17

Chng 1

Saccharide
L hp cht hu c c to nn t cc nguyn t: C, H, O c cng thc cu to chung Cm(H2O)n, thng m = n. Do c cng thc cu to nh trn nn saccharide thng c gi l carbohydrate - c ngha l carbon ngm nc. Tuy nhin c nhng saccharide c cng thc cu to khng ng vi cng thc chung ni trn v d: deoxyribose (C5H10O4). C nhng cht khng phi l saccharide nhng c cng thc cu to ph hp vi cng thc chung trn v d : acetic acid (CH3COOH). Saccharide l thnh phn quan trng trong mi sinh vt . thc vt, saccharide chim t 80 - 90% trng lng kh, saccharide tham gia vo thnh phn cc m nng , v d cellulose, hay tch tr di dng thc phm d tr vi lng ln, v d tinh bt. ng vt, hm lng saccharide thp hn nhiu, thng khng qu 2%, v d glycogen.

1.1. Monosaccharide
1.1.1. Cu to v danh php L cht c cha nhiu nhm ru v mt nhm kh oxy (nhm kh l nhm carbonyl l aldehyde hay ketone). - Nhm kh l aldehyde ta c ng aldose v c cng thc tng qut: CHO (CHOH)n CH2OH - Nhm kh l ketone ta c ng ketose c cng thc tng qut: CH2OH C= O (CHOH)n CH2OH

18 CHO - CH2OH c xem nh l monosacchariden gin nht. Trong thin nhin monosaccharide c cha t 2 n 7 carbon v c gi tn theo s carbon (theo ting Hy Lp) + ose V d: monosaccharide c 3C gi l triose. Tng t ta c tetrose, pentose, hexose, heptose. 1.1.2. ng phn quang hc Quy c Fischer: Fischer l ngi u tin nu ra nguyn tc biu din cc monosaccharide bng cng thc hnh chiu ca chng. Theo : hnh chiu ca cc nguyn t carbon bt i (C*) v cc nguyn t C khc nm trn mt ng thng, nguyn t C c s th t nh nht c hnh chiu nm trn cng. Cn cc nhm th c hnh chiu bn phi hay bn tri. V d : glyceraldehyde. V glyceraldehyde c 1 C* nn theo quy tc ca Vant Hoff c 2 ng phn (N = 2n) 1CHO 1CHO HO- 2C* -H
3CH2OH

H-2C*-OH
3CH2OH

D: -OH bn phi L: -OH bn tri

L glyceraldehyde D glyceraldehyde. Khi phn t monosaccharide c nhiu C* th cng thc c dng D hay L c cn c vo v tr nhm OH ca C* xa nhm carbonyl nht. V d : CHO CHO H-C-OH HO-C-H H-C-OH H-C-OH CH2OH D glucose H-C-OH HO-C-H H-C-OH HO-C-H CH2OH L glucose

19 Ch : monosaccharide t triose tr ln u c C* tr dihydoxy aceton CH2OH C=O CH2OH 1.1.3. Cng thc vng ca monosaccharide Cng thc thng theo Fischer nh trnh by trn khng ph hp vi mt s tnh cht ho hc ca chng nh: mt s phn ng ho hc thng xy ra vi aldehyde khng xy ra i vi monosaccharide . V vy c th ngh rng nhm -CHO trong monosaccharide cn tn ti di dng cu to ring bit no . Mt khc: monosaccharide c th to ether vi methanol to thnh mt hn hp 2 ng phn c cng nhm methoxy (- OCH3). iu chng t trong monosaccharide cn tn ti mt nhm -OH c bit. Qua nghin cu Kolle cho thy: s ng phn thu c ca monosaccharide thc t nhiu hn s ng phn tnh theo cng thc N=2n, do gii thch cc hin tng trn, Kolle cho rng ngai dng thng monosaccharide cn tn ti dng vng. S to thnh dng vng xy ra do tc dng ca nhm -OH cng phn t monosaccharide to thnh dng hemiacetal hay hemiketal.

