vmax 2

6.1.2. giai on: ES* E + P Khi ta c tc to ES l: V+1 = K+1([E] [S]) Tc phn li ES theo phn ng nghch V-1 = K-1[ES] Tnh tc phn gii phc h ES to E + P, hng s tc ny bng K+2 K+1 K+2 E+S ES E+P K-1 K-2 Khi nng c cht [S] cao hn nhiu so vi nng [E] th s t nhanh chng trng thi cn bng gia s hnh thnh ES v s phn gii phc h ES . Nh vy giai on ny ta c: K-1[ES] + K+2[ES] = K+1[E] [S]
Tc phn gii phc h ES E + S Tc phn gii phc h ES E + P Tc to phc h ES

Hay l: (K-1 + K+2). [ES] = K+1 [E] [S] T phng trnh (4): [E] = [Eo] - [ES] Thay vo phng trnh (6), ta c:

(6)

47

(K-1 + K+2). [ES] = K+1 ([Eo] - [ES]). [S] t :

K 1 + K + 2 = K +1
t

([E o ] [ES]).[S] [E o ][S] [ES][S] = [ES] [ES]

[E o ][S] [ES][S] K 1 + K + 2 = Km Km = [ES] K +1 Km c gi l hng s Michaelis (o bng mol/lit) (tnh theo khi m s to [ES] v phn li [ES] bng nhau) Km[ES] = [Eo] [S] - [ES] [S] Km[ES] + [ES] [S] = [Eo] [S] [ES](Km + [S]) = [Eo] [S]
[ES] =

. S] [Eo ][ K m + [S]

(7)

Mt khc tc phn ng tnh theo s hnh thnh sn phm P t [ES] bng: V = K+2[ES] t : [ES] =

V K+2

Thay vo (7) ta c: Vy V=

K +2 .[E o ][ . S] K m + [S]

V . S] [Eo ][ = K+2 K m + [S]

(8)

Phng trnh ny cho php tnh c s ph thuc tc phn ng vo nng ca enzyme v c cht. V t cc i khi [ES] = [Eo] ngha l khi E c kt hp hon ton vi S.

V [ES] = Vmax [E0 ]

t phng trnh 7 ta c: ES =

[E0 ][S] K m + [S]

48

Vy

[S] V 1 [E ][S] = . 0 = [E 0 ] Km + [S] Km + [S] Vmax

V = Vmax .

T :

K m + [S]

[S]

y l phng trnh Michaelis v Menten dng tnh tc phn ng t ES* E + P

* Vy

V=

1 .Vmax th Km = [S] 2

Lc hng s Michaelis c o bng mol/lt So snh gi tr Km v Ks ta thy: K 1 + K + 2 Km = K +1 m Ks = Vy

K1 K+1

K+2 K+1 Khi K+2 = K+1 th Km = Ks + 1 [S] c th vit di dng T phng trnh (8) V = Vmax. Km + [S] phng trnh: Vmax V= Km +1 [S]
Km = Ks + Theo phng trnh ny th khi [S] tng th tc phn ng tng. Nhng khi nng c cht [S] tng n mt mc no th t ti gi tr tc cc i:V = Vmax. Sau th V khng tng theo nng c cht [S] v V lun lun =Vmax. l phng trnh biu din s ph thuc ca tc phn ng vi nng c cht. C th biu din bng dng hiperbol:

Tc Vmax Vmax 2
A 0

C B

Km

[S]
49

C th gii thch ca ng biu din dng hiperbol nh sau: Trong phn ng c enzyme xc tc s phn t c cht ln hn s phn t enzyme rt nhiu (S>E). C cht ch c chuyn ho khi to thnh phc h enzyme. Do tc phn ng ph thuc vo nng phc h [ES]. Nng [ES] 3 im A, B, C khng ging nhau. im A v B cn vi phn t enzyme cha kt hp vi c cht, do nu tng hay gim nng c cht [S] s lm tng hay gim s va chm gia E v S ngha l lm tng hay gim nng [ES] v s nh hng ti tc phn ng v tc l hm s ca nng c cht [S]. Ti im C tt c phn t enzyme kt hp vi S nn khi tng nng c cht c th lm tng dn s va chm gia E v S nhng khng lm tng tc phn ng. V khng cn enzyme t do c th gn thm vo c cht v tc t c l cc i. im B ng tc =
1 s phn t enzyme kt hp vi c cht, 2 1 Vmax v nng c cht tng ng vi im B bng 2

hng s Km. T phng trnh Michaelis Menten V = Vmax . Nu V =

1 Vmax. 2 1 Vmax 2

K m + [S]

[S]

th Km = [S]. Nh vy ngha l hng s

Michaelis bng nng ca c cht khi m tc phn ng bng * Phng trnh Michaelis cng c th vit di dng khc theo ngh ca Lineweaver v Burk (1934) bng cch ly s nghch o:

K + [S] 1 = m Vmax.[S] V

1 Km 1 1 = . + Vmax S Vmax V

[]

50

Phng trnh c dng y = ax + b v ng biu din c dng ng thng nh sau:

1 V 1 Vmax 1 Km 1 [S]

Qua th c th xc nh c Vmax v Km.

6.2. ng hc ca cc phn ng enzyme trong trng hp c cht km hm

Trong trng hp c cht km hm cnh tranh hay khng cnh tranh th gi tr V ca phn ng s bao gm c hng s Ki (Ki l hng s phn li ca phc hp enzyme-cht km hm EI hay cn gi l hng s km hm). Hng s Ki c tnh nh phng trnh sau: Ki+1 E+I EI Ki-1

[E] [I] K i 1 = [EI] K i +1 * Cc cht km hm cnh tranh: l cc cht thng c cu to gn ging vi c cht. Cc cht km hm cnh tranh c tnh c hiu rt cao, ngha l ch km hm mt enzyme ring bit. V d: Malonic acid l cht km hm cnh tranh ca enzyme succinat-dehydrogenasa v n c cu to gn ging succinic acid.
Ki =

CH2 COOH COOH Malonic acid

CH2 COOH CH2 COOH Succinic acid

51

Khi tng nng c cht th mt km hm cnh tranh. * Cc cht km hm khng cnh tranh: l cc cht c tc dng km hm hot ng ca enzyme bng cch gn vo cc v tr khc vi cc v tr gn c cht ca enzyme. Trong trng hp ny ch lm gim V cc i ca phn ng ch khng nh hng g n hng s Km. V d: Cc ion kim loi nng Ag+2; Hg+; Trichloroacetic acid.
6.2.1.ng hc ca phn ng enzyme trong trng hp c cht km hm cnh trnh Vic tnh ton Vi s phc tp hn v ta c ng thi vi qu trnh gn E vi c cht, cn c c qu trnh gn E vo cht km hm I.

E+S

K+1 K-1

ES

K+2

E+P

E+I

Ki+1 Ki-1

EI

Bng cch tnh ton nh trn ta c : Vi =

Vi max .[S] Km K m + [S] + K .[I] i

Phng trnh ny ging phng trnh Michaelis nhng c thm Km .[I] phn nh qu trnh km hm bi cc cht km hm cnh tranh. Ki * Cng c th tnh Vi bng cch :

1 1 1 K m .[I] 1 = . . + K + m Vi Vi max Ki [S] Vi max

52

6.2.2.ng hc ca phn ng enzyme trong trng hp c cht km hm khng cnh tranh: Trong trng hp ny ngi ta thy rng cht km hm khng cnh tranh c th gn vo enzyme t do v c vo phc h ES theo cc phn ng sau:

E+S

K+1 K-1

ES

K+2

E+P

E+I

Ki+1 Ki-1 Ki+2 Ki-2 Ki+3 Ki-3

EI

ES + I

IES

EI + S

IES

Cch tnh phc tp hn: - Gi thit rng K+2 rt nh so vi K+1 v K-2, v phc hp IES khng to thnh sn phm tng ng v cc hng s cn bng c gi tr nh nhau (tng ng vi cc phn ng ghi trn), bng cch tnh nh trn s c:

Vi =

K +2 .[E o ][S] [I] (K s + [S]). 1 + K i

hoc tnh theo dng khc:

1 1 K [I] 1 = 1+ + m . V i K i Vmax Vmax [S]

Khi so snh tc phn ng khng b km hm vi tc phn ng c cht km hm khng cnh tranh ngi ta thu c t l sau:

53

[I] Vo = 1+ Ki Vi Vo l tc ban u ca phn ng enzyme khng c cht km hm. K .[E ][S] Vo = +2 o t (8) K s + [S] y Ks thay vo ch ca Km v K+2 rt nh so vi K-1 ngha l Km = Ks. Nh vy c th thy rng: S bin i tng i ca hot tnh enzyme c cht km hm khng cnh tranh ch ph thuc vo nng ca cht km hm m khng ph thuc vo nng ca c cht [S]. V * i vi cht km hm cnh tranh th t l o c dng: Vi Vo Km Ki [I] t phng trnh ny ta thy s bin i =1+ Vi K m + [S] ca hot tnh enzyme khi c cht km hm cnh tranh ph thuc c vo nng c cht v nng cht km hm. C th biu din tc phn ng enzyme trong trng hp c cht km hm cnh tranh v khng cnh tranh bng th:
V Vmax = Vimax 1 2 1 V Vmax 2 Vimax

Km Kmi 1 V 1

[S] 2 1

Km = Ki 1 V 1 Vi max 1 Vmax 2 1

[S]

1 = Vmax Vi max

1 1 1 [S] Km K mi 1 c cht km hm. 2 khng c cht km hm. C cht km hm cnh tranh

1 1 = Km K mi

1 [S]

C cht km hm khng cnh tranh 54

Tm tt: (Tt c nhng phng trnh sau y ch dng cho trng hp khi 1 c cht tham gia vo phn ng)
Khng c cht km hm Phng trnh C cht km hm cnh tranh C cht km hm khng cnh tranh

1 = V

1 Km 1 . + Vmax [S] Vmax

1 = Vi 1 K .[I] .Km + m . Vi max Ki 1 1 + [S] Vi max

[I] . 1 + K i 1 Km 1 + + V i max Vi max [S]

1 Vi

on thng trn trc tung ti im

1 Vmax
on thng ct trn trc honh ti im

1 Vmax

1 Vmax

1 Vmax

. 1 +

Ki

[I]

1 [S]

1 Km

[I] Km 1 + K i

1 Km

55

VII . CCH GI TN V PHN LOI ENZYME

Trong thi gian c ch tc dng ca enzyme cha c nu ra, ngi ta t cho mt s enzyme nhng tn ring bit nh pepsine, trypsine, chymotrypsine, papaine, bromeline, Khi con s cc enzyme c bit ngy cng nhiu tn enzyme c gi theo nguyn tc sau y: Tn enzyme = (tn c cht m enzyme xc tc + loi phn ng m enzyme xc tc + tip v ng ase).
V d:

AscorbatAscorbic acid + O2 oxidase Dehydroascorbic acid + H2O

CH2 COOH CH2 COOH Succinic acid

Succinat+ FAD dehydrogenase

CH COOH + FAD.H2 CH COOH Fumaric acid

Vic phn loi enzyme l cng vic kh khn v s enzyme m c ch xc tc ca n c hiu bit mt cch tng tn khng nhiu. Vic phn loi enzyme c da theo nguyn tc l: ly c s kiu phn ng do enzyme xc tc m Hip hi Ha sinh Quc t xut nm 1964. Nm 1973 h thng phn loi ny li tip tc c hon thin bi y ban danh php Ha sinh thuc Hip hi ha hc c bn v ng dng Quc t (IUPAC). Trn c s tt c cc enzyme c phn chia 6 nhm ch yu sau: 1. Oxydoreductase 4. Lyase 2. Transferase 5. Isomerase 3. Hydrolase 6. Synthetase (Ligase) Cc s thp phn trn bng phn loi c ngha nh sau: - S th nht ch nhm chnh (V d nhm 2: transferase) - S th hai qui nh mt s c tnh ca phn ng (V d: 2.1 cho bit enzyme vn chuyn gc 1 carbon) - S tip theo cho bit chi tit hn (V d 2.1.1: c ngha l enzyme vn chuyn nhm methyl) Tuy nhin cho n nay cc tn gi truyn thng ca enzyme vn cn c s dng.

VIII. CC NHM ENZYME RING BIT 8.1. Oxydoreductase (cc enzyme oxy ha kh)

56

Cc enzyme nhm ny lm nhim v xc tc cho cc phn ng oxy ha kh. Chng chuyn vn H+ hay cc in t v xc tc cho s oxy ha sinh hc. Cc enzyme ny c vai tr to ln trong qu trnh h hp v trao i nng lng trong c th. S tng qut ca cc phn ng c xc tc bi oxydoreductase nh sau: AH2 + B A + BH2 oxydoreductase Trong AH2 l c cht; B l cht nhn H no , c th l O2. Trong t bo oxydoreductase to nn cc h thng (c gi l chui enzyme oxy ha kh), trong vic vn chuyn cc nguyn t hydro (H+ v e-) qua nhiu giai on, t c cht n cht nhn cui cng l O2. Kt qu l cc nguyn t hydro c vn chuyn n O2 v s to thnh H2O. S h thng enzyme h hp v vic vn chuyn H+ v e- c trnh by tng qut sau y: 2H+, 2eAH2 NAD (NADP) 2H+ 2H+, 2eNADH2 (NADPH2) FAD FADH2 2H+, 2eQ QH2 2e2cytochrome b 2e2cytochrome c 2e2cytochrome a

H2O

O-2

O2

2e-

Ty theo kh nng ca oxydoreductase chuyn H2 va ly c t c cht (AH2) n O2 ca khng kh, ngi ta chia oxydoreductase thnh hai phn nhm: - Dehydrogenase ho kh (oxidase) - Dehydrogenase ym kh. a) Dehydrogenase ym kh: l nhng enzyme khng c kh nng vn chuyn H+ v e- t c cht oxy ha n trc tip cho O2 ca khng

57

kh, m chng s chuyn H+ v e- cho cc cht vn chuyn trung gian khc. Nhm ghp ca dehydrogenase ym kh l dn xut ca vitamin PP (B5) l NAD v NADP.

NH2 O C NH2 O H H (NAD+) N CH2 O (P)- O (P)- O CH2 H N O H N N

N(+)

H OH HO H

H OH HO H Nu l: O (P) th l NADP+

b) Dehydrogenase ho kh: l nhng enzyme c kh nng vn chuyn H+ v e- n trc tip cho O2 ca khng kh. Chng cng l nhng enzyme hai thnh phn. Nhm ghp ca chng l dn xut ca vitamin B2 (Riboflavin) l FMN v FAD. O N H3C H3C NH NH2 N CH2 (CH OH)3 CH2 O (P) O (P) CH2 H FMN N O H 58 N FAD N O N N

H OH HO H

Cc enzyme cha FMN, FAD thng khng oxy ha trc tip c cht m chng ly H2 t NADH2 (hoc t NADPH2) ri chuyn in t cho h cytochrome. a s enzyme cha FMN khi tc dng trc tip vi O2 khng kh s to thnh H2O2. V d: enzyme glycolatoxidase xc tc cho phn ng oxy ha glycolic acid theo s sau: COOH CH2OH Glycolic acid O CH COOH Glyoxilic acid Glycolatoxidase FMN FMNH2 H2O2 O2

8.2. Nhm transferase (cc enzyme vn chuyn)

Cc enzyme nhm ny lm nhim v vn chuyn cc nhm ha hc t hp cht ny sang hp cht khc m khng lm thay i ha tr ca cc nguyn t. S tng qut: Ax + B A + Bx transferase x: l nhm c vn chuyn. Ty thuc vo x m ta c cc tn enzyme vn chuyn khc nhau. V d: Acyltransferase: x l gc cc acid bo trong c gc acetic acid. y l enzyme hai thnh phn trong nhm hot ng l HS-CoA c cu to nh sau:
NH2 N N O N N H OH H H H O (P) H3C CH2 O (P) O (P) O CH3 O NH OH O NH SH

59

- Glycozyltransferase: Nu x l gc glycozyl. - Aminotransferase: Nu x l nhm amin ( NH2 ). - Methyltransferase: Nu x l nhm (CH3). - Aldotransferase: Nu x l nhm aldehyd. - Cetotransferase: Nu x l nhm cetone. - Phosphotransferase: Nu x l gc H3PO4. Enzyme ny cn c tn l: a) Kinase: Nu xc tc cho phn ng chuyn gc H3PO4 m ATP l nhm hot ng ca enzyme ny. V d: Glycerine + ATP Kinase b) Mutase: l enzyme vn chuyn gc H3PO4 trong ni ti phn t ca 1 cht. V d: Enzyme hexoso(P)-mutase xc tc cho phn ng sau: 6 CH2OH 5 O H 4 HO 1 OH H O (P) 3 2 H OH Glucoso-1(P) mutase 6 CH2 O (P) 5 O H H H 4 1 HO OH H OH 3 2 H OH Glucoso-6(P) Glycerol(P) + ADP

Hexoso(P) -

8.3. Nhm hydrolase (cc enzyme thy phn):

Cc enzyme nhm ny xc tc cho cc phn ng thy phn. S tng qut: R1 R2 + H2O Hydrolase R1 H + R2 OH

60

Da vo lin kt gia R1 v R2 m ta c tn cc enzyme thy phn khc nhau: - Peptidase: Thy phn lin kt peptide. - Glycosidase: Thy phn lin kt glycoside. - Esterase: Thy phn lin kt ester. - Amidase: Thy phn amide. R C NH2 + H2O O Amide
V d:

Amidase

R COOH + NH3

NH2 C = O + H2O NH2 Ure

Urease

CO2 + 2NH3

H2N CH COOH CH2 CONH2 Asparagin + H2O

NH3 Asparaginase

H2N CH COOH CH2 COOH Aspactic acid

8.4. Nhm Liase (cc enzyme phn ct)

Enzyme nhm ny phn ct mt nhm no ra khi mt hp cht m khng c s tham gia ca H2O. C trng hp kt qu ca phn ng l to lin kt i. V d: * Decarboxylase: xc tc cho phn ng sau: R CH COOH NH2 Aminoacid CO2 Decarboxylase R CH2 NH2 Amine

61

* Aldolase: xc tc cho phn ng thun nghch sau:

62

Fructosodi(P) CH2 O (P) C=O

Aldehyd(P)glyceric + Phosphodioxiacetone O CH Aldolase CH OH + CH2 OH C=O

(CH OH)3 CH2 O(P) Fructoso 1,6 di(P)

CH2 O(P) CH2 O(P) Aldehyd(P)glyceric Phosphodioxiacetone

* Hydratase (hydroliase): gm cc enzyme xc tc cho phn ng loi tr v kt hp H2O. CH2 COOH C(OH).COOH Aconitat-hydratase CH2 COOH Citric acid CH2 COOH CH COOH CH.OH.COOH Isocitric acid CH - COOH Cis-aconitic acid H2O Aconitat-hydratase H2O CH2 - COOH C COOH

8.5. Nhm Isomerase (cc enzyme ng phn ha):

Cc enzyme nhm ny xc tc cho phn ng chuyn ha cc cht hu c thnh ng phn ca chng. V d chuyn ha gia dng cetone v dng aldehyd. Gia dng L v dng D V d: Enzyme trioso(P)isomerase xc tc cho phn ng sau: O CH2 OH C=O CH2 O(P) Phosphodioxiacetone CH CH OH CH2 O(P) Aldehyd(P)glyceric 63

Enzyme alaninracemase xc tc cho phn ng bin i L-alanine thnh D-alanine v ngc li. COOH Alaninracemase H2N C H CH3 L-Alanine H C NH2 CH3 D-Alanine COOH

8.6. Nhm synthetase (cc enzyme tng hp)

Cc enzyme nhm ny xc tc cho cc phn ng tng hp cc cht hu c. Nhm ny ch xc tc khi c mt ca ATP v cc hp cht cao nng khc. V cc phn ng tng hp i hi phi cung cp nhiu nng lng. Mt s v d sau y: ATP * Aspactic acid + NH3 ADP + H3PO4

Asparagine

H2O Asparaginsynthetase

ATP * Glutamic acid + NH3

ADP + H3PO4

Glutamine

H2O Glutaminsynthetase

ATP * CH3 COOH + HS-CoA Acetic acid

AMP + H4P2O7

Acetyl-CoA-Synthetase

CH3 C ~ S.CoA O Acetyl-CoA

64

------ o0o ------

65

CHNG 4

CARBOHYDRATE V S TRAO I CARBOHYDRATE TRONG C TH THC VT

4.1 Cu to, tnh cht v vai tr ca carbohydrate Carbohydrate l nhm cht hu c ph bin kh rng ri trong c th sinh vt. Nhn chung hm lng carbohydrate thc vt cao hn ng vt. thc vt carbohydrate tp trung ch yu thnh t bo, m nng v m d tr. Tuy nhin hm lng carbohydrate thay i tu theo loi, giai on sinh trng, pht trin. Trong c th ngi v ng vt carbohydrate tp trung ch yu trong gan. Thc vt xanh c kh nng s dng nng lng nh sng tng hp carbohydrate t CO2 v H2O. Carbohydrate thc vt l ngun dinh dng quan trng ca ngi v ng vt. Trong c th sng carbohydrate gi nhiu vai tr quan trng nh: - Cung cp nng lng cho c th, carbohydrate m bo khong 60% nng lng cho cc qu trnh sng. - C vai tr cu trc, to hnh (v d: cellulose, peptidglican...) - C vai tr bo v (mucopolysaccharide) - Gp phn bo m tng tc c hiu ca t bo (polysaccharide trn mng t bo hng cu, thnh t bo mt s vi sinh vt). Da vo cu to, tnh cht carbohydrate c chia lm hai nhm ln: monosaccharide v polysaccharide. 4.2 Cc monosaccharide Monosaccharide l cc aldehyd hoc ketone c cha mt hoc nhiu nhm hydroxyl. S lng carbon trong phn t monosaccharide t nht l 3, l glyceraldehyd v dihydroxyacetone. Chng c cng thc nh sau:

56

CH2OH | C=O | CH2OH L-glyceraldehyd D-glyceraldehyd Dihydroxyaketone

Glyceraldehyd c cha 1 cacbon bt i (C*), c hai ng phn D v L, cn dihydroxyaketone khng cha carbon bt i. S ng phn lp th ca monosaccharide tnh theo cng thc X = 2n (n l s C* trong phn t). Nu s cacbon trong phn t monosaccharide l 4, 5, 6 hoc 7 chng c tn gi tng ng l tetrose, pentose, hexose v heptose. Cng thc cu to ca mt s monosaccharide thng gp c trnh by di y: CHO | HCOH | HCOH | CH2OH D-Erythrose CHO | HCOH | HCOH | HCOH | CH2OH D-Ribose CH2OH | C=O | HC-OH | HCOH | CH2OH D-Ribulose CH2OH | C=O | HO-CH | HCOH | CH2OH D-Xilulose CH2OH | C=O | HO-CH | HCOH | HCOH

CHO | HCOH | HOCH | HCOH | HCOH

CH2OH | C=O | HO-CH | HCOH | HCOH

57

| CH2OH D-Glucose

| CH2OH D-Fructose

| HCOH | CH2OH D-Sedoheptulose

Khi nh s th t cc nguyn t carbon trong phn t monosaccharide, bt u t carbon ca nhm carbonyl ca aldose, hoc carbon u gn nhm carbonyl ca cc ketose. Cn c vo v tr ca H v OH carbon bt i mang s th t ln nht (carbon bt i xa nhm carbonyl) ging vi D hoc Lglyceraldehyd xp monosaccharide vo dng D hoc L. Cc monosaccharide c kh nng lm quay mt phng nh sng phn cc v bn phi (k hiu du +) hoc bn tri (k hiu du -). Trong ho sinh hc ng 6C v 5C c ngha ln. Mt s ng v d 1,6 fructosediphosphate v ng ribose c nu trn. Hexose v pentose c th tn ti dng thng hoc vng. Theo ngh ca Harworth phn t ng c vit dng vng. y ngi ta tng tng mt mt phng nm ngang c 6 hoc 5 gc, nhng gc ca n sp xp pha trn hoc pha di nhng chui bn vung gc vi mt phng. dng vng xut hin mt nhm hydroxyl glycoside, nhm ny c kh nng phn ng cao, d dng to lin kt glycoside v c tnh kh.

58

. Nhm hydroxyl glycoside l c trng ca ng kh, nhm ny c th pha di hoc pha trn mt bng phn t. Nu n nm pha di th ngi ta gi l dng , v d -glucose, nu pha trn th c gi l dng . Glucose tn ti ch yu dng 6 cnh (pyranose), fructose v pentose dng vng 5 cnh (furanose). Mt lin kt glycoside xut hin khi mt phn t nguyn t H c thay th bng mt gc ng (glycosyl), v d adenine c thay th bi 1 ribosyl (gc ng ca ribose).

59

Hnh 4.1 Tng hp hexosephosphate t triosephosphate

Trong trng hp ny nguyn t N ni vi 2 thnh phn, v vy ngi ta gi N-glycoside. Tng t nh vy S-glycoside v O-glycoside cng c to nn. Trong trng hp th nht nguyn t H ca mt nhm SH,

60

trong trng hp th hai nguyn t H ca mt nhm OH c thay th bi mt gc glycosyl. Tnh cht chung ca cc monosaccharide Cc monosaccharide l nhng cht khng mu, phn ln c v ngt. Chng ho tan tt trong nc, khng tan trong dung mi hu c khng phn cc, tan trong dung dch ethanol 80%. Tnh cht l hc c trng ca monosaccharide l tnh hot quang ca chng, ngha l c kh nng lm quay mt phng ca nh sng phn cc sang phi hoc sang tri. C th o c gc quay ny bng my phn cc k. Ho tnh quan trng ca monosaccharide l nhng tnh cht ca nhm chc aldehyd hoc ketone, in hnh l tnh kh. a) Tu iu kin oxy ho monosaccharide b oxy ho thnh cc acid khc nhau - Khi oxy ho nh bng dung dch clo, brom, iod trong mi trng kim, hoc dung dch kim ca cc ion kim loi, nhm chc aldehyd ca monosaccharide b oxy ho. Ion kim loi b kh thnh dng c ho tr thp hn hoc n kim loi t do (vi d Cu2+ Cu+1, Ag+ Ag0).

CHO | HCOH | HOCH | HCOH | HCOH | CH2OH D-glucose -

Oxy ho nh

COOH | HCOH | HOCH | HCOH | HCOH | CH2OH D-gluconic acid

Khi cht oxy ho l HNO3 c, c hai nhm aldehyd v alcohol bc 1 ca monosaccharide u b oxy ho to thnh acid cha 2 nhm carboxyl gi l saccharic acid:

61

CHO | HCOH | HOCH HNO3 | HCOH | HCOH | CH2OH D-glucose

COOH | HCOH | HOCH | HCOH | HCOH | COOH saccharic acid

b) Di tc dng ca cc cht kh nhm carbonyl ca monosaccharide b kh to thnh cc polyol tng ng tin hnh phn ng kh c th dng dng kh hydro c cht xc tc l kim loi (hn hp Hg v Na). V d khi b kh fructose to thnh 2 polyol ng phn l sorbitol v mannitol. Sorbitol cng l sn phm ca phn ng kh glucose. CHO | HCOH | HOCH | HCOH | HCOH | CH2OH D-glucose CH2OH | HCOH | HOCH | HCOH | HCOH | COOH D-sorbitol

Trong c th sinh vt, glucose c th b kh to thnh polyol vng gi l inosite, v d mezoinosite l yu t sinh trng ca cc m thc vt nui cy. Estephosphoric ca n (fitin) l ngun d tr phospho trong ht.

62

c) Phn ng to este thng xy ra vi nhm alcohol bc 1, OH glycosite ca monosaccharide. Cc este phosphat ca monosaccharide l nhng sn phm trung gian quan trng nht ca nhiu qu trnh trao i cht trong c th sinh vt. d) Phn ng ca nhm OH glycosite to thnh cc hp cht glycoside Nhm OH glycoside c th phn ng vi ru to thnh este tng ng gi l glycosite. Tu theo v tr ca nhm OH ny m - hoc glycoside c to thnh. Trong phn t glycoside, phn khng phi carbonhydrate c gi l aglicon. C th phn bit cc kiu lin kt glycosite khc nhau: - Lin kt O-glycoside: (G-C-O-A) gc aglicon (A) kt hp vi carbonhydrate (G) qua O. Lin kt O-glycoside l dng lin kt ca i-, tri-, oligo- v polysaccharide. - Lin kt S-glycoside: (G-C-S-A) gc aglicon (A) kt hp vi carbonhydrate (G) qua S - Lin kt N-glycoside: (G-C-N-A) gc aglicon (A) kt hp vi gluxit (G) qua N - Lin kt C-glycoside: (G-C-C-A) gc aglicon (A) kt hp vi carbonhydrate (G) qua C Lin kt glycoside khng bn vi acid, tng i bn vi kim. Di tc dng ca acid hoc cc enzyme tng ng, glycoside b thu phn to thnh monosaccharide v aglicon. 4.3 Cc polysaccharide Polysaccharide do nhiu gc monosaccharide kt hp vi nhau, c khi lng phn t ln, do polysaccharide khng c tnh kh. Cc monosaccharide c th thuc mt loi hay nhiu loi khc nhau. Cc lin kt glycoside trong phn t polysaccharide c th l - hoc -glycoside. Tn polysaccharide da theo tn ca monosaccharide cu to nn n

63

nhng i ui ose thnh ui an. V d D-glucane, D-fructane, Dgalactane. Sau y ch cp chi tit hn mt s polysaccharide ph bin v quan trng. Tinh bt l polysaccharide d tr thc vt ph bin nht, l mt hn hp ca amylose v amylopectin, trong amylopectin chim khong 80%. Amylose l mt chui khng phn nhnh gm nhiu gc glucose. C khong 200-300 gc glucose kt hp vi nhau theo kiu -glycoside. Lin kt lun lun nm gia C1 v C4. Ngc li amylopectin l chui phn nhnh. S phn nhnh l do lin kt glycoside gia C1 v C6. Amylopectin c to thnh t amylose. Gc amylose (khong 40 gc glucose) c gn kt vi mt chui amylose khc bng lin kt -1-6-glycoside. Ngi ta cho rng chui amylose di v c nhng chui amylopectin ngn hn c cu to xon. C quan d tr tinh bt l v sc lp (amyloplast), tng t nh lc lp v pht trin t tin lc lp. C quan cha t bo d tr (ht ng cc, c khoai ty) gm cc v sc lp xp st nhau.

Tinh bt c tch lu trong v sc lp v lc lp dng ht tinh bt v to nn v tr tinh bt tp trung. Cu trc ny i vi tng loi thc vt l khc nhau nh hnh 4.2. Ngi ta c th nhn dng ht tinh bt c ngun gc t cc loi thc vt khc nhau. nhng v tr tinh bt tp trung

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th hin s kt hp ca cc ht tinh bt dy hn hoc tha hn. S kt hp dy hn th hin vo ban ngy, tha hn vo ban m. Tinh bt c nhiu trong cc loi qu v c khc nhau, nh trong ht ng cc (60-70 %) trong c khoai ty (15-20%).

Hnh 4.2 Cu trc ca cc ht tinh bt khc nhau

Trong lc lp tinh bt ch c tch lu tm thi v thc ra ch khi hot ng quang hp mnh, vo ban ngy. Ban m nhng ht tinh bt c phn gii thnh cc monosacaride, c bit l cc triose v c i ra khi lc lp. Glycogen l mt chui mch nhnh, i phn t c to nn t nhiu gc glucose. Glycogen (tinh bt ca gan) l carbohydrate d tr ca ng vt bc cao. N tn ti ch yu trong gan v trong m c. Nng glycogen trong gan cao hn trong m c, tuy nhin v trng lng ca m c ln hn trng lng ca gan nn m c c nng glycogen tng s cao hn gan. Glycogen nm trong t bo cht dng ht nh, c ng knh 10-40 nm. S tng hp glycogen thc hin t UDP-glucose.

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N l cht cho gc glycosyl i vi enzyme tng hp glycogensynthetase. Glycogen trong gan chu trch nhim iu khin nng ng trong mu. Nng ng trong mu nm gia 4,5-7mM. Khi cung cp ng qua thc n th glucose trong mu c tng hp thnh glycogen, khi cn glucose, v d hot ng ca c th glucose c gii phng t glycogen. Cellulose l mt thnh phn quan trng ca thnh t bo thc vt. thnh t bo s cp n chim khong 30%, t bo gi thnh phn ny cn cao hn. Cellulose l mt glucan, mt polysaccharide, c to nn t cc gc glycosyl. Lin kt nm gia C1 v C4 v tng t lin kt trong cellobiose. lin kt -glycoside nhm OH C1 nm pha trn, trong khi nhm OH C4 nm pha di (cng thc Harworth). Nh vy nhng nhm OH ca hai gc nh 2 pha i din nhau, ngi ta cho rng mt bng ca phn t t quay 1800. Sau phn t cellulose c mt hnh dng, m nguyn t C6 lc nm pha trn v lc th pha di ca phn t.

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C khong 36 chui cellulose ring bit kt hp vi nhau thnh 1 vi si, y cc chui lin kt vi nhau bng lin kt hydro. Cc vi si ny c nhng vng kt tinh v dng ny enzyme kh c th tc ng. Nhng vi si kt hp li thnh si, v b si, nh bng vi l dng si tinh khit. Cht cho glucosyl l UDP-glucose. Enzyme, cellulose synthetase, c gian bo. 4.4. Ho sinh quang hp 4.4.1 Pha sng trong quang hp Trong th gii sinh vt th quang hp l mt qu trnh c bn. Thng qua quang hp m nng lng ca nh sng mt tri chiu xung c bin thnh nng lng ho hc. Hu ht sinh vt ca hnh tinh chng ta sng trc tip hoc gin tip nh nng lng nh sng mt tri. Nh vy nng lng c tch lu l dng nng lng ho hc. Phn ng tng qut ca qu trnh quang hp thng c biu din bng phng trnh sau, mc d n th hin khng hon ton y . 6CO2 + 6H2O + Nng lng C6H12O6 + 6O2 Chnh xc l qu trnh quang hp nh nng lng nh sng mt tri m ng c to thnh v quang hp thc vt gn lin vi vic gii phng oxy. Qu trnh quang hp c chia lm hai giai on: - Phn ng sng - S ng ho CO2 Giai on th nht l qu trnh chuyn nng lng nh sng thnh nng lng ho hc. Giai on th hai l to ra dng nng lng ho hc bn vng v d d tr hn.

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Hnh 4.3 Cu trc ca dip lc a v ca farnesol. Farnesol l gc k nc ca mt s t bo vi khun

nh sng cn cho qu trnh quang hp c tip nhn bi hai nhm sc t: chlorophyll v carotenoid. mt s sinh vt bc thp hn cn c phycobillin. Cu trc c bn ca dip lc l vng porphyrin, c to nn t 4 vng pyrrol ring l (hnh 4.3). trung tm ca vng c mt nguyn t Mg lin kt vi cc nguyn t nit bi hai lin kt ng ho tr v hai lin kt phi tr. Nguyn t Mg c th c tch ra khi c tc ng ca axit long. Phn t dip lc khng cha Mg c gi l pheophytin cng c mt vai tr quan trng trong phn ng sng quang hp. Chui bn ca phn t dip lc c tnh k nc v nh vy m ton b phn t c c tnh k nc. Ngi ta bit nhng loi chlorophyll khc nhau: chlorophyll a, chlorophyll b v chlorophyll vi khun. S khc nhau ca nhng loi ny l do s khc nhau ca cc chui bn. chlorophyll b c nhm aldehyde, nh hnh 4.3, c thay th bng nhm methyl chlorophyll a. iu th v hn l chui bn di k nc c to nn t cc dn xut isopren. chlorophyll a v b chui bn l mt phytol (ru) lin kt ester vi mt chui bn ca vng porphyrine. mt s vi khun chlorophyll cha mt farnesol (alkohol) c lin kt ester vi chui bn ca vng. iu c ngha c bit hn l nhng lin kt i lin hp trong vng porphyrine. Chng c kh nng hp thu nh sng v v vy nhng lin i ny l yu t quan trng ca chlorophyll. Nhng lin kt lin hp

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ca carotenoide cng hp thu nh sng. Cc cht thuc nhm carotenoide c mu vng n mu da cam, thy r l cy vo ma thu, lc ny dip lc b phn hu nn khng cn che ph sc t carotenoid. hiu hn v qu trnh quang hp chng ta phi cp n c quan m trong quang hp xy ra, l lc lp. Lc lp c hnh trn n hnh bu dc, c ng knh khong 10 m. N c bao quanh bi mng trong v mng ngoi, nhng mng ny c ngha i vi vic i vo v i ra ca cc cht. Mng trong ca lc lp c to nn t h thng mng ko dn, mt s v tr c dng xp chng ln nhau.

Hnh 4.4 S biu din mt lc lp

Nhng mng ny xut hin di knh hin vi in t nh nhng ti dt. V vy ngi ta gi chng l thylacoid. Mng thylacoid bao quanh mt khng gian bn trong gi l c cht ca lp th v to nn s ngn cch gia bn trong thylacoid vi mi trng ngoi. S phn cch gia bn trong thylacoid vi stroma l cn thit cho phn ng sng ca quang hp. mt s v tr c nhiu thylacoid xp ln nhau thnh chng rt dy. Chng xut hin di knh hin vi nh nhng ht (grana), v vy ngi ta gi chng l grana-thylacoid. Chng c ni kt vi nhng thylacoid ring l nh mng kp xuyn qua c cht lc lp. Chng c gi l stroma-thylacoid (hnh 4.4).

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Mng thylacoid l ni m nh sng c tip nhn bi cc sc t m t trn v nh cc h thng oxy ho kh khc nhau m nng lng nh sng mt tri c bin i thnh nng lng ho hc v thm thu. Qu trnh ny phc tp v cha c gii thch chi tit. Mng thylacoid cha ba cu thnh siu phn t, chng chim ton b b rng ca mng v l thnh phn quan trng ca chui vn chuyn in t trong quang hp. Ba phc h l h thng quang ho I, h thng quang ho II v phc h cytochrome b/f (hnh 4.5).

Hnh 4.5 Cc phc h vn chuyn quan trng nht ca mng thylacoid: H thng quang ho II (HTQHII), phc h cytochrome b/f, h thng quang ho I (HTQHI)

H thng quang ho l v tr cha cc sc t hp th nh sng (nhng anten). H thng quang ho I cha t nht l 13 loi protein khc nhau, khong 200 phn t dip lc, mt s lng carotenoid cha bit v ba phc h Fe-S. Nhng polypeptide kt hp v nh hng cc phn t chlorophyll v carotenoid. H thng quang ho II cha t nht 11 phn t polypeptide khc nhau, khong 200 phn t dip lc a v 100 phn t dip lc b, hai phn t pheophytin, quinon, 4 Mn cng nh Ca v chlorid. C hai h thng tip nhn nh sng v chuyn ho chng trong s vn chuyn in t. H thng quang ho II cn c nhim v l tch in t ra t H2O, nh vy l oxy ho H2O. S oxy ho ny l qu trnh phc tp, gm 4 bc (mi bc 1e- c vn chuyn v y c s tham gia ca Mn vi s thay i ho tr. 2H2O 4e- + 4H+ + O2.

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Qu trnh tch nc trong quang hp c gi l quang phn ly nc, n gii phng ra O2 v c trng cho quang hp sinh vt nhn chun. H thng quang ho I v II khng ch cha sc t tip nhn nh sng m mi h thng cn cha mt phn t dip lc hon ton c bit th hin v tr ca n, phn t dip lc ny khi tip nhn nh sng c kh nng bn ra 1 in t. Qu trnh ny nh l s khi u cho s vn chuyn in t trong quang hp. Phn t dip lc c bit ca h thng quang ho I hp thu cc i nh sng c bc sng 700 nm. V vy gi l P700 (Pigment 700) v phn t dip lc c bit ca h thng quang ho II hp thu cc i 682 nm. bn ra 1 in t th phn t dip lc cn nng lng. Nng lng c hp th dng nng lng nh sng t nhng sc t khc (chlorophyll v carotenoid) ca mi h thng quang ho v c chuyn n P682 cng nh P700 qua s cng hng in t. Bng cch ny hai h thng quang ho to ra mt s vn chuyn in t khi c nh sng, s vn chuyn in t ny i t H2O nh l ngun in t, qua h thng quang ho II, plastochinon, phc h cytochrom b/f, plastocianin n h thng quang ho I v cui cng n NADP+ v n c kh thnh NADPH. NADP+ + H+ + 2e- NADPH Trong chui kh ny h thng quang ho II v h thng quang ho I khi ng k tip nhau. Plastoquinone cng nh phc h cytochrome b/f v plastocyanin c chc nng vn chuyn e- gia hai h thng quang ho ny v chng nh v nhng v tr khc nhau. S vn chuyn e- lm cho nng lng nh sng c bin i thnh nng lng ho hc, dng kh NADPH (1 mol NADPH = 220 kJ). Thnh vin vn chuyn in t c bn trong chui vn chuyn l phc cytochrome b/f, cng l protein c heme l nhm prostetic, v vn chuyn in t l do s thay i ho tr ca Fe. Bn cnh phc cytochrome b/f l plastoquinon, mt protein cha Cu (protein mu xanh) vn chuyn in t gia phc cytochrome b/f v h thng quang ho I. S vn chuyn ny c th l do s thay i ho tr ca Cu. H thng plastoquinon-plastoquinol vn chuyn H t h thng quang ho II n phc cytochrome b/f. Khong cch vn chuyn y ln hn. N khuch tn trong vng ui ca lp mng kp lipid. Thnh phn mng cha nhiu acid bo cha no th s khuch tn ca plastoquinone d dng hn. Sau y l phn t plastoquinone v s kh n thnh plastoquinol (plastohydroquinone). c tnh k nc ca phn t l do cc nhm CH3

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v chui bn di vi 9 gc isopren. Phn ng kh vng quinone thc hin qua s tip nhn ca 2e- v 2H+ v vng quinone chuyn sang mt h thng lin kt i ca bensoide.