20 V d : cu to vng ca glucose xy ra nh sau:

Do s to thnh hemiacetal vng m C1 tr nn C*, nhm -OH mi c to ra C1 l -OH glucoside. Tng t vi ketose th C2 tr nn C*, nhm -OH mi c to ra C2 l -OH glucoside khi to thnh hemiketal. Cch biu din cng thc vng nh trn da vo nguyn tc ca Haworth: C v cu ni vi oxy nm trn mt mt phng , cc nhm th cng thc thng nm bn phi th cng thc vng nm di mt phng v ngc li. Ring cc nhm th ca C c nhm OH dng to cu ni oxy th theo quy tc ngc li. 1.1.4. Hin tng h bin ca monosaccharide Nh ta thy, khng th gii thch c tt c cc tnh cht ca monosaccharide nu ta ch tha nhn mt dng cu to no ca monosaccharide. Nn ngi ta cho rng cc dng cu to c th chuyn ho ln nhau. pyranose pyranose Dng thng Furanose Furanose

21 1.1.5. Tnh cht ca monosaccharide 1.1.5.1. L tnh Cc monosaccharide tan trong nc, khng tan trong dung mi hu c, c tnh quay cc tr biose v khng c C*. 1.1.5.2. Ho tnh a. Monose l tc nhn kh Trong mi trng kim, kh cc ion kim loi nng c ho tr cao thnh ion c ha tr thp hay cc ion kim loi thnh kim loi. Tnh kh ny do nhm aldehyde hay nhm ketone to ra v cc monose bin thnh acid. V d: Cu2+ b bin i thnh Cu+ trong phn ng vi thuc th Fehling, Ag+ b bin i thnh Ag trong phn ng trng gng. b. Phn ng vi cc cht oxy ho Tu thuc vo cht oxy ho: - Cht oxy ho nh nh nc brom ng aldose s thnh aldonic acid, vi ketose phn ng khng xy ra. - Cht oxy ho mnh nh HNO3 m c c s oxy ho xy ra 2 u cho ta di acid. - Trng hp c bit nu ta bo v nhm -OH glucoside bng cch methyl ha hay acetyl ho trc khi oxy ho bng nc brom, sn phm to thnh l uronic acid. c. Phn ng vi cht kh D dng vng chim t l rt ln trong thnh phn, dng thng chim t l nh nhng cho ta thy r tnh cht ca mt carbonyl tht s. Khi b kh: monose s bin thnh polyalcohol. d. Phn ng to furfural Di tc dng ca acid m c, cc aldopentose to thnh furfural v aldohexose bin thnh hydroxymethylfurfural. Cc sn phm ny khi cho tc dng vi cc phenol cho mu c trng nh: naphthol cho vng mu tm (Molisch). y l phn ng phn bit ng vi cc cht khc. Nu ng 5C s cho mu xanh cm thch vi orcinol (Bial). e. Phn ng ester ho Cc gc ru ca monose c kh nng kt hp vi acid to thnh ester. Cc ester phosphate thng gp l: Glucose-6-phosphate, fructose6-phosphate...

22 1.1.6. Cc monose quan trng 1.1.6.1. Pentose

1.1.6.2. Hexose Cc hexose quan trng nh: * Glucose: cn gi l dextrose v lm quay mt phng nh sng phn cc v pha phi. Ph bin rng ri trong thc vt nht l trong qu nho, nn cn gi l ng nho, trong mu ngi c 0.8 - 1,1 g/l, nhng ngi b bnh i ng c th n 2g/l. Cc disaccharide quan trng l saccharose, lactose, maltose v cc polysaccharide quan trng l tinh bt, glycogen. Ngi ta s dng glucose trong y hc nh cht tng lc. * D - Mannose: t gp trng thi t do, thng gp trong polysaccharride v glucoside * D - Galactose: l thnh phn ca lactose c trong sa cn gi l ng no tu. Chng l thnh phn cu to ca raffinose, hemicellulose. pectine... * D - Fructose cn gi l levulose v lm quay mt phng nh sng phn cc v pha tri. Fructose cn gi l ng qu, c trng thi t do trong tri cy chn v mt ong. Chng l thnh phn ca disaccharide saccharose. Trong c th ta cn thy dng ester vi phosphoric acid ng vai tr quan trng trong trao i cht. Fructose c ngt rt ln, dng c ngt bng 1/3 dng .