H thng plastoquinone/plastoquinol vn chuyn H gia h thng quang ho II v phc h cytochrome b/f. Phc h dng oxy hoplastoquinon gn vo h thng quang ho II trc ht nhn 2e- v to thnh semiquinone (dng anion) v sau nhn 2H+ ca h thng quang ho II to thnh dng kh plastoquinol. Sau n tch ra khi h thng quang ho II v khuch tn n phc h cytochrome b/f. n y n b oxy ho khi nhng 2e- v 2H+, to nn plastoquinone v li khuch tn v h thng quang II (hnh 2.3). Chc nng c bn ca hai h thng quang ho l hp thu nh sng v tp trung nng lng ca chng n P682 v P700, ri dn n qu trnh quang ho, thc cht l bn ra 1e-. Chc nng ca 2 h thng tng t nhau, tuy nhin nhng qu trnh ring l hai h thng khc nhau, v v chi tit cng cha c gii thch ht. S nhng 1 in t ca P682 lm xut hin mt cht oxy ho rt mnh (phn t c kh nng oxy ho cht khc) c th nng khong 1200 mV. P682 dng oxy ho (P+682) c kh nng oxy ho H2O, ngha l tch e- ra khi H2O. Giai on quan trng qu trnh ny l s tip nhn e- t P682 bn ra.

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Ngy nay ngi ta cho rng, cht nhn tip nhn in t bn ra l nhng hp cht quinone. y quinone c kh thnh bn quinone (semiquinone mang in tch m). Semiquinone l 1 radical, tip tc phn ng nhanh, c th chuyn e- ca n n plastoquinone. Dng semiquinone c kh to thnh plastoquinone. Pheophytin cng tham gia vo s vn chuyn e-. in t c bn ra t P682 c th khng c tip nhn ngay, di tc dng ca nh sng to nn rt nhanh l hng in t ca P682. Dng oxy ho ca P682 (P+682) tn ti rt ngn vi thi gian khong 10-12giy. Nh c h thng quang ho II, di tc dng ca nh sng 1e- t H2O c th nng oxy ho kh cao (th nng oxy ho kh tiu chun l +800 mV) c chuyn n plastoquinone c th oxy ho kh thp (th kh tiu chun 100 mV). Nh vy kh nng kh ca e- c tng ln rt nhiu. Trong ch ra nng lng nh sng c tip nhn. Trong h thng quang ho I e- c bn ra t P700 c tip nhn bi 1 phc h Fe-S. N chuyn e- ny tip tc n ferredoxin dng ho tan nm trong stroma ca lp th. y cng thc hin mt phn ng ngc chiu ca e- ngha l in t c chuyn n ni c in th m ln hn, mt qu trnh cn nng lng, c thc hin nh nh sng mt tri tip nhn c. Th kh tiu chun ca P700 nm khong +400 mV, ca ferredoxin khong - 400 mV. Nng lng c nng ln r rt nh h thng quang ho I. Ferredoxin kh c to nn l mt cht kh mnh, c kh nng kh NADP+.

hnh 4.6 a ra s c gi l s Z, ch ra hai s nng nng lng c thc hin t h thng quang ho I v II trong chui vn chuyn e ca quang hp. y e- ca H2O c chuyn n NADP+. Nh vy NADPH c to nn l mt cht kh mnh cn cho nhiu qu trnh tng hp.

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Hnh 4.6 S Z biu din s vn chuyn in t t in th dng n in th m, s vn chuyn nng lng thc hin qua P682 v P700

Ferredoxin l mt cht kh mnh, c kh nng kh nhiu cht nht, c ch ra hnh 4.7. Ngoi ra NADP+ cn c ngha c bit trong vic kh nitrite, sulfate v oxoglutarate. Phn ng kh nitrite v sulfate l mt bc quan trng trong s ng ho cc ion v c. V vy phn ng sng ca quang hp khng ch l s ng ho CO2 m cn ng ho c N v S.

Hnh 4.7 Nhng kh nng phn ng ca ferredoxin

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Trong hnh 4.5 ch ra t ferredoxin e- khng nhng c th c chuyn n NADP-reductase m cn tr v plastoquinone. Nh vy to nn mt dng in t khp kn. y in t li i qua cytochrome f v plastocyanin tr v li P700 v t y li nh nng lng nh sng c a n ferredoxin. Dng in t khp kn ny cng to nn s phn tch proton v nh vy m tng hp ATP, dng in t khp kn ny c gi l quang phosphoryl ho vng. Quang phosphoryl ho vng ch sn xut ATP, quang phosphoryl ho khng vng to nn ATP v NADPH. Bng cch ny qu trnh quang hp c th t ph hp vi nhu cu ATP v NADPH. Qu trnh tng hp tinh bt ch yu cn ATP, trong khi tng hp acid bo cn tng i nhiu NADPH. ng ho CO2, cn ATP v NADPH theo t l 3:2. Ferredoxin cng c th chuyn e- ca n n O2 (hnh 4.5). Sau H2O xut hin, v O2 b kh lp tc phn ng vi H+ trong mi trng nc. 1/2O2 + 2e O2O2+ 2H+ H2O Phn ng ny c ngha l tiu hao e- v trit tiu gradient H+. V vy n lm quang phosphoryl ho tch ri s vn chuyn e- . Tng kt chuyn ho ca nng lng quang hp ngi ta cho rng 4 mol proton nh sng cn cho vic vn chuyn 2 mol e- v qu trnh quang hp chui vn chuyn e- to nn 1 mol ATP v 1 mol NADPH. Bng 4.1 Tng qut nng lng ca qu trnh quang hp u vo: 4 Einstein (= mol proton) 710 x 103J u ra: 1 mol NADPH 220 x 103J 1 mol ATP 32 x 103J Hiu sut: 3,5 % 252 x 103J Nng lng nh sng c hp thu khong 35% c chuyn sang dng nng lng ho hc. 4.4.2 Pha ti trong quang hp (chu trnh Calvin) S kt hp CO2 vo mt cht nhn hu c (s ng ho CO2) l mt trong nhng qu trnh sinh hc quan trng nht. Nh qu trnh ny m cacbon v c c a vo th gii sng. Phn ng c xc tc bi

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enzyme ribulosodiphosphate-carboxylase (RuDP-carboxylase), enzyme ny c nhiu trong l xanh v n chim khong 30-50% protein ho tan trong l. Enzyme ny gm 8 tiu n v ln v 8 tiu n v nh, trong ch c cc tiu n v ln c hot tnh xc tc, cc tiu n v nh c chc nng iu khin. Th v hn l thng tin di truyn ca cc tiu n v ln nh v trn DNA ca lp th (DNA l cht mang gen). xc tc enzyme cn nhng iu kin hon ton xc nh, l gi tr pH cao (pH 8,3-8,6) v nng Mg2+ tng i cao. C hai u b tc ng bi s vn chuyn e- qua mng thylacoid. V vy enzyme ch hot ng trong iu kin chiu sng. Phn ng ny thi nhit (G01 = -40 x 103J) v tng ng vi phng trnh sau: RuDP + CO2 + H2O 2 phosphoglyceric acid.

Ngi ta cho rng carboxyl ho ribulosediphosphate (RuDP) dn n to thnh mt hp cht trung gian c 6 nguyn t C vi enzyme, sau c tch ra thnh hai phn t phosphoglyceric acid. Nh s trn CO2 c tip nhn c mt trong phosphoglyceric acid pha trn v O t phn t H2O c tip nhn th trong phosphoglyceric acid pha di. Mt thi gian di ngi ta nh gi thp hot ng ca enzyme ny v khi phn lp enzyme thng mt hot tnh. Kh khn na l CO2 khng nhng l c cht m cn nh hng trc tip n xc tc. Ngy nay ngi ta bit hot tnh ca enzyme c c trng bi h s Km (CO2) t 1020m l s ng ho CO2 xy ra trong t nhin. Sn phm carboxyl ho, phosphoglyceric acid c kh to thnh aldehydphosphoglyceric. Qu trnh ny thu nng lng v cn ATP v

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NADPH. C hai coenzyme tn ti stroma ca lc lp nh phn ng sng. nng ATP v NADPH thp khi khng c nh sng th phn ng xy ra ngc li, ngha l aldehydphosphoglyceric b oxy ho thnh phosphoglyceric acid. Phn ng kh phosphoglyceric acid to ra sn phm trung gian l 1,3 iphosphoglyceric acid. qu trnh ny bn cnh ATP v NADPH cn c mt SH-enzyme- photphoglycerate-dehydrogenase tham gia, c biu din s sau:

Aldehydphosphoglyceric l ng u tin xut hin qu trnh ng ho CO2. N d dng chuyn sang dng ng phn phosphodioxyaketone (PDOA). Cn bng phn ng nghing mnh v pha phosphodioxyaketone (PDOA) v khong 90% triosophosphate nm dng PDOA. c mt s ng ho CO2 lin tc th iu cn thit l cht nhn CO2, RuDP phi c to nn mt nng nht nh. ng 3C c to ra l aldehydphosphoglyceric v phosphodioxyaketone c bin i mt phn thnh RuDP. Trnh t phn ng l mt qu trnh khp kn. S ng ho CO2 v bin i ca triosophosphate c gii thch bi Calvin v cng s ca ng. V vy chu trnh ny c gi l chu trnh Calvin hoc l chu trnh kh pentosophosphate v s bin i t phosphoglyceric acid thnh aldehydphosphoglyceric l phn ng kh.

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C ba loi phn ng c trng cho chu trnh Calvin: - Phn ng phosphatase - Phn ng aldolase - Phn ng transcetolase Phn ng phosphatase c cp chng enzyme. Nhng enzyme aldolase c th gn kt mt aldose vi mt cetose, v d aldehydphosphoglyceric v photphodioxyacetone. Phn ng ny ch c mt s bin i nh v nng lng, y l phn ng thun nghch, n c biu din nh sau:

phn ng transcetolase mt mnh 2C ca cetose c chuyn ln mt aldose (v d aldehydphosphoglyxeric). Bng cch tng t mt phn 2 nguyn t C ca fructosomonophosphate c chuyn ln aldehydphosphoglyxeric, xut hin mt pentosophosphate v phn cn li l mt tetrosophosphate.

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Chu trnh Calvin bt u vi mt phn ng aldolase (hnh 4.8). Khi tn ti nhiu triosophosphate th aldehydphosphoglyceric phn ng vi phosphodioxyacetone to nn fructose1,6 diphosphate.

Bng cch ny ng 6C u tin c to nn. T fructose1,6 diphosphate mt phosphate c tch ra nh enzyme phosphatase, lm xut hin fructosomonophotphate. Fructosomonophosphate cng vi mt aldehydphosphoglyceric khc c bin i thnh mt ng c 4C (erythrosophosphate) v mt ng c 5C (xylulosophosphate) bng phn ng transcetolase. Erythrosophosphate phn ng vi mt triosophosphate bng phn ng aldolase to thnh mt ng c 7C (sedoheptulosodiphosphate). T y mt phosphate c tch ra nh enzyme phosphatase lm xut hin sedoheptulosophosphate. Sedoheptulosophosphate phn ng vi mt triosophosphate khc nh enzyme transcetolase to thnh hai ng c 5C l xylulosophosphate v ribosophosphate. Trong phn ng tng qut th cn 5 ng triosophosphate to nn 3 ng pentosophosphate, trong 1 ribosophosphate v 2 xylulosophosphate. Ba ng pentosophosphate c bin i sang dng ng phn ca n l ribulosophosphate, ng ny c photphoryl ho nh ATP to thnh ribulosodiphosphate. Nh vy m cht nhn CO2 c ti to. Ba phn t ribulosodiphosphate c th kt hp vi 3 phn t CO2 v nh vy s to nn 6 phn t triosophosphate. y 5 phn t triosophosphate cn ti to nn 3 phn t ribulosodiphosphate. Mt phn t triosophosphate cn li cho mc ch tng hp. Tng ng c phng trnh nh sau:

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3CO2 + 9 ATP + 6NADPH 1 triosophosphate + 9 ADP + 6NADP+ T phng trnh trn ta thy kh mt phn t phosphoglyceric acid cn 1 ATP v 1 NADPH. to nn 1 phn t ribulosodiphosphate t ribulosophosphate th cn thm mt phn t ATP. Chu trnh tng qut cn 3 ATP v 2 phn t NADPH cho c nh 1 phn t CO2. Vi qu trnh quang phosphoryl ho vng th h thng s to c nhiu phn t ATP hn NADPH, ngha l t l yu cu theo l thuyt ATP/ NADPH ph hp l 3/2.

Hnh 4.9 Chu trnh Calvin, PGA: phosphoglyceric acid, PDOA: phosphodioxyacetone, FMP: fructosomonophosphate, FDP: fructosodiphosphate

4.4.3 H hp sng- Chu trnh Glycolate Cy xanh khi c nh sng cn tip nhn O2 v gii phng CO2. H hp ph ny c gi l h hp sng. S tip nhn O2 c th do nhiu qu trnh v d ferredoxin tham gia vo quang hp chuyn e- ca n trc tip n O2, nh vy H2O xut hin (hnh 4.7). Warburg ch ra rng O2 c ch s ng ho CO2. Enzyme xc tc cho ng ho CO2 l ribulosodiphosphate-carboxylase th hin khng nhng hot tnh carboxylase m cn hot tnh oxygenase. S th hin hot

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tnh no l ch yu ph thuc vo nng CO2 v nng O2. CO2 c ch oxygenase, O2 c ch carboxylase. Carboxyl ho v oxy ho c cng v tr hot ho mt gc lysine. Nhng iu kin kch thch carboxyl ho nh Mg2+ v gi tr pH cao kch thch s oxy ho.

Hnh 4.10 S cnh tranh gia CO2 v O2 vo cng mt v tr ca enzyme ribulosophosphate-carboxylase-oxygenase

Cui cng th c hai phn ng c cng c cht, l ribulosodiphosphate. S cnh tranh gia O2 v CO2 vi phn t enzyme c minh ha hnh 4.10. S cnh tranh ca CO2 v O2 cng mt v tr ca enzyme to ra mt s lin quan vi qu trnh quang hp. Khi cng quang hp cao th nng O2 trong t bo mesophyll tng ln do qu trnh quang phn ly nc, trong khi s ng ho CO2 th nng CO2 gim. Nhng iu kin ny thun li cho phn ng oxy ho. Nh vy nhng iu kin thun li cho quang hp nh nhit thch hp, cng nh sng mnh kch thch s oxy ho ribulosodiphosphate. Phn ng c biu din nh sau:

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 y phn t O2 c tch ra lm 2 nguyn t: mt nguyn t n gc 2C (phosphoglycolate), nguyn t oxy khc n gc c 3C. Phosphoglycolic acid l dn xut ca acetic acid. Bng phn ng phosphatase t phosphoglycolic acid glycolic acid s c to thnh. S oxy ho ribulosodiphosphate to nn chu trnh glyoxylate, y l nhng trnh t phn ng quan trng nht ca quang h hp. Phn ng to thnh phosphoglycolate v phn ng kh phosphoryl ho ca n c xc tc bi enzyme phosphatase xy ra trong c cht ca lc lp. Hot tnh ca glycolate-phosphatase c kch thch bi nng Mg2+ v pH cao v y l nhng iu kin thun li cho quang hp. Glycolate t lc lp i n peroxisome l vi c quan t nm st cnh lc lp, c bao bc bi mng v cha cc enzyme khc nhau nh transaminase, oxidase v catalase.

Trong peroxisome glycolate b oxy ho to thnh glyoxilic acid nh mt oxidase. H2O2 xut hin c phn gii thnh H2O + 1/2 O2 v glyoxilic acid b bin i thnh mt amino acid, l glycine nh mt

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transaminase. Transaminase s c cp k hn chng proteine, y ch nu ln nguyn l ca n l nhm amin ca mt amino acid c chuyn ln mt nhm ceto hoc nhm aldehyd. Amino acid tr thnh cetoacid.

Glycine i t peroxisome vo ty th, l nhng c quan t m trong h hp xy ra. Glycine b oxy ho di tc dng ca NAD+ v sau b kh amin ho (gii phng NH3). y xut hin li glyoxylic acid.

NADH xut hin phn ng kh v l c cht cho chui enzyme h hp. Glyoxilic acid b kh cacboxyl ho v phn ng vi gc =CH-OH t do.

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Tetrahydrofolic acid (ATHF) kt hp vi mt gc hydroxylmethyl, gc ny c chuyn n mt glycine. y xut hin aminoacid l serin v ATHF li tr v trng thi ban u ca n.

Serine b kh amin ho to nn hydroxylpyruvate v cht ny c th c bin i thnh phosphoglyceric acid bng phn ng kh v phn ng phosphoryl ha.

Phn ng cn 1 NADH. Phosphoglyceric acid l mt cht trao i ca chu trnh Calvin. Cht ny li i vo lc lp v n c s dng cho vic to nn triosophosphat. Nh trong hnh 4.11 ch ra, chu trnh glyoxylate xy ra trong cc c quan t khc nhau. Tng qut l c hai phn t glycolate c s dng tng hp nn mt phn t phosphoglyceric acid.

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Hnh 4.11 Nhng cht trao i quan trng ca chu trnh glycoxylate cc c quan t khc nhau

Chu trnh glyoxilate c mi lin h cht ch vi chu trnh Calvin qua ribulosodiphosphate-carboxylase-oxygenase. Tng qut ca chu trnh glyoxilate l 2 v tr O2 c tip nhn (phn ng oxygenase v phn ng oxidase) v CO2 v NH3 c gii phng ra. Nh vy chu trnh glyoxilate phn gii C v N hu c. ngha sinh l ca s phn gii ny vn cha c gii thch r rng. Quang h hp b nh hng bi nhiu yu t ngoi cnh. Nhng yu t kch thch quang hp nh cng chiu sng cao, nhit thch hp lm tiu hao nhiu CO2 trong m l v sn sinh nhiu O2 do quang phn ly nc lm cho h hp sng xy ra mnh m. CO2 v NH3 gii phng ra trong h hp sng li c ng ho. c bit l NH3. Cht km hm t nhin ca chu trnh glyoxilate l glutamate, aspactate v glyoxylate. nhng thc vt C4 h hp sng khng c ngha. y c th do c ch ng ho CO2 c bit m nng CO2 tng i cao trong lc lp, v vy m hot tnh oxygenase ca ribulosodiphosphat-carboxylase-oxygenase khng th hin. 4.4.4. Chu trnh Hatch v Slack Ribulosodiphosphate khng phi l cht nhn CO2 duy nht cho phn ng u tin ca s ng ho CO2. mt s loi thc vt, c bit loi cy h c nhit i (ng, cao lng, ma) cht nhn CO2 l phosphoenolpyruvate (PEP). Qu trnh bt u bng phn ng kt hp

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HCO3- vo PEP. Bng cch ny nh enzyme PEP-carboxylase, mt hp cht 4C- oxaloaxetic acid, c to thnh.

Nh ch ra s , HCO3- c tch ra thnh 1 gc hydroxyl v 1 gc carboxyl. Gc hydroxyl phn ng vi gc phosphoryl ca PEP to nn phosphate v c (H3PO4). Gc carboxyl kt hp vi cht c 3C (PEP) to nn cht c 4C (oxaloaxetic acid).

Hnh 4.12 S ng ho CO2 theo chu trnh C4 nh enzyme PEP-carboxylase

Phn ng ny gii phng nng lng, v lin kt cao nng trong PEP c phn gii. Oxaloaxetic acid c to nn (anion = oxalacetate) sau

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 tip tc b kh to thnh malate hoc b amin ho to thnh aspartate. V trnh t phn ng ny cha nhng hp cht c 4C, nn s ng ho CO2 nh PEP-carboxylase c gi l chu trnh C4. Nhng loi thc vt m ng ho CO2 u tin bng con ng ny, c gi l thc vt C4 ngc vi thc vt C3, l nhng cy ng ho trc tip CO2 qua chu trnh Calvin. Trnh t phn ng ca chu trnh C4 c ch ra hnh 4.12. Phn ng 1: HCO3- c kt hp vi PEP to thnh oxalacetate. Phn ng ny c xc tc bi enzyme PEP-carboxylase. Phn ng 2: Oxalacetate b kh to thnh malate nh NADPH Phn ng 3: Oxalacetate c bin i thnh aspartate bng phn ng chuyn amin ho. Phn ng 4: Nh cc malatenzyme m malate b kh carboxyl ho v oxy ho to nn pyruvate. CO2 c gii phng ra do kh cacboxyl ho c a n chu trnh Calvin. Phn ng 5: Pyruvate c bin i tr li thnh PEP bng phn ng kh carboxyl ho. Phn ng cn ATP v photphate v c. ATP c tch ra thnh AMP v pyrophosphate. Phn ng ny c mt gi tr m G01 cao, nn phn ng xy ra nhanh dn n cht nhn CO2 l PEP c to ra nhiu. Enzyme xc tc gi l pyruvate-phosphatdikinase, n gn vo enolpyruvate theo phn ng di y:

Trao i CO2 ca thc vt C4 c hiu khi ngi ta chia ra nhng qu trnh ring l nhng m l khc nhau v nhn bit nhng v tr ca t bo. Phn ln nhng thc vt C4 c kiu gii phu Kranz c trng, mt khi nim c Haberlandt (1904) ngh. y l nhng t bo xanh bao quanh b mch dng xon kp xp st xung quanh nhng t bo b mch, trong khi nhng t bo mesophyll nh hn sp xp lng lo hn (hnh 4.13).

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Hnh 4.13 S sp xp ca cc t bo tht l v cc t bo b mch cu trc Kranz ca thc vt C4

Trong nhng t bo mesophyll CO2 c ng ho nh PEPcarboxylase v oxaloacetate c to ra b bin i thnh malate hoc aspartate. Nhng t bo mesophyll ca thc vt C4 khng cha RuDPcarboxylase. Sn phm ng ho l malate v aspartate, loi ny l malate v loi kia l aspartate, c vn chuyn qua gian bo n cc t bo bao quanh b mch. y chng c kh carboxyl ho. Pyruvate xut hin li tr v t bo mesophyll. Qu trnh ny xy ra hai loi t bo v c ch ra hnh 4.14.

Hnh 4.14 S tng hp PEP, phn ng carboxyl ho, phn ng kh oxaloacetate t bo tht l, s kh carboxyl ho v chu trnh Calvin xy ra t bo b mch

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 loi cy malate pyruvate c to nn trc tip t malate bng cch kh carboxyl ho oxy ho.

phn ng do malat-enzyme xc tc bn cnh CO2 cn to nn NADPH, l cht cn trong chu trnh Calvin. Vi malate khng nhng cc nguyn t C m cn c cht kh c vn chuyn n chu trnh Calvin. loi cy aspartate aspartate c kh carboxyl ho to thnh alanine. Alanine c bin i thnh pyruvate nh phn ng chuyn amin ho. Phn ng chuyn amin ho xy ra trong t bo cht ca t bo tht l. Alanine c vn chuyn t t bo bao quanh b mch tr v t bo tht l.

S phn chia cc phn ng ring l t bo cht v lc lp ca t bo tht l tng ng vi t l nng lng. S phn chia ny c gii thch trong hnh 4.15. Pyruvate c a vo t bo tht l phi chui vo lc lp ca t bo ny, v y c mt ca enzyme pyruvat-phosphate- dikinase v ATP, ATP ny c to nn t quang hp. PEP to nn trong lc lp c vn chuyn ra t bo cht bng s trao i vi phosphat v c. y PEP cng vi CO2 l c cht ca enzyme PEP-carboxylase. Oxalacetat c

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to nn i vo lc lp ri b kh thnh malate nh NADPH. ATP v NADPH c to ra trong lc lp l nh qu trnh quang hp.

Hnh 4.15 S vn chuyn nng lng v carbon gia lc lp v t bo cht ca t bo tht l thc vt C4

Malate sau c vn chuyn n t bo bao quanh b mch. T bo tht l v t bo bao quanh b mch cha nhng lc lp c hnh thi khc nhau: Lc lp ca t bo bao quanh b mch ln hn v khng c grana v tch lu tinh bt. Lc lp ca t bo tht l nh hn v lun lun cha grana. u im ca s ng ho CO2 thc vt C4 l enzyme PEPcarboxylase c trong t bo cht tip nhn CO2 c trong m l v CO2 t qu trnh h hp tt hn s tip nhn CO2 thc vt C3, v thc vt C3 CO2 trc ht phi chui vo lc lp. Tng ng im b CO2 ca thc vt C4 c bn l thp hn thc vt C3, nm trong khong 0,5-1l/LCO2, im b CO2 ca thc vt C3 nm trong khong 3-5 l/LCO2. Ngi ta hiu im b quang hp l nng CO2 m khng c s tch lu cht hu c. Thc vt C4 c kh nng gim nng CO2 trong m l mnh hn thc vt C3. V vy chng c kh nng s dng CO2 trong khng kh c hiu qu hn. Hn na mt nng CO2 cao c to ra trong t bo bao quanh b mch nh qu trnh kh cacboxyl ho. Nh vy m im bo ho CO2

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ca RuDP-carboxylase gn t c v hot tnh oxygenase ca enzyme b c ch. V vy thc vt C4 c kh nng s dng tt nh sng c cng cao. V l do ny m chng thc t khng c im bo ho nh sng. c im ny c th hin trn hnh 4.16. y ng cong ng ho CO2 ca hai cy trng C4 l ma v ng tng cao hn ng cong ca thc vt C3.

Hnh 4.16 nh hng ca cng nh sng n s ng ho CO2 mt s cy trng

Tuy nhin thc vt C4 cn nhiu nng lng cho s ng ho CO2, v h thng ca chu trnh C4 khi ng trc chu trnh Calvin cn cho s kt hp 1 phn t CO2 hai phn ng phosphoryl ho (to nn hai lin kt cao nng) tng hp nn ATP t AMP theo phng trnh sau: AMP + ATP 2ADP 2 ADP + 2 phosphate 2ATP (quang phosphoryl ho) ------------------------------------------------------------------Tng qut: AMP + 2 phosphate 2ATP Phn ln nhng loi thc vt C4 c ngun gc t nhng vng c cng nh sng mnh, v vy ATP c to nn nhiu nh quang phosphoryl ho. i vi s ng ho CO2 th cng nh sng l yu t t hn ch hn so vi nng CO2. iu cng d hiu l trong qu trnh tin ho mt s loi thc vt pht trin mt c ch ng ho CO2, c ch ny s dng nhng iu kin thi tit c bit. Thuc vo s thch

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nghi ny l enzyme pyruvate-phosphate-dikinase mn cm khi nhit thp. V d iu kin kh hu Trung chu u trong ma xun th hin s ngng pht trin v s vng l ca ng, trong khi cc loi ng cc khc (la m, la mch, yn mch, kiu mch) pht trin tt, ch yu l do s mn cm ca enzyme pyruvate-phosphate-dikinase vi nhit thp. Khi thi tit m ln th ng bt u pht trin vi tc nhanh, vt nhiu so vi thc vt C3. Tc pht trin nhanh l do s ng ho CO2 c hiu qu, mt phn cng do thc vt C4 s mt mt sn phm ng ho do h hp sng thp hn s mt mt thc vt C3. Cu to gii phu kiu Kranz ca t bo l cng c mt ngha i vi trao i nc. Nhng t bo xp st b mch c s trao i kh thp hn v nh vy c s mt nc t hn do thot hi nc. iu ny gii thch ti sao thc vt C4 c nhu cu nc tng i thp hn. 4.4.5 Quang hp theo chu trnh CAM (Crassulaceae Acid Metabolism) Cc loi thc vt khc nhau ca h Crassulaceae v nhng i din ca Agavaceae, Bromeliaceae, Cactaceae, Euphobiaceae, Liliaceae v Orchideaceae c kh nng tip nhn CO2 vo ban m nh kh khng m v ban ngy th kh khng ng li trnh s mt nc. Tri vi a s cc loi thc vt khc, kh khng ng li ban m v m ban ngy, kh nng ny gn lin vi s trao i acid v CO2 c bit, trc ht c nghin cu loi Crassulaceae. Nhng thc vt c hnh thc trao i ny c gi thc vt CAM.

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Hnh 4.17 S ng ho CO2 v trao i acid thc vt CAM

Nguyn l ca s trao i cht c bit ny c minh ha trong hnh 4.17. Ban m CO2 c tip nhn nh kh khng m, enzyme PEPcarboxylase carboxyl ho PEP to thnh oxaloaxetate. Cht ny c kh to thnh malate. Sau malate c d tr trong khng bo. y nng malate c th ln n 100-200 mM, c bit nng ny cao trong nhng gi u bui sng, trc khi ngy bt u. Ban ngy khi kh khng ng, malate trong khng bo c huy ng, ngha l malate i vo lc lp v y n c kh carboxyl ho. CO2 t do v NADPH to ra i vo chu trnh Calvin. Bn cnh s ng ho CO2 c cung cp bi ATP v NADPH c to ra t quang hp. Sn phm ng ho ny mt phn c bin i thnh tinh bt, c tch lu trong lc lp. Ban m mt phn tinh bt ny c phn gii to thnh PEP, l cht tip nhn CO2 trong t bo cht. s kh carboxyl ho oxy ho ny m pyruvate c to thnh t malate, sau pyruvate c bin i thnh PEP nh enzyme pyruvate-phosphate-kinase. Bn cnh malate v oxaloaxetate th aspartate, glutamate, alanine, glycine bng phn ng chuyn amin ho to nn nhng sn phm trung gian gn lin vi s trao i cht ca thc vt CAM. S tch lu malate trong khng bo l mt qu trnh ch ng, c iu khin bi sc trng ca khng bo. Th nc ca cy iu khin s ng m kh khng. Ngi ta phn bit nhng thc vt CAM bt buc

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v khng bt buc. Thc vt CAM khng bt buc ch thc hin theo chu trnh CAM khi thiu nc, c bit l trong iu kin ngy di. Kh khng ng ban ngy th s mt nc gim rt ng k, nh vy m thc vt c kh nng vt qua c thi k kh hn. mt th nc trong t l 2200 kPa, vt xa im ho ca cc cy trng khc, th trong cc m ca loi cy ny vn duy tr c th nc t 500-1000 kPa. 4.4.6 Tng hp carbohydrate Vi mt s phn ng tng i n gin thc vt c kh nng tng hp cc hexosephosphate khc nhau, l nhng nguyn liu cn thit cho tng hp oligo- v polysaccharide. Nguyn l sinh tng hp l xy dng mt lin kt glycoside trc ht c gii thch bi Leloir. Nguyn l ny c ch ra v d tng hp glucan (polysaccharide ca glucose) (hnh 4.18). phn ng ny c hai enzyme tham gia: ADP-glucopyrophosphorylase v glucan- hoc ADPgluco--glucan-4-glucosyltransferase. Enzyme ADPglucopyrophosphorylase hot ho glucose1-phosphate nh ATP, vi s tch ra pyrophosphate xut hin dng glucose hot ho, l ADPglucose. phn ng ny nguyn t H ca nhm phosphoryl c thay th bng nhm acyl ca adenosinemonophosphate. Acyl tng ng gi l adenyl. Sau dng glucosyl hot ho c chuyn ln chui glucan v thc ra n thay th nguyn t H ca nhm OH v tr C4 (xem cng thc). Nh vy xut hin mt lin kt glycoside gia C1 ca glucosyl mi i vo v C4 ca glucosyl chui glucan. Lin kt nm pha di ca mt phng glucose v l lin kt glycosidic. Theo nguyn tc ny mt chui di ca cc gc glucosyl c to thnh. Chui ko di theo hng C1 C4. S tng hp polysaccharide phn ln bt u vi mt phn t m u (primer), y glycosyl u tin c nh vo. s tng hp tinh bt v glycogen th primer l 1 glycoprotein. Glucosyl l gc ng ca glucose, fructosyl ca fructose, ribosyl ca ribose.

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Hnh 4.18 Tng hp ADP-glucose nh enzyme ADP-glucosepyrophosphorylase v s chuyn glucosyl ln mt chui glucan

Bn cnh tinh bt, saccharose l sn phm quan trng khc ca qu trnh quang hp. Triosophosphate c to nn t chu trnh Calvin i t lc lp ra t bo cht, bin i thnh UDP-Glucose, Fructose 6-P v sau thnh saccharose 6-P nh enzyme saccharose-P- synthase. Cui cng phosphate c tch ra bng enzyme saccharose-P-phosphatase. iu khin enzyme saccharose-synthase thc hin bc fructose1,6-disphosphate bi nng ca fructose 2,6-P2 (xem gluconeogenese). Mt v tr iu khin khc l saccharose-P-synthase c hot ho bi c cht glucose 1-P v b c ch bng phosphate v c. Ngoi ra hot tnh ca enzyme gim xung do qu trnh phosphoryl ho v tng ln do s kh phosphoryl ho (tng t nh glycogen-synthase). Nng saccharose li iu khin s tng hp tinh bt.

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Hnh 4.19 S tng hp saccharosephosphate v saccharose

Saccharose va l mt dng carbonhydrate vn chuyn va l tin cht ca tinh bt (trong t bo, nm cch ra khi v tr quang hp) v ca cellulose. Enzyme saccharose-synthase c tc dng thun nghch to nn nucleosid - ng cn thit Saccharose + UDP = UDP-D-Glucose + Fructose

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Tng t ATP l coenzyme ca ADP-pyrophotphorylase, khi tng hp tinh bt UTP cung cp nng lng cho s tng hp saccharose, glycogen v cellulose. y UDP-pyrophosphorylase xc tc tng hp UDP-glucose t glucose1-phosphate v UTP. s tng hp saccharose mt glycosyl c vn chuyn t UDP-glucose n 1 fructosophosphate to nn saccharosephosphate, t cht ny saccharose v phosphate v c c tch ra. S tng hp saccharose sau khi ng ho CO2 cng nh s to nn saccharose t tinh bt trong ht ny mm thc hin theo con ng ny, c biu din hnh 4.19. Enzyme xc tc cho phn ng ny l saccharosephosphat synthetase. Hng phn ng dch v pha saccharosephosphate cng nh saccharose, v s to ra phosphate nh phosphatase, l mt qu trnh thun li v nhit ng hc. Khng nn nhm ln gia enzym saccharosephosphatsynthetase vi enzyme saccharosesynthetase. Enzyme saccharosesynthetase tng hp saccharose trong nhng trng hp ngoi l. Chc nng quan trng hn ca n l

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tng hp UDP-glucose t saccharose v UDP tng ng theo phng trnh phn ng sau: Saccharose + UDP UDP-glucose + fructose Cn bng phng trnh ny dch v pha to UDP-glucose. Enzyme xc tc l saccharose-UDP-glycosyltransferase. Phn ng ny ng mt vai tr quan trng trong m sinh trng v trong nhng c quan d tr ang ln (c, ht) m saccharose c chuyn n tng hp UDPglucose. Sau UDP-glucose c s dng ch yu cho vic tng hp cc cht ca thnh t bo, nh cellulose. C ch ca phn ng ca saccharose-UDP-glycosyltransferase c th hin s trn. Nhng polysaccharide m t trn c cha nhiu nhm OH, nn phn t c c tnh a nc v c kh nng to nn cc cu hydro. Cc nhm acid, carboxyl, nhm sulfate lm cho phn t c tnh acid. Trong iu kin pH sinh l (pH 5,5-7,5) cc proton c tch ra, v vy cc i phn t mang nhiu nhm anion, nn tch in m v c kh nng kt hp vi cc cht tch in dng, v d nhm amin b proton ho to lin kt ion. 4.5 Ho sinh h hp 4.5.1 nh ngha h hp H hp l qu trnh phn gii, oxy ho cc hp cht hu c phc tp thnh cc sn phm cui cng l CO2 v H2O, ng thi gii phng nhiu nng lng cn thit cho c th sng. Phng trnh tng qut ca h hp: C6H12O6 + 6O2 6CO2 + 6H2O + Nng lng V bn cht h hp l qu trnh oxy ho sinh hc c thc hin bng s tch cc nguyn t hydro ra khi c cht v vn chuyn n (H+ v e-) n nhng cht nhn khc nhau, cht nhn cui cng l O2. H hp xy ra hai v tr khc nhau trong t bo. Giai on th nht xy ra t bo cht (qu trnh ng phn), hoc bn trong ty th (chu trnh Krebs), l s tch hydro ra khi c cht v chuyn n n NAD v/hoc FAD to nn NADH2 v FADH2. giai on th hai NADH2 v FADH2 b oxy ho, l qu trnh chuyn H+ v e- cho cc cht nhn trung gian v cui cng n O2. Qu trnh ny xy ra mng trong ca ty th. Kt qu ca qu trnh ny to ra cc phn t ATP v H2O. H hp ho kh:

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c tnh ca h hp ho kh l gii phng CO2 v tip nhn O2. H hp ym kh theo nh ngha trc y l gii phng CO2 trong iu kin khng c O2. Ho sinh hc nh ngha h hp ym kh l s phn gii carbohydrate qua qu trnh ng phn v tip theo l s ln men ru. Ton b qu trnh ny khng cn O2, CO2 c gii phng ra trong qu trnh ln men bng s kh carboxyl ho pyruvate. H hp ho kh xy ra trong ty th. Khi qu trnh quang hp v h hp ho kh c gii thch cng chi tit th s ging nhau gia chng nh l my bin i nng lng cng nhiu. qu trnh quang hp ngi ta bit matrix cha nhng enzym ca chu trnh Calvin, h hp matrix ca ty th cha nhng enzyme ca chu trnh Krebs. H thng oxy ho kh ca quang hp (chui vn chuyn e- trong quang hp) nh v mng thylacoid ca lc lp, h thng oxy ho kh ca h hp nh v trn mng trong ca ty th. S ging nhau gia hai my bin i nng lng n l ngc nhin.

Hnh 4.20 Chui vn chuyn e- ca h hp vi nhng h thng oxy ho kh quan trng v cc phc h vn chuyn mng

Chui vn chuyn e- trong h hp, v s nh v ca chng trong mng trong ca ty th, c gii thch hnh 4.20. Chui vn chuyn e- gm c 4 phc h proteine vn chuyn ca mng v hai phc h protein nh hn, cuccinate dehydrogenase (FAD) v cytochrome. Ngoi ra cn c ubiquinol/ubiquinone trong chui vn chuyn. Cht kh tng ng l NADH v FADH2, c hai u c to ra trong chu trnh Krebs, c i vo chui h hp qua phc h FMN-protein cng nh qua phc h FAD-protein. Hai phc h vn chuyn H ca n n ubiquinone v kh n thnh ubiquinol. H

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thng ubiquinone/ ubquinol v h thng plastoquinone/plastoquinol c cu to v chc nng rt ging nhau (xem s ).

Ubiquinone/ ubiquinol c kh nng khuch tn rt nhanh vo mng nh c tnh a lipid ca n. H s khuch tn nm 10-8cm2/sec. Ubiquinol kh phc h cytochrome b,c1, y dn n s tch H thnh H+ v e-.

in t c tip nhn bi phc h cytochrome b,c1, trong khi H+ c thi ra pha ngoi mng. Phc h cytochrome b,c1, cn c gi cytochrome c-reductase, v n kh cytochrome c, in t ca n c chuyn cho cytochrome c. y cng l 1 phc proteine nh hn, c th khuch tn d dng ra pha mng ngoi. N chuyn e- n phc h

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cytochrome a, cn c gi l cytochrome c-oxidase, v n oxy ho cytochrome c. Phc h cytochrome a l thnh vin cui cng ca chui vn chuyn e. N chuyn e- n O2, ri phn ng vi H+ ca h thng to H2O. O2 trong chui h hp c nhim v tip nhn e- cui cng. Chui h hp trong hnh 4.20 lm r rng, s vn chuyn H v equa cc thnh vin ca chui dn n s chnh lch proton ca mng. S khc bit ny xut hin s kh ca phc h cytochrom b, c1 bi ubiquinol. V c s oxy ho NADH phc h FMN v s kh O2 phc cytochrome a lm cho matrix kim, v n lm gim H+, ngi ta c th nhn bit trn hnh 4.20. Bng cch ny m 1 th in ho c to nn. Matrix tch in m v c mt nng H+ thp (pH 8), ngoi mng tch in dng v c nng pH tng i cao. S chnh lch th nng gia 2 mng c ln l 200mV. Nng lng ca in th ho hc ny c s dng tng hp ATP. Bn phc h vn chuyn mng c cha ATP-synthetase (ATPase). Proton i qua cc knh vn chuyn ca phc h vn chuyn mng t ngoi mng vo matrix, thc hin tng hp ATP. y c s tng t nh sinh tng hp ATP trong quang hp. Trc khi gii thch nng lng ca chui h hp, th cn ni r hn h thng oxy ho kh. - Phc h FMN, NADH-dehydrogenase gm t nht 26 polypeptide khc nhau. N cha FMN l trung tm phn ng v c khong 20 nguyn t Fe v S linh ng/ FMN. Trng lng phn t ca n l 850 kDa. - Phc h FAD l cuccinate dehydrogenase, cha FAD v phc Fe-S ho tr 2 v ho tr 4 l trung tm phn ng. - Phc h cytochrome b,c1 cha cytochrome c1 v 2 loi cytochrome b, t nht 8 chui polypeptide ln hn v 1 phc Fe-S c ho tr 2. Antimycin l 1 cht km hm ca phc h cytochrome b,c1, trong n nh ch s vn chuyn e- gia cytochrome b v cytochrome c1. Cytochrome a, a3, cn c gi cytochrome c-oxidase gm 7 chui polypeptide khc nhau. ng thi n tip nhn t cytochrome c 4 e- v chuyn chng n 1 O2. s vn chuyn ny c s tham gia ca heme ca cytochrome a3 v ca cytochrome a cng nh nguyn t Cu. Oxy dng kh (O24-) phn ng vi 4H+ bt ngun t h thng nc to H2O. O24- + 4H+ H2O

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Phn ng oxy ho kh ng cui cng ca chui h hp ny c gi l s oxy ho cui cng. N s dng khong 90% nhu cu O2 ca t bo. S oxy ho cui cng ny b c ch bi CO2, cyanid v acid, trong nhng cht ny kt hp vo nhm heme v phong to s kt hp ca hem vi O2. Nhng cht ny l cht c i vi h hp. Phc h vn chuyn mng, cha ATP-synthetase gm nhiu chui polypeptide khc nhau. N cha 1 knh proton, thc hin s vn chuyn H+ t pha mng ny sang pha mng khc. ATP-synthetase ny rt ging ATP-synthetase trong mng thylacoid ca lc lp v ATP- synthetase ca vi khun. S vn chuyn e- trong chui h hp l s vn chuyn thun chiu, ngha l e- theo th oxy ho kh tiu chun, nh trong hnh 4.21. Nng lng gii phng ra y c bin i thnh nng lng thm thu, th hin ra trong s chnh lch nng H (gradient H+). S chnh lch nng H+ hai pha ca mng to nn mt th in ho, m ln ca n c tnh theo phng trnh sau y: P = -59 pH + P = lc pH = Grandient H+ = s khc nhau v in tch gia 2 mng

Hnh 4.21 S vn chuyn e- trong chui h hp ph thuc vo th oxy ho kh tiu chun

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Phng trnh ny ng vi mt nhit nht nh, v d nhit phng, m t lc di chuyn proton ngha l lc y 1 H+ t 1 pha mng nng H+ cao (pha ngoi mng) n 1 pha khc ca mng vi nng H+ thp (matrix). Lc ny c biu din mV l s chnh lch th in, y 1 n v pH tng ng vi 59 mV. V l do ny m trong phng trnh c h s 59. Lc di chuyn proton phi ln, mi nng lng tng hp ATP. y xut hin cu hi, bao nhiu H+ phi i vo matrix, mi tng hp c 1 ATP. Ngi ta cho rng 1 s chnh lch th (lc t pH v , xem phng trnh) l 220 mV s vn chuyn 1 mol H+ t mng ngoi n matrix th nng lng t do gii phng ra l 21.3kJ. Khi 2H t NADH trong chui h hp c a vo, n tch ra 5H+ m ngi ta c th rt ra t hnh 4.20. Ba H+ cn trong matrix, hai H+ c i ra ngoi mng. Khi 5 mol H+ tr v matrix, to ra 5 x 21,3 = 106.5 kJ nng lng t do vi iu kin chnh lch th l 220 mV. Gi s nng lng ny tng hp 3 mol ATP, v G0 ca 1 lin kt cao nng trong ATP l 30kJ. Hai nguyn t H, i vo chui enzyme h hp qua phc h FMN, i hi s oxy ho cui cng 1/2O2. mc th nng 220 mV c th tng hp t 2H (t NADH) 3 ATP vi s tiu hao 1/2 O2. Nh vy t l P/O l 3, l s phn t ATP c tng hp/nguyn t O. Tuy nhin gi tr thu c t thc nghim lch nhiu so vi t l P/O ny. iu ny d hiu l n ch ng khi s chnh lch th l 220 mV. Tuy nhin t khi nim bin i nng lng l chnh xc v 2H, i vo chui h hp qua phc FMN to ra nhiu ATP hn 2 H khi i vo chui h hp qua phc FAD. Trong trng hp th nht tch 5H+, trong trng hp qua phc FAD ch tch c 4H+, iu ny c th d dng thy trn hnh 4.20. s tnh ton nng lng bin i ngi ta cho rng c cht h hp NADH + H+ sn xut 3ATP, c cht l FADH2 l 2ATP. Tuy nhin t l P/O i vi NADH v FADH2 theo gii thch trn c th lch nhiu so vi gi tr thc t. Chng b lch l cn do 1 nguyn nhn khc. v d trn nng lng t do cho tng hp ATP c tnh trong iu kin tiu chun. Tuy nhin trong thc t phn ln cc trng hp khng phi trong iu kin ny. Nng lng cn cho tng hp ATP hng v trng thi cn bng phn ng: ADP + Pi ATP Nu hng v ATP th tng hp cn nhiu nng lng, ngc li cn bng phn ng v pha ADP, s tng hp ATP cn t nng lng. Nh vy t l ATP/ADP iu khin h hp rt hiu qu v c ngha. nng ADP cao, tng ng vi s thiu ATP th ATP c to nn nhiu v

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nhanh v vi tiu hao nng lng t, ngc li vi nng ATP cao th s tng hp ATP cn nhiu nng lng. y l mt s thch nghi tt vi hon cnh nng lng. ATP c s dng nhiu cho cc qu trnh sinh trng (tng hp protein, tng hp nucleic acid) hoc cho lao ng c bp, sau cng h hp tng ln, v nng ATP thp v nng ADP cao. Trong mt giai on c bp t hot ng nng ATP cao, tng ng vi cng h hp thp. mt nng ATP cao phn ng tng hp ATP c th theo chiu ngc li. y ATP c tch thnh ADP v Pi v nh nng lng gii phng ra H+ c bm ra pha ngoi mng. ATP-synthetase tr thnh mt bm proton. V l do ny ngi ta gi ATP-synthetase ngc cng l ATPase. Thc t l nhng bm proton rt ph bin trong t nhin. Ngc vi ATP-synthetase trong chui h hp ATP-synthetase c trong mng thylacoid khng th tch ATP. H hp c chia lm hai loi: H hp ym kh v h hp ho kh. C hai kiu h hp ny u tri qua mt qu trnh chung l qu trnh ng phn. 4.5.2 ng phn, s ln men ru v ln men lactic Nh nhng enzyme thu phn k trn m chui carbohydrate d tr c bin i cho n hexose, nu chng khng phi l glucose hoc fructose, th d dng bin i thnh hai dng monosaccharide ny. Glucose v fructose l nhng c cht trc tip ca qu trnh ng phn (glycolyse). Glycolyse c ngha l phn gii ng. Cc phn ng c gii thch bi Gustav Embden v Otto Mayerhof. V vy qu trnh ny c gi l s phn ng Embden- Mayerhof. S ny gm s phn gii ng v polysaccharide ym kh cho n pyruvate. Nhng enzyme ca qu trnh ny c mt t bo cht. Nh tc dng ca phosphorylase glucose1phosphate c tch ra l c cht trc tip ca qu trinh ng phn. Chui phn ng din ra nhng phn ng ring l nh sau: 1. 2. 3. T chui amylose nh phosphorylase m glucose1-phosphate c tch ra. Glucose1-phosphate c bin i thnh glucose-6-phosphate nh isomerase. Glucose-6-phosphate c bin i thnh fructose-6-phosphate nh enzym isomerase.

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Fructose-6-phosphate c phosphoryl ho nh ATP v enzyme phosphofructokinase to thnh fructose-1,6diphosphate 5. Fructose-1,6-diphosphate nh enzyme aldolase c tch ra thnh aldehydphosphoglyceric v phosphodioxyacetone. 5a. Aldehydphosphoglyceric (AlPG) v phosphodioxyacetone (PDOA) trng thi cn bng, khi AlPG c ly i v l c cht c s dng trong cc phn ng tip theo th cn bng phn ng dn n, AlPG lun lun c to thnh t PDOA. Trong trng thi cn bng c hn 90% dng PDOA. 6. Aldehydphosphoglyceric (triosephotphate) c kt hp vi mt HS-enzyme. Phc h HS-Enzyme ny cn kt hp vi NAD+ 7. Bng vic nhng 2H cho NAD+, AlPG c oxy ho thnh phosphoglyceric acid. S thay i trong phn t lm cho nhm acyl giu nng lng c gn vo S ca phc h enzyme. 8. NADH gn vo phc SH-Enzyme c oxy ho, trong H v e- ca n c chuyn ln mt NAD+ t do. 9. Nh gc phosphate v c m phosphoglycerinacyl c tch ra khi phc h. y nhm SH li xut hin khi to thnh 1,3 phosphoglyceric acid. qu trnh ny phosphoric acid (P-OH) c tch ra 1 nguyn t H v 1 gc P-O- (xem s ). Gc P-O- c gn kt vo acyl ca phosphoglyceric acid, trong khi H c kt hp vi nguyn t S ca enzyme. Enzyme li tr li trng thi ban u.

4.

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10. 11. 12. 13. 14.

Nhm phosphoryl mang nhiu nng lng ca 1,3 diphosphoglyceric acid c chuyn n ADP, tng hp nn ATP. 3-phosphoglyceric acid c bin i thnh 2-phosphoglyceric acid nh enzyme isomerase Nh enzyme hydratase H2O c tch ra t 2-phosphoglyceric acid. Phosphoenolpyruvate c to thnh. Cu trc phn t ca lin kt ny to nn nhm phosphoryl giu nng lng. Nhm phosphoryl ca phosphoenolpyruvate c chuyn n ADP. ATP c tng hp. Enolpyruvate trong 1 trng thi cn bng vi dng ceto ca pyruvic acid (s ).

Hexose (glucose, fructose) cng nh triose (aldehydglyceric) c phosphoryl ho nh kinase, trc khi chng tham gia vo phn ng ca qu trnh ng phn. Ton b qu trnh ng phn c chia lm 3 giai on: 1. Cc carbohydrate c bin i thnh triosephosphate do s oxy ho ng. 2. Triosephosphate c bin i do s oxy ho thnh cc acid hu c (glycerate). 3. Phosphoglycerate c bin i thnh pyruvate. Vi s gii phng ra nng lng t do, ATP c tng hp.

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Tng qut: khi a vo 1 glucosephosphate v kt qu to ra 2 phn t triosephosphate: u vo u ra + 1 glucosephosphate +1ATP + 2 NAD ADP + 2NADH + 2H+ + 2 HS-Enzyme + 2HS- Enzyme +4ATP +2H2O 2 P-OH + 4 ADP + 2 pyruvic acid + Tng qut: glucosephosphate + 2 NAD + 3 ADP + 2 P-OH 2 pyruvic acid + 2 NADH + 2H+ + 3ATP +2 H2O S tng qut ny lm r vai tr sinh l ca qu trnh ng phn hexosephosphate c phn gii thnh apyruvic acid. Nng lng gii phng ra c dng tng hp ATP v kh NAD+. V vy qu trnh ng phn v ngha sinh l tng ng vi phn ng sng ca qu trnh quang hp. Qu trnh quang hp cn nng lng nh sng mt tri tng hp ATP v NADPH, trong khi qu trnh ng phn th nng lng ho hc c s dng tng hp ATP v NADH. S tng hp ATP qu trnh ng phn xy ra trong iu kin ym kh. V vy ngi ta gi phosphoryl ho ym kh, ngha l phosphoryl ho ADP tng hp ATP khng cn O2. 4.5.3. Cc loi h hp Pyruvate c coi l sn phm cui cng ca qu trnh ng phn. N c th ln men ru to ethanol hoc ln men lactic to lactic acid hoc c phn gii qua chu trnh Krebs to CO2 v H2O. s ln men ru trc ht pyruvate c kh cacboxyl ho thnh acetaldehyde, sau di tc dng ca NADH n c kh thnh ethanol.

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NADH c ra trong qu trnh ng phn c s dng oxy ho acetaldehyde, nh vy s ti to NADH l do oxy ho aldehydphosphoglyceric (phn ng 8 ng phn) cung cp cho phn ng ny. Qu trnh ln men ru gm qu trnh ng phn t carbohydrate cho n pyruvic acid, t cht ny n acetaldehyde v ethanol l sn phm cui cng. ng hexose (glucose, fructose) l nguyn liu ban u ca qu trnh ln men, tng qut c biu din nh sau: 1 Hexose + 2ATP fructose-1,6-diphosphate Fructose-1,6-diphosphate 2 aldehydphosphoglyceric 2 aldehydphosphoglyceric + 2 NAD+ 2 pyruvic acid + 4ATP +2NADH 2 pyruvic acid + 2 NADH 2 ethanol + 2 NAD ++ 2 CO2 Cui cng: 1 hexose 2 ethanol + 2 CO2 + 2 ATP Nhiu c quan nh r ca thc vt bc cao, c kh nng thc hin s ln men ru trong iu kin ym kh. i vi cng ngh quan trng l cc men ru v y c bit l Saccharomyces cerevisiae. Saccharomyces cerevisiae l mt loi men k kh khng bt buc, ngha l n c th pht trin trong iu kin c hoc khng c O2. Khi khng c O2 s phn gii glucose xy ra nhanh hn. Ngi ta gi hiu ng ny l hiu ng Pasteur. Pasteur pht hin ra rng, s phn gii ng trong iu kin ym kh xy ra nhanh hn 7 ln so vi iu kin ho kh. Ngy nay chng ta bit rng, hiu ng ny l do s iu khin bin i cu trc (bin cu) ca enzyme fructosephosphatkinase, c m t trn. s phn gii ho kh th ATP c to ra tng i nhiu, v khi nng ATP cao, n c ch hot tnh ca fructosephosphatkinase lm cho s i vo ca glucosephosphat b gim xung. Ngc li khi phn gii ng trong iu kin ym kh, nng AMP cao, n kch thch hot tnh ca fructosephosphatkinase. ADP c to ra nhiu phn ng do kinase xc tc, c bin i thnh ATP v AMP nh sau: 2 ADP ATP + AMP Ln men lactic cng gm qu trnh bin i t carbohydrate n pyruvate, cht ny c kh to thnh lactic acid nh NADH c to ra trong qu trnh ng phn.

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Qu trnh ny c tng qut nh sau: 1 glucose 2 lactic acid + 2 ATP Nhng vi khun lactic quan trng thuc nhng chng loi sau: Streptococcus khng mn cm vi O2 Lactobacillus khng mn cm vi O2 Clostridium ym kh nghim ngt Nhng vi khun streptococus v lactobacillus l khng mn cm vi O2, ngha l chng c enzyme superoxiddismutase, enzyme ny c kh nng lm mt c tnh ca gc O2 . Streptococcus v lactobacillus c kh nng s dng O2 nhng khng phi cho s phn gii ho kh (h hp), v chng khng c kh nng tng hp heme. N khng c cytochrome, l thnh phn quan trng ca h hp. Clostridium sng nghim ngt trong iu kin ym kh, i vi chng O2 l c. Clostridium botulinum tng hp c cht c t nhin mnh nht, l botulinus-toxin. Mt lng rt nh 0.03 ng /kg trng lng c th gy cht. Hu ht nhng vi khun lactic khng mn cm i vi nhit v pH thp. qu trnh ln men lactic nhanh th pH trong mi trng gim nhanh v v nhiu vi sinh vt mn cm vi pH thp, nn chng khng c c hi cnh tranh vi vi khun lactic. iu ny c bit i vi s cnh tranh vi cc vi khun gy thi, nh vy m nng sn c bo v. Khi sn xut thc n chua th s gy chua nhanh nh vi khun lactic l mt iu kin c bn cho thnh cng ca ln men lactic v bo qun. ln men ch yu l lactobacillus plantarum, loi vi khun ny sng trn l cy v bng cch ny m c a vo cng vi nng sn. ln men lactic nhanh iu quan trng l thc vt c carbohydrate d dng b phn gii, c bit l saccharose. Nhng bp ci trng gi tho mn iu kin ny. Thc phm xanh, non th hm lng carbohydrate t, v cy

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thc n giu m th s ln men lactic xy ra chm, pH ch c gim rt t v nhng vi khun khc, nh vi khun butyric c th phn gii thc phm xanh. Vic cho thm mt vo c th hn ch nhc im trn, v mt cha nhiu saccharose v raffinose, nhng cht ny sau khi thu phn c phn gii nhanh bi vi khun lactic. Ng chn sa l nguyn liu ln men l tng, v trong qu ca n cha nhiu carbohydrate d phn gii. Khi sn xut phomat s ln men lactic cng c ngha quan trng. Sa chua c gy ra phn ln l do streptococcus lactis v streptococcus cremonis. Gi tr pH thp dn n s kt ta protein (sa c li), c gi l phomat trng. Phomat ny khng nhng cha protein m cn c cht bo ca sa. Phomat ny l nguyn liu cho vic sn xut cc loi phomat khc nhau. sn xut phomat cn c s tham gia ca nhng vi khun lactic, streptococus, lactobacillus, nhng loi vi khun ny khng nhng to nn lactic acid m cn thu phn protein, ngha l phn gii thnh peptide v aminoacid. Trong phomat lactic acid cn c chc nng bo qun. Nguyn l ca s phophoryl ho ym kh c nghin cu k. Nh hnh 4.22 phn t c cht c bin i cho n nhm phosphoryl ca n dng lin kt giu nng lng. Nhm phosphoryl ny c chuyn n ADP, lm xut hin ATP. V vy ngi ta gi s phosphoryl ho ny l phosphoryl ho c cht. So snh hai giai on ca phn gii carbohydrate (h hp ho kh) th nng lng thu c (ATP) t qu trnh ng phn ch chim khong 20% tng nng lng d tr trong phn t glucose. Phn nng lng ln nht c to ra s giai on phn gii tip theo. Qu trnh ng phn c iu khin bn vng bi 1 enzyme gn nh khi bt u chui phn ng, l enzyme fructosephosphatkinase (phn ng 4 trong s ). cng l enzyme quan trng ca qu trnh ng phn. Enzyme c 4 tiu n v b c ch theo c ch tc ng bin cu bi ATP v citrate, ngc li c hot ho bi AMP. V vy t l AMP/ATP cao kch thch v t l ATP/AMP cao th c ch hot tnh ca enzyme. C ch ny rt c ngha, v khi trong h thng c nhiu ATP th khng cn sn sinh nhiu qua qu trnh ng phn. Ngc li khi t l AMP/ATP cao biu hin s thiu ATP, xy ra s tng hp ATP. Mt nng citrate cao l cht trao i cung cp cho chu trnh Krebs. V qu trnh ng phn cung cp cc cht trung gian cho chu trnh Krebs, nn s iu khin qu trnh ng phn qua hiu qu bin cu ca citrate n fructosephosphatkinase l mt c ch rt c ngha.

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4.5.4 H hp ho kh (chu trinh Krebs) Chu trnh tricarboxylic, hay l chu trnh Krebs c gii thch bi Hans Krebs, v Schueler Otto Warburg. Nh pht minh ny m Krebs nhn c gii thng Nobel vo nm 1953. Chu trnh ny xy ra trong ty th. C quan t ny c th so snh c vi lc lp, thc ra n nh hn lc lp (ng knh 0.5-1m), tuy nhin v cu to v chc nng chng tng t nhau (hnh 4.23).

Hnh 4.23 S cu to ca ty th: hnh pha trn l s ct ngang qua ty th, hnh pha di l ct mt phn ty th

Ngi ta cho rng cng nh lc lp, cc ty th xut hin trc y khong 109 nm trong nhng c th c lp, trong qu trnh tin ho chng c sinh vt nhn chun bt v ho nhp. Nhng t bo c s bin i nng lng ln cha nhiu ty th, nh t bo m c hoc t bo gan, chng c hn 1000 ty th trong mt t bo. Ty th xut hin di knh hin vi l nhng ht nh hoc si. Tuy nhin, ngi ta thy r n di knh hin vi in t, n c bao bc bi mng ngoi v bn trong l mt h thng mng ko dn vi nhiu s un cong vo. S gp khc nhiu ca mng trong lm cho n c din tch b mt ln, din tch ca n ln hn mng ngoi khong 5

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ln. Mng ngoi cho nhng phn t ln thm qua, nhng phn t c trng lng ln cho n 10.000 Da, mng ny cha nhng protein vn chuyn vi nhng knh ln cha nc. Mng trong kh thm qua hn v vn chuyn cc cht c tnh chn lc. Mng trong bao quanh stroma, trong stroma cha cc enzyme ca chu trnh Krebs v enzyme phn gii cht bo (hnh 4.23). Ngc li trn mng trong ty th c cc enzyme v h thng oxy ho. y tng t nh lc lp, chui vn chuyn e- nh v trong mng thylacoid v cc enzyme ca chu trnh Calvin trong matrix. C cht ca chu trnh Krebs l sn phm ca qu trnh ng phn, pyruvate. N c vn chuyn chn lc t t bo cht qua mng trong ty th n stroma v y n c kh carboxyl ho oxy ho nh enzyme pyruvate dehydrogenase. Pyruvate dehydrogenase l mt v d v a enzyme, l mt phc h cha 1 carboxylase, 1 transacetylase v 1 dehydrogenase. Bn cnh trong phn ng cn c cc nhm prostetic l thyaminpyrophosphate, liponic acid, coenzyme NAD+ v Coenzyme A . Sau y l cc bc c th ca chu trnh Krebs. y cng c s kh carboxyl ho oxy ho, phn ng ny khng k trc tip vo chu trnh. Tng t qu trnh ng phn chu trnh Krebs l mt v d ni ln rng trong c th sng cc phn ng phn gii cc cht cha nhiu nng lng thc hin theo tng bc. chu trinh Krebs mi vng quay ly vo chu trnh 1 acetyl CoA. Cc bc ca chu trnh nh di y (hnh 4.24). 1. Pyruvic acid c chuyn thnh acetyl CoA bng phn ng kh carboxyl ho oxy ho. phn ng oxy ho nng lng gii phng ra c lin kt trong NADH v trong lin kt cao nng ca acetyl-CoA.

2. Nng lng cha trong lin kt cao nng ca acetyl-CoA gn acetate vo oxaloacetate to thnh citrate.

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3. Qua s ngng t ny 1 phn t c 6 nguyn t C c to nn. Phn ng ny c xc tc bi enzyme citrate synthetase. Bng phn ng 3 v 4 citric acid c bin i thnh isocitric acid qua aconitic acid.

4. Di tc dng ca aconitase (hydratase) xy s kt hp vi nc. trng thi cn bng citric acid chim 91%, aconitic acid % v isocitric acid 6%.

5. hnh 4.24, phn ng 5 v 6 c thc hin bi enzyme isocitrate dehydrogenase. Trc ht isocitric acid c oxy ho thnh

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oxalosuccinic acid v sau c kh carboxyl ho thnh axit cetoglutaric acid. 6. C hai dng isocitrate dehydrogenase, 1 dng cng vi NAD+ v 1 dng cng vi NADP+. ty th isocitrate dehydrogenase dng NAD+ quan trng hn. 7. Bng phn ng kh carboxyl ho oxy ho - cetoglutaric acid c bin i thnh succinyl-CoA. Enzyme xc tc l 1 phc h cetoglutarate dehydrogenase. Phn ng v enzyme tng t s kh carboxyl ho oxy ho pyruvic acid.

8. T succinyl-CoA succinic acid c to thnh. Lin kt giu nng lng ca succinyl-CoA c b gy, nng lng gii phng ra dng tng hp guanosintriphosphate (GTP) t guanosindiphosphate (GDP) v phosphate v c. Trong c th thc vt GTP c to thnh tng t nh ATP.

9. Di tc dng ca FAD succcinate b oxy ho to thnh fumarate.

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Enzyme ny l mt flavoprotein, c nhm prostetic l FAD c gi l succinate dehydrogenase. s oxy ho succinate thnh fumarate nng lng gii phng khng kh NAD+. Tuy nhin n kh FAD (xem th kh tiu chun). 10. Di tc dng ca H2O, fumarate bin thnh malate

11. Malate dehydrogenase kh malate thnh oxaloacetate, l cht khi u ca 1 chu trnh mi.

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Nhng phn ng c bn trong chu trnh Krebs l s kh carboxyl ho v phn ng oxy ho. Nu tnh phn ng t pyruvate n acetyl-CoA vo chu trnh Krebs th c 3 v tr khc nhau CO2 c gii phng (phn ng 1,6,7) v 5 v tr khc nhau c cht b oxy ho (phn ng 1,5,7,9 v 11). nhng phn ng oxy ho lun lun c 1 coenzyme (NAD+) hoc 1 nhm prostetic (FAD) tham gia. Chng chuyn thnh dng kh bng phn ng oxy ho. Nng lng c gii phng ra do s oxy ho c cht c s dng cho vic kh nhm coenzyme cng nh nhm prostetic. Quan trng l nng lng phn ng 8 c dng tng hp 1 phn t GTP, tng ng vi 1 ATP, v GTP c th phn ng vi ADP nh sau: GTP + ADP ATP + GDP T hnh 4.24 rt ra rng, trong chu trnh, CO2 c tch ra nhng O2 khng c tip nhn vo. Nguyn t O c trong CO2 khng phi l oxy cho h hp. ngha ca chu trnh Krebs l cung cp nhng coenzyme kh (NADH, FADH2), nhng cht kh ny l cn thit cho h hp. Ngoi ra cc acid khc nhau ca chu trnh cn tham gia vo nhng qu trnh trao i cht khc.

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Hnh 4.24 Chu trnh Krebs

Qua chu trnh Krebs chui carbon t carbohydrate n trao i lipid v trao i ca aminoacid v nh vy l n protein.

4.5.5 Chu trnh pentosophoptphate- oxy ho trc tip glucose Bn cnh qu trnh ng phn trong t bo cht cn xy ra mt con ng khc, ngi ta gi l chu trnh pentosephotphate v qu trnh phn ng c phn tng t chu trnh Calvin. Phn ng bt u vi vic oxy ho trc tip glucose-6-phosphate, v vy gi l oxy ho trc tip glucose. Enzyme xc tc cho phn ng u tin l glucose-6-phosphatedehydrogenase, c Otto Warburg pht hin nm 1931. Cc bc oxy ho c ch ra trong hnh 4.26. - Phn ng 1: T glucose-6-phosphate 2 nguyn t H nh C1 c tch ra nh enzyme glucose-6-phosphate dehydrogenase, xut hin dng lactone. Glucose-6-phosphate dehydrogenase c hiu i vi NADP+. Ai lc ca n i vi NADP+ cao hn 1000 ln so vi NAD+.

Hnh 4.25 Chu trnh pentosephosphate (oxy ho trc tip glucose)

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Phn ng 2: Dng lactone c thu phn, to nn 6 phosphogluconic acid. Phn ng 3: 6-phosphogluconate c kh carboxyl ho oxy ho. Enzyme xc tc l 6-phosphogluconate dehydrogenase.

Trong chu trnh ny 2 v tr (phn ng 1 v 3) NADP+ c kh. NADPH c to ra c th s dng cho phn ng tng hp, v d tng hp cc acid bo. Ribulose-1-phosphate khng phi l sn phm cui cng ca chui phn ng ny. N c th c bin i d dng thnh cc dng ng phn nh ribose-5-phosphate hoc xylulose-5-phosphate, theo nh sau. Chu trnh pentosephosphate cung cp nguyn liu cho cc qu trnh tng hp khc. Ribose-5-phosphate tham gia tng hp nucleotide, nh vy l tng hp cc nucleic acid v nhiu enzyme. Xylulose-5-phosphate cng vi ribose-5-phosphate nh phn ng transcetolase to nn sedoheptulosephosphate v aldehydphosphoglycic.

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Transcetolase vn chuyn 1 phn gm 2 nguyn t C t ng ceto n 1 aldose, nhm prostetic ca chng l thiaminpyrophosphate, vng thiazol ca n ng ceto c kt hp vo khi bt u phn ng. Sedoheptulosephosphate phn ng vi aldehydphosphoglyceric trong mt phn ng transcetolase to thnh erythrose-4-phosphate. phn ng ny 1 phn 3 nguyn t C c vn chuyn n aldehydphosphoglyceric.

Fructose-6-photphate l cht trao i ca qu trnh ng phn, c th i vo cc qu trnh khc. Erythrose-4-phosphate phn ng vi xylulose-5-photphate to thnh fructose-6-photphate v aldehydphosphoglyceric.

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Phn ng ny cng vn chuyn 1 phn 2C (transcetolase). Hai sn phm ca phn ng l cht trao i ca qu trnh ng phn, c th c phn gii tip tc trong qua trnh ny. Ton b cc phn ng ca chu trnh pentosephosphate c biu din trong hnh 4.26.

Hnh 4.26. Chu trnh pentosephosphate

Cc bc oxy ho kh u tin dn n 2 v tr kh NADP+ thnh NADPH. Dng kh ca coenzyme cn cho phn ng tng hp v 1 H c i vo cu to 1 cht hu c. NADPH ng 1 vai tr quan trng

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trong tng hp cht bo. Nhng t bo trong acid bo c tng hp nhiu th trao i cht theo chu trnh pentosephotphate xy ra mnh m, v d trong cc m tuyn ca ng vt b sa. Ribulose-5-photphate l tin cht ca ribose-5-photphate, l thnh phn ca nhiu nucleotide v nh vy l ca nucleic acid. c bit nhng m sinh trng, tng hp nucleic acid thng xuyn cn ribosephosphate, y trnh t phn ng ca chu trnh cho n ribulose-5-photphate thc hin vi tc ln. Erythrose-4photphate l tin cht ca cc aminoacid thm (phenylalanine, tyrosine) c s dng tng hp protein c bit trong cc m sinh trng. Khi c nhu cu ln v NADPH th phn ng xy ra vi tc ln cho n qu trnh ng phn. T 6C ca 1 phn t glucose th 5C i vo qu trnh ng phn. Chu trnh pentosephotphate c kh nng thch nghi vi nhu cu sinh l, tng t chu trnh Krebs cung cp cho cc con ng trao i cht khc nhau vi chui carbon hu c v cht kh tng ng.

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CHNG 5

LIPID V S TRAO I LIPID TRONG C TH THC VT

Lipid l nhm cht hu c c cc c tnh ha l ging nhau, chng khng tan trong nc, ch tan trong cc dung mi hu c nh ether, cloroform, benzene, acetone, Khng phi tt c lipid u ha tan nh nhau trong cc dung mi hu c ni trn m mi lipid ha tan trong dung mi hu c tng ng ca mnh, nh c tnh ny ngi ta c th phn tch ring tng loi lipid. V mt ha hc lipid l nhng ester gia ru v acid bo, in hnh l triglycerid. CH2 O CO R1 R1, R2, R3 c th ging nhau, c th khc nhau, c th bo CH O CO R2 ha hoc cha bo ha. CH2 O CO R3 Ngoi ru v cc acid bo, cc lipid phc tp (lipoid), trong phn t ca chng cn cha cc dn xut c phospho, nit, Vai tr ca lipid: - L cht d tr nng lng, khi oxy ha mt gam lipid c th thu c 9,3 Kcal. - Lipid cu t ca t bo cht l thnh phn cu to ca t bo v cha trong t bo vi s lng n nh. Lipid l thnh phn cu trc ca mng t bo, mng ty lp th, Trong mng sinh hc lipid trng thi kt hp vi protein to thnh hp cht lipoproteid. Chnh nh hp cht ny to cho mng sinh hc c c tnh thm thu chn lc. - Lipid di da ng vt c tc dng gi m v gi m cho c th. - Lipid l dung mi cho nhiu vitamin quan trng (nh A, D, E, K). - i vi loi ng vt ng ng, ng vt di c, cc loi su kn, lipid cn l ngun cung cp nc, v khi oxy ha lipid cho mt lng nc sinh ra. - Cc ht cy trng khc nhau c hm lng lipid khc nhau. V d: u tng (20 30%); go (2,2%); ng (4,9%); la m (1,9%); cao lng (3,9%); lc (44 56%); thu du (50 60%).

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I - CU TO V TNH CHT CA LIPID 1.1. Cu to: Du, m c tng hp cc c th sng v ty theo ngun gc m chng c phn ra du thc vt v m ng vt. Glycerine l mt ru c 3 chc, do c th hnh thnh mono; di hay triester. Cc ester ny c bit t lu vi cc tn mono, di v triacylglycerol. Du, m c ngun gc t nhin lun l hn hp cc triglycerid. Cc acid bo ca du, m c ngun gc t nhin u c s nguyn t carbon chn. Bi v cc acid bo u c tng hp t cc n v 2C (gc acetyl). Bn cnh cc acid bo bo ha, mt s acid bo khng bo ha c tm thy trong du, m. Sau y l mt s acid bo bo ha thng gp: - Caproic acid (6C): CH3 (CH2)4 COOH - Caprilic acid (8C): CH3 (CH2)6 COOH - Caprinic acid (10C); lauric acid (12C); miristic acid (14C); panmitic acid (16C); stearic acid (18C); arachidic acid (20C). * Cc acid bo cha bo ha thng gp l: - Oleic acid: CH3 (CH2)7 CH = CH (CH2)7 COOH - Linoleic acid: CH3 (CH2)3 - [CH2 CH = CH]2 (CH2)7 COOH - Linolenic acid: CH3 [CH2 CH = CH]3 (CH2)7 COOH - Eruxic acid: CH3 (CH2)7 CH = CH (CH2)11 COOH

1.2. Tnh cht

a. Nhit nng chy: Ty thuc vo lng acid bo bo ha hay cha bo ha chim u th trong thnh phn ca du m m nhit nng chy khc nhau. Nu trong thnh phn ca du, m c nhiu acid bo bo ha th nhit nng chy cao, nu nhiu acid bo cha bo ha th nhit nng chy thp v trng thi lng. a s du thc vt dng lng nhit thng. b. Ch s acid: l lng mg KOH cn thit trung ha cc acid bo t do c trong 1 gam du, m. Ch s acid cng cao th lng acid bo t do cng nhiu. Cht bo lu ngy, khng bo qun cn thn s c nhiu acid bo t do. Ch s ny cho ta bit c cht lng ca cht bo. c. Ch s x phng ha: l lng mg KOH cn thit trung ha cc acid bo t do v cc acid bo kt hp vi glycerine khi x phng ha 1 gam cht bo. Ch s ny c trng cho phn t lng trung bnh ca glyceride c trong du, m. d. Ch s iod: l s gam iod c th kt hp vi 100 gam du, m.

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Ch s ny dng m t mc khng bo ha ca cc acid bo c trong thnh phn du, m. Ch s iod cng cao th du m cng long, chng cng b oxy ha nhanh hn v cc acid bo b oxy ha d nht v tr cc lin kt i. S gn iod vo acid bo cha bo ha xy ra theo s sau: C=C + I2 CC I I

Ch s iod ca m ng vt dao ng trong khong 30 70; cn ca du thc vt trong khong 120 160. e. S i ha du, m: Du, m lu ngy s c v hi, ng. Nguyn nhn l do tc dng ca O2. Trng hp ny thng xy ra khi du m cha nhiu acid bo cha bo ha. Oxy kt hp vo cc ni i ca acid bo cha bo ha to thnh peroxid: C=C + O2 CC OO Peroxid hoc O2 kt hp vi nguyn t C bn cnh lin kt i to thnh hydroperoxid: C=CC + O2 C=CC O OH Hydroperoxid Peroxid v hydroperoxid c to thnh lp tc b phn gii to thnh aldehyd v cetone l nhng cht c mi v kh chu. II. S PHN GII TRIGLYCERID Du m l nhng cht dinh dng c gi tr nng lng cao, trong ht cc cy ly du vn c t carbohydrate th du l cht d tr chnh v chng l ngun nng lng v ngun vt liu xy dng cho mm ang pht trin. Khi phn gii du ngi ta thy hm lng carbohydrate tng ln.

133

2.1. Phn ng thy phn triglycerid

CH2 O CO R1 CH O CO R2 CH2 O CO R3 Triglycerid + 3H2O lipase CH2 OH CH OH CH2 OH Glycerine + R1 COOH R2 COOH R3 COOH Cc acid bo

2.2. Phn gii glycerine

CH2 OH CH OH (1) CH2 OH Glycerine ATP ADP CH2 OH CH OH CH2 OH NAD NADH2 C=O

(2) CH2 O(P) CH2 O(P) Glycerol(P) (P)dioxiacetone Phn ng 1: do enzyme glycerolkinase xc tc. Phn ng 2: do enzyme dehydrogenase xc tc. - Phosphodioxiacetone di tc dng ca enzyme trioso(P)-isomerase s chuyn thnh aldehydphosphoglyceric. CH2 OH C=O isomerase CH2 O(P) (P)dioxiacetone CH OH CH2 O(P) Aldehyd(P)glyceric Trioso-(P) O CH

* Glycerine c quan h gn vi carbohydrate hoc l c s dng tng hp fructose v carbohydrate khc hoc b phn gii nh carbohydrate. Sau y l s cc ng hng trao i glycerine: Glycerine glycerol(P) (P)dioxiacetone Aldehyd(P)glyceric Fructoso 1,6di(P) Pyruvic acid Acetyl-CoA

Carbohydrate khc

Chu trnh Krebs

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2.3. Phn gii acid bo

Mt acid bo mun c oxy ha phi tri qua mt s phn ng sau: 2.3.1. Hot ha acid bo:Nh h thng enzyme Acyl-CoA-Synthetase, gm 2 bc sau: Bc 1: R CH2 CH2 COOH + ATP Bc 2: R (CH2)2 CO AMP + HS-CoA R CH2 CH2 CO ~ S.CoA Acyl-AMP -AMP Acyl-CoA Qu trnh ny c thc hin ngoi ty lp th (bo tng) 2.3.2. Gn Acyl-CoA vo carnitine to thnh acylcarnitine: Cht ny i qua mng ty th. Trong ty th cc gc acyl ca acid bo c vn chuyn li cho HS-CoA. R CH2 CH2 CO ~ S.CoA Acyl-CoA + (CH3)3 N+ CH2 CH OH CH2 - COOH Carnitine Transferase -HS.CoA (CH3)3 N+ CH2 CH O CO CH2 CH2 - COOH CH2 R Acylcarnitine 2.3.3. To Acyl-CoA tr li: qu trnh ny ngc li bc gn acyl vo carnitine. Carnitine c gii phng v tr li mt ngoi ca ty th. 2.3.4. Qu trnh -oxy ha acid bo: Trong c th sinh vt, s oxy ha acid bo xy ra bng cch oxy ha nguyn t carbon v tr so vi nhm carboxyl, do qu trnh ny cn c gi l qu trnh -oxy ha. Kt qu ca s -oxy ha l tng i nguyn t carbon c tch ra di dng acetyl-CoA v acid bo mi to -H4P2O7 R CH2 CH2 CO AMP Acyl-AMP

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thnh c mch carbon ngn hn trc 2 nguyn t carbon. S oxy ha khng th t xy ra, c th tham gia phn ng, acid bo phi c hot ha nh nng lng ca ATP, nhng y nng lng chuyn t ATP ti cht bo khng thng qua con ng phosphoryl ha nh trong trng hp oxy ha glucose m thng qua s to thnh hp cht acyl-CoA. Qu trnh -oxi ha c Knoop (ngi c) a ra 1904, qu trnh ny xy ra trong gian bo ty th. Phn ng tng qut ca s to thnh acyl-CoA nh sau: Acyl-CoA-synthetase R CH2 CH2 COOH + HS-CoA ATP (1) R CH2 CH2 C ~ S.CoA AMP + H4P2O7 O Acyl-CoA

Sau acyl-CoA b oxy ha bi enzyme acyl-CoA-dehydrogenase c nhm hot ng l FAD: Acyl-CoA-dehydrogenase R CH2 CH2 C ~ S.CoA O Acyl-CoA FAD (2) FADH2 R CH = CH C ~ S.CoA O Enoyl-CoA

Di tc dng ca enzyme Enoyl-CoA-hydratase, phn t H2O kt hp vo ni i v to thnh -oxyacyl-CoA: Enoyl-CoA-hydratase R CH = CH C ~ S.CoA + H2O O Enoyl-CoA (3) R CH CH2 C ~ S.CoA OH O -oxyacyl-CoA

* -oxyacyl-CoA li b oxy ha ln th 2 di tc dng ca enzyme -oxyacyl-CoA-dehydrogenase c coenzyme l NAD to thnh -cetoacyl-CoA:

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NAD R CH CH2 C ~ S.CoA OH O

(4)

NADH2

-oxyacyl-CoAdehydrogenase

R C CH2 C ~ S.CoA O O -cetoacyl-CoA

* -cetoacyl-CoA li phn ng vi HS-CoA to thnh acyl-CoA mi v acetyl-CoA di tc dng ca enzyme -cetoacyl-CoA-thiolase: (5) R C CH2 C ~ S.CoA + HS-CoA O O -cetoacylCoA-thiolase R C ~ SCoA + CH3 C ~ S.CoA O Acyl-CoA(mi) O Acetyl-CoA

Acyl-CoA mi to thnh ny cha gc acid bo c t hn acid bo ban u 2 nguyn t carbon, n li c th tip tc tham gia cc phn ng 2, 3, 4, 5 to thnh acyl-CoA mi c mch carbon ngn hn 2 nguyn t carbon v tch ra mt phn t acetyl-CoA na v qu trnh -oxy ha c tip tc lp li nhiu ln cho n khi ton b phn t acid bo c chuyn thnh cc acetyl-CoA. S -oxy ha c Lynen m t di m hnh xon c. Acetyl-CoA to thnh do kt qu ca s -oxy ha acid bo c th i vo chu trnh Krebs, chu trnh glyoxilate hoc tham gia vo nhiu phn ng khc.

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Chu trnh Krebs

R CH2 CH2 COOH ATP HS-CoA AMP 1) H4P2O7 R CH2 CH2 C ~ S.CoA

CH3 C ~ S.CoA O Acetyl-CoA O Chu trnh glyoxilate

Acyl-CoA O 2)

FAD FADH2 R CH = CH C ~ S.CoA O Enoyl-CoA

Acyl-CoA R C ~ S.CoA 2C 2C 2C 2C HS-CoA 5)

R C CH2 C S.CoA O O -cetoacyl-CoA NADH2

3) H2O 2CH3 C ~ S.CoA Acetyl-CoA O R CH CH2 C ~ S.CoA 4) NAD OH O -oxyacyl-CoA

Hnh 5.1 - S ca s -oxy ha acid bo (theo Lynen) Hiu qu nng lng trong qu trnh -oxy ha acid bo: V d -oxy ha hon ton palmitic acid (16C) t bo thu c ngun nng lng nh sau: 7 vng quay to ra 7FADH2 v 7NADH2 v c 8 phn t acetyl-CoA. 7FADH2 7NADH2 7 x 2ATP = 14ATP 7 x 3ATP = 21ATP Tng cng : 35ATP

Nhng v phi tiu tn 1 ATP hot ha acid bo nn s ATP to ra khi -oxy ha palmitic acid l: 35ATP 1ATP = 34ATP. Nu 8 phn t acetyl-CoA i vo chu trnh Krebs s to ra: 8 x 12ATP = 96ATP Tng cng: 34ATP + 96ATP = 130ATP.