23

24

1.2. Oligosaccharide
1.2.1. Disaccharide S to thnh disaccharide l do s kt hp ca 2 monose cng loi hay khc loi nh lin kt glucosidic. Lin kt glucosidic c th c to thnh gia -OH glucoside ca monose ny vi -OH glucoside ca monose kia, hay gia mt nhm -OH glucoside ca monose ny vi -OH ( khng phi -OH glucoside) ca monose kia. Disaccharide ch c tnh kh khi t nht mt trong 2 nhm -OH glucoside trng thi t do. Ngha l disaccharide s khng c tnh kh khi 2 nhm -OH glucoside lin kt vi nhau. Cc disaccharide quan trng * Maltose do 2 phn t - D-glucose lin kt vi nhau v tr C1 C4 to thnh. Cng thc cu to:

Maltose c nhm -OH glucoside trng thi t do nn c tnh kh. Maltose c nhiu trong mm la v mch nha (maltum) nn gi n l maltose. * Lactose (ng sa) do mt phn t D-galactose lin kt vi mt phn t D- glucose v tr C1- C4.

* Saccharose do mt phn t D-glucose lin kt vi mt phn t D-fructose v tr C1-C2. Do n khng c tnh kh, cn gi l ng ma v c nhiu trong ma. D b thy phn khi un nng.

25

1.2.2. Trisaccharide L oligosaccharide c cha 3 monosaccharide, ph bin trong thin nhin l raffinose. Cng thc cu to nh sau: -D-galactopyranosyl 1-2 -D glucopyranosyl 1-2 -D fructofuranose. Do c cng thc nh trn nn khng c tnh kh oxy. D b thy phn, di tc dng ca fructofuranosidase s to thnh fructose v melobiose vi galactosidase s to thnh galactose v saccharose.

1.3. Polysaccharide
Cn gi l glycan, ty thnh phn monose c trong polysaccharide ngi ta chia chng ra lm: homopolysaccharide (ch cha mt lai monosaccharide) v heteropolysaccharide (c t nht 2 lai monosaccharide).

26 Polysaccharide ng vai tr quan trng trong i sng ng vt, thc vt. Mt s polysaccharide thng gp nh tinh bt, glycogen, cellulose... 1.3.1. Polysaccharide thc vt 1.3.1.1. Tinh bt L polysaccharide d tr ca thc vt, do quang hp to thnh. Trong c v ht c t 40 n 70% tinh bt, cc thnh phn khc ca cy xanh c it hn v chim khong t 4 n 20%. Tinh bt khng ha tan trong nc, un nng th ht tinh bt phng ln rt nhanh to thnh dung dch keo gi l h tinh bt. Tinh bt c cu to gm hai phn: amylose v amylopectin, ngoi ra cn c khong 2% phospho di dng ester. T l amylopectin/amylose cc i tng khc nhau l khng ging nhau, t l ny go np l ln hn go t. *Amylose Chim 15 n 25% lng tinh bt, do nhiu gc D- glucose lin kt vi nhau thng qua C1-C4 to thnh mch thng khng phn nhnh. Trong khng gian n cun li thnh hnh xon c v c gi bn vng nh cc lin kt hydro. Theo mt s ti liu trong amylose cn c cha cc D- glucopyranose dng thuyn.

Amylose bt mu xanh vi iodine, mu ny mt i khi un nng, hin mu tr li khi ngui. Mt c trng ha l khc cn ch l n b kt ta bi ru butylic.

27

Ht tinh bt trong lc lp amylose

* Amylopectin Cu to do cc phn t D- glucose lin kt vi nhau, nhng c phn nhnh. Ch phn nhnh l lin kt C1-C6 glucosidic.

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1.3.1.2. Cellulose c cu to bi nhng phn t D-glucose lin kt vi nhau bng lin kt 1-4 glucosidic. Chng l thnh phn ch yu ca vch t bo thc vt. i vi ngi th cellulose khng c gi tr dinh dng v cellulose khng b thy phn trong ng tiu ha. Mt s nghin cu cho thy n c vai tr trong iu ha tiu ho. ng vt n c thy phn c cellulose nh enzyme cellulase. Cellulose khng tan trong nc, tan trong dung dch Schweitzer. Khi un nng vi H2SO4, cellulose s b thy phn thnh cc phn t D-glucose. Cellulose c dng hnh si di, nhiu si kt hp song song vi nhau thnh chm nh cc lin kt hydro, mi chm (micelle) cha khang 60 phn t cellulose. Gia cc chm c nhng khong trng, khi ho g khong trng ny cha y lignin v ta xem lp lignin ny nh l mt lp cement. Lignin l cht trng hp ca coniferylic alcohol

Cc gc -OH ca cellulose c th to ester vi acid v d: to nitro cellulose vi HNO3 , to acetyl cellulose vi CH3COOH.