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Ta c th p dng cng thc tnh sau: n n A = 5 1 1 + x12 2 2 *---------------* * -------* (-oxy ha) (Krebs) A l s phn t ATP. n l s nguyn t C (n: chn).

III. S PHN GII CC ACID BO CHA BO HA

i vi acid bo c mt hoc nhiu lin kt i nh oleic acid, linolenic acid qu trnh -oxy ha din ra bnh thng, cc phn t acetyl-CoA c tch dn ra, cho ti gn mch kp. Ti y, ty theo v tr ca mch m s phn gii c th c cc ng hng sau: 3.1. nhng acid bo cha bo ha m c lin kt i khng gia C v C th nh tc dng ca enzyme isomerase y lin kt i v ng gia v tr C v C sau chu s -oxy ha. Cng c nhng th nghim chng minh rng: nhng acid bo cha bo ha m c lin kt i ng v tr gia C v C th n chu s -oxy ha bnh thng. 3.2. Cc acid bo cha bo ha trc khi b oxy ha th chng chuyn thnh acid bo ha bng cch gn thm H2 v sau li chu s -oxy ha nh bnh thng.

IV. S PHN GII CC ACID BO C S NGUYN T CARBON L

S phn gii cc acid bo ny vn tin hnh bnh thng bng s -oxy ha cho n khi hnh thnh CH3 CH2 C ~ S.CoA (propionyl-CoA), O sau propionyl-CoA ( ty th thc vt v vi sinh vt) giai on u ging nh s -oxy ha cho n khi hnh thnh -hydroxipropionyl-CoA. Sau nguyn t C b oxy ha 2 ln (cht nhn H2 l NAD v NADP) to malonyl-CoA. Sau malonyl-CoA b kh carboxyl ha to acetyl-CoA. Cht ny c th i vo chu trnh Krebs hoc i vo chu trnh glyoxilate.

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Hnh 5.2 - S ca s phn gii acid bo c s nguyn t carbon l:

-oxy ha Acid bo c C l CH3 CH2 C ~ S.CoA O Propionyl-CoA (1) FAD FADH2 CH2 = CH C ~ S.CoA H2O (2) O Propionolyl-CoA

CH2 CH2 C ~ S.CoA OH (3) O -hydroxipropionyl-CoA NAD NADH2 CHO CH2 C ~ S.CoA O Aldehyd malonyl-CoA H2O NADP

(4)

NADPH2 HOOC CH2 C ~ S.CoA O (5) CO2 Malonyl-CoA CO2

KREBS

CH3 C ~ S.CoA O Acetyl-CoA

H2O

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V. CHU TRNH GLYOXILATE

So vi chu trnh Krebs th chu trnh Glyoxilate khng c ngha my v mt nng lng v phi tiu tn mi vng mt 2 phn t Acetyl-CoA. ngha ca chu trnh ny l s to thnh succinic acid, ngha l c s chuyn ha cht bo thnh carbohydrate. V t succinic acid s hnh thnh nn oxaloacetic acid v theo qu trnh ngc ng phn s to nn carbohydrate. Ngoi ra t chu trnh ny s hnh thnh glyoxilic acid. Bng cch kh carboxyl ha acid ny s chuyn thnh aldehyd formic l cht tham gia vo hng lot qu trnh trao i cht khc. T glyoxilic acid s tng hp c glycine tham gia to nn protein. Hnh 5.3 - S chu trnh Glyoxilate Acetyl-CoA H2O Oxaloacetic acid (1) NADH2 NAD (6) Malic acid (2) H2O HS.CoA Citric acid

Cis-Aconitic acid (3) H2O

HS-CoA Acetyl-CoA

(5)

CH2 COOH CH COOH CH.OH.COOH Isocitric acid (4) CH2 COOH CH2 COOH Succinic acid Carbohydrate

H2O

O C

H COOH Glyoxilic acid

Phn ng 1: Enzyme citratesynthetase xc tc. Phn ng 2: Enzyme aconitate-hydratase xc tc.

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Phn ng 3: Enzyme aconitat-hydratase xc tc. Phn ng 4: Enzyme isocitrat-liase xc tc. Phn ng 5: Enzyme synthetase xc tc. Phn ng 6: Enzyme malatdehydrogenase xc tc.

VI. SINH TNG HP TRIGLYCERID

Do khng tan trong nc nn du khng th vn chuyn c trong cy. V vy thc vt vic tng hp cht bo (du) phi tin hnh ti ch trong tt c cc c quan ca thc vt. Nguyn liu tng hp cht bo l t nhng cht ha tan c a vo cc c quan ny. D ng vt hay vi sinh vt , thc vt, cht bo u c to thnh t carbohydrate. Trong c th cht bo c to thnh t glycerol(P) v cc acid bo, bi vy mun tng hp c cht bo trc ht phi tng hp c 2 cu t ny.

6.1. Tng hp glycerolphosphate

Nhng cht ban u to thnh glycerolphosphate l aldehyd(P)glyceric hoc (P)dioxiacetone. Nhng cht ny c hnh thnh trong qu trnh quang hp (sn phm ca chu trnh Calvin) hoc t s phn gii carbohydrate m c (v d t qu trnh ng phn hoc t chu trnh Pentoso(P)). C ch hnh thnh glycerol(P) nh sau: O CH2 OH C=O isomerase CH2 O(P) Phosphodioxiacetone CH Trioso(P)CH OH NADH2 NAD Dehydrogenase CH2 O(P) Aldehyd(P)glyceric CH2 - OH CH - OH CH2 O (P) Glycerol(P)

6.2. Tng hp acid bo theo vng xon Lynen-Wakil

- Acid bo c tng hp t acetyl-CoA trong bo tng. - Qu trnh ny c s tham gia ca phc hp multienzyme, cu trc ca phc hp ny nh sau: + gia l phn t protein-ACP (acyl-carrier-protein) khng c hot tnh enzyme, c nhim v mang nhm acyl trong qu trnh tng hp acid bo. ACP u tin c phn lp t E.coli do 77 gc aminoacid to nn. ACP c coi nh mt ci nhn nm chnh gia 6 phn t enzyme

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ca phc hp multienzyme. ACP gn vi mt phn t pantoten (vitamin G) to thnh mt b phn hot ng ging nh b phn hot ng ca CoA c nhim v mang nhm Acyl trong qu trnh sinh tng hp acid bo. Gc acyl trung gian c mc vo ACP qua lin kt thioester vi nhm SH ca protein. Gc phosphopantoten hot ng c v nh cnh tay di ng vn chuyn gc acyl t enzyme ny n enzyme khc. - Phc hp multienzyme c 2 nhm SH + Nhm SH ca ACP(SH-gia) + Nhm SH ca phn t enzyme(SH-bin) sinh tng hp acid bo cn nhiu phn t acetyl-CoA t cc ngun khc nhau v tr mt acetyl-CoA ban u i vo dng khng i, cn tt c nhng acetyl-CoA sau u phi di dng carboxyl ha l malonyl-CoA. HOOC CH2 C ~ S.CoA O Theo Lynen-Wakil, qu trnh tng hp acid bo xy ra qua ba giai on nh sau: * Giai on 1: Hot ha acetyl-CoA. T acetyl-CoA to thnh malonyl-CoA vi s tham gia xc tc ca carboxylase cng vi s c mt ca ATP v Mn+2. ATP carboxylase CH3 C ~ S.CoA + CO2 O Acetyl-CoA Biotin, Mn+2 ADP+H3PO4 HOOC CH2 C ~ S.CoA O Malonyl-CoA

* Giai on 2: Ni di chui carbon trong acid bo: Bc 1: Gn acetyl-CoA u tin vi ACP hnh thnh acyl-ACP SH-bin ACP-acyl transferase CH3 C ~ S.CoA + E.ACP O SH-gia E.ACP HS.CoA S C CH3 O SH-bin

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SH-bin ACP-acyl transferase E.ACP S C CH3 O E.ACP

S C CH3 O SH-gia Acyl-ACP

Bc 2: Acyl-ACP kt hp vi malonyl-CoA hnh thnh diacyl-ACP. S C CH3 E.ACP O + HOOC CH2 C ~ S.CoA SH-gia O Malonyl-CoA E.ACP S C CH3 O + CO2 + HS-CoA

S C CH3 O Diacyl-ACP

Bc 3: Ngng t diacyl-ACP thnh -cetoacyl-ACP. S C CH3 E.ACP O S C CH3 O SH-bin -cetoacyl-ACP-synthetase E.ACP S C CH2 C CH3 O O

Bc 4: -cetoacyl-ACP b kh nh dehydrogenase c coenzyme l NADPH2 hnh thnh -oxybutiryl-ACP. SH E.ACP dehydrogenase S C CH2 C CH3 O O NADPH2 NADP E.ACP S C CH2 CH CH3 O OH -oxybutiryl-ACP 144 SH

Bc 5: Loi nc nh enzyme hydratase hnh thnh Crotonyl-ACP. SH-bin E.ACP H2O Hydratase S C CH2 CH CH3 O OH E.ACP S C CH = CH CH3 O Crotonyl-ACP SH-bin

Bc 6: Crotonyl-ACP b kh nh enzyime dehydrogenase c coenzyme l FMNH2 hnh thnh Butiryl-ACP. SH-bin E.ACP Dehydrogenase S C CH = CH CH3 O FMNH2 FMN E.ACP S C CH2 CH2 CH3 O Butiryl-ACP SH-bin

* Phn ng hon nguyn ca FMN: FMN + NADPH2 FMNH2 + NADP + Khi hnh thnh butiryl-ACP, qu trnh li tip tc nhn mt phn t malonyl-CoA v tip tc nh trn. Mi vng quay li ni di thm mt cp carbon cho n khi phn t acid bo c di p ng vi phn t acid bo m t bo cn tng hp. * Giai on 3: Kt thc tng hp acid bo: Giai on ny nh s xc tc ca enzyme acyltransferase. SH-bin E.ACP S C (CH2)n CH3 Acyl-ACP O HS-CoA acyltransferase CH3 (CH2)n C ~ S.CoA O Acyl-CoA 145 E.ACP SH-gia SH-bin

Hnh 5.4 - S SINH TNG HP ACID BO THEO LYNEN-WAKIL

SH-bin E.ACP NADPH2 S CO (CH2)2 CH3 Butiryl-ACP CH3 CO ~ S.CoA Acetyl-CoA SH-bin E.ACP SH-bin NADP FMNH2 E.ACP HS-CoA SH-gia SH-bin E.ACP S CO CH = CH CH3 Crotonyl-ACP H2O SH-bin E.ACP S CO CH2 CH CH3 OH -oxybutiryl-ACP NADP NADPH2 E.ACP S CO CH2 C CH3 O -cetobutiryl-ACP SH-bin E.ACP SCOCH3 diacyl-ACP CO2 SCOCH3 E.ACP S COCH3 SH-gia SCOCH3 CH2 CO ~ S.CoA Malonyl-CoA Acetyl-CoA CH3 CO ~ S.CoA ATP ADP + H3PO4 COOH CO2

FMN

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6.3. Tng hp triglycerid

Qu trnh tng hp acid bo ni trn to ra cc acid bo di dng cc dn xut acyl-CoA. Trong t bo c sn glycerol(P). Qu trnh gn acid bo c thc hin tng phn t ln lt v to thnh cc sn phm mono, diglycerid(P) v cui cng mi to triglycerid. CH2 OH CH OH CH2 O (P) Glycerol(P) + 2R C ~ S.CoA (1) O Acyl-CoA CH2 O (P) Diglycerid(P) (2) H2O H3PO4 CH2 O CO R CH O CO R HS-CoA CH2 O CO R3 Triglycerid (3) R3 C ~ S.CoA O Acyl-CoA CH2 O CO - R CH O CO - R CH2 OH Diglycerid CH2 O CO R CH O CO R

2HS.CoA

VII. SINH TNG HP V PHN GII PHOSPHOLIPID

Phospholipid l cc diglycerid lin kt vi phosphoric acid bng lin kt ester v tr nhm hydroxyl th 3. Sau gc phosphoric acid b ester ha bi ru cha nhm amin (choline, ethanolamine,). Cng thc tng qut ca phospholipid nh sau: CH2 O CO R1 CH O CO R2 OH CH2 O P = O O (X) (X) hp cht ph R1; R2 gc acid bo.

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7.1. Sinh tng hp phospholipid

Nguyn liu cn c l: glycerine; cc acid bo v cc hp cht ph(x)-Qu trnh tng hp i hi chi ph mt lng nng lng do ATP v CTP cung cp. V d: Sinh tng hp phosphatidyl-choline (tn c l lecithin). S sinh tng hp nh sau: Glycerol(P) R1 CO ~ S.CoA Choline: (CH3)3N CH2 CH2 OH ATP OH ADP Choline(P) CTP H4P2O7 Cytidyldi(P)choline

2HS-CoA

R2 CO ~ S.CoA Diglycerid(P) H2O H3PO4 Diglycerid CMP CH2 O CO R1 CH O CO R2

CH2 O (P) O Choline Phosphatidyl-Choline(lecithin)

7.2. Phn gii phospholipid (V d phn gii lecithin)

Qu trnh phn gii phospholipid xy ra mnh m khi ht ny mm, s phn gii chng bng cch thy phn vi s tham gia ca H2O di tc dng ca nhm enzyme c gi l phospholipase. u tin phospholipase A xc tc cho s loi b bng cch thy phn acid bo khi nguyn t C2 ca glycerine; sau phospholipase B loi acid bo khi nguyn t C1 ca glycerine. Phospholipase C tch phosphoryl-Choline. Cn phospholipase D thy phn Cholinephosphate. Kt qu ca s tc dng hp nht ca cc phospholipase l to thnh cc acid bo; glycerine t do; H3PO4 v Choline.

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S c trnh by nh sau: Phospholipase B CH2 O C R1 O Phospholipase A CH O C R2 O OH CH2 O P = O O CH2 CH2 N(CH3)3 Phospholipase C OH Phospholipase D

------o0o------

149

TI LIU THAM KHO

1. V Kim Bng ng Hng L Khc Thn: Bi ging ha sinh NXBH v GDCN H ni 1991. 2. L Don Din: Ha sinh thc vt NXBNN H ni 1993. 3. ng Hng V Th Th: Ha sinh hc cy trng nng nghip NXBNN H ni 1993. 4. V Vn Quang: Bi ging sinh ha thc vt i hc Nng lm Hu 1996. 5. Hong Vn Tin L Khc Thn L Don Din: Sinh ha hc vi c s cng ngh gen NXBNN H ni 1997. 6. Nguyn Duy Minh: Nhng bi tp trc nghim chn lc v tr li sinh ha NXB i hc Quc gia H Ni 2001. 7. Nguyn nh Huyn Mai Xun Lng: Ha sinh hc hin i theo s NXB y hc TP HCM 1983. 8. Hong Minh Tn - Nguyn Quang Thch: Cht iu ha sinh trng i vi cy trng NXBNN 1993. 9. Nguyn Vn Uyn: Phn bn l v cc cht kch thch sinh trng NXBNN TP HCM 1995. 10. Stryer L. Biochemistry NY 1987. 11. BreskiW. Practikum z Biochemi Warszawa 1971.

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Chng 6

NUCLEIC ACID V S CHUYN HO CA NUCLEIC ACID

6.1 Cu trc ca nucleic acid Nucleic acid c vai tr quan trng trong sinh hc. Chng l nhng cht mang thng tin di truyn v chuyn nhng thng tin di truyn ny n trao i cht. Cu to ca chng gm nhiu n phn c ba thnh phn: bas nit, phosphate v ribose hoc desoxyribose. Base nit l dn xut ca pyrimidine hoc purine. Pyrimidine c tng hp t carbamylphosphate v asparagine. Purin c tng hp t asparagin, glycine, glutamine, CO2 v formiate c hot ho bi tetrahydrofolic acid.

Adenine v guanine l nhng dn xut ca purine. Cytosine, thymine v uracil l dn xut ca pyrimidine.

trng thi cn bng nguyn t H ca nhm hydroxyl dch chuyn n nguyn t N.

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 dng ny nguyn t H ng cnh nguyn t N c th to mt lin kt N-glycosid vi mt ribose.

Ribose l thnh phn ng ca ribonucleic acid (RNA). Desoxyribose l thnh phn ng ca desoxyribonucleic acid (DNA). Desoxyribose khc vi ribose ch v tr carbon th 2: desoxyribose c 1 nguyn t H thay cho nhm OH. Desoxyribose c to nn t ribose sinh vt nhn chun bng phn ng kh nh enzyme nucleosidiphosphate-reductase, phn ng kh c thc hin nh 2 nhm SH ca enzyme. Phn ng tng qut c biu din nh sau:

ng (ribose, desoxyribose) lin kt vi base nit bng lin kt glycoside. Ngi ta gi chng l nucleoside. S biu din nhng base nit quan trng nht v nucleoside nh sau: Base nit Nucleoside Adenine Adenosine

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Guanine Guanosine Cytosine Cytidine Thymine Thymidine Uracil Uridine S biu din cc nucleotide v nucleotidphosphate cng theo quy lut ny, v d: thymidinmonophosphate hoc adenosintriphosphate. nucleoside nguyn t H ca nhm OH nh v tr carbon th 5 c thay th bng nhm phosphate, nh vy ta c mt nucleotide. Nucleoside: Ribose- Base nit Nucleotide: Phosphate - Ribose - Base nit ng lin kt vi base nit bng lin kt glycoside, vi phosphate bng lin kt ester. nucleotide ny, mt nucleotide th 2 kt hp vo, y H ca nhm OH nh v tr carbon th 3 c thay th bng mt nucleotidyl, c biu din nh sau:

Nguyn t H ca nhm hydroxyl v tr C3 ca nucleotide th hai c thay th bng mt nucleotidyl tip theo, ngha l chui ko di theo hng ca ribose v tr C3. Nucleotidyl l gc ca mt nucleotide, l nucleosidphosphoryl.

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Hnh 6.1

Cu to chung ca mt ribonucleic acid

Theo nguyn tc ny c th to nn mt chui rt di. mt u n mang mt nhm phosphate v u kia mt nhm OH t do ca ng ribose. hnh 6.1 biu din cu trc ca 1 si n ca RNA, y n l mt s c ln 102 n104 . Nucleic acid l mt polyme, c to nn rt h thng, vi mt nhm phosphate mt u v mt nhm OH v tr carbon th 3 ca ribose hoc desoxyribose u kia. Gc phosphate ca nucleic acid cn cha mt nhm OH t do, nhm ny c kh nng phn ly H+. V vy kha cnh ny phn t c tnh acid v ngi ta gi l nucleic acid. Cc base nit, cng vi mch bn ngn ca nhng si rt di c kh nng kt hp vi proton mt i in t n c ca cc nguyn t nit ca chng. Trng hp ny phn t c tnh kim. Cc si n ca DNA c to nn tng t RNA. Trnh t ca cc base nit ring l v thnh phn ca n trong ton b phn t l nhng tiu chun quan trng nht ca nucleic acid. DNA cha ch 4 base nit khc nhau: adenine, thymine, guanine v cytosine. Trnh t ca chng trong phn t DNA gn lin vi ton b yu t di truyn ca sinh vt. Sinh vt cng pht trin cao th si DNA ca chng cng di v trng lng cng ln. Trng lng phn t ca DNA ca vi sinh vt khong 109, ca ngi khong 3,8. 1010. C khong hng nghn nucleotide trn mt phn t. Phn t di nht c bit c 108 nucleotide, tng ng vi di l 3mm. Bnh thng th phn t DNA gm 2 si chy ngc chiu nhau v qun vo nhau. Hai si chy i song v ngc chiu nhau vi theo hng t C5 n C3. Nhng base nit hng v trc helix c bo v trc nhng nh hng bn ngoi nh chui ribosephosphate. Tng t polypeptide cc phn t DNA c cu trc bc mt l trnh t cc nucleotide v cu trc bc hai l s xon ca mch kp. Trn mt vng xon c 10 base

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nit. y mi base nit c mt base i din xc nh. Adenine i din vi thymine, guanine vi cytosine. S i din ny l do cc cu hydro c to nn gia dn xut pyrimidine (thymine, cytosine) v dn xut purine (adenine, guanine). Adenine ni vi thymine bng 2 lin kt hydro v guanine vi cytosine l 3 lin kt ( hnh 6.3). Watson v Krick a ra m hnh DNA gm 2 si b sung. M hnh ny c ngha i vi cc qu trnh sinh hc c bn s c gii thch r hn di y. Lin kt gia cc base b sung thc ra l khng bn vng nhng rt chn lc. Chng to iu kin cho si b sung c th c tng hp t mt si ring l. Hai si ny ging nh nh v phim.

Hnh 6.2

S biu din mt phn t DNA: a) mch thng, b) mch xon kp, Bn phi cc si chy theo hng i song C5 C3

Hnh 6.3 S cp i ca adenine v thymine bng 2 lin kt hydro v ca guanine v cytosine bng 3 lin kt hydro

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Hnh 6.4 S biu din cu trc ba chiu ca desoxyribosephosphate c base nit l thymine v adenine

Mch kp ca DNA c ng knh khong 2 nm, di c th n nhiu mm. DNA ca Escherichia coli l mt si c 4,2 .106 cp base. Mch kp ca DNA l tay phi ngha l nhng vng xon theo hng quay ca kim ng h, to nn rnh ln v nh. Rnh ln hay nh l nhng im tc ng c bit ca enzyme. Enzyme desoxyribonuclease I (DNAase I) kt hp tnh in vng xon nh, y c bit nhm NH2 tch in dng ca arginine v lysine ca enzyme kt hp vi nhm phosphate tch in m ca DNA. DNAase I l enzym thu phn, c kh nng thu phn tch ra mt trong hai si. Nhng enzyme endonuclease kt hp vi mt trnh t xc nh c 6 base nit, thu phn hai mch ca si kp. Nhng nuclease ny l nhng enzyme gii hn, ng mt vai tr quan trng trong cng ngh sinh hc.

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Hnh 6.5 a) Mch kp DNA vi rnh ln v rnh nh b) Xon "tay phi", mi tn ch hng xon theo kim ng h

Dng B-DNA c th chuyn sang dng A-DNA bng s kh nc, chng cng xon theo hng kim ng h. Nhng nhm phosphate ca chng hp th nhng phn t nc t hn dng B-DNA. Dng A-DNA rng hn v ngn hn dng B-DNA. 6.2 Phn loi nucleic acid Bng 6.1 Gii thiu cc nucleic acid K hiu Ngun Trng lng phn t DNA RNA mRNA rRNA tRNA cRNA nhn, ty th, lp th nhn, t bo t bo cht, ribosome t bo cht nhn 109-3,8 x 1010 108 108 25 000 106-107 S lng nucleotide 4 x 106-108 4 x 103 4 x 103 80 -100 103

ribonucleic acid c 4 loi khc nhau (bng 6.1). mRNA chuyn thng tin di truyn t DNA n trung tm tng hp ca t bo. Trng lng phn t khong 106, bng 1/1000 n 1/10 000 trng lng ca DNA, v vy n c di ngn hn. Nhng base nit quan trng nht l guanine, cytosine, adenine v uracil. Ngc li DNA cha base nit thymine thay cho uracil v b sung cho adenine. mRNA c tng hp trong nhn v mc nht nh c tng hp trong ty th v lp th. rRNA l RNA ribosome, l ni tng hp protein ca t bo. Thnh phn ribosome gm c cc phn t protein v cc rRNA. Cc rRNA c trng lng phn t khong 106

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Da. Bn cnh adenine, uracil, guanine v cytosine rRNA cn cha nhng base nit khc m phn ln c methyl ho. Nhng rRNA ca thc vt c methyl ho nhiu hn rRNA ca ng vt. Khong 80% RNA tng s ca mt t bo l rRNA, phn cn li l tRNA, trong khi mRNA v cRNA ch chim mt thnh phn nh. tRNA l RNA vn chuyn aminoacid n trung tm tng hp protein. tRNA ch c khong 80 nucleotide, v vy c trng lng phn t thp, ch khong 25 000 Da. Bn cnh 4 base nit k trn chng cn cha nhng base nit khc c methyl ho. V l do ny m n khng th tn ti trng thi mch kp hon chnh. Trong phn t ca n c nhng on khng kt hp vi nhau, hnh thnh nn cu trc l c tam ip (c ba nhnh), cu trc ny rt quan trng i vi chc nng sinh l ca chng (hnh 7.20). RNA chromosome, ngi ta cn gi l hnRNA (heterogen nuclear RNA) ch c sinh vt nhn chun. l dng tin cht ca cc loi RNA. 6.2 Sinh tng hp nucleic acid 6.2.1 Sinh tng hp nucleotide dng purine Qu trnh sinh tng hp nucleotide dng purine c bt u t -D-ribose 5-P, sn phm trung gian ca chu trnh pentosephosphate. Cc nguyn t ca base c ngun gc t nhng hp cht khc nhau v c gn ln lt vo gc ribose.

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Hnh 6.6 Cc phn ng tng hp purine nucleotide

Ribose 5-P c hot ho bng vic tip nhn mt nhm pyrophosphate, to nn 5phosphoribosyl-1-1pyrophosphate (PRPP). Sau nhm pyrophosphate ny c thay th bng mt nhm amin c ngun gc t glutamine, cho sn phm l 5phosphoribosylamine. Phn ng ny c thc hin do s thu phn pyrophosphate. ng thi ribose dng chuyn sang dng . Enzyme ribose-P-pyrophosphokinase v amido-P-ribosyltransferase xc tc cho hai phn ng, l nhng im tip cn ca cc qu trnh iu khin. Phn ng tip theo l gn thm glycine trc tip vo nhm NH2, c thc hin do thu phn ATP. Sau N10-formyltetrahydrofolat gn vo mt nhm formyl. Trong mt phn ng ph thuc vo ATP v glutamine nhm oxo C-2 c thay th bng mt nhm imin. ATP cung cp nng lng cho vic ng vng v bin i vng imidazol thnh vng thm 1(5-phosphoribosyl)-5-aminoimidazol (AIR). Mt phn ng carboxyl ho xy ra khng c biotin, tip nhn CO2 to nn dn xut carboxy. Phn t ny gn vi aspartate bng mt lin kt amide. Sau khung aspartate- carbon c gim tr li, gii phng fumarate. Aspartate a vo cht to thnh 1(5-phosphoribosyl)-5amino-4-imidazol-carboxamid (AICAR) ch nhm amine ca n. S formyl ho tip theo bi N10-formyltetrahydrofolate cung cp thnh phn cui cng ca vng 6 cnh. Vng c ng li bng cch loi H2O, xut hin inosin-5-phosphate. IMP khng tch lu trong t bo, n c bin i nhanh thnh adenosine- v guanisinemononucleotide.

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6.2.2

Sinh tng hp nucleotide dng pyrimidine

Sinh tng hp uridin-5phosphate (UMP)

232

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Hnh 6.7 S tng hp uridine-5phosphate (UMP)

s tng hp purine thnh phn ca vng purine tng bc c gn vo nhm ribose, cn s tng hp pyrimidine trc ht xut hin vng base, sau ribose c gn vo. Nhng nguyn t ca vng c gn vo t aspartate v carbamylphosphate. sinh vt nhn chun carbamylphosphate l mt thnh phn ca chu trnh ornithine, nhng hai ng hng phn ng c tch ra trong khng gian. s tng hp pyrimidine, carbamylphosphate c to nn trong t bo cht (nhng phn ng tip theo cng xy ra y), phn ng tng ng ca chu trnh ornithine xy ra ty th. Carbamyl-synthase tham gia vo hai qu trnh c nhng c im khc nhau: Synthase II tham gia vo tng hp pyrimidine s dng glutamine l ngun nit v khng c hot ho bin cu bi N-acetylglutamate, synthase ca chu trnh ornithine phn ng v c hot ho bi NH3. vi khun ch c carbamyl-synthase typ II. Phn ng tip theo c xc tc bi enzyme aspartate-carbamyl-transferase, l bc xc nh ca sinh tng hp pyrimidine, y c tc ng ca c ch iu khin. Bc tip theo l ng vng v gii phng H2O. S oxy ho tip theo to thnh orotate. sinh vt nhn chun dihydroorotate-oxidase, nh v pha ngoi ca mng ty th, chuyn cht kh n quinone. vi khun enzyme gn kt vi mng v phn ng vi cc quinone. Phn ng vi 5-phosphoribosylpyrophosphate (PRPP) dn n s kt hp vi ribose, PPi c gii phng ra. Bc cui cng ca sinh tng hp uridin-5-P l kh carboxyl ho orotidine-5-P. Pyrimidine-mononucleotide c bin i thnh di- v triphosphate bng s phosphoryl ho. Uridin-5-triphosphate (UTP) c amine ho bng glutamine ( ng vt c v) hoc NH4+ ( E.coli) trong mt phn ng cn ATP, to thnh cytidin-5-triphosphate (CTP). Phn ng amine ho ny tng t phn ng bin i ca xanthosin-5phosphate, tuy nhin Pi c gii phng thay v PPi. 6.3 Sinh tng hp DNA: DNA c kh nng t tng hp (t nhn i)

cho qu trnh sinh tng hp DNA xy ra cn phi c cc iu kin sau y - C DNA lm khun - C mt 4 loi desoxyribonucleotide di dng triphosphate (Nguyn liu cho qu trinh tng hp DNA l nhng desoxynucleotidtriphosphate) Desoxyadenosintriphosphate = dATP

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Desoxyguanintriphosphate = dGTP Desoxythymidintriphosphate = dTTP Desoxycytosintriphosphate = dCTP Chng l nhng cht tng t ATP, GTP, TTP v CTP v cha desoxyribose thay cho ribose. : - C cc enzyme DNA-polymerase xc tc Nhng enzyme quan trng nht l DNA- polymerase, DNA-ligase v mt helicase. Khun l hai si ca mch xon kp. S tng hp bt u bng vic thu phn lin kt hydro ca mch kp nh mt enzyme nuclease, tch lin kt ester gia ribose v gc phosphate ca hai si, nh vy t v tr ny mch kp c th c dui xon. V mt nng lng y khng phi l qu trnh n gin v nh trn cp hai si c ni vi nhau bng cc cu hydro. Trong ng nghim ngi ta ct lin kt ny khi a nhit ln 90 0C. dui xon cn mt phc h enzyme c bit l helicase, l mt phc h nh v ch ch ba ca mch kp v lm dui xon DNA, trc khi c hai si dui xon lm khun cho s tng nhng si DNA mi. nhng sinh vt khc nhau ngi ta thy helicase c nhng c tnh khc nhau: E.coli, nm men, ng vt v thc vt bc cao. V nguyn tc c ch tc ng ca n nh sau: si kp c dui xon v helicase y ch ch ba tip tc dch chuyn. Qu trnh m xon thc cht vn cha c gii thch cn k. Nhng lin kt hydro gia cc base nit phi c ct t. Rt c th mt si DNA c ct ra bng thu phn lm cho xon c m ra, sau c ni li vi nhau. Ton b qu trnh cn nng lng. Tu theo loi helicase m nng lng cn l ATP hoc cc nucleosidtriphosphate khc, v cc desoxynucleosidtriphosphate cng c s dng y ch ch ba dch chuyn. Nh c cc nucleosidtriphosphate m cu hnh ca protein enzyme thay i do qa trnh phophoryl ho enzyme, qu trnh ny tng t nh co tht c.

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Hnh 6.8 S biu din s sao chp. S m xon kp nh helicase v s tng hp DNA hai si khun nh DNA-polymerase

Nh s 6.8 mt phn mch m xon c mt u l gc phosphate v mt u l mt nhm OH. Nhng on tch ra ny l khun cho vic to nn mt si b sung. Si mi ny chy theo hng ngc li vi si khun. Ngi ta phn bit gia chng 1 si leading strand l si trc v 1 si lagging strand l si sau. Si sau c ct ra bng thu phn v sau c ni kt li v to thnh ng cong ln c thc hin nh phc h helicase bao gm nhiu phn t protein theo hnh 6.9. ng cong ny c tc dng lm cho si DNA mi to thnh t khun l si sau ko di ra hng theo ch ba, tuy nhin hng ko di vn m bo t 5 3. V vy khi nim lagging c ngha l nhng base nit b sung cho si lagging strand c c chm hn. dui mch xon kp th mt on ca si lagging strand c ct ra, m xon v cc on c gi l Okazaki c tng hp. Trc khi nhng on ny ni li vi nhau nh enzyme ligase chng phi c hot ho ngha l chng c kt hp vi primer. Primer ny l mt RNA. Enzyme xc tc cho phn ng ny l primase. Phn ng ca primase xy ra sau khi dui xon, tip theo l phn ng polymerase, ngha l lp rp cc nucleotide vo si mi v cui cng l phn ng ligase. Ch ch ba chuyn dch v pha trc theo hng 5 3 ca si lagging strand. Vn tc dui xon l 700 cp baz trong 1 giy nhit 30 0C. Polymerase to lin kt ester gia OH v tr carbon th 3 ca si mi vi phosphate ca dnucleosidtriphosphate mi i vo. y pyrophosphate c tch ra v nhanh chng c thu phn to photphate v c.

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Hnh 6.9 Si trc v si sau phc h helicase-primase-polymerase

Nng lng cn to lin kt ester cha trong hai lin kt cao nng trong dnuclotidtriphosphate. S kt hp vo khun theo nguyn tc b sung tin hnh theo cch H ca nhm OH v tr carbon th 3 ca ribose c thay th bng 1 dnucleotide, chnh xc hn nhm OH c thay th bi acyl ca 1 dnucleotide (nucleosidphophoryl) (xem hnh 6.10). Chui cng ko di v tr 3 theo hng C-5 C-3 hoc t u cui phosphate n OH ca v tr carbon th 3. S kt hp vo khun theo nguyn tc b sung, ngha l thymine s kt hp vi adenine, guanine kt hp vi cytosine. Bng cch ny mt bn sao trung thnh ca si khun c tng hp. Nu trong qu trnh kt hp m xut hin mt li, v d hai base nit i din nhau khng b sung to nn khng gian m 1 dnucleotide mi vo khng kt hp c vi nhm OH ca ribose. Polymerase c kh nng sa cha nhng li ny, thc cht l ct nucleotide lp sai ra v kt hp nucleotide chnh xc vo. C ch sa cha quan trng ny lm cho t l li qu trnh nhn i l rt thp, thc ra khong 109 cp base th mi c mt li. Nhng li trong qu trnh ny l nhng t bin dn n nhng ri lon rt ln trong trao i cht. Mt khc n cng cn thit v to nn tnh mm do thch nghi vi mi trng sng.

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Hnh 6.10 S kt hp mt nucleotide vo si DNA bng vic to lin kt ester. Nucleosidphosphoryl thay th H OH ca d'ribose

Nhng si c to nn gm khong 1000 nucleotide, c kt hp nh cc ligase. Enzyme ny xc tc to lin kt ester gia gc phosphate ca carbon th 5 v nhm OH v tr carbon th 3 ca dribose. Phn ng ny gn lin vi s phosphoryl ho, gc phosphate do ATP cung cp. Nhng si mi c tng hp to xon vi si khun, nh vy mch kp xut hin. Ngi ta gi s nhn i ny l bn bo tn v trong mt mch kp mi th mt si c t trc (bo th) v mt si mi to thnh. Hai mch kp mi hon ton ging vi mch kp ban u. Thng tin di truyn tng ln gp i. Vic sao chp thng tin di truyn l mt qu trnh c ngha sinh hc v trnh t cc base trong DNA l trnh t cc ch ci, m vi nhng ch ci ny thng tin di truyn c biu din. Trc khi t bo phn chia (mitose) DNA c nhn i, nh vy nhng t bo con c h gen ging nh t bo m.

6.5 Sinh tng hp RNA (s phin m - transcription)

V nguyn tc tng t qu trnh tng hp DNA. y mt si DNA l khun v cc nucleosidtriphosphate l nguyn liu v kt qu l mt phn t RNA c tng hp. sao chp cn mt si DNA khun l si giu base pyrimidine, cc nucleosidtriphosphate v cc enzyme, ch yu l RNA-polymerase. Nguyn liu l ATP, GTP, CTP v UTP thay th cho TTP. Bas nit uracil b sung vi adenine trn khun. nhm OH v tr C3 ca ribose c kt hp vi nucleoside tip theo c s

238

tch ra ca pyrophosphate. Chui cng ko di u C3 ca ribose. Phn ng do RNApolymerase xc tc tng t s tng hp DNA. Nng lng cn cho s tng hp ny l s tch ra ca pyrophosphate. Trong hnh 6.11 ch ra s kt hp ca base b sung GTP vi cytosine trn mch khun vi s tch ra ca pyrophosphate. Chui RNA ko di ngc hng vi si DNA khun. s tng hp ny s to nn mt mch kp tm thi, gm DNA v RNA. V vy ngi ta gi hin tng ny l lai, mch kp l phn t lai. V RNA c bn l nh hn mch DNA, nn ch mt on tng i ngn ca DNA c c tng hp RNA. Nh c tnh ny m mt on c c k tip nhau bi nhiu enzyme polymerase. RNA mi c to thnh khng ch l 1 m c t 30 n 100 phn t. RNA-polymerase trt trn on DNA v tng hp nn si RNA. bt u v kt thc on DNA cn c nhng tn hiu ho hc, nhng tn hiu ny cho n nay cha c bit nhiu. Tuy nhin nhng on ring bit ca DNA th hon ton xc nh, n mang thng tin di truyn cho 1 phn t RNA xc nh.

Hnh 6.11 S tng hp RNA si giu pyrimidine

Nu si khun DNA c li, v d thiu 1 base nit, s c mt c ch sa cha. C nhng enzyme lm nhim v nhn bit nhng v tr li trn si DNA. C ch ny rt quan trng, nu khng th nhng thng tin v ngha c c. sinh vt nhn s th mRNA c c trc tip t DNA. nhng on DNA m ho cho rRNA v tRNA th trc ht nhng on RNA c tng hp nn, sau chng s c bin i to rRNA v tRNA trng thnh. S bin i ny ch yu l thay i cc base nit. Mt phn c methyl ho, nhng nhm OH c thay th bi cc nhm methyl. Qua nh hng n kh nng to cu hydro ca cc base nit. S methyl ho cc base nit xy ra sau khi sao chp c ngha i vi cu trc bc hai ca rRNA v tRNA. DNA sinh vt nhn s nm t do trong t bo cht v ch yu dng vng. trong ty th v lp th c cha mt lng nh DNA, DNA ny tng t DNA ca sinh vt nhn s. Mt phn t DNA dng vng khng c im bt u v im kt thc. c chng phi c 1 im kt hp ca polymerase. Vng bt u ny (initiation sites) c c trng bi mt trnh t base c bit. S di y c nhng ch in m l 4 im khc nhau cho s sao chp DNA t ty th ca ng.

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 sinh vt nhn chun s tng hp RNA khng thc hin trc tip trn DNA. a s on sao chp tng i di, c gi l hnRNA hoc cRNA, sau chng c bin i thnh mRNA, rRNA v tRNA. Qu trnh ny gi l processing (hinh 6.13). Khi 20 nucleotide u tin c lp rp tng hp mRNA th 1 m (cap) c gn vo u si RNA v tr C5 ca ribose. M ny gm guanine methyl ho, c kt hp vi RNA nh ba gc phosphate. Qu trnh tng hp mRNA tip tc cho n khi to nn trnh t base adenin-adenin-uracil-adenin-adenin. y l tn hiu cho s kt thc qu trnh sao chp, c khong 20 base nm sau tn hiu kt thc. u cui ny mt si polyadenine c gn vo, c t 150 -200 adenylnucleotide. Phn ny bo v cho mRMA trc s tn cng ca enzyme trong t bo cht. M guanosine kch thch qu trinh sao chp. Bn sao ny cha mt s on c ct ra. Nhng on c ct ri li gn kt vi nhau theo th t. Bng cch ny mRNA c tng hp. Nhng trnh t trn phn t mRNA c gi l exon, nhng on c ct ra gi l intron (hnh 6.12). Qu trnh processing xy ra trc tip sau khi sao chp v ch yu l qu trnh methyl ho. y nhm OH hoc nhm NH2 ca base nit c thay th bi nhm methyl. Bng cch ny kh nng to cu hydro b hn ch. Qu trnh processing ca tRNA bn cnh methyl ho cn c nhng thay i cc base nit, v vy m cc base nit mi nh dihydrouracil v pseudouridine xut hin. rRNA v tRNA c tng hp trong nhn, RNA-polymerase II xc tc cho tng hp mRNA, v RNA- polymerase III cho tRNA. Hu ht rRNA c tng hp trong hch nhn (nucleus) nh enzyme RNA polymerase I. C ba loi RNA u c tng hp trong nhn t bo v c a ra t bo cht, ni m chng tham gia vo qu trnh tng hp protein. Ba loi RNA trong t bo cht chim t l nh sau: rRNA chim 80%, tRNA chim 15% v mRNA chim 5%.

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Hnh 6.12 S ct ra cc exon t bn sao v s gn li to thnh si mRNA

Hnh 6.13 S biu din "processing" bn sao to mRNA, rRNA v tRNA

6.6 S chuyn ho nucleic acid Trong qu trnh trnh trao i cht cc phn t DNA v RNA khng cn thit na s b phn gii di tc dng ca cc nuclease thnh cc mononucleotide. Cc mononucleotide c th c dng tng hp cc phn t anucleic acid mi, hay b phn gii tip tc. Nuclease xc tc cho s ct lin kt phosphodiester ca DNA (DNAse) ca RNA (RNAse). DNAse ng mt vai tr quan trng trong s tng hp, sa cha v ti t hp ca DNA hoc l h thng bo v, trong chng phn gii DNA ngoi lai. RNAse tham gia vo qu trnh bin i tin mRNA thnh mRNA (RNA-procossing), v iu khin s phin m do s phn gii mRNA. Nuclease c th tch ra cc nucleotid u cui (exonuclease) hoc ct bn trong phn t (endonuclease).