29 1.3.1.3. Hemicellulose Tn gi chung cho lp polysaccharide thng i theo vi cellulose trong thc vt. Hemicellulose khng tan trong nc, tan trong dung dch kim v thy phn bng acid d hn cellulose. Khi b thy phn hemicellulose to thnh mt hn hp gm cc hexose v pentose hay ch mt mnh hexose m thi. Trong hemicellulose khi monose no chim a s th hemicellulose c tn tng ng vi monose : Xylose chim a s th hemicellulose c tn l Xylan, Arabinose chim a s th hemicellulose c tn l Araban, Galactose chim a s th hemicellulose c tn l Galactan... Xylan c nhiu trong rm r, trong mt s c quan ca thc vt, galactose c nhiu trong rm, g v cc loi ht. 1.3.1.4. Inulin L polysacchride d tr ca thc vt c trng lng phn t khong 5000-6000, do nhng phn t D- fructose lin kt vi nhau bng lin kt 1-2 v tn cng bng mt phn t saccharose. Inulin c tm thy trong c thc dc khong 40%. Ngi ta x dng inulin sn xut fructose. xc nh inulin ngi ta thy phn n v xc nh bng phn ng nh tnh Seliwanoff. 1.3.1.5. Pectin L loi polysaccharide c nhiu trong qu , c v thn cy, thnh phn chnh l galacturonic acid c nhm -COOH b methyl ha. Ngi ta s dng rng ri pectin trong sn xut keo. 1.3.2. Polysaccharide ng vt 1.3.2.1. Glycogen L polysaccharide d tr ng vt c tm thy trong gan v c, hin nay cn tm thy trong mt s thc vt nh ng, nm. C cu to ging amylopectin nhng phn nhnh nhiu hn, b thu phn bi phosphorylase ( c coenzyme l pyrydoxal phosphate), ct lin kt 1-6 cn enzyme debranching. Sn phm cui cng l cc phn t glucose-1-P.

30 Pha ngoi glucose lin kt 1-6

Mch chnh

ht glycogen t bo gan

1.3.2.2. Hyaluronic acid C cng thc cu to c lp li t n v sau:

Hyaluronic acid c trng lng phn t rt ln, c th ln n nhiu triu, hyaluronic acid rt ph bin v l thnh phn quan trng ca m lin kt, c tm thy trong dch khp xng, trong thy tinh th mt, n tc dng nh mt lp cement bo v bn trong t bo chng li s xm nhp ca vi khun cng nh cc cht l khc. khp xng n lm

31 cho dch c tnh trn gip c ng khi b au. Hyaluronic acid b thy phn bi hyaluronidase, enzyme ny c tm thy trong vi khun gy bnh, trong tinh trng. Hyaluronidase to d dng cho tinh trng i vo non ca bun trng, mt khc n cng l yu t gip cho cc cht khc v vi khun gy bnh i vo cc m trong c th. 1.3.2.3. Chondroitin L heteropolysaccharide, thnh phn khng th thiu c m xng sn.

1.3.2.4. Heparin Heteropolysaccharide c tc dng chng li s ng mu v ngn chn s bin i prothrombin thnh thrombin.

GlcA2S hoc IdoA2S

1.3.3. Mt s polysaccharide ph bin khc 1.3.3.1. Chitin L homopolysaccharide, c v s, c, cc loi cn trng v nm mc. N c cu to nh sau:

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1.3.3.2. Dextran c tm thy vi khun v nm men, cu to mch chnh l Dglucose1-6, nhnh l 1-3 v thnh thong c nhnh 1-2 hay 1-4. Do c cu to 1-6 nn i vi ng vt, dextran khng b phn gii hay b phn gii rt chm.

33 Dextran c di v hnh dng ging albumin, ngi ta thng dng nhit thy phn khng hon ton dextran nhm thay th protein ca huyt tng , dung dch 10% ca n hon ton trong sut. Trong cng ngh ngi ta tng hp dextran v c gi l sephadex s dng trong tch tng phn protein.

TI LIU THAM KHO

Ti liu ting Vit
1. Phm Th Trn Chu, Trn thi ng. 1999. Ho sinh hc, NXB Gio dc, H Ni. 2. Qu Hai. 2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 3. Trn Thanh Phong.2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 4. L Ngc T (ch bin), L Vn Ch, ng Th Thu, Phm Quc Thng Nguyn Th Thnh, Bi c Hi, Lu Dun, L Don Din, 2000. Ha sinh Cng nghip, Nxb KH&KT, H Ni.

Ti liu ting Anh

1. Gilbert H. F. 1992. Basic concepts in biochemistry, Copyright by the Mcgraw- Hill companies, Inc. 2. Lehninger A. L. 2004. Principles of Biochemistry, 4th Edition. W.H Freeman.

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Chng 2

Lipid
Cng nh saccharide, protein, lipid l cht hu c phc tp, ta c th nh ngha nh sau: * nh ngha rng: Lipid l cht tan c trong dung mi hu c, khng tan trong nc, nh ngha ny khng phn nh ht tnh cht ca cc lipid v: - C lipid khng tan c trong dung mi hu c nh phospholipid khng tan trong aceton. - Nhng cng c cht khng phi lipid nhng tan c trong dung mi hu c. * nh ngha hp: Lipid l ester ca ru v acid bo. Tuy nhin c nhng lipid do acid bo lin kt vi ru bng lin kt peptide. * nh ngha dung ho: Lipid l nhng cht chuyn ho ca acid bo v tan c trong dung mi hu c. Lipid rt ph bin ng vt cng nh thc vt v tn ti di 2 dng m nguyn sinh cht (dng lin kt) v dng d tr (dng t do). - M nguyn sinh cht: thnh phn ca mng t bo cng nh cc bo quan khc v d: ty th, lp th... dng ny khng b bin i ngay c khi con ngi b bnh bo ph hoc b i. - Dng d tr (dng t do) c tc dng cung cp nng lng cho c th, bo v cc ni quan, l dung mi cn thit cho mt s cht khc. Cn c vo thnh phn nguyn t c mt, ngi ta chia lipid ra lm 2 loi * Lipid n gin: trong phn t ch cha C, H, O. * Lipid phc tp: ngoi C, H, O cn c mt s nguyn t khc nh N, P, S.

2.1. Lipid n gin

2.1.1. Glycerid Glycerid l ester ca ru glycerol v acid bo, l m d tr ph bin ng vt v thc vt.

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1- Stearoyl, 2- linoleoyl, 3-palmitoyl glycerol, mt triacylglycerol hn tp

2.1.1.1. Glycerol L triol khng mu, v ngt nhn. Khi t glycerol hay lipid c cha glycerol vi cht ht nc s to acrolein c mi kht. 2.1.1.2. Acid bo Acid bo thng gp l nhng acid bo c s carbon chn, mch thng, c th no hay khng no v chui C xp theo hnh ch chi. Tuy nhin cng c nhng acid bo ngoi nhm chc acid cn cha nhng nhm chc khc nh ru, ketone, mch carbon c vng hay nhnh. a. Acid bo chn, thng, no: CH3(CH2)nCOOH C4 CH3 -(CH2)2 COOH butylic acid c nhiu trong c. C6 CH3 -(CH2)4 -COOH caproic acid c trong b, sa d. C8 CH3 - (CH2)6-COOH caprylic acid c trong b, sa d. C10 CH3-(CH2)8 COOH capric acid c trong b, sa d.

36 C12 n=10 lauric acid c trong du da. C14 n=12 myristic acid c trong du da. C16 n=14 palmitic acid c trong du ng vt,thc vt. n=16 stearic acid c trong du ng vt,thc vt. C18 C20 n=18 arachidic acid c trong du lc. b. Acid bo chn, thng, khng no - Cha mt ni i (C): 10 9 C 16 (9-10): CH3-(CH2)5 -CH = CH- (CH2)7COOH Palmitoleic acid : Tm thy trong du thc vt. C18 (9-10): CH3-(CH2)7 -CH = CH- (CH2)7 COOH Oleic acid: acid ny c ba ng phn. C18 (6-7): Petroselenic acid C18 (11-12): Vaccenic acid. C18 (12-13): Heparic acid - Acid bo c 2 ni i (C): C18 (9-10,12-13): Linoleic acid CH3-(CH2)4 -CH = CH- CH2-CH=CH-(CH2)7-COOH C th khng tng hp c acid ny m ly t ngoi vo. Ngy xa ngi ta quan nim acid ny l vitamin v gi l vitamin S. Nhng thc cht l mt acid bo m c th cn vi mt lng ln. - Acid bo c cha 3 ni i (C): C18((9-10,12-13,15-16): Linolenic acid, c th khng tng hp c acid ny. - Acid bo c 4 ni i (C): C20 (5-6,8-9,11-12,14-15): Arachidonic acid. Ngoi ra cn c cc acid bo c cha ni ba nhng khng quan trng. c. Acid bo c cha chc ru Thng gp trong lipid phc tp v cha nhm ru gn chc acid nn c tn l - hydroxy... R-CH-COOH OH