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Exodesoxyribonuclease (Exo-DNAase) Exo-DNAase phn bit hng phn gii, trong c cht l si n hoc si kp v mono- hoc oligonucleotid (him hn) c tch ra. Mt s enzyme c th c nhiu chc nng. Ty theo c ch phn ng m cc nuclease tch ra khi c cht hoc l gn vi c cht qua nhiu chu k phn ng. Enzyme exonuclease III ca E.coli c nhiu chc nng: ct c hiu nhng v tr nht nh nh DNA-nuclease hoc l nh 3-phosphatase. Endodesoxyribonuclease (Endo-DNAase) Thng th hin ch yu i vi si DNA n v kp. Endonuclease sa cha nhng v tr li, nhn bit DNA li v ct DNA mt pha ca v tr li tch ra. Mt c tnh c bit khc ca nhng enzyme ny l nhn bit cc trnh t base. Mt v d l T4-endonuclease IV ct c hiu cytosine. Enzyme ct gii hn (restrictionendonuclease) tn ti s lng ln trong vi sinh vt. Hn 2000 loi enzyme ny c pht hin, chng nhn bit trnh t c di t 4 n 8 nucleotide trn si kp vi chnh xc cao. Ribonuclease (RNAse) Ribonuclease cng phn bit hot tnh exo v endo, tc ng vo nhng u cui nht nh (exo-enzyme) hoc nhng trnh t c hiu (endo-enzyme). Mt s nuclease ct c DNA cng nh RNA. Mt s phn ng dng RNAase c xc tc bng trnh t RNA (ribozyme). Cc mononucleotide b thu phn bng cc enzyme (phosphatase, nucleosidase) to nn cc sn phm cui cng l cc base purine v pyrimidine. Cc cytosine c bin i thnh cc uridine tng ng bng phn ng kh amin ho. S phn gii uracil v thymine bt u bng phn ng kh lin kt i gia v tr th 5 v 6 v tip theo l s m vng bng phn ng thu phn. S kh amin ho v kh carboxyl ho to nn -alanine cng nh 3-aminoisobutyrate. Cc base purine b phn gii thnh uric acid c o thi ra ngoi c th. Qu trnh ny in hnh cho cc ng vt c xng sng (ngi, vn ngi, chim v b st). i vi cc ng vt khc sn phm phn gii cui cng ca cc base purine l allantoin. Trong c th allantoin b phn gii thnh ur. Cc base pyrimidine thng b phn gii thnh ur v NH3.

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Chng 7

Protein v s trao i Protein trong c th thc vt

7.1 i cng v protein Protein c to nn t cc gc aminoacid. Cc gc ny ni vi nhau bng lin kt peptide. Lin kt peptide l lin kt gia nhm carboxyl ca 1 aminoacid v nhm amin ca mt aminoacid tip theo, v thc ra y 1 H ca nhm NH2 c thay th bi acyl ca aminoacid. Trong 1 dipeptide c hai, trong 1 tripeptide c ba v trong 1 oligopeptide c t 3 cho n 10 gc aminoacid lin kt vi nhau bng lin kt peptide. Polypeptide cha nhiu gc aminoacid. Nhng hp cht gm nhiu gc aminoacid ni vi nhau bng lin kt peptide c gi l polypeptide. Mi aminoacid cha 3 nhm nguyn t (NH, CH v CO) cu to nn theo s nghim ngt sau:

Ba nhm nguyn t ny l lun lp li trong chui polypeptide, phn cn li ca cc gc aminoacid (R) l chui bn.

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Trong cc protein t nhin c mt 20 loi aminoacid khc nhau. Nu mt chui ngn c 4 gc aminacid (tetrapeptide) th t c 204 = 160 000 kh nng, ngha l c 160 000 tetrapeptide khc nhau. Nu di 1 chui gm 300 gc aminoacid, mt di m thng gp i vi nhiu polypeptide, th ngi ta c mt s lng khng tng tng c l 10300 kh nng. Nhng trong t nhin ch tn ti mt s lng nh cc kh nng trn. Nh trn nu, tt c polypeptide c cng mt trnh t CO-CHNH. c tnh ca tng polypeptide v ca protein c xc nh do c tnh ho hc ca cc gc aminoacid, n c th tch in m, dng, a nc hay k nc. Trong t nhin, tu thuc vo trnh t v thnh phn, to nn rt nhiu i lc ho hc khc nhau, chng c ngha ln trong cc phn ng ho hc v i vi s nhn bit trnh t aminoacid. Trong c nhng vng gc aminoacid ch yu l acid hoc base, nhng vng khc l nhng gc aminoacid a nc hay k nc. Nhng protein ca mng thng c nhng gc k nc, nhng gc hng ra ngoi hoc nh vo t bo cht l a nc. S hnh 7.1 l mt v d.

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7.2 Chc nng ca protein Chc nng ca protein c chia ra cc nhm nh sau: 1. Chc nng enzyme 2. Chc nng cu trc 3. Chc nng d tr 4. Chc nng vn chuyn 5. Hormone 6. Chc nng bo v 7. Chc nng co rt 8. Cht c V chc nng enzyme ca protein c cp nhiu ln, c bit trong cu to v c ch tc ng ca enzyme. Protein l cht xy dng ng mt vai tr c bit quan trng trong c th ng vt. Cht khung c to nn t protein. Thuc loi ny gm c da, tc, sng, lng v collagen l thnh phn c bn ca gn v c khong 18% tham gia vo cu to ca xng. Mt chc nng ph bin khc ca protein l cu to nn mng sinh hc. Mng sinh hc c chc nng l gii hn nhng vng trao i cht v tham gia vo vic vn chuyn cc cht. Mng sinh hc cng c kh nng chuyn i nhng tn hiu. Protein mng cng c th l cc enzyme. Chc nng ny c th hin mng trong ca ty th v lp th. Mng sinh hc bao gm lp kp lipid vi nhng protein phn b trong . Nhng thnh phn lipid quan trng nht l phosphoglyceric, glycoglyceric, sterol v sphingolipid. Lp kp lipid ca mng sinh hc l nhng phn t lipid c nhng ui k nc quay li vi nhau, trong khi nhng gc ng v phosphate hng ra pha ngoi (hnh 7.2). y thnh phn glycerine l trc vung gc vi mt phng ca mng. Lp kp lipid khng i xng, ngha l hai pha ca mng c nhng phn t lipid khc nhau. Nhng phn t protein nm trong lp kp lipid lm bn vng mng v nh ra hai pha mng (hnh 7.2). Thnh phn protein nm trong lp kp lipid, c c tnh ch yu l k nc, thnh phn protein m nh ra pha ngoi, c c tnh ch yu l a nc. Si protein nh ra pha t bo cht thng kt hp vi mt protein ngoi

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vi, nhng phn t protein hng v pha mng t bo th thng kt hp vi mt chui carbohydrate (hnh 7.2).

trong lp kp lipid th phn t protein c nhng lin kt k nc v lin kt ion. Nhng lin kt ion nh lin kt gia nhm NH3+ ca protein v ca gc phosphate ca phospholipid. Nhng lin kt ny nhy cm vi pH. V vy mng sinh hc b nh hng, thm ch b ph hu bi gi tr pH thi qu. Mc qunh c nhiu hay t ca mng sinh hc b nh hng ln bi nhit . nhit thp mng sinh hc c cu trc ging nh tinh th. Khi nhit tng ln n nhit chuyn pha cu trc rn s chuyn sang cu trc tinh th lng. Nhit chuyn pha tng theo di v no ca acid bo. Ca2+ lm n nh mng, n lin kt vi nhng nhm phn u ch yu tch in m, v d gc phosphate ca phospholipid. Ca2+ c kh nng to lin kt vi hai nhm tch in m. V vy Ca2+ s lm gim tnh thm ca mng, trong khi nhng ion ho tr mt, c bit l H+ lm tng tnh thm. Tnh thm cng b nh hng bi c tnh ca lipid. Thnh phn steroid v nhng gc acid bo no v c mch carbon di lm cho mng st li, nhng gc acid bo cha no lm cho mng lng lo ra, do vy lm tng tnh thm ca mng. Khi ngi ta ni n tnh thm ca mng ch yu l ni n tnh thm ca nhng cht a nc. Nhng cht ny thm qua mng rt t, trong khi nhng phn t k nc c th thm qua hoc khuch tn qua mng. Hu ht nhng cht trao i c c tnh a nc, i vi chng mng sinh

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hc l ci chn ng k v y chng cn mt c ch c bit hn vn chuyn qua mng. Protein d tr c tch lu trong m d tr v khi cn s dng th c huy ng. Protein d tr in hnh ng vt l protein trng v sa. l protein cn thit i vi ng vt cn non, tng t protein ca ht v qu, l ngun dinh dng i vi cy con.

Cht lng protein c nh gi theo thnh phn cc aminoacid khng thay th. l nhng aminoacid m c th ngi v ng vt khng c kh nng tng hp c. Hm lng cc aminoacid khng thay th ca cc loi protein khc nhau c trnh by bng 7.1 v 7.2 Bng 7.1 Nhng aminoacid khng thay th i vi ngi -------------------------------------------------------------------Valine Lysine Phenylalanine Leucine Methionine Tryptophan Isoleucine Threonine Bng 7.2 Hm lng aminoacid khng thay th ca cc loi protein khc nhau (%) -----------------------------------------------------------------------------------------Protein trng 51 Protein u tng 40 Protein sa 50 Protein go 39 Protein m c 47 Nm 35 Collagen 17 Protein la m 33 Protein l cy 40 Protein ht lc 32 -----------------------------------------------------------------------------------------Nhng protein ng vt, v c protein ca l cy phn ln l giu aminoacid khng thay th. Protein ca ht v ng cc cha t aminoacid

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khng thay th. protein ng cc l lysine, protein ht v u cc loi l methionine c hm lng thp v n gii hn gi tr sinh hc ca nhng loi protein ny. s gii c cc kim loi nng nh nhng polypeptide n gin theo kiu phytochelatin c ngha quan trng. V d polypeptide n gin c ngun gc t glutation v c cng thc chung nh sau: (-glutamyl-cysteinyl)n-glycine Do c nhiu nhm SH chng c kh nng kt hp cht vi cc kim loi nng, lm cho nhng kim loi nng ny khng th gy ri lon trao i cht. S tng hp phytochelatin c kch thch bi nhng kim loi nng nh Cd, Cu, Ag, Bi v Au. Protein bo v c mt vai tr ln trong sinh hc min dch. ng vt c xng sng c mt c ch phc tp, pht trin cao, vi c ch ny chng ngn nga nhng tc nhn vi sinh vt gy bnh (virus, vi khun, nm, cht c vi khun). Chc nng ny c phn lin quan n c tnh ca chui polypeptide. H thng t v ton b, sinh hc min dch l mt lnh vc khoa hc pht trin c lp. Mt protein l (virus, vi khun, nm) xm nhp vo mu hoc vo m th c ch t v c huy ng rt nhanh. Protein l c gi l antigen. N c mt vng gm mt trt t xc nh cc nguyn t, vi vng ny n kt hp vi t bo lympho v kch thch t bo ny sn sinh ra khng th. Nhng t bo lympho tn ti trong h thng min dch vi s lng 109 v c trn b mt ca n nhng vng nhn, ni m antigen c kt hp vo. Nhng vng nhn ny rt khc nhau v ph hp mi vng cho 1 antigen xc nh. Nhng tc nhn khc nhau c nhng t bo lympho xc nh khc nhau vi nhng vng nhn ph hp. Khi mt antigen kt hp vi t bo lympho th n bt u sn sinh khng th c hiu i vi tc nhn gy bnh. Nhng t bo lympho khc khng c kch thch cho vic sn sinh ra khng th. C sn mt s lng ln cc t bo lympho khc nhau, chng c th tng hp c rt nhanh nhng khng th khc nhau khi khng nguyn xut hin. Nhng loi khng th khc nhau ny l xc nh, tn ti vi s lng khng m c, c th mt vi triu, y mi mt loi c mt v tr kt hp duy nht c trng. Kh nng ln khng th tng tng c ca h thng min dch lm cho protein l, protein ca tc nhn gy bnh tr thnh v hi. Nhng khng th ny c gi l globulin min dch. Chng chim khong 20% protein tng s trong mu. Globulin min dch Ig c chia lm 5 nhm khc nhau. Tuy nhin thnh vin c bn Ig cc nhm u c dng ch Y. Cu trc ny gm 2 si

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polypeptide ngn, xc nh, mi si c khong 220 gc aminoacid (hnh 7.4).

Hnh 7.4: S biu din khng th v khng nguyn. a) Khng th gm 4 chui polypeptide b) Khng th kt hp vi khng nguyn c) Kt hp gia khng nguyn v khng th

C ch m khng th nhn bit v tr khng nguyn ca protein l l c bit th v. S kt hp gia khng th v khng nguyn l thun nghch v c th so snh vi s kt hp gia enzyme v c cht. Lin kt ny khng phi ng ho tr m l nhng lin kt hydro v lin kt ion. iu lm r rng trnh t aminoacid ca v tr kt hp xc nh tnh c hiu ca lin kt. T rt ra rng chui polypeptide ngn hay di ca khng th l mt on siu bin c t 20-30 gc aminoacid,

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ngi ta cho rng, trong vng siu bin ny cha v tr lin kt c hiu i vi khng th. Ch kt hp thc s ch gm 5-10 aminoacid. V tr kt hp rt chn lc i vi khng nguyn. V d protein vn chuyn l albumin v hemoglobin trong mu, myoglobin trong m c v leghemoglobin trong mng ca Rhizobium. Albumin vn chuyn acid bo. Hemoglobin, myoglobin, leghemoglobin c kh nng kt hp lng lo vi O2 heme ca chng. Chng ging nhau ch u l phng tin vn chuyn O2. S kt hp ny ch xy ra dng Fe kh (Fe2+), ngha l kt hp vi heme. Hemoglobin vn chuyn O2 t phi qua mu n nhng c quan v m rt khc nhau. Leghemoglobin iu khin s cung cp O2 cho chui enzyme h hp ca vi khun. Myoglobin vn chuyn O2 trong m c, c nhiu trong m c ca ngi ln v l cht d tr O2. Trong khi ln (khi dng th) th O2 c kt hp vi myoglobin, c gii phng ra ty th. Fe2+ ca heme c th kt hp vi CN- v CO lm cho v tr hp th O2 b ng li, hu qu l O2 khng c vn chuyn, dn n cht ngt. Nng tng i cao CO trong khng kh cng nh s t chy khng hon ton trong ng c, v khi ht thuc l, cn tr s vn chuyn O2 trong mu. Ngi ta gi l ht b ng, l nguyn nhn gy hi n sc kho V d v chc nng hormone ca protein l insulin, gm 1 chui A vi 21 aminoacid v 1 chui B vi 30 aminoacid, hai chui ny c ni vi nhau qua hai cu disulfide. Insulin iu khin nng ng glucose trong mu. Khi khng insulin th s tip nhn ng trong t bo b hn ch. V vy mc ng trong mu tng v dn n s thi ng mnh m qua nc tiu (bnh i ng). V vy nhng t bo ny thiu ng lm cho ton b c th suy yu. Mt s peptide quan trng trong t nhin: - Glutation (tripeptide): glutamyl-cysteinyl-alanine: l mt tripeptide c chc nng sinh l quan trng. N c to nn t 1 gc glutamyl, 1 gc cysteine v glycine.

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Hai nguyn t glutatione c kh nng phn ng tch 2H to thnh 1 disulfide. y l h thng m sulfhydryl, chc nng quan trng nht ca n l gi nhm SH ca enzyme v coenzyme trng thi kh. Glutathione c trong tt c cc c th sng, tham gia vo nhiu phn ng oxy ho kh, v d s gii c cc gc c. Ngoi ra trong thc vt glutathione cn l dng d tr v dng vn chuyn lu hunh dng kh. Trong nhng loi c c nhng tripeptide tng t nh glutatione v trong cy h u 1 glutamyl-cysteinyl-alanine c tm thy.

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Nhng dng tng t c chc nng nh glutatione. Cysteine v cystine v nguyn tc to nn h thng oxy ho kh tng t glutatione, biu din nh sau:

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Oxytocin v vasopressin: Oxytocin v vasopressin l nhng oligopeptide, chng c cu trc rt ging nhau, u bao gm 9 gc aminoacid khc nhau, trong phn t c mt cu disulfide. Hormone oxytocin kch thch phn c phng ca rut gi, bng quang, ti mt v t cung. Hormone vasopressin nh hng n co rt ca phn c phng ca mch mu, lm tng p huyt. Vasopressin c vai tr quan trng trong vic lm c nc tiu. Nu nng thp th ng vt c v thi ra 1 lng ln nc tiu long, l mt bnh c gi l bnh tho nht.

7.3 Cc bc cu trc ca protein Phn ln cc lin kt c trong chui polypeptide c th quay t do v trc ca 1 chui polypeptide rt linh ng. Tu thuc vo lc tc ng

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m chng c nhng dng rt khc nhau. S bin dng ca mt chui peptide, s sp xp ca nhng phn si, s un cong v nhng np gp c trong mt chui peptide, c gi l cu trc. thun tin ngi ta thng phn bit cu trc ca phn t protein thnh 4 bc nh sau: bc I, bc II, bc III v bc IV. Cu trc bc I l trnh t sp xp cc gc aminocid trong chui polypeptide. Cu trc ny c gi vng nh lin kt peptide (lin kt cng ho tr). V mi mt aminocid c gc khc nhau, cc gc ny c nhng c tnh ho hc khc nhau, nn mt chui polypeptide cc iu kin khc nhau c nhng c tnh ho hc rt khc nhau. Tuy nhin v tng qut th tt c cc si polypeptide c xy dng mt cch c h thng t cc nhm nguyn t CO, CH v NH. S xy dng c h thng ny l c s to nn cu trc bc hai. Cu trc bc II l tng tc khng gian gia cc gc aminoacid gn nhau trong mch polypeptide. Cu trc c lm bn ch yu nh lin kt hydro c to thnh gia cc lin kt peptide k gn nhau, cch nhau nhng khong xc nh. Cu trc bc hai ca phn t protein: xon (-helix), phin l gp v xon collagen. Loi -helix l si dng xon c, qun xung quanh mt trc, mi vng xon c 3,6 gc aminoacid (hnh 7.5). y do nhm CO v nhm NH ca aminoacid th t trn chui gn nhau, gia hai nhm ny c th to nn mt cu hydro. Cc lin kt H tng i lng lo v c th c ct t khi nhit cao. Phn t protein mt cu trc bc hai th enzyme mt hot tnh xc tc. -helix l cu trc ca hu ht nhng protein mch thng (protein si). Dng si tn ti trong -keratin ca tc, lng, mng v da t chui polypeptide c cu trc . Cu trc ny trong phn tch Rnghen l nhng chu k 0,5-0,55 nm. Khi x l bng hi nc ngi ta c th ko di chui, lc ny xut hin chu k l 0,7 nm. Cu trc tn ti trong -keratin c gi l cu trc phin l gp. Nh trong hnh 7.6 chng gm nhng si polypeptide i song, gn vi nhau bng cc lin kt hydro. Khc vi -helix, nhng lin kt hydro c to nn khng phi trong mt si m gia si ny vi mt si khc. Nh vy c th nhiu si kt hp vi nhau to phin l gp. Cu trc phin l gp tn ti trong si la, nhng cng c trong nhng vng ca protein hnh cu.

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Hnh 7.5 S biu din mt -Helix. Mi tn ch lin kt H gia CO v H

Thuc protein thng cn c collagen, gm 3 si polypeptide xon vo nhau, chng li kt hp nhiu si li vi nhau bng lin kt ng ho tr (hnh 7.7). i phn t ny ch c mt s cu hydro c to nn, v ngoi glycine n c to nn ch yu t proline v hydroproline. Khi mt aminoacid i vo to lin kt peptide, trong chui peptide cn li 1 N l yu t cu to ch khng phi l -NH-, nh c ch ra trong s di . y 1 H ca nhm NH c thay th bng 1 aminoacyl, ngha l thiu H cho vic to lin kt hydro v v l do ny m si peptide khng th t xon c.

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Nhng si collagen chy song song to nn nhng b si dai ca gn. Collagen cng c trong xng v trong cc m ni. Elastin l mt protein, gm nhng si protein tng i ngn, gn kt vi nhau nh lin kt ng ho tr (hnh 7.7). Nhng si polypeptide quay theo dng xon c, t dui xon khi c p lc (hinh 7.8). Vt liu ny to nn mt khi dng keo dnh, cho php quay trong khng gian ngoi t bo v s co rt ca cc m.

Hnh 7.6 Nhng chui polypeptide chy i song trong cu trc -keratin (phin l gp)

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Hnh 7.7 S biu din mt si collagen bao gm ba si ring l

Hnh 7.8 Cu trc Elastin khi khng c v c lc tc ng

Nhng v d ny ni ln rng polypeptide c nhiu c tnh v chc nng khc nhau. Vic xc nh cu trc bc I ca phn t protein c ngha quan trng: L bc u tin quan trng xc nh c s phn t hot tnh sinh hc, tnh cht ho, l ca protein. L c s xc nh cu trc khng gian ca phn t protein da vo cu trc khng gian ca cc phn t protein tng ng. Cu trc bc I l bn phin dch m di truyn. V vy cu trc ny ni ln quan h h hng v lch s tin ho ca th gii sinh vt. L yu t gp phn quan trng trong nghin cu bnh l phn t. Kt qu nghin cu cho thy: khi thay i th t aminoacid, thm ch ch mt gc aminocid trong phn t protein c th lm thay i hot tnh sinh hc, gy nhng bnh c trng. V d in hnh l bnh thiu mu hng cu hnh li lim, l do cu trc bc I ca hemoglobin thay i: gc aminocid v tr th 6 trong chui ca hemoglobin A (bnh thng) b thay th bng gc aminoacid valin. L c s tng hp nhn to protein bng phng php ho hc hoc bng phng php cng ngh sinh hc. Frederick

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Sanger (1953) ra phng php v s dng phng php ny thnh cng xc nh trnh t sp xp cc aminocid trong phn t insulin (polypeptide c hot tnh hormone). n nay rt nhiu protein c xc nh cu trc bc I. Insulin l protein u tin c tng hp bng phng php ho hc vo nm 1966. Ngy nay bng phng php cng ngh sinh hc ngi ta s dng E.coli tng hp insulin. Cu trc bc III l tng tc khng gian gia cc gc aminacid xa nhau trong chui polypeptide, l dng cun li trong khng gian ca ton mch polypeptide. Nhiu si polypeptide trong c th sng tn ti khng phi dng thng m gp khc v qua m to nn cu trc khng gian ba chiu. Tuy nhin cu trc ny hon ton xc nh, ch yu l do trnh t cc aminoacid v mi trng. Khi mt si polypeptide tch ra khi ribosome sau khi tng hp v c thi ra trong t bo cht nh l mi trng to hnh th n hnh thnh nn cu trc t nhin rt nhanh, c bit i vi cu trc hnh cu, mang li cho protein nhng c tnh sinh l quan trng. C th do chuyn ng nhit ca cc si polypeptide m cc nhm ca cc gc aminoacid tip xc vi nhau, dn n c th kt hp vi nhau. Trong nhiu protein hnh cu c cha cc gc cysteine, s to thnh cc lin kt disulfide gia cc gc cysteine xa nhau trong chui polypeptide, lm cho mch b cun li ng k. Cc lin kt khc, nh lin kt Val de Waal, lin kt tnh in, phn cc, k nc v hydro gia cc mch bn ca cc gc aminoacid u tham gia lm bn cu trc bc III, nh protein hnh cu biu din hnh 7.9.

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Hnh 7.9 S biu din mt protein hnh cu cha nhng on c cu trc -helix v phin l gp

Cu trc hnh cu ca protein c gi l cu trc bc ba, l cu trc ca enzyme. chng mt nhm prosthetic c th kt hp ng ho tr, v d heme. Nhng nhng nhm gc aminoacid ring l cng c th l cc nhm hot ng trong phn ng enzyme. S ho tan ca protein c xc nh mt mc nht nh nh cu trc bc ba. nhiu protein hnh cu cc nhm k nc nh hng vo bn trong, nhng nhm a nc hng ra ngoi. Nhng nhm a nc ny kt hp vi nc nh l cht ho tan bng cc cu hydro, phc h ny tn ti trong dung dch. Trnh t aminoacid ca protein ca nhng enzyme cng chc nng cho bit mc quan h h hng ca cc loi. Mc quan h h hng cng gn, th mc tng ng v trnh t aminacid cng ln. iu rt th v y l nhng on trnh t quan trng nht, v d nhng vng phn ng, trong qu trnh tin ho hu nh khng thay i, c bo tn v nh vy cu trc bc hai, bc ba c duy tr. Cu trc bc IV: Cu trc ny c hnh thnh cc phn t protein gm 2 hay nhiu chui polypeptide hnh cu. Tng tc khng gian gia cc chui ny trong phn t gi l cu trc bc IV. Mi chui polypeptide ny gi l phn di n v (subunit). S kt hp gia cc phn t ny lng lo v ch yu l do lin kt hydro v k nc. Bng cch ny hai phn t xc nh c th kt hp vi nhau to thnh 1 dimer. Mt th d v s kt hp ny l hemoglobin, c to nn t 2 chui vi mi chui c 141 v 2 chui vi mi chui l 146 gc aminoacid. i phn t dng ny bn cnh protein cn l thnh phn cu to ca nucleic acid, cc ribosome. Cc ribosome ca sinh vt tin nhn gm 55 si polypeptide khc nhau v 3 nucleic acid khc nhau kt hp li. Nhiu virus c lp v bn ngoi, c cu to t nhiu phn t protein xc nh v bao quanh nucleic acid xon c bn trong. Cc i phn t trn kt hp vi nhau t ng trong mi trng ph hp thnh dng tn ti trong t nhin. c tnh vt l ca protein ph thuc vo c tnh ho hc ca cc gc aminoacid protein hnh cu cng nh s gp khc ca chng. Chim u th trong s kt hp ca cc aminoacid c tnh acid th protein c tnh acid. Ch yu l aminoacid c tnh kim l protein kim. Nhng si protein c nhiu nhm a nc l do nhng aminoacid phn cc v hydroxyacid th c ho tan ln. Ngc li, thnh phn ca gc k nc (valine, leucine, isoleucine) th ho tan ca nhng protein ny thp. Trng lng phn t protein ph thuc vo di ca tng chui polypeptide v s lng cc si polypeptide cu to nn 1 phn t protein.

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V kha cnh ny bng 7.3 ch ra s khc nhau ng k gia cc phn t protein ring l. Bng 7.3 Trng lng phn t v s lng si polypeptide ca mt s phn t protein MG S lng S lng gc aminoacid chui polypeptide Insulin 5 733 51 2 Ribonuclease 12 640 124 1 Myoglobin 16 890 153 1 Hemoglobin 64 500 574 4 Glutamate dehydrogenase 106 8 300 40 20 000 21 Synthetase tng hp acid bo 2,3 x 106 -----------------------------------------------------------------------------------------Cu trc ca mt hoc nhiu chui polypeptide c ngha quan trng i vi ho tan v chc nng ca chng. Cu trc protein c hiu l s sp xp ca nhng si ring l hoc nhiu si. Chng ph thuc nhiu vo pH ca mi trng. Protein v chui polypeptide ho tan tt khi nhng nhm a nc hng ra pha ngoi, nhm k nc hng vo bn trong. Khi mt protein thay i cu trc th nhng nhm k nc quay ra ngoi, protein mt kh nng ho tan trong nc, v d trng hp kt ta khng dng tinh th ca protein sa trong mi trng chua. Acid lactic c sn sinh do vi khun lm gim pH sa, lm thay i protein sa. Nhiu nhm k nc c hng ra bn ngoi, protein mt kh nng tan trong nc. V v s thng xuyn duy tr gi tr pH trong t bo cht rt quan trng, v ch c nh vy chc nng hot ng ca cc enzyme trong t bo cht mi c m bo. Bn cnh H+ cn c nhng cation khc c ngha i vi cc phn ng enzyme v d K+. Nng ca n trong t bo cht cao hn so vi cc loi cation khc v nm trong khong 100 n 150 mM. Nng K+ cao c ngha i vi s tng hp protein. Nng Ca2+ trong t bo cht rt thp, tuy nhin n c vai tr quan trng i vi cu trc ca cc protein khc nhau. Calmodulin l mt polypeptide kt hp vi Ca2+ lm cho cc vng k nc c hng ra ngoi, cc vng ny kt hp vi cc enzyme v hot ho cc enzyme nh phosphodiesterase, adenylate-cyclase, photoarylase v ATPase. Bng cch tng t troponin C v paralbumin hot ho nhng enzyme khc bng vic thay i cu trc khi kt hp vi Ca2+. Nhng Ca-protein ny thay i

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phn ng enzyme. Nhng lin kt thc hin nhng v tr hon ton xc nh, thc cht l helix-di ni-helix (hnh 7.10).

Hnh 7.10 S biu din lin kt Ca trong chui polypeptide ni hai cu trc helix li vi nhau

Dng ny ca mt lin kt Ca tn ti tng i v c trng i vi calmodulin, troponin C v paralbumin. Cu trc ny bao gm mt chui polypeptide c 12 gc aminoacid, trong 7 nhm carboxyl c nguyn t O kt hp vi Ca2+. Cu trc ny tng ng vi mt hnh thp i 5 mt, ngha l mt din tch c bn vi 5 mt, pha trn v pha di cn c mt CO. trung tm cu trc khng gian ba chiu ny c nguyn t Ca2+, mi mt gc c 1 nhm CO (hnh 7.11). Lin kt vi Ca2+ trung tm lm thay i cu trc ca protein.

Hnh 7.11 Mt phng c hnh 5 gc ca mt khi chp i vi Ca2+ trung tm

Troponin C c tng s 4 v tr kt hp vi Ca2+, trong 2 lin kt c i lc mnh v 2 lin kt c i lc yu. S bo ho Ca2+ ph thuc vo nng Ca2+ trong h thng. Cng nh vy i vi calmodulin. Nng Ca2+ cng nh hng n nhng qu trnh enzyme bng nhng Ca-protein.

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Vai tr quan trng nht ca troponin C l iu khin s co rt ca m c. Paralbumin cng c mt trong m c, c ngha i vi th gin m c. V tr kt hp ca n vi Ca2+ cng c i lc cao vi Mg2+. 7.4 Aminoacid Phn ln nht ca nhng cht nit hu c d ho tan trong thc vt l aminoacid, amide v amine. Trong trao i cht th aminocid c vai tr rt quan trng. Chng l vt liu to nn protein. Cng thc chung ca aminoacid: COOH | H2N -CH | R Cng thc tng qut ca L--aminoacid Nhm amine nh nguyn t C2, theo tn c l nguyn t C. V vy ngi ta gi l nhm -amine. Cc aminoacid tn ti ch yu trong t nhin c nhm amine ng bn tri trc, c gi l aminoacid dng L. Dng D-aminoacid tn ti ch ring bit, v d trong thnh t bo vi khun. Mi mt aminoacid t nht c c trng bi 1 nhm carboxyl v 1 nhm amine. Hn 100 loi aminoacid khc nhau m ngi ta bit, c khong 20 loi l thnh phn ca protein. Di y l nhng aminoacid quan trng v cng thc ho hc ca chng. n gin v theo thi quen nhm carboxyl ca aminoacid proton ho. Tuy nhin dng ny khng c aminoacid no tn ti, v nh nu trn, khi nhm carboxyl phn ly, th nhm NH2 phi phn ly. Proton ho hay kh proton ho lun lun ph thuc vo pH ca mi trng. Cc aminoacid ring bit c nhng c tnh khc nhau l do phn gc aminoacid. nhng aminoacid trung tnh c tnh trung tnh. Khi chiu di chui tng th c c tnh k nc. Protein c cha nhiu aminoacid nh valine, leucine, isoleucine c c tnh k nc l ch yu. Nhng aminoacid c tnh acid trong phn gc c 1 nhm carboxyl. Protein cha nhiu aminoacid c tnh acid l nhng protein acid. Tng t nh vy i vi protein, ch yu c to nn t nhng aminoacid c tnh kim l nhng protein kim. Phn gc ca aminoacid c ngha quyt

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nh i vi c tnh ca protein m chng to nn. iu ny khng nhng c ngha i vi tnh cht ho hc m c cu trc ca protein. 7.4.1 Phn loi aminoacid Thy phn hon ton protein, thu c ch yu cc L--aminoacid. Mc d protein rt a dng nhng hu ht chng u c cu to t 20 L--aminoacid. Da vo c tnh ca mch bn (gc R) aminoacid c chia lm 7 nhm chnh sau y: 1. Aminoacid trung tnh mch thng Nhm ny gm 5 aminoacid: glycine, alanine, valine, leucine, isoleucine. Cc aminoacid ny u c 1 nhm amine v 1 nhm carboxyl. Glycine l axit amine n gin nht, nhm R ca n l nguyn t H, l aminoacid duy nht khng cha carbon bt i. Bn aminoacid cn li c mch bn khng phn cc.

2. Cc hydroxyl aminoacid mch thng Thuc nhm ny c 2 aminoacid l serine v threonine. Chng ging vi nhm trn ch ch c 1 nhm amine, 1 nhm carboxyl v cng l mch thng nhng c cha 1 nhm OH.

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3. Aminoacid cha lu hunh mch thng Nhm ny c 2 aminoacid l cysteine v methionine. Khi oxy ho 2 nhm -SH ca 2 phn t cysteine to thnh cystine c cha cu (-S-S-).

4. Cc aminoacid acid v cc amide Thuc nhm ny c aspartic acid v glutamic acid . Trong phn t ca chng c 1 nhm amine v 2 nhm carboxyl. pH sinh l (6-7) cc aminoacid ny tch in m. Amine ho nhm carboxyl mch bn ca aspartate v glutamate to thnh cc amide tng ng l asparagine v glutamine. s tng hp glutamic acid mt nhm amine t 1 cht cho c chuyn n cetoacid. Ngi ta gi qu trnh ny l chuyn amine ho do

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enzyme transaminase xc tc. Enzyme ny c nhm prosthetic l pyridoxalphosphate.

5. Cc aminoacid kim Trong nhm ny lysine v arginine tch in dng ( pH =7), cn histidine cha nhm imidazol c tnh base yu pH = 7

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6. Iminoacid (proline) Prolin khc vi tt c cc aminoacid khc ch nhm amine bc 1 C kt hp vi mch bn to thnh vng pyrrolidine. Do proline l mt iminoacid cha nhm amine bc 2.

7. Cc aminoacid thm v d vng Cc aminoacid thuc nhm ny l phenylalanine, tyrosine v tryptophan. Do c cha vng thm nn cc aminoacid ny c mt s phn ng c trng. Nhng aminoacid thm l nhng tin cht ca nhiu alcaloid, phytohormone nh indolacetic acid.

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Nhng gc aminoacid ca histidine, lysin v cysteine l nhng nhm hot ho enzyme. Nhng aminoacid thm tyrosin c h hng gn vi thyronine, y l tin cht ca hormone tuyn gip trng thyrocine. Thyrocine c to nn trong tuyn gip trng t thyronin nh s kt hp vi iod.

7.4.2 Tnh cht ca aminoacid Tnh cht lng tnh ca aminoacid Trong cu to ca aminoacid va c nhm amine (-NH2) th hin tnh base, va c nhm carboxyl (-COOH) th hin tnh acid, ngha l chng mang tnh lng tnh v c th phn ly thnh ion H+ v OH-. Trong dung dch nc, cc aminoacid c th ng thi phn ly nh sau:

190

R - CH - COOH + HOH R - CH - COOH R - CH - COO| | | + H+ NH3OH - HOH NH2 NH2 Nhng cng trnh nghin cu v quang ph khuch tn xc nhn rng: phn t aminoacid khng c nhm carboxyl t do, cng nh khng c nhm amine t do v chng u l mui ni c dng nh sau: R - CH - COOR - CH - COO| | NH3+ NH3+ Do c tnh lng tnh nn aminoacid l nhng cht m tt. V aminoacid c tnh lng tnh trong dung dch nc nn tu theo pH ca dung dch m s phn ly ca nhm carboxyl hoc nhm amine s b km hm v aminoacid th hin tnh kim hoc tnh acid. Trong mi trng acid, khi nng H+ cao, s phn ly ca nhm carboxyl s b km hm v phn t aminoacid tch in dng, trong in trng n s dch chuyn v pha cc m. R - CH - COO| + NH3+ R - CH - COOH | NH3+

Trong mi trng kim, di tc dng ca cc ion OH- s phn ly ca nhm amine s b km hm, nhm carboxyl s b phn ly, phn t aminoacid tch in tch m v dch chuyn v cc dng trong in trng. R - CH - COOR - CH - COOH R - CH - COO| + OH | | + H2O NH3+ NH3OH NH2 im ng in pI (isoelectric point) phn t aminoacid trung ho v in v n khng dch chuyn trong in trng. Tu theo s nhm carboxyl v nhm amine trong phn t aminoacid cng nh tu theo hng s phn ly ca cc nhm ny m im ng in pI ca cc aminoacid s khc nhau. Cc aminoacid monoamine dicarboxylic (aspartic acid, glutamic acid) c im ng in trong mi trng acid, cc aminoacid diamine

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monocarboxylic (acginine, histidine, lysine) c im ng in trong mi trng kim. Bng 7.4 im ng in ca cc aminocid Tn aminoacid im ng in Tn aminoacid im ng (pI) in (pI) Alanine 6,00 Lysine 9,74 Arginine 10,76 Methionine 5,74 Asparagine 5,41 Hydroproline 5,83 ng dng tnh lng tnh ca aminoacid Nh c tnh lng tnh m aminoacid c vai tr l cht m trong t bo v trong trao i khong ca c th thc vt. Da vo tnh cht ny nh tnh v nh lng cc aminoacid trong dung dch nghin cu bng phng php in di, sc k trn giy, sc k ct... Cng do tnh cht lng tnh m cc aminoacid c th to mui ni ng thi vi acid cng nh base: R - CH - COO| NH3+ R - CH - COOH | NH3+

HCl

Cl-

R - CH - COO| NH3+

+ NaOH

R - CH - COONa | NH2

+ H2O

Ngoi ra phn t aminoacid cn to mui ni vi cc ion kim loi nng nh Fe2+, Cu2+ , Zn 2+ ,... c bit vi Cu 2+. Cu hnh ca mui phc ny c th biu din nh sau: R R - CH - NH2 O-C=O | | Cu | 2+ 2 CH- NH2 + Cu O = C - O H2N - CH - R | COOH Phn ng to ester

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Vi ru, aminoacid c th to thnh ester R - CH - COOH | NH2 + C2H5OH R - CH - COOC2H5 | NH2 + H2O

Cc ester ca aminoacid u l nhng cht lng d bay hi, c tnh kim, d iu ch bng php ct chn khng. Phn ng ca gc R Gc R l nhm nguyn t trong phn t aminoacid vn lin kt vi nguyn t C v khng tham gia trong vic hnh thnh lin kt peptide. Bn cht ho hc ca gc cho php aminoacid thc hin nhiu phn ng khc nhau: Phn ng to mui vi acid, base v kim loi nng (do nhm NH2 v nhm -COOH c trong gc R) nh trnh by trn. Phn ng oxy ho kh (do nhm -SH v nhm -S-S-) SH CH2 - S - S - CH2 | | | CH - NH2 CH - NH2 CH2 | - 2H | | COOH COOH 2 CH - NH2 | COOH + 2H Cysteine v cystine ng vai tr quan trng trong vic vn chuyn e+ v H ca qu trnh oxy ho kh ni ring v qu trnh trao i cht ni chung Phn ng to amide (do nhm -COOH) COOH CO - NH2 | | CH2 CH2 | | CH - NH2 + NH3 + ATP CH - NH2 + H3PO4 + ADP | | COOH COOH Phn ng xy ra tng t vi glutamic acid. Phn ng kh amine ho bi HNO2 (do nhm -NH2) R - CH - COOH R - CH - COOH

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| + HNO2 | + N2 + H2O NH2 OH Ngoi ra vng thm ca aminoacid tham gia phn ng nitro ho, halogen ho v mt s phn ng c trng ca aminoacid: phn ng vi aldehyd formic, vi ninhydrin... 7.5 Sinh tng hp aminoacid vi khun v lc lp ca thc vt con ng quan trng nht tng hp glutamate l t 2-oxoglutarate v glutamine. Phn ng ny c xc tc bi enzym glutamat-synthase. ng vt glutamate xut hin ch yu bng phn ng chuyn amine gia 2-oxoglutarate v aminoacid. Phn ng ny xy ra ch yu gan, l c quan trung tm ca s bin i aminoacid. s chuyn amine ho glutamate v aspartate l nhng cht cho nhm NH2. Mt s vn chuyn amine t glutamate n pyruvate c ch ra nh sau:

Enzyme xc tc l glutamat-pyruvat-aminotransferase. cetoglutaric acid c to nn li tr thnh cht nhn NH3. Nh s chuyn amine m nhiu aminoacid c to thnh. Nhng v d quan trng nht l: -cetoglutaric acid glutamic acid

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oxaloacetic acid aspartic acid glyoxylic acid glycine pyruvic acid alanine hydroxy pyruvic acid serine -ceto hydroxybutyric acid threonine Glutaminsemialdehyde ornithin Tuy nhin khng phi tt c cc aminoacid c trong t nhin c tng hp bng con ng chuyn amine ho. Cn c nhng con ng phn ng quan trng khc tng hp nn cc aminoacid. Ngi ta bit cc con ng phn ng tng ng cho 5 h aminoacid khc nhau (bng7.5). V d hnh 7.12 ch ra con ng tng hp ca h glutamic acid. 1. Glutamic acid c phosphoryl ho nh ATP nhm -carboxyl to thnh glutamylphosphate 2. Cht ny c kh nh NADPH to thnh aldehydglutamic acid ng thi tch ra phosphate. 3. Cht ny c amine ho nh enzyme transaminase v tr carbon, to nn ornithine. 4. Cht ny c kh amine ho nh 1 enzyme transaminase v tr -carbon lm xut hin -ceto--amino-valeric acid. 5. Qua s ngng t mt vng 5 cnh c to nn (pyrrolidincarbonic acid) 6. Cht ny c kh thnh proline nh NADPH.

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Hnh 7.12 Cc con ng phn ng ca h glutamic

Bng 7.5 Nhng aminoacid quan trng nht ca cc h aminoacid H glutamic Glutamate Ornithine Arginine Proline Hydroxyproline H aspartate Aspartate Lysine Methionine Threonine Isoleucine H pyruvate Alanine Valine Leucine H serine Serine Glycine Cysteine H aminoacid thm Phenylalanine Tyrosine Tryptophan

Arginine cng thuc vo h glutamic acid c th c tng hp nn t ornithine. Hydroxyproline c tng hp t proline nh 1 oxidase c chc nng hn hp. Cht cho H l ascorbic acid (vitamin C). Nh s di y nhm OH nh v C3 ca ascorbic acid c kh nng phn ly 1 H+. Phn t ny c c tnh acid. Gi tr pK ca n l 4,2. Ascorbic acid tham gia vo phn ng vi proline khi tch ra phn t O2, trong n cung cp nguyn t H cn thit cho s to H2O tng ng.

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S hydroxyl ho proline xy ra khng phi nhng phn t proline ring bit, m xy ra sau khi phn t proline tham gia cu to protein. dng ny hydroxylproline l thnh phn quan trng trong collagen v elastin. Nu thiu aminoacid ny s dn n s ri lon trong cc m lin kt, m chng (xng, rng) v gn. Khi thiu vitamin C th vic xy dng cc m ny b tn thng (bnh hoi huyt). S ri lon ny c gii thch l thiu cht cho H, ascorbic acid, tng hp hydroxylproline t proline.

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Proline v hydroxylproline l nhng aminoacid d vng. y nguyn t N dng NH. Mc d vy ngi ta khng gi n l iminoacid, v n th hin nh aminoacid. Thc t nguyn t H b thiu c thay th bi 1 cht khc. iminoacid c nhm imine, y nguyn t N kt hp vi C bng 1 lin kt i (HN=C). S tng hp cc aminoacid thm i t phosphoenolpyruvate v erythrosophosphate. Con ng tng hp phc tp ny khng nu ln y. Mt sn phm trung gian quan trng ca qu trnh ny l shikimic acid.

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 tng hp cysteine, mt aminoacid cha S, cn kh sulfate. Qu trnh ny ch xy ra thc vt v cc vi sinh vt khc nhau, tng t qu trnh kh nitrate, khng xy ra ng vt. Sn phm u tin ca s kh sulfate l aminoacid cha S, cysteine. N cung cp S cho tng hp nhng aminoacid quan trng cha S khc: methionine. Qu trnh phn ng ch ra hnh 7.13. 1. Phosphohomoserine, mt hp cht tng t nh serine, phn ng vi cysteine to thnh cystathionine v tch ra phosphate v c 2. Cystathionine c tch ra thnh serine v homocysteine bng phn ng thu phn 3. Nh c cht cho nhm CH3 nguyn t H ca nhm SH ca homocysteine (cht ny ging cysteine) c thay th bng 1 nhm methyl. Methionine c to thnh.

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Hnh 7.13 Tng hp methionine

7.6 Qu trnh kh nitrate Nhng nguyn t c bn ca carbohydrate v lipid l C, H v O. Ngoi ra cn c cc nguyn t S v P, v d trong phospholipid v sulfolipid, nhng chng khng phi l cc nguyn t c bn. Trong protein v nucleic acid, N l nguyn t khng th thiu c. Trc khi cp k hn trao i cht ca chng, chng ta cn tm hiu cc i phn t, cc cht trao i, cc cht hu c c trng lng phn t thp c to nn t cc cht v c nh th no. S sau y a ra 4 con ng phn ng: tng hp polysaccharid, lipid, protein v nucleic acid.

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Polysaccharid, lipid, protein v nucleic acid c kh nng to nn nhng phc hp ln l nhng thnh phn ca t bo nh thnh t bo, mng t bo, enzyme v ribosome. Thc vt v mt s sinh vt tin nhn c kh nng tng hp nhng phn t hu c t nhng cht v c ngho nng lng. Cng nh vy i vi s c nh nit v c.
-

H2O CO2

Phosphoglyxerate Oxaloacetate Acetate Malonate Axit -oxoacid

Monosaccharide Glycerine Acid bo Aminoacid Nucleotide n phn MG: 100 - 300

Polysaccharide Lipid Protein Nucleic acid Hp cht cao phn t MG: 103 - 108

Thnh t bo Mng sinh hc Enzyme Ribosome Cc kt hp i phn t MG: 106 - 109 T bo

N2 NH3 NO3-

Ribose Carbamylphosphate Cc hp Hp cht trung cht v c gian MG: 50 - 250

Nit dng NH3, NO3- v mt phn dng N2 c th c ng ho. S ng ho trc tip thc cht l s kt hp NH3 vi mt phn t hu c. Nitrate v N2 trc ht c chuyn sang NH3. Hai qu trnh ny s c cp phn sau y. Qu trnh kh nitrate v nitrite l chuyn NO3- thnh NH3 qua NO2-, c xc tc bi 2 enzyme khc nhau l nitratreductase v nitritreductase. Nitratreductase l 1 enzyme ph bin trong vi khun, nm, thc vt bc cao v thc vt bc thp. nhng nhm sinh vt khc nhau th enzyme ny c nhng c tnh khc nhau. Chc nng ch yu l kh NO3thnh NO2-. Nhm prosthetic ca enzyme ny thc vt bc cao l FAD, cytochrome b v Mo. Trnh t phn ng c phn tng t chui enzyme h hp, c biu din trong hnh 7.14. NADH, mt s loi sinh vt l NADPH chuyn e- n FAD, to thnh FADH2, sau in t ny c chuyn n cytochrome b, cytochrome b trng thi kh, kh Mo6+ thnh Mo4+. Mo4+ c kh nng ko 1 nguyn t O ca NO3- v kh to thnh NO2-. Oxy dng kh phn ng trc tip vi proton to thnh H2O.

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Trong s 2H+ c gii phng ra trong qu trnh oxy ho kh gia FADH2 v cytochrome b (hnh 7.14).

Tng qut: NAD(P)H + H + NO3- NO2- + H2O + NAD(P)+ Hnh 7.14 Trnh t phn ng ca nitratreductase

Enzyme ny nh v trong t bo cht v dng kh ca n c to nn t qu trnh ng phn (kh aldehydphosphoglyxeric thnh phosphoglyxeric acid), thc vt C4 bng s oxy ho malate thnh oxaloacetate. Hot tnh ca enzyme nitratreductase b nh hng mnh bi nh sng, nguyn nhn l do cht kh ca nitratreductase c gii phng trc tip t lc lp. y lc lp tip nhn oxaloacetate hoc phosphoglyxerate, trong qu trnh quang hp c kh thnh malate v aldehydphosphoglyxeric. Hai cht ny c a ra li t bo cht. y chng b oxy ho c s tham gia ca NAD+ to thnh NADH. C ch ny c bit c ngha thc vt C4, v chng mt lng ng k cht kh c to thnh t malate i ra t lc lp. Hot tnh ca enzyme nitratreductase b nh hng bi cc cht khc nhau. Nitrate hot ho mnh m, ngc li NH4+ v cc aminoacid km hm enzyme ny. Enzyme ny c na thi gian tn ti l mt s gi, v vy n phi lun lun c tng hp mi. S tng hp ny ph thuc vo cng nh sng. Bn cnh nitratreductase ng ho nh nu trn, ngi ta cn bit mt loi nitratreductase h hp. Loi ny c mt ch yu trong vi sinh vt v c to nn c bit trong iu kin ym kh. Chc nng ca n l khi khng c O2, in t bt ngun t chui enzyme h hp kt hp vi nguyn t O ca NO3-. Trong trng hp ny NO3- thay th nguyn t oxy trong chui h hp. Nitratreductase h hp ny gn cht vi mng v cng tn ti vi cytochrome, b c ch bi O2. Ngc vi nitratreductase ng ho khng kt hp v mn cm vi O2, nitratreductase h hp l 1 enzyme xc tc qu trnh phn nitrate ho . Phn nitrate ho c hiu l kh NO3-

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cho ti N2O v N2. Qu trnh ny xy ra ch yu trong t, nc trong iu kin ym kh v c vai tr quan trng trong t nhin. S kh NO3theo kiu h hp to thnh NO2- xy ra trong h thng tiu ho ngi v ng vt. NO2- c to ra nh enzyme nitratreductase trong t bo cht ca t bo thc vt l 1 anion ca 1 acid yu. V vy mt phn NO2- to nn c proton ho v th hin cn bng phng trnh sau: NO2- + HOH HNO2 + OH-

NO2- kh i qua mng ngoi ca ty th v lp th, tuy nhin d dng i vi HNO2. Lc lp v ty th tip nhn ch yu HNO2 v nh vy lm cho t bo cht b kim ho. S kim ho ny lm tng s tng hp cc anion hu c. Trong ty th xy ra s kh HNO2 thnh NH3. Enzyme nitritreductase gm 1 chui polypeptide vi 1 siroheme (cng tn ti Fe v S) v mt phc h 4 nguyn t Fe-S. Trnh t phn ng ca nitritreductase c ch ra trong hnh 7.15. Trong lc lp cht kh trc tip l ferredoxin. y c mt s kt hp cht ch gia quang hp v kh nitrate. Phn ng xy ra nh sau: HNO2 + 2e- + 2H+ NH3 + O2

Trong nhng c quan thc vt khng c mu xanh, v d nh r cy, NO2- cng c kh n NH3. Mt thi gian di qu trnh ny khng c gii thch, v nhng b phn khng c mu xanh, c bit l r cy khng c ferredoxin, cho n khi nhng kt qu mi ch ra rng, trong ty th ca r c 1 ferredoxin tng t h thng oxy ho kh c kh nng kh HNO2 thnh NH3. H thng kh ny cha cht kh l NADH hoc NADPH.

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Tng qut: HNO2 + 2e- + 2H+ NH3 + O2 Hnh 7.15 Trnh t phn ng ca nitratreductase

Hot tnh ca nitritreductase c kch thch bi nitrite v nitrate. Nng nitritreductase cao hn nng nitratreductase, l nguyn nhn ti sao khi cung cp nhiu nitrate cho cy dn n s tch lu nitrate m khng tch lu nitrite. S tch lu nitrate xy ra khi thc vt c bn qu nhiu nitrate hoc khi nh sng cho quang hp yu. Trong trng hp nh sng yu dn n s thiu cht kh. l nguyn nhn ti sao i vi ging rau ba lng nhng gi bui sng th c nng nitrate cao hn bui chiu. Ban m ch yu l phosphoglyxerate v mt t triosephosphate i ra khi lc lp. Ban ngy th ngc li. Triosephosphate c kh nng kh NAD+ thnh NADH khi phn gii theo qu trnh ng phn cung cp cht kh cho qu trnh kh nitrate. Ngc li phosphoglyxerate khng th kh NAD+ v nh vy khng cung cp NADH cho qu trnh kh nitrate. Nng nitrate cao thng gp trong nhng b phn sinh trng dinh dng v y c th nh hng n sinh l dinh dng. c bit i vi nhng sn phm thc vt c dng n sng nh x lch, bp ci, c rt. Thc ra nitrate khng c hi nhng trong qu trnh bo qun vi sinh vt bin i thnh nitrite v trong c th ngi, c bit trong tuyn nc bt nitrate c kh thnh nitrite. Trong mi trng acid to thnh HNO2, n c th c tch thnh nitrosyl v hydroxyl. HNO2 = HO N= O OH- + NO+

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Cation nitrosyl (NO+) c kh nng oxy ho Fe2+ ca hemoglobin bng cch ly i 1e- ca Fe Fe2-heme + NO+ Fe3-heme + NO

Hemoglobin dng oxy ho khng th kt hp v vn chuyn O2, dn n s ngt th. Hin tng ny gp ngi trong mt s thng u v sau c th pht trin enzyme c kh nng kh hemoglobin dng oxy ho. 7.7 Qu trnh c nh nit phn t Ch sinh vt tin nhn mi c kh nng c nh N2, l 11 trong 47 h vi khun v 6 trong 8 h vi khun lu hunh. Mt s sng t do, mt s sng cng sinh vi thc vt bc cao. Nhng vi sinh vt c nh m quan trng nht l: - Rhizobium, sng cng sinh vi cy h u. - Bradyrhizobium, cng sng cng sinh vi cy h u, chng khc vi rhizobium ch l sinh trng chm hn. - Loi frankia, vi khun sng trong t. - Vi khun lu hunh heterocysten sng cng sinh vi cc loi thc vt khc nhau nh vi bo hoa du (azolla) l. Nhng loi vi sinh vt sng cng sinh v t do c cha gen Nif trong h gen (Ni l vit tt ca nitrogen v f l fixing). Gen ny m ho cho enzyme nitrogenase, l mt a enzyme, xc tc cho phn ng c nh N2, kh N2 thnh NH3. Nitrogenase gm 2 protein cha Fe-S, protein nh hn c trng lng phn t 60 kDa, cha 1 chm Fe-S c 4 nguyn t, phn t protein ln hn c trng lng phn t 220 kDa v cha nguyn t S linh ng, 36 nguyn t Fe v 2 nguyn t Mo. Phn t protein nh hn c chc nng vn chuyn e-, trong e- ca ferredoxin hoc flavodoxin vn chuyn ln phc h Fe-Mo (hnh 7.16). S vn chuyn e- ny cn nng lng, do ATP cung cp. ATP kt hp vi protein cha Fe qua 1 cu Mg v sau c thu phn thnh ADP v Pi. S thu phn lm thay i hnh dng ca protein enzyme, e- c vn chuyn ln phc h Fe-Mo. vn chuyn 1e- cn 2 ATP. Phc h Fe-Mo thc cht l 1 N2-reductase. Phc h nh hn c gch cho l 1 protein cha Fe, phc ln hn l protein cha Fe/Mo, phc ln ny thc cht l dinitrogen-reductase (hnh 7.16).

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Hnh 7.16 Cc phn ng xy ra bacteroid ca vi khun rhizobium

vn chuyn e- t phc Fe/Mo n N2 th s thay i ho tr Mo c mt vai tr quan trng, nh hnh 7.17. 1. 2e- v 2H+ c kt hp vi nguyn t Mo 2. Nguyn t H2 ang gn vi enzyme b y ra bi N2 3. N2 c kh thnh -N=N do tip nhn H+ + e4. -N=N c kh thnh =N-NH2 do tip nhn H+ + e5. =N-NH2 c kh thnh NH3 do tip nhn H+ + e-, NH3 c tch ra khi phc h enzyme 6. Nguyn t N ang gn vi enzyme c kh thnh NH3 do tip nhn 3e- v 3H+ v sau NH3 c tch ra khi phc h. S gn N2 vi phc h Fe-Mo cn 1 lin kt trc ca phc h ny vi H2. V vy kt qu l bn cnh to ra NH3 ngoi ra cn xut hin H2, phn t ny c th c tch ra thnh H+ v e- v e- ny c a tr li h thng (hnh 7.16).

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Tng qut: 8H+ + 8e- + N2 H2 + 2NH3 Hnh 7.17 S kh N2 thnh NH3 phc h Fe-Mo ca nitrogenase rhizobium v bradyrhizobium h thng nitrogenase c biu din hnh 7.15, h thng ny nh v bacteroid c tm thy trong khng bo nhim ca nhng t bo nt sn. Bacteroid tng t mt c quan v c th so snh vi ty th. N c cung cp cc hp cht carbon hu c, ch yu l anion hu c (succinate, malate, pyruvate) bi t bo cht ca k ch, nhng cht ny c tip tc phn gii trong chu trnh Krebs. y xut hin NADH, c s dng mt phn l cht cho in t ferredoxin v flavodoxin, mt phn c s dng lm nguyn liu ca h hp. H thng oxy ho kh ca chui h hp cu to nn mng. Chng cung cp ATP v ATP cn cho s hot ho enzyme nitrogenase. Chui h hp c cung cp O2 bi leghemoglobin. Phc h protein cha heme c trong khng bo nhim ny c th so snh vi myoglobin trong m c hoc hemoglobin trong mu. N cung cp O2 cho chui enzyme h hp v n kim sot O2 i vo bacteroid. S kim sot ny rt quan trng, v enzyme ny b c ch bi O2. NH3 c to nn bi nitrogenase c a n t bo cht ca t bo ch v y c s dng tng hp nn aminoacid. S tng hp enzyme nitrogenase c iu khin bi enzyme glutamate synthetase, xc tc cho tng hp glutamin t NH3. Nu trong h thng c t NH3 th

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glutamate synthetase kch thch tng hp nitrogenase, nng NH3 cao th c ch s tng hp nitrogenase. C ch iu khin ny rt c ngha, tuy nhin khi cung cp NH3 d tha th nitrogenase khng c s dng, v c NH3. V vy bn phn m c ch s to thnh nt sn ca cy h u. vi khun rhizobium sng cng sinh, NH3 to thnh c a nhanh n t bo ch, nn s tng hp nitrogenase hu nh khng b km hm. Hn na vi khun trong t bo ch lun c cung cp bi nhng anion hu c. l nhng nguyn nhn quan trng nht, ti sao s cng sinh vi khun/cy h u c t l c nh N2 cao. V d cy c (trifolium pratence) sng cng sinh vi vi khun trong iu kin sinh trng tt c th c nh 300-400 kg N/ha/nm. Vi khun sng t do t l c nh nit b nh hng do s tch lu NH3 trong t bo vi khun v s cung cp khng y carbon hu c. Loi vi khun c nh nit sng t do cn cho s trao i cht ca chng nhng hp cht hu c d phn gii, nh ng v cc acid hu c, thng c trong t vi lng khng . V vy kh nng c nh m ca vi khun sng t do c bn l thp. Nhng vi khun lu hunh t dng thuc loi anabaena, nostoc v rivularia t ng ho CO2 bng quang hp v c nh N2. Kh nng c nh N2 b nh hng rt nhiu khi nng O2 cao. 7.8 Cc ng hng bin i NH3- chu trnh ornithine S kh nitrate-nitrite v h thng nitrogenase sn sinh NH3, sau NH3 kt hp vi mt phn t hu c, thng l -cetoglutaric acid hoc glutamic acid. Xc tc cho phn ng gn NH3 ny thc vt bc cao v bc thp l glutmate synthetase (GS), enzyme ny thng hot ng trong mt nhm. Glutmate synthetase l mt protein c 8 tiu n v vi trng lng phn t t 350 n 400 kDa. Ngi ta bit c 2 loi enzyme ng phn, mt loi c trong t bo cht (GS1) v mt loi c trong lc lp. Phn ng do glutmate synthetase xc tc nh sau:

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Cht nhn NH3 l glutamate. Trong phn ng, enzyme c phosphoryl ho (kinase) v sau c kh phosphoryl ho (phosphatase). C hai phn ng v tng quan l s thu phn ATP. Sn phm phn ng l glutamine, mt amide. Mt phn t tng t glutamine l asparagine, c to nn tng t glutamine. Tuy nhin n khng phi l cht nhn NH3 quan trng. Vai tr trung tm ca glutmate synthetase i vi s ng ho NH3 c gii thch glutamine l cht cho nhm NH2 khi tng hp glutamate. y -cetoglutaric acid l cht nhn, c ch ra nh sau:

Cc cht tham gia phn ng trao i 1 nhm NH2 vi nguyn t O. mi sn phm to thnh thiu 1 nguyn t H, c ferredoxin hoc NAD(P)H cung cp. Bng cch ny t glutamine v -cetoglutaric acid 2

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phn t glutamate c to nn. Qu trnh ny gm s chuyn nhm amine v s kh v phn ng c xc tc bi glutamate synthetase. Glutamate synthetase c hai loi isoenzyme c cu to khc nhau. Glutamate synthetase ph thuc vo ferredoxin l 1 flavoprotein cha Fe-S c trng lng phn t 140-230 kDa, enzyme ny c trong lc lp, nhn cht kh qua ferredoxin trc tip t quang hp. Glutamate synthetase c trong t bo cht c trng lng phn t khong 240 kDa. Cht kh ca n l NADH (t ng phn) hoc NADPH (t oxy ho trc tip glucose). C 2 enzyme glutamate synthetase v glutamine synthetase hot ng trong 1 nhm, lm cho qu trnh phn ng thnh 1 vng trn, nh hnh 7.18. Tng qut chu trnh ch ra rng -cetoglutaric acid c amine ho nh NH3 vi s tiu tn 1 ATP v 2e-. -cetoglutaric acid (oxoglutarate) c ngun gc t chu trnh Krebs. Sn phm ca phn ng l glutamate, c ngha quan trng, v nh chuyn amine ho m nhm amine c chuyn n mt cht khc (cetoacid, aldehyd). Hu ht nit c trong thc vt xy ra qua phn ng ny, mt ln c trong nhm amine ca glutamate. Glutamate synthetase l 1 glutamine-oxoglutarate-aminotransferase, c gi tt l GOGAT. V vy ngi ta gi chu trnh tng hp glutamate l con ng phn ng GOAGT. Nhng enzyme ca chu trnh tng hp glutamate c bit c nhiu trong nhng t bo ca nt sn. y nhng phn t NH3 c cung cp bi bateriod ca vi khun nt sn c ng ho qua chu trnh ny. Con ng GOAGT cng c ngha trong lc lp ca thc vt C3. NH3 c gii phng ra mt lng ln trong h hp sng, nu NH3 ny bay ra khng kh th y l mt s mt mt ln i vi thc vt. NH3 gii phng ra c tip nhn bi chu trnh tng hp glutamate v nit ca n c s dng li trong trao i cht ca thc vt dng aminoacid.

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Tng qut: NH3 + ATP +2e- + 2H+ + Oxoglutarate Glutamate + ADP +Pi Hnh 7.18 Chu trnh tng hp glutamate (con ng GOGAT)

lu ngi ta cho rng enzyme glutamate dehydrogenase (GDH) cng c ngha i vi s ng ho NH3 trong thc vt. Glutamate dehydrogenase xc tc cho phn ng sau:

y l s kh amine ho oxy ho, l mt qu trnh gii phng NH3, ngc vi qu trnh ng ho NH3. Tuy nhin khi nng NH3 cao th n s gy c cho thc vt. i vi nhiu loi vi sinh vt s ng ho NH3 nh GDH c ngha. Trong thc vt GDH c ch yu trong ty th. NADH m n to ra c th i vo chui h hp, NH3 c th c s dng cho tng hp asparagine nh enzyme asparaginsynthetase theo phng trnh sau:

Cht nhn NH3 l aspartate. Phn ng ny tng t phn ng tng hp glutamine. Glutamate dehydrogenase c tng i nhiu trong m r

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v nhng m gi. V vy ngi ta cho rng, y n sn sinh NH3 v l iu kin tng hp asparagine, l dng nit vn chuyn v c vn chuyn ra khi nhng m ny. 7.9 Cc ng hng tng qut ca s bin i aminoacid Cc aminoacid v c bn c s dng tng hp protein song cng c th chu nhng s bin i khc. l phn ng theo nhm amine carboxyl v phn ng theo gc R ca aminoacid. - Cc phn ng theo nhm -amine: l cc phn ng phn gii aminoacid thnh NH3 v cc sn phm tng ng theo 4 hng c bn sau: a. Kh amine ho bng cch thu phn R - CH - COOH | NH2 Aminoacid + H2O R - CH -COOH | OH Hydroxyacid + NH3

b. Kh amine ho bng cch kh R - CH -COOH | NADH2 NADPH2 NH2 Aminoacid R - CH2 - COOH + NH3 NAD NADP Acid hu c

c. Kh amine ho bng cch oxy ho R - CH - COOH R - C -COOH | + 1/2 O2 || + NH3 NH2 O Aminoacid Cetoacid Phn ng ny tin hnh qua 2 giai on: trc ht aminoacid b loi hydro to thnh iminoacid tng ng vi s tham gia ca enzyme dehydrogenase R - CH -COOH R - C - COOH | NAD NADH2 || NH2 NADP NADPH2 NH

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Aminoacid

Iminoacid

Tip iminoacid kt hp vi phn t H2O, NH3 b loi tr v gii phng cetoacid R - CH - COOH || NH Aminoacid + HOH R - C -COOH || O Cetoacid + NH3

d. Kh amine ho bng cch chuyn ho ni phn t. R - CH2 - CH -COOH | NH2 Aminoacid R - CH = CH - COOH + NH3 Acid cha no

Trong cc phn ng trn th phn ng amine ho bng cch oxy ho l ph bin trong thc vt so vi ng hng khc. Cc -cetoacid u l sn phm ch yu ca s kh amine ho cc aminoacid. - Cc phn ng theo nhm carboxyl: din ra ch yu theo hai qu trnh: s kh carboxyl ho v s to thnh aminoacyl adenylate a. Qu trnh kh carbocyl ho cc aminoacid Qu trnh ny c thc hin trong cc ng vt, thc vt v c bit l ph bin trong c th sinh vt. S kh carboxyl tin hnh theo s sau: R - CH - COOH | NH2 Decarboxylase CO2 R - CH2 - NH2 Amine

Sn phm phn ng l cc amine. Mi mt aminoacid ring bit c xc tc bi mt enzyme decarboxylase tng ng.

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Sn phm ca s kh carboxyl ho cc aminoacid l amine, diamine, CO2, NH3 v cc hp cht khc. Tr CO2, NH3, tt c cc hp cht khc u b phn gii tip tc. Nu trong iu kin bt li th cc amine v diamine tch t li gy c cho cy, trong iu kin bnh thng chng s b oxy ho di tc dng ca enzyme oxydase to thnh NH3 v cc aldehyd. Monoaminoxydase R - CH2 -NH2 + H2O + O2 R - CHO + NH3 + H2O2 Aldehyd dehydrogenase R - CHO + H2O R -COOH Cc acid v aldehyd li b oxy ho tip tc to thnh CO2, H2O v NH3. Cc sn phm ny chnh l sn phm cui cng ca s phn gii aminoacid. b. Phn ng theo nhm carbocyl l qu trnh to thnh phc hp aminoacyl-adenylate. Chi tit c ch ca phn ng ny trong phn sinh tng hp protein. - Nhng chuyn ho ca phn t aminoacid c lin quan n gc R trong phn t. Do tnh mun mu mun v ca gc R nn cc phn ng ho hc ca chng v cng phong ph v a dng. Nhiu phn ng c thc hin trong qu trnh trao i aminoacid, trong quan trng nht l cc phn ng bin i t mt aminoacid ny thnh mt aminoacid khc. Nh ng hng ny m kh nng tng hp cc aminoacid tng ln rt nhiu. Sau y ch nu ln mt s v d: Khi oxy ho phenylalanine s to thnh tyrosine Thu phn arginine th ornithine c to thnh v t ornithine s thu c proline hoc glutamic acid. H2N - C = NH | NH | (CH2)3 | CH - NH2 | COOH

Arginase + HOH

NH2 | (CH2)3 + O = C | CH - NH2 | COOH

NH2

NH2

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Arginine

Ornithine

Ur

Khi oxy ho nhm -SH ca cysteine s thu c cystine SH | NAD NADH2 CH2 - S -S - CH2 CH2 Cystine reductase | | | CH-NH2 CH-NH2 CH - NH2 | | | NAD NADH2 COOH COOH COOH Cysteine Cystine T threonine nu loi tr gc acetaldehyd s thu c glycine. CH3 | CHOH | CH - NH2 | COOH Threonine 7.10 Sinh tng hp protein Tham gia vo s tng hp polypeptide v protein c mRNA, tRNA, rRNA, trong rRNA l thnh phn ca ribosome. Vi s tng polypeptite thng tin di truyn c chuyn t DNA n polypeptide. Trc khi nghin cu qu trnh tng hp protein cn phi cp n tng nucleic acid v ribosome.

CH3 | + CHO Acetaldehyd

CH2-NH2 | COOH Glycine

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Hnh 7.19 S tng hp protein

1. Nguyn liu tng hp polypeptide l cc aminoacid. tham gia vo phn ng chng phi c hot ho nh ATP, to nn AMPaminoacyl = aminoacid hot ho 2. Aminoacyl c chuyn ln tRNA. 3. tRNA vn chuyn aminoacyl n ribosome. y aminoacyl c th to lin kt peptide. 4. mRNA c tng hp trong nhn t bo ri i n ribosome, l ni tng hp polypeptide. mRNA mang thng tin cho chui polypeptide c tng hp, xc nh trnh t ca aminoacid trong chui polypeptide. Thng tin di truyn trong mRNA th hin trnh t base ca chng v ba nucleotide ng cnh nhau m ho cho mt aminoacid xc nh. Cc b ba ny c gi l codon. Di y l cc codon ca mt on mRNA m ho cho alanine, aspartic acid, cysteine v serine.

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Hnh 7.20 Mt on DNA vi nhng codon m ho cho alanine, aspartic acid, cysteine v serine

Ngy nay nhng codon m ho cho cc aminoacid cu to nn protein c bit. iu rt th v l cc m di truyn cho tt c mi sinh vt t sinh vt nhn s n gin nht cho n nhng ng vt c v l nh nhau. Tuy nhin cng c mt s ngoi l nh ty th ngi. Qua bng m di truyn ta thy phn ln cc aminoacid c m ho bi hai codon, v cc codon c hai base nit u tin ging nhau. Ribosome c to nn t rRNA v protein. s kt hp ny c s tham gia ca Mg+. Ribosome c to nn t 1 tiu phn ln v 1 tiu

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phn nh. ln ca cc tiu phn c c trng bi h s lng S ( h s Svedberg). dx/dt S = --------2 x X l khong cch ca trc ly tm = vn tc gc t l thi gian dx/dt, khong cch m tiu phn di chuyn trong n v thi gian. Khong cch ny cng di khi tiu phn cng nng. H s lng S i vi protein c ln t 1 x 10-13 n 200 x 10-13. S bin i n v h s lng ca Svedberg l nhn vi 1013, tng ng h s lng ca protein l 1-200. S cng ln th tiu phn cng ln (cng nng). Bng 7.7 ch h s lng ca cc tiu phn ribosome khc nhau v ca nucleic acid. Bng 7.7 H s lng ca cc phn t khc nhau v cc kt hp (S) ------------------------------------------------------------------------------------tRNA 4 mRNA 6-25 rRNA 5-23 Tiu n v nh ribosome 30 (sinh vt nhn s) Tiu n v ln ribosome 50 (sinh vt nhn s) Ribosome 70 (sinh vt nhn s) Tiu n v nh ribosome 40 (sinh vt nhn chun) Tiu n v ln ribosome 60 (sinh vt nhn chun) Ribosome 80 (sinh vt nhn chun) ------------------------------------------------------------------------------------Bng 7.8 Cc rRNA ca ribosome ca tiu phn ln v nh ca sinh vt nhn chun v nhn s: ------------------------------------------------------------------------------------Sinh vt nhn s Sinh vt nhn chun ------------------------------------------------------------------------------------Tiu n v nh 16S rRNA Tiu n v nh 18S rRNA Tiu n v ln 23S rRNA Tiu n v ln 5,8S rRNA Tiu n v ln 5S rRNA Tiu n v ln 28S rRNA Tiu n v ln 5S rRNA --------------------------------------------------------------------------------------

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Bng 7.8 cho bit ln ca ribosome v cc tiu n v ribosome. Da trn n v S ny ta thy ribosome ca sinh vt tin nhn nh hn sinh vt nhn chun. Ribosome trong ty th v lp th tng t v ln v chc nng nh ribosome sinh vt tin nhn. Hai tiu phn ca ribosome c gn vi mRNA, ngha l khi khng c mRNA th hai tiu phn khng kt hp vi nhau m trng thi ring l. Cu trc khng gian hai chiu tRNA tng t l c tam ip. Tuy nhin tRNA c cu trc khng gian ba chiu, v vy phc tp hn v c biu din theo s hnh 7.21. Trong tRNA c mt s base nit l nh cytosine v guanine methyl ha l pseudouridine, dihydrouracil v hypoxanthine.

Pseudouridine lin kt vi ribose khng phi N-glycosid m l Cglycosid ( v tr C6 ca uracil). V y l nhng base nit l nn cc v tr khc nhau ca tRNA khng c s cp i cc base. ngha c bit hn v chc nng ca tRNA l c v tr anticodon v ch kt hp ca aminoacyl.

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Hnh 7.21 Cu trc ca tRNA vi anticodon, ch kt hp vi aminoacyl, dihydrouridine v pseudouridine

Ch kt hp ca aminoacyl ca tt c cc phn t tRNA l nh nhau, l trnh t cytosine-cytosine-adenine. Aminoacyl c kt hp vo ribose ca adenine. Anticodon c trng cho tng loi aminoacid, l nhng base b sung cho codon trn mRNA. Anticodon c c t phi sang tri. c tnh ca anticodon l hai base u tin c hiu i vi aminoacid. Base th ba c th l khc nhau, n c th linh ng, c ngha l khng to cu hydro vi coon. bng sau anticodon C-A (codon = G-U) m ho cho aminoacid valine. Inosine l mt nucleoside c to nn t hypoxanthine v ribose. Mt aminoacid c nhiu tRNA c hiu. Trnh t ca mt tRNA c hiu cho mt aminoacid nht nh c th l khc nhau. S khc bit trong cu trc bc mt ca tRNA gia nm men v chut c khng nh.

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Trc khi aminoacid c s dng tng hp protein chng phi c hot ho nh ATP, ATP kt hp vi aminoacyl v pyrophosphate c tch ra (hnh 7.22). y xut hin mt adenosinphosphataminoacyl (aminoacid hot ho). ATP cung cp nng lng cn thit cho qu trnh tng hp. Adenosinphosphataminoacyl mang gc acyl ribose cui cng ca tRNA. N kt hp vi aminoacyl v tr carbon th 3 ca ribose ng thi gii phng AMP. Nh vy aminoacyl lin kt vi cht mang c hiu v vn chuyn chng n v tr tng hp protein. Ton b qu trnh hot ho aminoacid v kt hp aminoacyl vi tRNA c xc tc bi mt enzyme aminoacyl-tRNA-synthetase. Ribosome, mRNA v tRNA-aminoacyl to iu kin cho vic to lin kt peptide. Theo kt qu th nghim vi khun, c bit l E.coli th qu trnh xy ra nh sau: Tiu n v nh ca ribosome kt hp vi gc phosphate ca mRNA cng vi mt tRNAaminoacyl. Chng c gi l phc h khi u v sau mi c kh nng kt hp vi tiu phn ln ca ribosome, to ra b my tng hp polypeptide. Tip theo mt phn t tRNA-aminoacyl th hai kt hp vi ribosome v m bo tRNA-aminoacyl ng v tr ca mnh (hnh 7.23). V tr chnh xc l base nit ca codon c kt hp b sung vi base nit anticodon.

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Hnh 7.22 Hot ho aminoacid v s kt hp ca aminoacyl vo tRNA

- Amino acid c hot ho bng cch gn vi tRNA ring ca n. Qu trnh ny gm 2 phn ng, c xc tc bi cng mt enzym c hiu i vi mi amino acid, l cc amino acid-tRNA-synthetase. Nh vy c 20 amino acid th cng c 20 loi enzyme khc nhau. Trong phn ng th nht amino acid phn ng vi ATP to nn AMPaminoacyl v 1 pyrophosphate (hnh 7.22, phn ng 1). Hp cht c to nn bi nhm carboxyl ca amino acid v mt gc phosphoric acid v gn vi enzyme nn c kh nng phn ng cao. Trong phn ng th hai, n s phn li, cho php aminoacyl t AMPaminoacyl c vn chuyn n tRNA. y aminoacyl thay th H ca nhm OH v tr carbon th 3 to thnh aminoacyl-tRNA.

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- giai on khi u tng hp chui polypeptide cn c hai iu kin: Mt l trn mRNA c mt khu vc khng m ho, l du hiu kt hp vi ribosome m u cho vng m ho. Hai l c b m khi u AUG lm im xut pht. vi khun i khi thy m khi u GUG thay cho AUG. Nh ta bit b m AUG m ho cho methionine. vi khun, ngi ta thy c hai loi tRNA i vi methionine: mt l tRNA nhn gc methionine avo thnh phn ca chui polypeptide Met-tRNA, hai l tRNA nhn gc formyl-methionine (fMet-tRNA) c vai tr quan trng trong vic khi u tng hp chui polypeptide. sinh vt nhn s, nhm amine ca methionine c th c formyl ho bi N10-formyltetrahydrofolic acid, nh enzyme transformylase c hiu. Do nhm amine b gc formyl bao vy nn khng cho php n tham gia vo qu trnh ko di chui, m ch tham gia vo qu trnh khi u tng hp protein. sinh vt nhn s anticodon c nh v gia ca 16SrRNA. Base b sung kt hp vi base ca codon mRNA khong cch t 0,3 n 0,4 nm, nh vy s xut hin lc hp dn Van der Waal. Bng cch ny trc ht tRNA-aminoacyl kt hp vi tiu phn nh ca ribosome v thc cht l vi mRNA. Phc h ny phn ng vi nhn t ko di. Nhn t ny c trn tiu phn ln ca ribosome. sinh vt nhn s, ngoi cc yu t tham gia khi u tng hp protein nh fMet-tRNA, mRNA, cc tiu n v ribosome 30S v 50S, GTP, cn c 3 protein na l cc yu t khi u: IF1 (M 9000), IF2 (M 65000-80000) v IF3 (M 29000). Kt qu ca giai on ny l to thnh phc hp khi u: fMet-tRNA-mRNA-ribosome 70S. Qu trnh c bt u bng s kt hp gia yu t IF3 vi tiu n v 30S. Nh yu t ny m tiu n v 30S kt hp vi mRNA v tr khi u. Trong giai on hai fMet-tRNA gn vi phc IF2-GTP to thnh IF2-GTP-fMet-tRNA (t hp 3). giai on ba v bn phc hp IF3-30S kt hp vi t hp 3. Nh yu t IF2 m fMet-tRNA c a vo khu vc P. Ngi ta cha bit chnh xc vai tr ca IF1, c th n tham gia vo vic i mi chu trnh bng cch gp phn gii phng yu t IF2 ra khi phc hp. Cui cng tiu n v 50S kt hp vi tiu n v 30S to ra phc hp ribosome 70S, ng thi gii phng c 3 yu t khi u cng nh gii phng c GDP v Pi. sinh vt nhn chun, qu trnh tng hp polypeptide trong t bo cht c tRNA khi u cng mang methionine nhng khng c formyl ho. y cng c cc phn ng vi cc yu t khi u eIF1, eIF2 v eIF3 tng t nh sinh vt nhn s.

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Trn tiu phn ln c ba vng quan trng: vng A: l ni tRNAaminoacyl c kt hp, vng P: ni gn tRNA-peptidyl v vng E: thc hin s ko di chui peptide (hnh 7.23).

Hnh 7.23 Nhng vng phn ng quan trng ca ribosome A: v tr gn ca tRNA-aminoacyl, P: v tr gn ca peptidylaminoacyl

- giai on ko di chui polypeptide, amino acid th hai (do m tip theo ca mRNA quy nh) gn vi tRNA ring ca n s c a vo phc hp khi u vng A. Giai on ny cn GTP v cc yu t ko di chui. sinh vt nhn s yu t ko di l EF-T (gm 2 b phn hp thnh l EF-Tu v EF-Ts). Yu t ny sinh vt nhn chun l eEF-1. Cc yu t ny l nhng protein: EF-Tu (M 43000), EF-Ts (M 74000). Chng c vai tr a aminoacyl-tRNA vo vng A ca ribosome thng qua cc bc sau: EF-Tu + GTP EF-Tu. GTP (t hp hai) EF-Tu.GTP + amonoacyl-tRNA aminoacyl-tRNA-EF-Tu-GTP (t hp ba). T hp ba ch kt hp vi vng A trong ribosome v vng P c fMettRNA ( giai on sau l peptidyl-tRNA) chim ch. y l giai on quyt nh bo m nh hng ng to ra lin kt peptide gia aminoacyl-tRNA v peptidyl-tRNA. Trc khi aminoacyl c gn kt vo peptidyl th n kt hp vi GTP vng E ca ribosome to nn mt phc gm ba yu t. Sau khi GTP thu phn th lin kt peptide c to thnh. Qu trnh ny c xc tc bi enzyme peptidyltransferase, chuyn peptidyl n tRNA-aminoacyl va kt hp vo (hnh 7.24). Enzyme ny c cu to ch yu t rRNA. Phn phi protein 50S rRNA th hin y hot tnh ca peptidyltransferase. Nucleic acid c hot tnh enzyme c gi l ribozyme. Protein ca ribosome tham gia qu trnh enzyme l do s thay i cu hnh. i vi hnh th ca tng thnh phn ribosome ion Mg+ c mt ngha quan trng.

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Sau khi kt hp vi ribosome, GTP b thu phn ng thi gii phng t hp EF-Tu-GDP ra khi ribosome. Phc hp EF-Tu-GDP khi tc ng tng h vi EF-Ts v GTP s li bin thnh EF-Tu-GTP. Do n c th lai tng tc vi aminoacyl-tRNA tip theo. Cc nghin cu cho thy s lng yu t EF-Tu bng khong s lng phn t aminoacyltRNA. T ngi ta cho rng phn ln cc aminoacyl-tRNA l nm trong thnh phn t hp ba ch khng phi trng thi t do. Hnh 7.24 biu din s hnh thnh lin kt peptit. V tr A ribosome l vng nhn, v tr P l vng cho. vng nhn kt hp vi tRNA-aminoacyl mi i vo (aminoacyl vi R0 hnh 7.24). vng cho th tRNA-aminoacyl kt hp vi ribosome cng nh tRNApolypeptidyl. tRNA-polypeptidyl mang mt chui polypeptide vi hai aminoacid (R1 v R2). v tr ny c hai thnh phn tRNA-aminoacyl v tRNA-polypeptidyl tham gia phn ng: nguyn t H ca nhm amine ca aminoacid gn kt vi polypeptidyl ribose ca tRNA bng lin kt ester. Chui polypeptidyl c chuyn t cht cho n cht nhn. tRNA gn vng cho by gi trng thi t do, ngha l ribose cui cng ca n c nhm OH v tr C3 t do. Lc ny chui polypeptide di thm mt n v (aminoacyl) (hnh 7.24, bn phi). y l phn ng vn chuyn peptidyl. Bng cch ny lin kt peptide c to nn v tRNA c gn kt trc tr nn t do. Phn ng ko di chui cn c s tham gia ca GTP v mt yu t ko di khc l EF-G ( sinh vt nhn chun l eEF-2). Yu t EF-G l mt trong nhng yu t c bn ca t bo, c khi lng phn t 72000. Yu t ny khi kt hp vi ribosome gp phn to cho ribosome chuyn dch mt khong cch bng mt b ba trn phn t mRNA, khi peptidyl-tRNA chuyn t vng A sang vng P. Sau khi kt hp ribosome dch chuyn n codon tip theo, tRNA mang aminoacid tip theo c kt hp vo. Ribosome l h thng c ho hc c kh nng chuyn ng tng t nh protein c. - Giai on kt thc: Qu trnh tng hp si polypetide kt thc khi trn mRNA c mt codon kt thc xut hin hoc l n u cui ca mRNA. Nhng codon kt thc l UAA, UAG v UGA. Si polypeptide tch khi ribosome v t ng hnh thnh cu trc bc bn. Sau ribosome b phn ly thnh cc tiu n v 30S v 50S ri nhp vo kho d tr ribosome. S nhn bit cc b m kt thc c thc hin bi cc yu t kt thc. l cc protein, sinh vt nhn s l RF1 (M 4000) nhn bit b m UAA v UAG v RF2 (M 47000) nhn bit m UAA v UGA. sinh vt nhn chun c mt yu t kt thc l eRF. thc hin phn

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ng gii phng chui polypeptide th peptidyl-tRNA phi nm vng P, cn cc yu t kt thc tng tc vi vng A ca ribosome.