37 V d: - hydroxy lynoceric acid CH3 -(CH2 )21- CH-COOH OH Ricinoleic acid CH3-(CH2)5 - CH - CH2 - CH = CH- (CH2)7 - COOH
10 9

OH d. Gc R trong phn t acid c nhnh v c s C l Phocenic acid: Undecylonic acid : e. Acid bo c vng * Chaulmoogric acid: CH3 CH CH2 COOH CH3 CH2 = CH - (CH2)8 - COOH - (CH2)12 - COOH

* Sferculic acid:

CH3 - CH2 - C = C - (CH2)7 - COOH

CH2 2.1.1.3. Tnh cht ca acid bo v triglyceride * Tnh cht vt l: a. im tan chy im tan chy ph thuc vo s C ca acid bo, acid bo c chui C di th im tan chy cao v ngc li. Nhng acid bo c C l c im tan chy thp hn acid bo c s C nh hn n 1 n v . Ngoi ra tan chy cn ph thuc vo s ni i trong phn t acid bo, acid bo cha nhiu ni i th im tan chy cng thp. b. si Acid bo c chui C di th si cng cao, thng p dng tnh cht ny tch cc acid bo ra khi nhau. c. Tnh ho tan - Trong nc: acid bo c chui C ngn (4,6,8) d tan, C10 kh tan, C12 khng tan. Nu acid bo dng mui th d ha tan hn. - Trong dung mi hu khng phn cc nh benzen, ether, ether du ho acid bo d tan.

38 - Trong dung mi hu c phn cc nh aceton, acid bo kh ho tan hay ho tan rt t. * Tnh cht ho hc: a. S hydrogen ho Acid bo cha no c th kt hp vi H2 to thnh acid bo no R - (CH2)n - CH =CH- (CH2)n - COOH + H2 R - (CH2)n -CH2 - CH2 - (CH2)n - COOH Ngi ta dng phn ng ny ch to thc phm nh margarin. b. S halogen ho Acid bo khng no kt hp vi cc nguyn t thuc h halogen (F, Cl, Br, I) to thnh acid bo no. R - (CH2)n - CH = CH- (CH2)n - COOH + I 2 R - (CH2)n - CH - CH- (CH2)n - COOH I I C th dng phn ng ny xc nh s ni i trong phn t acid bo. Phn ng d dng hay kh xy ra tu thuc vo v tr ni i i vi nhm carboxyl, ni i cng gn nhm carboxyl phn ng cng kh xy ra. xc nh s ni i ngi ta cn c vo ch s Iod. Ch s Iod: L s gam Iod cn thit tc dng ln 100gam cht bo. Do ch s iod cng ln th s ni i cng nhiu. c. S thu phn: Ester nn khi thu phn s to thnh ru glycerol v acid bo. Tc nhn thy phn l acid, kim, nc hay enzyme. * Thy phn bng nc cn nhit v p sut cao. * Thy phn bng kim: NaOH hay KOH Ch s x phng ho: s mg KOH cn thit trung ho 1g cht bo Do ch s x phng cng ln th di mch cng ngn, nn c dng xc nh di ca mch C. xc nh tnh cht ca cht bo ngi ta cn cn c vo mt s ch s khc nh ch s acid. Ch s acid: s mg KOH dng trung ho tt c acid bo t do c trong 1g cht bo.

39 * Thu phn bng enzyme: trong c th lipid b thu phn bng enzyme lipase. - Lipase dch trng tc dng vo v tr . - Lipase ty tng tc dng vo v tr v . d. S i ha: Du m lu c mi v v kh chu gi l s i ha, mt trong nhng nguyn nhn gy ra l do oxy khng kh kt hp vo ni i to thnh peroxide. Nu oxy kt hp vo nguyn t carbon ng cnh lin kt i th s to thnh hydrogen peroxide. Sau peroxide v hydrogen peroxide s b phn gii to thnh aldehyde v ketone. Cc aldehyde v ketone ny u l nhng cht c mi v v kh chu. 2.1.2. Cerid Cng l ester ca ru v acid bo, nhit thng th rn, c ng thc vt, thc vt n thng to thnh mt lp mng ph ln l, thn, qu ca cy. Cng thc tng qut: R O CO R Ru trong cerid l ru cao phn t, ch cha mt nhm OH , mnh C khng phn nhnh, rt t khi mch C c vng V d: Ru cetol:CH3 - (CH2)14-CH2OH. Sp ong, sp c voi (spermaceti) l ester ca ru cetol v palmitic acid.