Hnh 7.24 S to thnh lin kt peptide ribosome a) tRNA-amioacyl kt hp vi tRNA-peptidyl b) Aminoacyl c chuyn ln peptidyl, tRNA gn vng cho tr nn t do, si peptide di hn 1 aminoacid ang vng cho

Bng cch ny hng tng hp ca si polypeptide c xc nh, v aminoacid ch c th c kt hp vi nhm carbonyl m khng kt hp vi nhm amine. sinh vt nhn chun s tng hp polypeptide cng bt u bng methionine, khng c formyl ho. Methionine v methionine c formyl ho s tch ra bng thu phn khi qu trnh tng hp polypeptide kt thc. Qu trnh tng hp protein xy ra ribosome ca t bo cht v ty th, lp th. Chng c h thng tng hp protein ring v c nhng enzyme cn thit cho qu trnh . Tuy nhin trong ty v lp th khng c tt c cc loi enzyme.

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Hnh 7.25 Cc tiu n v ca ribosome, polysome v mRNA xuyn qua cc ribosome

Trong nhiu trng hp mt si mRNA thc hin gii m trn nhiu ribosome, nh vy cng mt si mRNA c t 20 n 40 protein c tng hp. S tp hp mt lot ribosome trn mt si mRNA c gi l polysome. hnh 7.25 biu din s sp xp ca cc ribosome. Trn s cho thy nhng ribosome no trt xa nht trn si mRNA th si polypeptide c to nn l di nht.

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Mt s khng sinh c th nh hng n qu trnh sao chp v gii m. Qu trnh sao chp b c ch bi actinomycin D v rifamycin. Cht th nht phong to base nit guanine, cht th hai c ch RNApolymerase. Qu trnh gii m b c ch bi puromycin, streptomycin v chloramphenicol v thc ra l chng kt hp vi vng phn ng ca ribosome. Khng sinh cng km hm qa trnh tng hp protein ca vi sinh vt. T cc vn nu trn ta c nhn xt rng: qu trnh sinh tng hp protein v cng phc tp, c s tham gia ca nhiu yu t. Ngi ta xc nhn c ti hng trm yu t tham gia vo qu trnh sinh tng hp protein trong t bo. V mt nng lng, qu trnh sinh tng hp l mt qu trnh thu nng lng. vi khun cng nh sinh vt nhn chun, nhu cu nng lng cho qu trnh l rt ln. a mt amino acid vo chui polypeptide cn chi ph 3 lin kt cao nng: - 1 ATP cho hot ho amino acid giai on u - 1 GTP a amonoacyl-tRNA (trong t hp ba) vo vng A ca ribosome - 1 GTP cn ribosome di chuyn mt b ba trn mRNA Do giai on khi u cn cn 1 GTP tham gia, nn tng hp lin kt peptide u tin phi s dng ti 4 lin kt cao nng. Ngi ta cho rng phn t GTP tham gia vo qu trnh ny c hai chc nng: mt l to cu hnh thch hp cn thit cho s tng tc tip theo, hai l bo m nng lng ribosome di chuyn trn mRNA. V mt tc , qu trnh tng hp protein xy ra rt nhanh v ph thuc vo nhit . vi khun nhit 370C trong mt giy chui polypeptide tng t 12 n 17 aminoacid. V vy, tng hp 1 phn t protein trung bnh c ln 300 aminoacid mt khong 20 giy. Trong t bo sinh vt nhn chun, tc tng hp chm hn, v d trong t bo hng cu li 370C tc ko di chui peptide l 2 aminoacid trong 1 giy. So vi tc giai on ko di chui th giai on khi u v kt thc xy ra chm chp hn. 7.11 S phn gii cc hp cht cha nit S phn gii protein, nh trong h tiu ho ca ng vt, ht ny mm hoc trong t khi phn gii cc hp cht hu c nh vi sinh vt, bt u bng s thu phn. Cc lin kt peptide b ct ra do cc peptidase, xut hin cc aminoacid t do. Tu thuc vo loi peptidase m chui peptide c thu phn gia hoc mt u chui.

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 qu trnh ny mm cc aminoacid c to ra c s dng tng hp nn cc protein mi. S phn gii protein nh vi sinh vt trong t v trong h tiu ho c thc hin tip theo l cc aminoacid b kh amine ho, gii phng ra NH3. Nhng vi sinh vt sng trong t c kh nng thc hin phn ng ny v vy c gi l vi sinh vt to amn.

Hnh 7.26 Chu trnh Ornithine

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Nguyn tc ca s kh amine ho l phn ng oxy ho v s tch NH3 (kh amine ho oxy ho). trong ng vt, c bit l glutamate c s kh amine ho bng cch oxy ho. Enzyme xc tc cho phn ng ny l glutamate dehydrogenase. Trong iu kin ym kh nhng aminoacid b kh amine ho bng phn ng kh (xem s ), mt qu trnh xy ra trong t ngp nc, nh t la.

Trong c th ng vt c v ur c to ra t NH3 qua chu trnh c gi l ornithine (hnh 7.26). Trong chu trnh ny gc carbamyl v guanidyl ng vai tr quan trng. C hai u xut pht t ur, nh cng thc sau:

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Chu trnh bt u vi s tng hp carbamic acid (xem s ). y l phn ng cn bng t nhin. Cht phn ng c cung cp do qu trnh h hp (CO2) v qu trnh kh amine ho bng cch oxy ho (NH3). Carbamic acid c phosphoryl ho nh ATP to thnh carbamylphosphate:

Carbamylphosphat phn ng vi ornithine to citrulline v phosphate v c c tch ra.

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 y nguyn t H ca nhm amine cui cng c thay th bng nhm carbamyl. Citrulline phn ng vi aspartic acid to thnh argininosuccinic acid. Phn ng cn ATP.

Qua s bin i ni phn t cht ny c tch thnh arginine v fumaric acid.

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Arginine l 1 dn sut ca guanidyl. Nh arginase (hydrolase) arginine c tch ra thnh ornithine v ur.

Sau khi to thnh ornithine chu trnh mi li bt u. Cht i vo chu trnh: CO2, NH3, asparagine v ATP. Chu trnh to nn ur v fumaric acid. Hai nguyn t N ca ur bt ngun t NH3 v asparagine. Nh vy NH3, dng c c bin i thnh dng khng c, v c thi ra cng vi nc tiu. Fumaric acid l cht trao i ca chu trnh Krebs v cng c tip tc bin i . Vic s dng ATP y ni ln rng, vic gii c NH3 cn nng lng. Khung carbon ca fumaric acid bt ngun t asparagine. Nh vy mt aminocid trong qu trnh phn gii ca chu trnh Krebs ni vi chu trnh ornithine. Nhng enzyme ca chu trnh ornithine phn th nh v trong c cht ca ty th l glutamate dehydrogenase v citrullinsynthetase, nhng enzyme cn li ca chu trnh nh v trong t bo cht. Trong thc vt bc cao cng c cc enzyme ca chu trnh ornithine. Trong ht ny mm arginase v ornithin-transcarbamylase c tm thy. Trong enzyme cui cng xc tc cho phn ng tng hp citrulline t carbamylphosphate v ornithine. V vy ngi ta cho rng, nhng enzyme k trn c vai tr quan trng i vi vic huy ng cc protein d tr. Trong cy nho arginine l mt dng d tr nit, c tch lu vo ma thu trong cnh (g). Trong nhiu trng hp ur c tch thnh CO2 v NH3 trong t nh urease, nh vy nit c bn dng ur c tip nhn dng NH4+ hoc sau c bin i nh vi sinh vt sang dng NO3- .

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S bin i NH3 trong t thnh NO3- c thc hin nh vi khun, chng oxy ho (t chy) NH3 cng nh HNO2 thu c nng lng cho qu trnh trao i cht. Nhng loi vi khun quan trng nht oxy ho NH3 l loi nitrosomonas, nitrosocystus, nitrospira. S oxy ho cn c O2 tng ng theo phng trnh sau: NH3 + 3/2 O2 HNO2 + H2O

S oxy ho HNO2 thc hin nh loi nitrobacter v tng ng vi phng trnh sau: HNO2 + 1/2 O2 HNO3

qu trnh ny mt acid mnh, HNO3 c to thnh. l nguyn nhn thc t s phn gii cc cht hu c cha nit lm cho t chua. Qu trnh to nitrite (HNO2-) v nitrate trong t c gi l qu trnh nitrate ho. Nhng loi vi khun cn cho qu trnh ny c nhiu trong t. t trng thng thong vi pH trung tnh l iu kin l tng i vi nhng vi khun ny. Trong iu kin ym kh c th xy ra s kh NO3- (kh nitrate h hp hoc l d ho). Qu trnh ny c gi l phn nitrate ho. Ngy nay ngi ta bit 23 loi khc nhau c kh nng ny. l Alcaligenes, Agrobacterium, Azospirillum, Holobacterium, Pseudomonas v Rhizobium. Chng c kh nng to ra nhng enzyme kh trong iu kin thiu O2, s dng O trong NO3- cho h hp ho kh, l cht nhn e- ca chui h hp. qu trnh ny NO3- b kh thnh N2O v N2. Trong qu trnh phn nitrate ho cng xut hin NO. Trnh t phn ng c gii thch nh sau:

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 qu trnh ny c 4 enzym kh tham gia: NO3- -Reductase, NO2Reductase, NO-Reductase v N2O-Reductase. Enzyme quan trng nht l NO2--Reductase, xc tc cho qu trnh chuyn ion NO2- thnh dng cht kh (NO). N2O-Reductase khng c trong tt c vi khun phn nitrate ho. mt s vi khun qu trnh ny kt thc vi chui phn ng N2O. T qu trnh phn ng rt ra rng, s phn nitrate ho to tnh kim, v chng cn H+ mi bc ca phn ng. S phn nitrate ho c mt vai tr ln trong t nhin. T hp cht nit v c v hu c N2 c gii phng li i vo khng kh. S phn nitrate ho i lp vi s ng ho N2. Trong qu trnh u tin sn sinh N2, qu trnh th hai cn N2. Nh vi khun c nh m m hng nm t 100-200 x 106 t N t khng kh c a vo t. Bng qu trnh phn nitrate ho m hng nm c khong 200-300 x 106 t N dng N2 c gii phng ra trong khng kh. N2O c to ra trong qu trnh phn nitrate ho l vn sinh thi kh khn. l cht rt bn vng, bay ln n tng tnh kh v tham gia vo s phn gii tng ozon.

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Chng 8

Mi lin quan gia cc cht trong trao i cht

Trn y chng ta nghin cu s trao i cht ca tng hp cht c bn trong c th sng. Tuy nhin s phn chia nh vy ch gip lm n gin ho nhng vn mun trnh by. Trong thc t gia cc cht c nhiu mi lin h tng h. iu buc chng ta phi nghin cu s trao i cht ca t bo trong mt tng th cc mi lin h. Tht vy, trong c th khng th tm thy s trao i cht ca mt cht no xy ra tch ri vi s trao i ca cc hp cht khc. Mi lin quan tng h gia s trao i cc hp cht ny th hin trn hai mt c bn: nguyn liu v nng lng. Mi lin quan v mt nguyn liu l kh nng chuyn ho mt cht ny thnh mt cht khc thng qua mt s sn phm trung gian. V d, carbohydrate c th chuyn ho thnh aminoacid bng cch amine ho mt s cetoacid. Ngc li mt s aminoacid c th chuyn thnh carbohydrate bng cch loi nhm aminoacid thnh cc cetoacid, ri t tng hp carbohydrate. Mi lin quan v mt nng lng th hin ch: khi phn gii mt hp cht no nng lng c tch lu trong ATP. Ngun ATP ny c s dng cho cc phn ng tng hp. V d, ATP c to thnh trong qu trnh ng phn, quang phosphoryl ho (quang hp) v ch yu c to thnh trong qu trnh phosphoryl ho oxy ho (h hp). S phosphoryl ho oxy ho qua chu trnh Krebs v chui vn chuyn in t cho thy rng: bt k acetyl-CoA c ngun gc t carbohydrate, hoc acid bo hay aminoacid cng u b oxy ho v tng hp ATP. Nh kh nng chuyn ho tng h gia cc cht m c th sinh vt thch ng vi mi trng. V d vo ma ng, cy trng xy ra s chuyn ho tinh bt thnh ng v cht bo, nh kh nng chu rt ca cy trng c nng cao. Hoc i vi mt s ng vt ng ng, do d tr lipid ln, nn m bo cung cp nng lng v cc cht cn thit cho c th s dng trong sut thi gian di ma ng. nghin cu mi lin quan ny chng ta cn xem xt s trao i ca tng cp hp cht sau:

8.1 Mi lin quan gia carbohydrate v lipid Carbohydrate d dng chuyn thnh lipid thng qua hai hp cht trung gian l dihydroxyacetonphosphate v acetyl-CoA dihydroxyacetonphosphate glycerolphosphate carbohydrate glucose Glyceraldehydphosphate acetyl-CoA acid bo Glycerine hoc glyceraldehydphosphate c th bin i thnh fructose-1,6- diphosphate (xem chng lipid). - Acetyl-CoA thng qua chu trnh Krebs khng th tng hp c carbohydrate v hai nguyn t carbon ca n b loi thnh CO2 trc khi to ra hp cht oxaloacetic l cht c vai tr tng hp mi carbohydrate (qu trnh gluconeogenese). Khi m c b mt mi c hin tng l chu trnh Krebs khng th tip nhn tt c cc phn t pyruvate to ra t qu trnh ng phn. Pyruvate c kh thnh lactate, cht ny c tch lu v khi m c ngh ngi chng li c oxy ho thnh pyruvate. -

Sau pyruvate vo trong ty th v c carboxyl ho nh enzyme pyruvatcacboxylase to thnh oxaloacetate. Pyruvate vo trong ty th v c th tham gia vo 2 ng hng phn ng khc nhau (hnh 8.1). N c th c bin i nh enzyme pyruvatdehydrogenase v sau i vo chu trnh Krebs. Pyruvate cng c th c carboxyl ho to oxaloacetate v chu s bin i theo con ng gluconeogenese. Theo ng hng no l tu thuc vo nng acetyl-CoA, cht c kh nng hot ho enzyme pyruvatcarboxylase theo c ch bin cu, trong khi ADP li c ch enzyme ny.

Trong t bo cht malate c oxy ho thnh oxaloacetate nh NAD+

Oxaloacetate c bin i tip tc thnh photphoenolpyruvate nh enzyme quan trng ca gluconeogenese l phosphoenolcarboxykinase. Phn ng ny gm 1 phn ng kh carboxyl ho v 1 phn ng phosphoryl (phn ng kinase). Phn ng cn 1GTP, tng t ATP.

PEP l 1 cht c to ra trong qu trnh ng phn. T cht ny i ngc li nhng phn ng ring l ca qu trnh ng phn cho n fructozo-1,6-diphotphate. Cht ny c kh phosphoryl ho v ng phn ho cho n glucose cng nh glucose-1-phosphate. Cc nguyn t C trong lactate c s dng tng hp nn glucose hoc glycogen nh ATP v NADH. S phosphoryl ho trc tip pyruvate to thnh PEP l khng thc hin c v l do nng lng, ngha l mc nng lng ca pyruvate thp hn nhiu so vi mc nng lng ca PEP. Nh hnh 8.1 s bin i ny c thc hin qua oxaloacetate, nhm i quanh dc ng gia pyruvate v PEP. Con ng ny i hi mt nng lng b sung, l 1 ATP cho carboxyl ho pyruvate v 1 GTP cho to thnh PEP. Hai ngun nng lng ny tng hp PEP, lin kt cao nng ca chng gii phng 62 kJ/mol.

Hnh 8.1 S Gluconeogenes

Nh trn nu enzyme pyruvatcarboxylase c iu khin theo c ch bin cu. S iu khin ny l mt c ch c ngha. Khi nng acetyl-CoA cao th khng cn to acetyl-CoA b sung theo con ng kh carboxyl ho bng cch oxy ho. Trong trng hp ny pyruvatcarboxylase c hot ho nh acetyl-CoA v pyruvate c s dng to oxaloacetate. Ngc li khi nng ADP cao, ngha l nng ATP thp th pyruvatcarboxylase b c ch. Pyruvate c thc hin kh carboxyl ho bng cch oxy ho cung cp acetyl-CoA cho chu trnh Krebs, l chu trnh cng vi chui enzyme h hp ti to ATP.

 thc vt v mt s vi sinh vt c chu trnh glyoxilate xy ra glyoxisome, cu trc ny cha enzyme -oxy ho peroxesome. y acetyl-CoA c ngun gc acid bo s to thnh oxaloacetic acid, sau l phosphoenolpyruvate, ri t to glucose. C mi vng chu trnh glyoxilate (xem chng lipid) t 2 phn t acetyl-CoA to ra c 1 phn t succinic acid. Acid ny b oxy ho to ra oxaloacetic acid. Oxaloacetic acid s b kh carboxyl ho bin thnh phosphoenolpyruvic acid. Cht ny s chuyn thnh glucose-6phosphate. Nh vy t 4 phn t acetyl-CoA to ra 2 phn t succinic acid, sau 2 phn t oxaloacetic acid. Cc bin i tip theo s cho ra 2 phn t phosphoenolpyruvic acid. Cui cng thu c 1 phn t glucose. 8.2 Mi lin quan gia carbohydrate v protein S phn gii carbohydrate to ra mt s -cetoacid, khi amine ho chng s to thnh cc aminoacid tng ng. Cc -cetoacid l: pyruvic acid alanine -cetoglutaric acid glutamic acid oxaloacetic acid aspartic acid Cc aminoacid ny nh kh nng chuyn ho ring l s to thnh cc aminoacid khc. Chng hn t glutamic acid c th tng hp thnh proline (hnh 7.12). Aspartic acid c th to thnh lysine, threonine, isoleucine v methionine.

8.2 Cc ng hng tng hp aminoacid trong thc vt

Mt sn phm khc ca s phn gii carbohydrate l 3phosphoglycerate. Cht ny to thnh serine bng cch sau:

COOH NAD NADH +H+ COOH Glucose NH3 H - C - OH C =O dehydrogenase transaminase CH2O-P CH2O-P 3-phosphoglycerate 3-phosphohydroxypyruvate COOH H2O H-C-NH2 photphatase CH2OP 3- phosphoserine COOH H- C-NH2 CH2OH serine

T serine c th to thnh cysteine hoc glycine (nh phn ng loi b nhm gc). Ngc li mt s aminoacid (alanine, phenylalanine, tyrosine, histidine, tryptophan, serine, cysteine , glutamic acid , proline, aspartic acid) c coi l cc aminoacid to glucose. Trong qu trnh trao i cht cc aminoacid ny to thnh pyruvic acid hoc mt hp cht trung gian ca chu trnh Krebs nh -cetoglutarate, oxaloacetate. T oxaloacetate c th to ra phosphoenolpyruvate, ri t s tng hp nn glucose. Ngoi ra gia qu trinh d ho gluxit (chu trnh Krebs) v qu trnh d ho aminoacid (chu trnh ornithine) c nhng giai on to ra nhng sn phm trung gian ging nhau, l aspartic acid, glutamic acid, fumaric acid. iu chng t s trao i carbohydrate cng nh hng qua li i vi s trao i protein. 8.3 Mi lin quan gia lipid v protein Acid bo l tin cht ca mt s aminoacid thng qua cc qu trnh sau: hoc qua chu trnh Krebs, mt s aminacid c tng hp t cetoglutaric acid, hoc qua chu trnh glyoxilate, to ra oxaloacetic acid. Oxaloacetic acid b loi carboxyl to thnh pyruvate t 2 cht ny mt s aminoacid cng c tng hp (hnh 8.2).

Ngc li mt s aminoacid (leucine, isoleucine, tryptophan) khi phn gii s to thnh acetyl-CoA, t tng hp nn acid bo. Mt s aminoacid khc (alanine cysteine, serine) b phn gii thnh pyruvic acid. Theo con ng tng hp mi glucose, pyruvic acid s to thnh 3phosphoaldehydglyceric (ngc qu trnh ng phn). S chuyn ho protein thnh lipid c th khi qut trong s sau: Protein

aminoacid leucine, isoleucine, tryptophan acetyl-CoA acid bo alanine, cysteine, serine pyruvic acid 3- phosphoaldehydglyceric glycerine

Lipid

CHNG 9

CC CHT C NGUN GC TH CP
Ngoi cc cht carbohydrate, lipid, protein, nucleic acid, trong thc vt cn tn ti nhng hp cht khc nhau do qu trnh trao i cc cht ni trn to thnh. Mc du chng ch chim mt t l rt t trong cy, nhng cc cht ny quy nh tnh c th ca s trao i cht trong tng loi thc vt. Cc hp cht ny c gi l cc cht c ngun gc th cp. Mt s trong chng v d nh cc acid hu c va mi c to thnh c t bo s dng vo cc qu trnh tng hp khc nhau, do vy chng khng c tch ly vi lng ln m l cc sn phm trung gian ca qu trnh trao i cht. Mt s cht khc v d nh cc hp cht phenol, tannin, alkaloid, tinh du li c tch ly vi lng ln, do c tnh c th v trao i cht. Trong phm vi gio trnh ny, chng ti xin gii thiu mt s nhm hp cht quan trng. 1. TANNIN: Tn gi tannin c xut vo nm 1796 k hiu nhm hp cht cha trong mt s cy vn dng thuc da ng vt. T tan c ngun gc t ting latinh ca cy si, v t v cy si ngi ta thu c cht thuc da. Cc tannin c chia thnh 2 nhm: Cc tannin c th thy phn c v cc tannin khng th thy phn c. 1.1. Cc tannin c th thy phn c: C c tnh ester. Chng l dn xut ca gallic acid v protocatechic acid. Hai gallic acid kt hp vi nhau to thnh digallic acid. Acid ny c vai tr trong vic to thnh tannin. OH OH HO OH OH HO OH CO O OH COOH Gallic acid OH COOH Digallic acid HO COOH Protocatechic acid

248

1.2. Tannin khng th thy phn c: Trong thnh phn c catechin v gallocatechin. Hai cht ny khi phn ng vi gallic acid s cho ta catechingallate v gallocatechingallate. Theo Kursanov th tannin ca ch xanh c 12% gallic acid t do; 78% catechingallate v mt t catechin t do. 2. CC ACID HU C: c hnh thnh do qu trnh trao i cht, chng tham gia vo chu trnh Krebs v nhng bin i trung gian khc. Tuy nhin mt s thc vt cng tch ly mt s acid hu c khc nhau cc c quan khc nhau nh l, qu, thn,chnh nhng acid ny gy nn pH khc nhau ca t bo. C 2 nhm acid hu c: - Nhm d bay hi:thng c mi hc nh acetic acid, butyric acid - Nhm khng bay hi:thng l cc acid hu c cha nhm -OH hoc nhm cetone nh pyruvic acid, glycolic acid, oxaloacetic acid, malic acid, Sau y l mt s acid hu c quan trng: 2.1. Citric acid: Acid ny c ph bin trong cy trng. Trong qu cc cy thuc h cam, qut, citric acid l acid hu c ch yu. chanh hm lng citric acid t ti 9% trng lng kh. Citric acid c s dng rng ri trong cng nghip thc phm v nh l cht bo qun trong truyn mu. 2.2. Malic acid (HOOC - CH2 CHOH - COOH): Malic acid chim u th qu to ty v qu cc cy c ht. Malic acid c ht v l cc cy ng cc, cc cy u . Trong thuc l v thuc lo, hm lng malic acid t 6,5%. Malic acid cha nhiu cc cy mng nc (l bng, xng rng), c chua, Malic acid c hng v d chu, khng c i vi c th ngi. N c s dng rng ri sn xut nc qu v bnh ko. 2.3. Oxalic acid (HOOC - COOH): C ph bin trong cy trng dng t do cng nh dng mui. qu, oxalic acid thng c lng rt nh 0,005-0,06% nhng trong l mt s cy nh chua me, oxalic acid li c mt hm lng ln. 2.4. Tartric acid (HOOC - CH(OH) - CH(OH) - COOH): Tartric acid thng c trong cy di dng D-tartric acid, cng c th dng L-tartric acid. N cha nhiu trong qu nho. Tartric acid v mui Kali ca n c s dng trong sn xut nc qu, trong cng nghip ho cht v cng nghip dt. 3. GLYCOSIDE: L nhng dn xut ca carbohydrate (thng l dn xut ca monose) vi cc hp cht c bn cht ho hc rt khc nhau

249

khng phi l carbohydrate. Phn khng phi carbohydrate c gi l aglycon (Aglycon c th l ru, cc hp cht thm, cc alkaloid, ) Ty thuc vo kiu lin kt gia phn carbohydrate v aglycon m ngi ta chia ra cc nhm sau: 3.1. S-glycoside: Aglycon kt hp vi carbohydrate qua lu hunh. V d: Sinigrin c ci ng, c mi hc CH2 = CH CH2 NC Sinigrin 3.2. N-glycoside: Aglycon kt hp vi carbohydrate qua N. V d: cc nucleotide 3.3. C-glycoside: Aglycon kt hp vi carbohydrate qua C. 3.4. O-glycoside: Aglycon kt hp vi carbohydrate qua Oxi (O) V d: - Solanin: rt ph bin trong mm khoai ty, trong thn l khoai ty. - Saponin: khng cha nit trong phn t ca chng. Saponin c kh nng ph hy hng cu, lm ho tan hng cu. Saponin c nhiu cy Offininalis, hm lng Saponin t cc i thi k bt u ra hoa v cc tiu thi k chn qu. sn c loi glycoside sinh ra HCN. l linamarine v lotostraloside. Cc glycoside ny tan trong nc. Sn phm ca s phn hy ny l glucose v aglycon. Phn hy aglycon ny s to ra HCN: C6H11O5 O R C C N + H2O CH3 OH C6H12O6 + R C C N glucose Aglycon CH3 R Nu R l CH3- th l linamarine Nu R l C2H5- th l lotostraloside HCN + C = O Acetone CH3 S.C6H11O5 OSO2OK

250

4. ALKALOID: L nhng cht d vng cha nit, c c tnh kim v c tc dng sinh l mnh; nhiu alkaloid l nhng cht c. i b phn alkaloid tc ng n h thn kinh. liu lng thp chng c tc dng kch thch, cn liu lng cao chng c tc dng c ch. V d: Cocaine c s dng trong y hc nh l cht gim au ti ch, tc ng n nhng u cm gic ca h thn kinh ngoi bin. Trong qu c c dc cha atropine l cht tc ng mnh n cc dy thn kinh vn ng ca mt, lm cho ng t dn ra. Sau y l mt s alkaloid in hnh: 4.1. Hordenine: C trong ht i mch lc ny mm vi mt lng khong 0,2%. Tc dng ca hordenine l lm tng huyt p ca ngi v ng vt. Trong thc vt, hordenine c to thnh t aminoacid l tyrosine, theo s sau:
CH3 H2N CH COOH CH2 CO2 H2N CH2 CH2 2CH3 - 2H OH Tyrosine OH Tyramine OH Hordenine CH2 CH2 N CH3

4.2. Ricinin: l alkaloid c cy thu du (Trong ht cha khong 0,15%, trong l non gn 1%). Ricinin c c tnh v cha nhm CN. Do hm lng ricinin c nhiu trong b kh thu du (n 0,2%) nn ngi ta khng dng kh du thu du lm thc n cho gia sc. O CH3 CN O N CH3 Ricinin

251

4.3. Piperin: c nhiu trong t, trong ht h tiu (t 5 9%). Piperin khng c, n ch gy cm gic cay nng tng b phn. O O

N Piperin CO - CH = CH CH = CH 4.4. Atropin: C nhiu trong qu c c dc v trong ht cy h c. Atropin c tc dng mnh ln h thn kinh hot ng ca mt. Atropin c th c dng lm cht gii c khi b ng c bi nicotin. Cc liu lng iu tr cao ca atropin l 0,001-0,003 gam. liu lng cao hn th atropin l mt cht c.

CH3

O CO CH C6H5 CH2OH Atropin

4.5. Cocaine: l alkaloid ch yu ca loi cy coca min Nam M. Cy ny hin nay cng c trng n v trn o Java. Hm lng cocaine trong l t t 1-2%. CO O CH3

N - CH3

O CO C6H5 Cocaine

252

Cocaine lm t lit h thn kinh cm gic v c dng lm cht gy t b phn. ngoi ra n cng c tc dng ln h thn kinh trung ng v gy nn cm gic say rt t trng. Khi ung nhiu ln cocaine, c th s quen vi cocaine v s b bnh nghin cocaine. Liu lng c ca cocaine i vi ngi lm l 0,2gam. 4.6. Morphin: Morphin v methylic ether ca n tc l codein c ngha nht. HO H3C O

O N CH3 HO Morphin

O N CH3 HO Codein

- Morphin l mt cht an thn l lm gim au. N c tc dng ln h thn kinh trung ng v thn kinh ngoi vi. 4.7. Cafein: C trong ht c ph (1-3%), trong l ch (n 5%), cn c trong cacao v mt s cy khc. O H3C N O N CH3 Cafein kch thch h thn kinh trung ng v hot ng ca tim. Di tc dng ca cafein th huyt p s cao. Cafein c tc dng thng tiu. Cafein c dng trong vic iu tr cc bnh v tim v c dng lm thuc li tiu. 253 N Cafein N CH3

4.8. Nicotin: Trong thuc l (1 10%) trung bnh 4 %. Nicotin l mt cht rt c. N c tc dng mnh ln h thn kinh trung ng v h thn kinh ngoi vi. Di tc dng ca nicotin xy ra s co mch mu do huyt p tng mnh. Con ngi c th cht do t lit h hp khi ung vo c th mt lng khong 0,01 hay 0,04 gam nicotin. V c c tnh cao nn trong y hc ngi ta khng dng nicotin. Tuy nhin n cng c ng dng rng ri trong th y chng cc bnh ngoi da hoc ng dng trong vic chng su b ph hoi ma mng.

N - CH3 N Nicotin

Nicotin c pht minh t r. R pht trin tt th nicotin cng nhiu. Trng tha, rung khng nc, ngt ngn sm th thuc l hm lng nicotin cng cao. t tt, bn nhiu m, nicotin cng nhiu. 5. TERPEN: L thnh phn ch yu ca tinh du. S c mt ca terpen quy nh mi v ca nhiu thc vt (hoa hng, hoa nhi, l chanh, l s, ) Mt s v d v terpen: - Limonen: c trong tinh du thng, tinh du h chanh. - Cimol c trong du khuynh dip, bch n. - Terpinene, xivestren c trong du thng. - Pinen c trong du thng. - Menthol: c trong du bc h CH3 CH3 CH3

C = CH2 CH3 Limonen

CH CH3 CH3 Cimol

CH CH3 CH3 Terpinene 254

CH3

CH3

C = CH2 CH3 Xivestren

H3C

CH3

OH CH3 CH3 Menthol

Pinen

6. CAO SU V NHA KT (GUTTA)

- Trong thc vt c to thnh t isopren: CH2 C = CH CH2 CH3 n = 500 5000: gi l cao su n = 100 : gi l gutta-percha * Cao su l hp cht cao phn t polyisopren c hnh thnh trong m cc cy thuc 300 tc khc nhau. Trong s nhng cy ny ch c cy cao su Hevea Brasiliensis c s dng vi quy m cng nghip. M cha cc ht cao su c tch ly trong cc t bo chuyn dng l ng nha. cy cao su, m c hnh thnh v tch ly trong v cy v cc ng nha vng. Nh c cc ng ni gia cc ng dn cnh nhau trong cc ng nha vng m m c th chy ra t mt vng ln ca v cy trong khi khai thc m. * Nha kt (gutta) c hnh thnh trong cc cy tc Palaquium (Sapotaceae) mc nhiu Malaysia. M cc cy ny khng chy ra d dng nh m cao su, do vy thu hoch m phi cht cy. iu ny dn n vic tiu dit dn ngun nha kt ban u Palaquium gutta. Nha kt l cht do v nhit. Thut ng chuyn mn gi l cht nhit do. nhit di 65oC n khng do v cng nhng nhit 65oC th chuyn thnh dng mm, n hi nhng vn cha do. n

255

7. CC CHT IU HO SINH TRNG THC VT

Cc cht iu ho sinh trng thc vt l mt nhm cht c bn cht ho hc khc nhau nhng u c mt tc dng iu ho qu trnh sinh trng, pht trin ca cy v m bo mi lin h gia cc c quan, b phn ca cy. Cht iu ho sinh trng thc vt c chia ra lm 2 nhm: a. Cc cht kch thch sinh trng. b. Cc cht c ch sinh trng. Hai nhm ny c tc dng i khng vi nhau v mt sinh l. - Cc cht kch thch sinh trung bao gm cc cht m nng sinh l c tc dng kch thch qu trnh sinh trng ca cy (gm cc nhm cht auxin, gibberellin v cytokinin). - Cc cht c ch sinh trng gy tc dng c ch ln qu trnh sinh trng ca cy (gm cc cht nh abcisic acid (ABA), ethylene; cc cht phenol, cc cht lm chm sinh trng, cc cht dit c, ) * Cc cht iu ho sinh trng do cy t tng hp gi l cht t nhin, cn cc cht do con ngi tng hp bng con ng ho hc gi l cht nhn to. Bng phn loi cc cht iu ho sinh trng thc vt
Cht iu ho sinh trng t nhin Cht iu ho sinh trng nhn to

A. Cht kch thch sinh trng (Stimulator)

1. Auxin (IAA, IAN, PAA) 2.Gibberellin (GA1, GA2, GA3,, GA54)

Auxin nhn to (IBA, -NAA; 2,4D; 2,4,5T, MCPA)

3. Cytokinin (Zeatin, diphenylurea) Cytokinin nhn to (kinetin, BA, ) B. Cht c ch sinh trng (Inhibitor)

1. Abcisic acid (ABA) 2. Ethylene 3. Phenol

Cht lm chm sinh trng(CCC, MH, TIBA) CEPA

256

Cc danh php quc t: IAA: -indol acetic acid. IAN: -indol acetonitril. PAA: Phenyl acetic acid. IBA: -indol butyric acid. NAA: -Naphtyl acetic acid. 2,4D: 2,4 dichlorophenoxyacetic acid. 2,4,5 T: 2,4,5 trichlorophenoxyacetic acid MCPA: 4 chloro, 2 methyl phenoxyacetic acid. ABA: Abcisic acid. BA: Benzyladenin. CCC: Chlorocholine chlorid. MH: Malein hydrazide. TIBA: Triiodobenzoic acid. CEPA: Chloroethylenphosphoric acid.

257

A. CC CHT KCH THCH SINH TRNG THC VT

1. Auxin:
1.1. Lch s pht hin: Nm 1880, Dacwin pht hin ra hin tng hng quang th gii thc vt. Hng quang l kh nng hng v ngun nh sng chiu t mt hng nh cc cy trng ca s lun lun vn ra ngoi ca s. Hin tng hng quang c bit r rt nht i vi bao l mm cc cy ho tho (Coleoptyl), v vy chng l i tng c dng nhiu trong vic nghin cu tnh hng quang thc vt. Nu chiu sng t mt hng n ngn bao l mm th s gy nn s un cong hng v ngun sng, nhng nu chng trong ti hoc loi tr nh ngn ca bao l mm th hin tng khng xy ra. ng cho rng nh ngn ca bao l mm l ni tip nhn kch thch ca nh sng. Sau Boyen Jensen pht hin ra rng nh ngn b loi tr nu t tr li trn b mt ca bao l mm th n c kh nng gy phn ng hng quang bnh thng nh trng hp cy nguyn vn. Nh vy th mt cht gy hng quang no sn sinh trong nh ngn v vn chuyn theo hng gc gy nn s sinh trng khc nhau ca cc m pha di. Paal (1919) ct ri nh bao l mm v t li ln b mt ct nhng lch sang mt bn v trong ti. Hin tng un cong hng ng xy ra mnh m nh trng hp c chiu sng mt hng. ng cho rng nh ngn hnh thnh nn mt cht sinh trng no y, cn nh sng xc nh s phn b ca cht v hai pha ca bao l mm. Nm 1933 Kogh (H Lan) bo co rng ng tch c auxin A v auxin B t nc tiu ngi. ng cng b trng lng phn t ca auxin A l 328. Cc cht ny gy ra phn ng un cong mnh ca bao l mm cy la mch. Nm 1934, Kogh v Kosterman tch c chng t dch chit nm men v nm 1935 tch c chng t dch ca mi trng nui cy nm Rhizopus. Tip theo ngi ta xc nh bn cht ho hc ca auxin. chnh l indolylacetic acid (IAA). V lc u ngi ta cha xc nh c s tn ti ca auxin trong cy nn ngi ta gi cht l heteroauxin. Nhng nm v sau ngi ta ln lt tch c auxin t cc i din ca thc vt thng ng khc nhau v chng minh rng y l mt phytohormone quan trng nht tn ti trong ton b th gii thc vt. ng thi ngi ta cng tng hp c rt nhiu cc hp cht khc nhau nhng c hot tnh sinh l tng t nh IAA v thm ch nhiu cht c hot tnh mnh hn nhiu so vi IAA v chng c s dng rng ri trong sn xut nhm iu chnh sinh trng ca cy trng c li cho con ngi. Chng thng c dng mch vng. l cc dn xut ca indol nh -indol butyric acid (IBA); -indol propionic acid (IPA), ; Cc dn xut cu naphtalen nh -naphtylacetic acid (-NAA); -naphtylacetic acid (-NAA); Cc dn xut ca chlorophenoxyaxetic acid nh 2,4 dichlorophenoxyacetic acid (2,4D); 2,4,5 trichlorophenoxyacetic acid (2,4,5T)

243

CH2 - COOH (IAA) NH CH2 COOH

CH2 CH2 CH2 COOH (IBA) NH

CH2 COOH

(-NAA) (-NAA) O CH2 COOH Cl (2,4D) Cl Cl Cl O CH2 COOH Cl (2,4,5T)

1.2. Cc dng auxin trong cy: Auxin c tng hp ch yu nh sinh trng ngn, t vn chuyn n cc c quan khc nhau theo hng gc. Ngoi nh sinh trng ngn, auxin cn c tng hp mt phn cc c quan cn non nh l non, chi non, qu non. Trong cy auxin tn ti hai dng: dng t do v dng lin kt. * Auxin dng t do: l dng c hot tnh sinh hc. Chng tn ti nh sinh trng ngn, sau n chi bn, u r v cc c quan non. Cng xa nh sinh trng nng auxin t do cng gim dn. Dng auxin t do ch yu trong cy l IAA. Auxin t do trong cy chim mt t l rt thp. Nhiu ti liu cng b hm lng auxin t do trong cy chim khong 5% auxin tng s. * Auxin dng lin kt: l dng auxin ch yu trong cy. Hm lng ca dng lin kt ny c th chim trn 90% lng auxin c trong cy. IAA c th lin kt vi rt nhiu cc hp cht khc nhau nn chng khng c hot tnh sinh hc hoc c hot tnh rt thp. IAA c th lin kt vi ng; cc aminoacid v amide to nn cc auxin lin kt nh IAA-glycoside; IAA-glucan; indolacetamide, indolacetylaspartate; indolacetylglutamate, indolacetylinositol; indolacetylarabinose, 244

COOH CH2 C = N CH O H CH2 COOH NH Indolacetylaspartate * Chc nng ca auxin lin kt trong cy ht sc quan trng. Trc ht chng l ngun d tr auxin trong cy. S lin kt ca IAA vi ng, vi cc aminoacid v cc cht khc lm cho chng bn vng vi cc tc nhn c th lm ph hy auxin v l phng thc tt nht chng li s tc ng ca cc peroxidase phn hy auxin. Ngi ta chng minh rng auxin lin kt l dng auxin vn chuyn ch yu trong cy. V vy, s bin i thun nghch ca IAA t do IAA lin kt di tc nhn kch thch ca mi trng gp phn iu chnh s cn bng auxin trong cy v do iu chnh qu trnh sinh trng ca cy. 1.3. S tng hp auxin trong cy: C quan tng hp auxin l nh sinh trng ngn. T m phn sinh nh, IAA c vn chuyn xung cc c quan pha di; auxin cn c tng hp t hn trong cc c quan ang sinh trng nh l non, qu non, r non, Ngi ta xc nhn cht tin thn tng hp nn IAA l aminoacid tryptophan. S tng hp auxin trong cy:

245

CH2 CH COOH NH2 NH Tryptophan

NH3

O2

CH2 C COOH O NH Indolpyruvic acid

CO2 CH2 - CH2 NH2 NH Tryptamine O2 NH3

CO2 CH2 C = O H NH Indolacetaldehyde O2

CH2 COOH

NH

(IAA)

1.4. S phn gii auxin trong cy: S phn gii auxin trong cy xy ra trong cc trng hp sau: Khi c tha auxin do s tng hp qu nhiu auxin trong cy v sau khi auxin gy hot tnh sinh l vi cy xong. Vic lm mt hot tnh auxin c th xy ra bng hai con ng: S oxy ho bng enzyme IAA-oxidase v s quang oxy ho. * S oxy ho bng enzyme IAA-oxidase: y l enzyme oxy ho IAA, enzyme ny hot ng rt mnh trong h thng r thc vt lm mt hot tnh ca IAA vn chuyn xung l ni cui cng. Phn ng oxy ho ca IAA xy ra nh sau:

246

CH2 COOH + H2O2 + O2 NH IAA

IAA-oxidase - H2O - CO2

CH2 O NH Methylenoximdol

* S mt hot tnh ca IAA c th xy ra do nh sng. Ngi ta nhn thy rng nh sng t ngoi c th lm mt hot tnh ca IAA, bi v cu trc vng ca phn t IAA hp thu nh sng t ngoi = 280 nm v gy nn s phn gii IAA. 1.5. S vn chuyn ca IAA trong cy Auxin c tng hp trong nh ngn v t y vn chuyn xung cc c quan pha di. S vn chuyn ca auxin trong cy c tnh phn cc r rt, theo hng gc, tc l t ngn xung r m khng vn chuyn ngc li. - C ch ca s vn chuyn phn cc ca IAA trong cy c ngi ta a ra mt s da theo thuyt ho thm ca Goldsmith (1977) nh sau: IAAo Trn IAAo IAATrong t bo IAADi Theo thuyt ho thm ca Goldsmith th: Cht mang IAAIAAo +oxi ho mi khuy ch tn qua mng t bo pha trn v - Ch c IAA khng b H (Protein) IAAo thm qua c mng lipit. - Bn trong t bo, pH cao IAA hn ov c cc bm H+ hot ng, do IAAo b ion ho thnh IAA- . Nng IAA- tng ln v khng vn chuyn ra khi t bo c v khng tan c trong lipid. - Do nng ca IAAo trong t bo gim nn tn ti mt gradient IAAo gia bn ngoi v bn trong t bo, IAAo s khuych tn vo t bo. - IAA- c vn chuyn ra khi t bo nh cc cht mang protein c hiu nm mng t bo pha di. Khi ra khi t bo, do pH thay i nn IAA- IAAo v tip tc vn chuyn sang t bo di n. Ngoi s vn chuyn phn cc ny, bng nguyn t nh du ngi ta cn xc nhn c mt s vn chuyn ngc li (hng ngn) nhng t l thp. S vn chuyn 247 Bm H+ H+ Ngoi t bo

phn cc l qu trnh cn nng lng ATP, cn s vn chuyn ngc li l qu trnh khuych tn. Tc vn chuyn ca auxin trong cy rt thp, khong 1 cm/h trong r v thn. 1.6. Hiu qu sinh l ca auxin i vi cy Cc c quan khc nhau c hm lng auxin khc nhau. Hm lng ny cn ph thuc vo tui cy, vo iu kin ngoi cnh. Cc c quan cn non ang sinh trng c hm lng auxin cao hn cc c quan trng thnh v c quan gi. Tuy nhin, phn ng sinh trng ca cc c quan khc nhau ph thuc vo hm lng auxin l rt khc nhau. Ni chung thn cy cm ng kch thch vi nng auxin cao hn chi v r. Sau y l th minh ho mi quan h gia nng auxin v s sinh trng:

+ 200% 100 0

10-6

10-4

10-2

102

104

IAA

a) Auxin v tnh hR ng h -100 quang,Ch ing a: Thn * Hng quang (phototropism) l s sinh trng ca cy hng v ngun sng kch thch mt hng. * Tnh hng a (geotropism) l phn ng sinh trng ca cy di tc ng ca lc ht tri t (trng lc). T khi pht hin ra auxin ngi ta gii thch tnh hng quang v hng a ca cy trn c s s phn b ca auxin hai pha ca c quan khc nhau dn n s sinh trng khc nhau hai pha ca c quan v kt qu l gy nn s un cong hng ng. C s ca s phn b ca auxin khc nhau l do s khc nhau v in th (in sinh hc). Khi c chiu sng t mt hng th pha khut sng s tch in dng v pha chiu sng s tch in m. Mt c quan nm ngang th mt trn s tch in m v mt di s tch in dng. Ngi ta c th o in th ca c quan trong cc trng hp . _ + + + 248 nh sng

 trong t bo IAA thng dng IAA- v do m phn b v pha mang in dng. Khi c nh sng chiu mt chiu th auxin phn b v pha khut sng v hm lng ca IAA cao hn gy s sinh trng mnh hn pha khut sng v lm cho cy sinh trng un cong v pha chiu sng. Trong trng hp hng a th vi c quan nm ngang, auxin s phn b pha di nhiu hn, gy nn s sinh trng ca mt di nhanh hn mt trn lm cho chi ngn sinh trng un cong ln trn. Tuy nhin do r phn ng kch thch vi nng auxin thp hn nhiu so vi thn, nn nng auxin cao s gy c ch v do m s sinh trng ca mt di chm hn mt trn lm cho r m vo t. b) Hin tng u th ngn v auxin: Hin tng u th ngn l hin tng ph bin ca th gii thc vt, khi m s sinh trng ca chi ngn, ca r chnh s c ch s sinh trng ca chi bn v r ph. Nu loi tr chi ngn v r chnh th lp tc chi bn v r ph s sinh trng mnh. Trong thc t khi ngi ta bm ngn, ta cnh, n cy, tc l loi b s c ch ca chi ngn, to iu kin cho cc chi bn mc ra mnh m. y l c s ca k thut to hnh cho cy n qu, cy cnh, cy cng nghip v ci to c qun th cy trng. Khi pht hin ra auxin, ngi ta cho rng auxin l tc nhn gy nn hin tng u th ngn. Nu loi b chi ngn th lp tc chi bn mc ra, nhng khi loi b chi ngn m x l IAA vt ct th chi bn cng khng mc ln c, tng t nh trong trng hp c chi ngn tn ti. C hai gi thuyt gii thch vai tr ca auxin i vi hin tng u th ngn: * Gi thuyt c ch trc tip ca IAA: Cho rng chi ngn l ni tng hp IAA v do nng ca auxin trong chi ngn lun lun cao. Trn con ng vn chuyn ca IAA xung di th gy nh hng c ch ln s sinh trng ca chi bn v phn ng ca chi bn ln hm lng auxin thp hn chi ngn. Trong trng hp ct b chi ngn tc l lm gim hm lng IAA v do cc chi bn c gii phng khi trng thi c ch. * Gi thuyt c ch gin tip ca auxin: Cho rng auxin khng gy nh hng trc tip nhng auxin gy nn s tng hp mt cht c ch sinh trng no y nh ethylene chng hn; v n lt ethylene li gy c ch sinh trng ca chi bn. Tuy nhin d gi thuyt no th cng phi tha nhn vai tr ca auxin ln hin tng u th ngn. Thc ra, hin tng u th ngn c iu chnh bi s cn bng hormone ca auxin/cytokinin. Auxin tng u th ngn, c ch chi bn, cn cytokinin ngc li, lm gim u th ngn, kch thch chi bn. Auxin c tng hp nh ngn v c vn chuyn xung r, nn cng xa nh ngn th nh hng c ch ca auxin cng yu. Ngc li, cytokinin c tng hp trong r v c vn chuyn t r ln ngn, nn cng xa r nh hng kch thch ln chi bn cng yu dn. V vy, t ngn xung gc th hin tng u th ngn cng gim dn, chi bn cng mc nhiu hn. c) Auxin kch thch s gin ca t bo: Auxin lm tng kch thc ca t bo v do m lm cho cy sinh trng c. Tc dng ca auxin c hai mt: Va gy s gin n ca thnh t bo lm cho t bo

249

phnh to ra v va hot ho cc qu trnh tng hp cc vt cht cu to nn cht nguyn sinh. Auxin kch thch s gin n ca t bo theo chiu ngang mnh hn l chiu dc. d) Auxin vi s hnh thnh v s phn ha r : Trong s hnh thnh r, c bit l r bt nh, auxin c tc dng rt c trng. V vy ngi ta xem auxin nh l cht hnh thnh r. S hnh thnh r bt nh ca cnh gim, cnh chit c th chia ra lm 3 giai on: - u tin l s phn phn ho t bo mnh m vng tin tng tng, tr b, ni xut pht ca r bt nh thnh mt m t bo ln xn. - Giai on tip theo l xut hin mm r. - Giai on cui cng, r sinh trng, m thng v v ra ngoi thnh cc r bt nh. giai on u tin l giai on khi xng s hnh thnh r cn hm lng auxin cao cho s phn phn ho t bo. Cc giai on sau cn hm lng thp hn v i khi khng cn. Ngun auxin c th l ngun ni sinh v cng c th b sung t ngoi vo. Thng xc tin qu trnh hnh thnh r nhanh chng cnh chit, cnh gim ngi ta thng x l auxin ngoi sinh. Vai tr ca auxin vi s hnh thnh r c chng minh r trong nui cy m t bo. Nu ch c mt ca auxin trong mi trng nui cy, m ch pht sinh r m thi. to cy hon chnh th cn phi c chi na, tc l phi b sung cytokinin vo mi trng nui cy. e) Auxin vi s hnh thnh qu, s sinh trng v to qu khng ht: Ht phn l ngun rt giu auxin, kch thch s ny mm ca ht phn v s sinh trng ca ng phn. Ngoi ra nm nhy cng tit ra cc cht kch thch s sinh trng ca ng phn v ng thi cng sn sinh ra cc cht c ch s ny mm ca ht phn v s sinh trng ca ng phn khc loi, gy nn s tuyt giao gia cc loi. Khi th tinh xong, hp t s pht trin thnh phi v chnh phi cng l ngun tng hp auxin mnh m ri khuych tn vo bu kch thch s ln ln ca bu thnh qu. V vy kch thc v c hnh dng ca qu hon ton ph thuc vo hm lng v s khuych tn ca auxin trong phi ht. Hu ht cc trng hp s pht trin ca qu ch xy ra sau khi th tinh xong, tc l hnh thnh phi v bt u sn sinh auxin ni sinh. Nhng cng c trng hp qu c hnh thnh khng c th tinh v to nn qu khng ht. Nm 1934, Yasuda thnh cng trong vic gy nn qu khng ht bu b bng cch x l dch chit ca ht phn ln hoa. Sau ngi ta phn tch v thy trong dch chit ca ht phn c cha nhiu auxin. T ngi ta x l trc tip auxin ngoi sinh cho hoa th cng c th loi tr s th tinh v to nn qu khng ht v rng auxin trc tip khuych tn vo bu, kch thch sinh trng ca bu thnh qu khng ht. f) Auxin v s rng l, hoa, qu, : Laibach (1933) ch ra rng trong dch chit ca ht phn phong lan c cha mt cht c th km hm s rng. Cht l IAA. Trc khi rng, c mt tng ri c hnh thnh cung ca c quan thc vt. y l mt vi lp t bo nhu m c thnh mng, hon ton thiu lignin v suberin. Khi hnh thnh tng ri th c 3 trng hp xy ra: 250

- Lp v gia ca t bo b phn hy, cn lp v s cp vn gi nguyn. - Tt c thnh b phn hy. - Tt c t bo b phn hy. Khi nghin cu hm lng auxin lin quan n s hnh thnh tng ri cho thy rng l non c hm lng auxin cao hn l gi, bn l c hm lng auxin cao hn cung l. Khi hm lng auxin cao s ngn s hnh thnh tng ri. V vy, nu x l auxin ngoi sinh s lm tng hm lng auxin trong l nn c th ngn nga c s rng. Thc ra s rng ca c quan gy ra do s tch ly cc cht c ch sinh trng (abcisic acid, ethylene). Do s cn bng gia auxin / (abcisic acid+ethylene ) c nh hng n iu chnh s rng cy. Auxin cn nh hng n qu trnh bin i sinh l-sinh ho trong cy: S vn ng ca cht nguyn sinh, cc qu trnh trao i cht, tng qu trnh quang hp v h hp, tng qu trnh vn chuyn vt cht trong cy, 1.7. C ch tc ng ca auxin: Hiu qu c trng nht ca auxin chnh l kch thch s gin ca t bo. S gin ca t bo xy ra do hai hiu ng: s gin ra ca thnh t bo v s tng th tch v khi lng ca cht nguyn sinh. Khi x l auxin, khong sau 10 15 pht th phn ng sinh trng xy ra. Vy th auxin ng vai tr nh th no trong s sinh trng nhanh chng nh vy? Trc ht, chng ta cp n mt hin tng c pht hin t nm 1930 l hin tng sinh trng acid. Trong mi trng acid th s sinh trng ca thc vt tr nn nhanh chng hn. Vy mi quan h gia hiu qu acid v auxin ln sinh trng ca t bo l nh th no? Hai nh hng c phn bit vi nhau l: - nh hng ca acid l ln thnh t bo, cn nh hng ca auxin l ln t bo. - nh hng ca acid xy ra nhanh hn, cn nh hng ca auxin th chm hn nhiu. Hai nh hng c th lin kt vi nhau nh sau: Auxin gy nn acid cho thnh t bo, cn acid th gy nn lng lo, khng bn vng ca thnh t bo v chnh nh lng lo ca thnh t bo m thnh t bo mi gin ra c. Cng trong thi gian ngi ta pht hin ra nhng bm trao i cht trong cht nguyn sinh. Mt trong nhng bm l bm ion H+. Bm H+ t cht nguyn sinh vo thnh t bo, lm gim pH trong thnh t bo. IAA l tc nhn hot ho bm proton . Nh vy khi c mt ca IAA th bm proton hot ng v pH ca thnh t bo gim xung. Ti sao pH thp li lm gin thnh t bo? tr li cu hi chng ta cn xem xt cu trc ca thnh t bo: Thnh t bo c cu to t cc si cellulose, gia chng c ni vi nhau bng cc cu ni polysaccharide, to nn mt cu trc rt bn vng. cho cc si cellulose lng lo, tch ri nhau th cn phi phn hy cc cu ni polysaccharide. Enzyme phn hy cc cu ni ny l pectinmethylesterase hot ng pH thp (= 4) v khi hot ng s methyl ho nhm carbocyl v ngn chn s hnh thnh cu ni ion vi canxi to nn pectatcanxi, nn cc si cellulose tch ri nhau, chng c th trt ln nhau m gin ra, Nh kh nng ht nc thm thu ca t bo m to nn sc trng tc ng ln thnh t bo, lm cho thnh t bo phi gin n ra. Tuy nhin, do t bo tng kch thc nhanh chng nh vy nn cn c s tng hp mi v nhanh chng cc thnh phn cu to nn thnh t bo cng nh cht nguyn 251

sinh (cellulose, hemicellulose, pectin, protein, nucleic acid, ). y, auxin ng vai tr hot ho cc gen tng hp nn cc enzyme cn thit cho s tng hp nn cc vt cht . DNA mRNA Auxin S v c ch tc ng ca auxin ln s gin ca t bo: DNA mRNA IAA Protein Enzyme tng hp cc cu t ca thnh t bo v nguyn sinh cht Protein-enzyme

Sinh trng

Enzyme gin thnh t bo Bm H+ H

+

Enzyme h hp Cellulose, pectin, Protein, ATP H2O Tng th tch t bo T bo gin, th tch, kch thc t bo tng

Thnh t bo lng lo, gim tnh bn vng

1.8. ng dng cc hp cht auxin trong trng trt. a) Nhn ging v tnh cy trng: Hin nay c hai phng php x l auxin cho cnh chit, cnh gim: * Phng php x l nng hay phng php x l nhanh. Nng auxin dao ng t 1000 n 10000 ppm (1ppm = 0,1 mg% hoc = 1x10-4% tc 0,0001% hay = 1 mg/l). Vi cnh gim th nhng phn gc vo trong dung dch trong 3-5 giy ri cm vo gi th. Cn vi cnh chit th sau khi khoanh v, tm bng bng dung dch auxin c ri bi ln trn khoanh v, ni xut hin r bt nh. Sau b bu bng t m. Phng php ny hiu qu cao, n gin v ho cht tiu tn t hn. * Phng php nng long-x l chm: Nng auxin t 20 n 200 ppm ty thuc vo loi v mc kh ra r ca cnh gim. i vi cnh gim, ngm phn gc

252

vo dung dch auxin trong 10 24 gi ri cm vo gi th. Vi cnh chit th trn dung dch vi t b bu. Cc auxin s dng y l IBA; -NAA; 2,4D;...trong hiu qu ca IBA > NAA > 2,4D. b) ng dng auxin tng u qu, sinh trng ca qu v to qu khng ht: S hnh thnh qu l mt qu trnh din ra c lin quan n s th phn v th tinh. S th phn l qu trnh m ht phn ri trn nm nhy. Ht phn c cha nhiu auxin nn khi gp dch nm nhy, chng s ny mm, ng phn mc v sinh trng ko di xung tn non. ng phn mang cc tinh t xung th tinh cho t bo trng to nn hp t v sau hp t s pht trin thnh phi, v sau phi pht trin thnh ht. Bu hoa s sinh trng thnh qu. Qu ch c hnh thnh sau khi c qu trnh th tinh xy ra, cn nu khng th tinh th hoa s b rng. iu c gii thch rng phi ht l ngun tng hp nn cc cht kch thch sinh trng trong c auxin. Cc cht ny s c vn chuyn vo m ca bu kch thch bu ln ln thnh qu. V vy, hnh dng v kch thc ca qu hon ton ph thuc vo hm lng cc cht ni sinh t phi ht. Chnh v l do m ta c th s dng auxin v gibberellin ngoi sinh thay th cho ngun ni sinh. Nu chng ta x l cho hoa cha xy ra th tinh th auxin ngoi sinh s khuych tn vo bu v kch thch s ln ln ca bu thnh qu m khng qua qu trnh th tinh. Trong trng hp ny, qu to nn khng qua th tinh v do s khng c ht. Mt s cy trng nh c chua, bu b, cam chanh,...ngi ta thng x l auxin di dng -NAA (10 20 ppm); 2,4D (5 10 ppm). Cn cc cy trng khc nh nho, anh o,.... th x l gibberellin (20 50 ppm) li c hiu qu hn. c) Phng nga rng qu: S rng l do s hnh thnh tng ri cung l, cung qu. Auxin l cht km hm s rng. Chnh v vy m ngi ta x l auxin cho cy v cho qu non c th km hm s rng ca chng. V d nh phun -NAA (10 20 ppm) cho l hoc qu non c th ko di thi gian tn ti ca chng trn cy. d) Ko di s chn ca qu: S chn ca qu c iu chnh bng t l auxin/ethylene. Mun km hm s chn ta cn tng hm lng auxin trong m qu. Phun dung dch auxin ln qu xanh hoc qu sp chn ang trn cy c th ko di thi gian tn ti ca qu. Trc y ngi ta hay s dng 2,4D (10 15 ppm). Hin nay ngi ta s dng -NAA(10 20 ppm) cng c hiu qu tt m khng c hi. e) Dit tr c di: Khi s dng nng cao c tc dng dit tr c di hi cy trng. Cc cht nh 2,4D; 2,4,5T trc y c s dng nhiu dit tr c di, nhng ngy nay ngi ta to ra rt nhiu cht dit c mi c hiu qu cao m khng c hi cho mi trng.

2. Gibberellin
2.1. Lch s pht hin: Vic pht hin ra gibberellin gn lin vi nhng nghin cu v bnh la von m cc nh khoa hc Nht quan tm t lu. Triu chng in hnh ca bnh l cy la tng trng chiu cao qu mc, lm yu cy, gim nng sut trn 40%. Bnh la von l do nm k sinh cy la c tn l Gibberela fujikuroi gy nn. Nm ny tit ra mt cht no kch thch s sinh trng chiu cao ca cy la gy nn bnh l. 253

Nm 1926, Kurosawa thnh cng trong vic gy bnh nhn to cho cy la v cy ng. Nm 1935, Yabuta tch c hai hp cht di dng tinh th t dch nui cy ca nm G.fujikuroi v ng gi l gibberellin A v B nhng cha xc nh c bn cht ha hc ca chng. Nm 1955 hai nhm nghin cu ca Anh v M ng thi nghin cu vn ny. Brian (Anh) v Stodole (M) xc nh c cng thc ha hc ca chng. Sau cc nh khoa hc chit tch v xc nh GA t cc thc vt bc cao khc nhau v xc nh gibberellin l mt phytohormone quan trng v ph bin ca ton th gii thc vt. Ngy nay ngi ta xc nh c trn 50 loi gibberellin khc nhau v c k hiu l GA1; GA2; GA3, trong GA3 (Gibberellic acid) l c hot tnh mnh nht. O

C=O HO OH

GA3 (Gibberellic acid)

2.2. S vn chuyn ca GA trong cy: CH COOH CH2 Khc vi 3auxin, gibberellin vn chuyn khng phn cc trong cy: c th theo hung gc hoc hng ngn ty theo nhu cu v v tr ca c quan cn GA. 2.3. S tng hp GA: Cht tin thn ca s tng hp GA l acetyl-CoA: O CH3 C ~ S.CoA Qu trnh tng hp GA cn c s tham gia ca ATP v NADPH2, cng nh s xc tc ca Mg+2 v Mn+2. Qu trnh tng hp GA xy ra ch yu trong lc lp. C th tm tt qu trnh tng hp GA nh sau:

254

3Acetyl-CoA

2NADPH2

2NADP CH2 - COOH C(OH) CH3 CH2 CH2 OH 2ATP Mevalonic acid

3HS-CoA

2CO2 CH3 C CH2 CH2 O (P) O (P) CH2 Isopentenylpyrophosphate CH2 Gibberellin Kaurenic acid CH3 COOH

CH3

Qu trnh tng hp GA dc xc tc bi cc enzyme c hiu. Tn ti mt s t bin ln m c th thiu mt s gen nht nh iu khin s tng hp nn cc enzyme xc tc cho s hnh thnh cc cht trung gian no v kt qu l GA khng c hnh thnh, cy khng th sinh trng chiu cao c. Vi cc t bin ny, nu x l GA ngoi sinh c th ly li c dng bnh thng. 2.4. Hiu qu sinh l ca GA a) Cc t bin ln ca thc vt: Cc t bin ny do c s sai lch nht nh trong qu trnh tng hp enzyme cn cho s tng hp nn GA; do GA khng c hnh thnh nh bnh thng. b) GA vi s gin v s sinh trng ca t bo: Cng vi auxin, GA c nh hng quan trng ln s gin ca t bo. GA kch thch gin ca t bo lm cho t bo gin v chiu dc. V vy m t bo ko di rt nhanh chng. Kt qu kch thch s gin ca t bo bi GA lm cho cy sinh trng nhanh v chiu cao. Mt s cy cn chiu cao nh cy ay, cy gai, C th s dng GA ngoi sinh iu khin chiu cao ca chng. c) GA vi s ny mm: Trong ht, ni nh ng vai tr th ng trong qu trnh ny mm, n ch cha cht d tr. Khi ny mm th c cc qu trnh thy phn cc cht d tr trong chng. S thy phn ny nh cc enzyme thy phn trong amylase ng vai tr quan trng. 255

S hnh thnh nn cc enzyme thy phn ny do s hot ha ca GA c tng hp hoc c gii phng t trng thi lin kt trong phi ht. Chnh v vy m ngi ta c th kch thch cho ht ny mm bng cch x l GA ngoi sinh. Vi cc loi c: S ny mm ca chng cng c kch thch bi GA c trong c. X l bng GA ngoi sinh cng ph b c trng thi ng ngh ca c. d) GA vi s sinh trng v to qu khng ht: Hiu qu ca GA trong trng hp ny cng tng t nh ca auxin. V vy nu x l GA ngoi cng c th thay th c GA ni sinh trong phi ht v to nn c nhng qu khng c ht. Trong mt s trng hp, auxin khng gy hiu qu th GA li c hiu qu tt nh trng hp i vi nho, anh o, to, e) GA vi s ra hoa v s phn ha gii tnh: Trailakhian xem GA l mt trong hai thnh phn ca hormone ra hoa (florigen) trong hc thuyt ra hoa ca ng. Khi x l GA cho cy ngy di c th lm cho chng ra hoa trong iu kin ngy ngn v trong iu kin ngy di s tng hp GA rt kh khn. - GA cng c th thay th cho x l lnh v lm cho cy hai nm ra hoa ngay trong nm u. - GA cng c tc dng trong vic phn ha c quan sinh sn c bit l s phn ha gii tnh c v ci. Trong trng hp ny th GA c tc dng i khng vi cytokinin, tc l kch thch s biu hin v hnh thnh gii tnh c: kch thch s hnh thnh ca hoa c v c ch s hnh thnh hoa ci. Chnh v vy ngi ta s dng GA iu khin s lng hoa c ca cc cy h bu b. 2.5. C ch tc ng ca GA Mt trong nhng tc ng c trng ca GA l lm tng hot ng thy phn ca ht, c, trong qu trnh ny mm. V d nh trong cc ht la, ng, la m, phi ht l ni tng hp nn GA. GA c gii phng ra khi phi v khuych tn vo lp t bo aleuron v ti y, GA kch thch s hnh thnh v gii phng cc enzyme thy phn trong cc t bo aleuron. Cc enzyme ny khuych tn vo t bo ni nh v ti y chng xc tc cho s thy phn cc hp cht hu c phc tp thnh cc cht n gin cn cho qu trnh ny mm. GA ng vai tr l nhn t m gen m cc gen ny b c ch hon ton trong qu trnh ng ngh ca ht, c. Trong qu trnh ng ngh, cc gen ny b bao vy bi mt loi protein gi l histone. GA l nhn t hot ha gen, gii phng gen ny khi trng thi c ch v do cc enzyme thy phn mi c hnh thnh.

256

S tc ng ca GA ln s ny mm: DNA GA Amylase Tinh bt n H3PO4 H hp T bo ht nc Nguyn liu ATP n.glucoso-(P) mRNA Protein-enzyme

* S gin ca t bo cng c th do c ch hot ha bm H+ ca GA v gy nn hiu ng mm t bo. S c mt ca GA cng gy nn s tng cc cht hot ng thm cho YM M thu v p suS t thtr m u ca t bo to iu kiN n t bo ht nc, sc trng tng c th ng ln, t bo gin ra. * Cc tc ng khc ca GA nh s ra hoa, s phn ha gii tnh: c th c thc hin thng qua c ch hot ha gen ca GA. 2.6. ng dng ca GA a) Kch thch tng chiu cao, tng sinh khi: Vi cy ay, ngi ta phun GA (2 5 ppm) s lm chiu cao cy tng ln nhiu, tng nng sut si m cht lng si m bo. Vi cc cy rau nh bp ci, xu ho, x lch vic x l GA (2 5 ppm) lm tng nng sut cht xanh. b) Tng nng sut v to qu khng ht. c) Ph b s ng ngh ca ht, c. X l GA (2 ppm) c th ngm hoc phun s lm cho c khoai ty ny mm nhanh. d) iu chnh gii tnh: X l GA cho bu, b, cc loi da, lm tng t l hoa c. iu ny c ngha quan trng trong vic sn xut ht lai ca cc cy h bu b. Ngi ta c th bin mt hng thnh cy mang hoa ci v xen mt hng l cy mang hoa c. Chng th tinh cho nhau to nn ht lai. e) ng dng trong sn xut malt bia: kch thch sn xut amylase trong cng ngh sn xut bia ngi ta x l GA (2 5ppm) cho ht i mch trong qu trnh ny mm s tng nng sut malt v phm cht malt bia.

3. Cytokinin
3.1. Lch s pht hin:

257

T 1913, Haberlandt chng minh rng: Cc m thng tn c kh nng sinh ra mt cht kch thch sinh trng v khuych tn vo m khng thng tn kch thch s phn chia ca n. Nm 1941, Overbeek chng minh l nc da c kh nng tn ti mt cht kch thch s phn chia v phn ha t bo. Nm 1955, Miller v Skoog tch c mt hp cht t vic hp mu DNA ca tinh dch c mi, cht ny c kh nng kch thch s phn chia t bo rt mnh trong nui cy m thuc l gi l Kinetin. Cytokinin trong cy c chit tch t 1963(Letham v Muller) dng kt tinh t ht ng non gi l Zeatin. Sau ngi ta chng minh rng Zeatin c mt trong tt c th gii thc vt. Trc ngi ta cng pht hin ra ngun cht giu c trong nc da c hot tnh cytokinin l diphenylurea. Cng thc cu to tng qut ca cytokinin nh sau: NH - R N N

N Nu R l: CH2 O

NH

th l Kinetin. th l Zeatin.

Nu R l CH CH = C CH OH 2 2 CH3

NH C=O NH Diphenylurea 3.2. S trao i cytokinin trong cy: R l c quan tng hp cytokinin. T r cytokinin c vn chuyn ln cc b phn trn mt t. Cytokinin c tng hp mnh nht l giai on cy con. Cytokinin phn b nhiu nht nhng ni c hot ng trao i cht v sinh trng mnh nht. Do vy cy no sinh trng mnh, chi non nhiu th hm lng cytokinin cao. Cytokinin tn ti trong cy di dng t do c hot tnh sinh hc v dng lin kt khng c hot tnh hoc hot tnh yu. Cytokinin c th lin kt vi ng to nn 258

glycoside. Ngoi ra cytokinin c mt trong RNA vn chuyn cho serine, phenylalanine, tryptophan, tyrosine, cysteine. Cytokinin b phn gii nh h thng enzyme c hiu v sn phm phn gii cui cng l ur. 3.3. Hiu qu sinh l ca cytokinin: * Cytokinin v s gin n ca t bo * Cytokinin v s phn ha chi. * Cytokinin v s ha tr: Cytokinin l hormone ha tr. Chng c tc dng ko di tui th ca l v ca c quan, ca ton cy. Tc dng c trng nht ca cytokinin l km hm s ha gi ca l. Khi cc l gi th chng c th rng i. Trong l s phn hy nhanh protein trong bn l, tch cc cht phiprotein, lipid, nucleic acid qua mng t bo gn l, cung l v dn theo s ph hy ca lc lp. Do vy vic x l cytokinin c tc dng km hm s phn hy protein nn km hm c s phn hy ca dip lc v ko di tui th ca l. Trong thc t nu b r pht trin mnh, tr lu l v r l c quan tng hp cytokinin. V vy mun ko di tui th ca c quan v ca ton cy th cn phi iu khin s sinh trng ca b r tht tt. Ngc li mun rt ngn thi gian sinh trng, nhanh chng ra hoa kt qu th phi hn ch sinh trng b r, cht bt r. * Cytokinin cng c nh hng n s ny mm ca ht. V vy trong mt s trng hp nht nh, cytokinin c tc dng kch thch s ny mm, ph b s ng ngh ca ht, c. 3.4. C ch tc ng ca cytokinin: Cytokinin tc ng n s phn chia t bo, s phn ha c quan, s tr ha c quan v ca ton cy. C ch tc ng ca cytokinin ln s sinh trng ca cy c th tm tt nh sau: - Khi thiu cytokinin th t bo khng phn chia c mc d DNA vn tip tc c tng hp, nhng m s tng hp protein khng xy ra. V vy c th cho rng cytokinin kim tra s tng hp protein giai on gii m (translation): Transcription DNA mRNA Cytokinin - C th c c ch th hai l cytokinin c mt trong RNA vn chuyn nn n nh hng n s tng hp protein. Cytokinin trong RNA vn chuyn v tr lin kt vi anticodon trn ribosome. V vy chc nng iu chnh ca cytokinin c l bng c ch ngn chn s nhn mt sai lm ca cc codon trong qu trnh tng hp protein. - Hiu ng ca cytokinin trong vic ngn chn s ha gi ca cy lin quan nhiu n kh nng ngn chn s phn hy protein, nucleic acid v dip lc. 3.5. ng dng ca cytokinin Cytokinin c ng dng ch yu trong vic nui cy m t bo thc vt. Cc cht cytokinin nhn to nh benzyladenine (BA), kinetin cng nh cytokinin t nhin Translation Protein-Enzyme

Sinh trng

259

trong nc da c ng dng rng ri trong nui cy m. Vic c mt ca chng trong mi trng quyt nh s phn chia v phn ha chi trong m nui cy.

NH CH2 N N Benzyladenine (BA) N NH

B. CC CHT C CH SINH TRNG THC VT

1. Abcisic acid (ABA)

1.1. Lch s pht hin: * 1961 - Luc Carn (M) tch c mt cht di dng tinh th t qu bng gi v khi x l ln l bng non th gy nn hin tng rng ca l v t tn l abcisin I. 1963 Ohkuma v Eddicott tch c mt cht khc t l bng tr cng gy nn hin tng rng, h gi l abcisin II. ng thi Waring v cng s bng phng php sc k tch c mt cht c ch t l cy Betula v khi x l cho cy gy nn hin tng ng ngh ca chi. H gi cht l dormin. Nm 1966 ngi ta xc nh c bn cht ha hc ca cht . Nm 1967 ngi ta gi cht l abcisic acid (ABA).

CH3 CH3 CH = CH C = CH COOH OH CH3 CH3 Abcisic acid (ABA)

* ABA c tng hp hu ht cc b phn ca cy, tch ly nhiu nht trong cc c quan sinh sn, c quan d tr, trong cc c quan gi, c quan ang ng ngh. - ABA gy nn s rng (l, hoa, qu, ), hot ha s hnh thnh tng ri ch yu l s hnh thnh cc enzyme thy phn v t bo. ABA c ch s sinh trng ca chi; ca cy. Gy nn s ng ngh. - ABA l cht i khng ca auxin. - ABA l hormone ha gi. - ABA iu chnh s ng m kh khng: Vic pht hin ra vai tr ca ABA trong vic iu chnh ny xut pht t vic pht hin ra mt t bin ho c chua. Cy c 260

chua lun lun b ho ny v kh khng ca n lun lun m m khng ng c. Phn tch hm lng ca cc cht trong t bin ny th thy thiu ABA. Khi x l ABA cho n th kh khng ng li. t bin ny l do thiu gen tng hp ABA nn ABA khng hnh thnh c. Vai tr ca ABA l c ch s hp thu K+ bi t bo kh khng. Mt dng vt cht khc c lin quan n s ng m ca kh khng di tc ng ca ABA l malic acid. Acid ny c gii phng cng vi K+ khi c mt ca ABA v to nn mt s thiu ht K+ t bo kh khng, thiu sc trng, t bo mt nc v kh khng ng li. - ABA c th xem l hormone stress: Cc iu kin bt thun ca mi trng nh hn, ng, nng, lnh, mn, su bnh, gi l iu kin stress ca mi trng. Khi gp mt tc nhn stress bt c no th hm lng ABA lp tc tng ln rt mnh. l phn ng thch nghi ca cy chng li cc iu kin bt thun ca mi trng. Khi gp hn, hm lng ABA trong l tng ln mnh (c l do s tng hp mi) v do kh khng ng li, chng c s thot hi nc. Ngi ta xc nhn rng ABA c tng hp trong lc lp v c gi trong lc lp khi t bo nc. Khi t bo mt nc th tnh thm ca mng lc lp tng ln v ABA c thm ra ngoi t bo cht, vn chuyn vo t bo kh khng gy nn s ng ca kh khng. Khi ht stress th tnh thm ca mng lc lp vi ABA gim xung v do ABA c d tr trong lc lp. V vy, ABA c vai tr nht nh trong tnh chng chu ca cy i vi cc iu kin bt thun ca mi trng. 1.2. C ch tc ng ca ABA a) Tc ng ca ABA ln tnh thm ca mng: ABA lm thay i in th qua mng v do m iu tit c s tit ion K+ qua mng. C ch ny lin quan n s ng m ca kh khng. C ch v vai tr ca ABA ln s ng m kh khng nh sau: Khi thiu ABA, bm H+ khng b c ch, malate v c K+ vn chuyn vo khng bo v do t bo ht nc kh khng m. Khi x l ABA, bm H+ b c ch nn H+ khng vo thnh t bo; malate v K+ khng qua mng vo khng bo m vn chuyn th ng ra khi t bo, lc t bo mt nc v kh khng ng li. Thnh t bo Cht nguyn sinh Glucose ABA c ch bm H+, c ch ATP-ase c ch thm K+ v malate vo khng bo CO2 H+ K+ PEP Malate K+ Khng bo

K+

b) ABA tc ng ln c ch tng hp protein v nucleic acid: Nm 1964, Dorothy v Bonner cho rng trong cc t bo ang ng ngh th vt liu di truyn (DNA) hon ton b trn p v qu trnh tng hp protein khng din ra; qu trnh sinh trng b ngng tr. 261

Nu x l cc cht i khng ca ABA nh GA chng hn lm gim v c ch tc dng ca ABA ln h thng v do vy m qu trnh sinh trng din ra. Hiu qu c ch ca ABA l hiu qu ng gen (kha gen) do qu trnh sao chp (Transcription) v gii m (Translation) khng din ra c. Gibberellin (GA) s lm mt tc dng c ch gen ca ABA, lm cho cc qu trnh din ra bnh thng. C l ABA tc ng ln giai on transcription: 1.3. ng dng nhng hiTranslation u bit v ABA trong trng trt: Transcription Sinhtrng pht trin Trong cy ABA th ng c ch ln qu trnh sinh DNA mRNA gy hiu qu Protein tr ng ta thng s khc nhau. loi b tc dng c ch c a ABA i cy trng chng (Enzyme, ci uv trc) dng cc tc nhn i khng vi ABA lm thay i s cn bng hormone trong cy theo hng lm mt tc dng ca ABA: - ph b ng ngh, kch thch s ny mm th hoc l lm gim lng ABA hoc lm tng gibberellin (GA) hay c hai. - Km hm s rng ca l, qu bng cch tng hm lng auxin, gim hm lng ABA. Cytokinin - iu chnh s ha gi: T l quy nh gi ha ca cy. ABA Mun chng s gi ha c th tin hnh bng cch phun cytokinin ngoi sinh, cho cytokinin vo mi trng nui cy hoc bng cch tng cng s pht trin ca b r tng hp nhiu cytokinin cung cp cho cc c quan trn mt t ko di tui th ca chng. L sn phm ca s trao i cht, n l mt cht kh n gin. Ethylene kch thch s chn ca qu. Ethylene cng tng tc vi ABA gy nn s rng ca l, qu. Ethylene kch thch s ra hoa ca mt s thc vt nh kch thch cho da ra hoa tri v. Trong ngh trng da vic s dng cht to ra ethylene ( l ethrene) hoc t n (cho acetylene) lm cho da ra thm mt v qu. Ethylene l mt cht kh nn trong thc t khng s dng trc tip ethylene m ngi ta s dng cc cht to ra ethylene nh CEPA: O Cl(CH2)2 P OH H2O HCl H3PO4 CH2 = CH2 Ethylene

2. Ethylene (CH2 = CH2)

OH CEPA (Chloroethylenephosphoric acid)

- Trong vic phn ha gii tnh ca cc cy thuc h bu b th ethylene c nh hng ln s hnh thnh gii tnh ci. X l ethrene s lm cho chng pht trin ch yu hoa ci.

3. Cc cht phenol
262

L sn phm ca s trao i cht trong cy. Chng c tc dng c ch hot ng trao i cht v c ch sinh trng. Cc phenol kch thch hot ng ca enzyme phn hy auxin; km hm s gin ca t bo v lm cho t bo ha g nhanh. Mt s phenol thng gp nh: O COOH O O O Urgon Cumarin Oxybenzoic acid HO

4. Cc cht lm chm sinh trng (Retardant)

Retardant l mt nhm cht nhn to, c bn cht ha hc khc nhau, chng gy nh hng c ch gin ca t bo, lm cy thp, m rng phin l, gy s ra hoa sm. Mt s v d in hnh: 4.1. CCC (Chlorocholine chlorid): [(CH3)3N CH2 CH2 Cl]+ Cl- CCC l cht khng GA, n lm cho cy thp i. Lm tng s hnh thnh dip lc. Khi x l CCC ln cy, tc thm vo cy rt nhanh, tn ti vi tun ri b phn hy, mt hot tnh. 4.2. MH (Malein hydrazide) MH l cht khng auxin v n lm tng hot tnh enzyme phn hy auxin, l IAA-oxidase O NH NH O MH l cht c ch sinh trng, n c ch s ny mm v ko di thi gian ng ngh; c ch s sinh trng chi bn ca cy nn c s dng ngn chn s mc chi nch ca thuc l, trnh c vic nh ta nhnh bng tay rt tn cng lao ng. 4.3. TIBA (Triiodobenzoic acid) I COOH I I 263

TIBA cng l cht khng auxin, n km hm s vn chuyn auxin trong cy; lm gim s sinh trng ca chi ngn, hn ch chiu cao, xc tin s phn cnh; TIBA cn xc tin s ra hoa v s hnh thnh c.

264

TI LIU THAM KHO

1. V Kim Bng ng Hng L Khc Thn: Bi ging ha sinh NXBH v GDCN H ni 1991. 2. L Don Din: Ha sinh thc vt NXBNN H ni 1993. 3. ng Hng V Th Th: Ha sinh hc cy trng nng nghip NXBNN H ni 1993. 4. V Vn Quang: Bi ging sinh ha thc vt i hc Nng lm Hu 1996. 5. Hong Vn Tin L Khc Thn L Don Din: Sinh ha hc vi c s cng ngh gen NXBNN H ni 1997. 6. Nguyn Duy Minh: Nhng bi tp trc nghim chn lc v tr li sinh ha NXB i hc Quc gia H Ni 2001. 7. Nguyn nh Huyn Mai Xun Lng: Ha sinh hc hin i theo s NXB y hc TP HCM 1983. 8. Hong Minh Tn - Nguyn Quang Thch: Cht iu ha sinh trng i vi cy trng NXBNN 1993. 9. Nguyn Vn Uyn: Phn bn l v cc cht kch thch sinh trng NXBNN TP HCM 1995. 10. Stryer L. Biochemistry NY 1987. 11. BreskiW. Practikum z Biochemi Warszawa 1971.