Ngoi ra trong sp ong v sp c voi cn c ru t do, acid bo t do v hydrocarbon. 2.1.3. Sterid L ester ca ru sterol v acid bo. Ru sterol c vng v trng lng phn t rt ln, sterol tiu biu l cholesterol, acid mt. Acid bo thng l palmitic, oleic, ricinoleic.

40 2.1.3.1.Cholesterol Cholesterol bao gm nhn phenanthrene kt hp vi cyclopentan to thnh cyclopentanoperhydrophenanthrene. Cholesterol c mang nhm ru C3, ni i C5 - C6 v 2 gc CH3 C10, C13 v mt nhnh isooctan C17. Cholesterol ch c ng vt, trong mu c khong 2.10-3, c nhiu trong c, nhng m l lch, gan, da cng c cha cholesterol hay cc cht chuyn ho ca n. Cholesterol c tm thy u tin sn mt, sn mt l do s dn mt n rut non b nghn, mt cha nhiu cholesterol nn kt ta li thng sn mt. Cholesterol l cht quan trng trong s sinh tng hp acid mt, vitamin D v nhiu cht khc. Cholesterol + acid bo cholesterid

Trong thin nhin, cc sterol trng thi t do nhiu hn trng thi sterid. c th ngi, ch 10% sterol b ester ha to thnh sterid. T l sterol v sterid cc m khc nhau l khng ging nhau. * L tnh ca cholesterol: kt tinh di dng vy ng nh nh x c, dng kt tinh cng khc nhau tu theo mi trng kt tinh. * Ho tnh: - Phn ng vi acid bo do nhm -OH C3. - B hydrogen ha hay halogen ho C5 - C6.

41 - Phn ng mu: + Phn ng Liebermann: Cholesterol cho mu xanh lc, mu ny rt bn trong nhiu gi, phn ng ny c dng xc nh cholesterol bnh vin. + Phn ng Salkowski: Cholesterol cho vnh mu . 2.1.3.2. Acid mt: Acid mt c tm thy trong ng vt c v gm 3 dng sau: cholic acid, deoxycholic v chenodeoxycholic acid. Acid mt l cht c i vi ngi. V vy trong mt, acid mt lin kt vi acetamin to thnh mt cht t c hn. Ngoi cholesterol v acid mt cn c cc sterol khc cng c ngun gc ng vt nh hormone nang thng thn, hormone tuyn sinh dc, cc sterol c ngun gc thc vt nh ergosterol, stigmasterol...

2.2. Lipid phc tp

Khc vi lipid t do c nhim v cung cp nng lng , hm lng lun thay i. Lipid phc tp c nhim v tham gia xy dng cc cu t ca t bo, hm lng khng thay i hay rt t thay i. 2.2.1. Glycerophospholipid (phosphatid) Chng ta c th hnh dung cu to chung ca glycerophospholipid nh sau:

Glycerophospholipid l diester ca phosphoric acid. Mt pha phosphoric acid lin kt vi glycerol, pha kia lin kt vi X. Ty cu to ca X ta c cc loi glycerophospholipid khc nhau:

42

Lecithin: Lc X l choline nn lecithin cn c gi l choline phosphatid

Lecithin c nhiu trong lng trng g, trong u nnh, trong mu, trong cc dy thn kinh. Qua cu to ta nhn thy n gm 2 phn - Phn phn cc bao gm phosphoric acid v base nitrogen a nc.

43 - Phn khng phn cc bao gm cc gc acid bo, ru glycerol ght nc. Do c cu to nh trn nn lecithin trong nc s to thnh dung dch gi l dung dch gi. Nh c tnh va a nc, va ght nc m hnh nh phospholipid tham gia trong vic bo m tnh thm mt chiu ca cc mng cu trc di t bo. Lecithin c th b thu phn bng acid, kim hay enzyme: * Thu phn bng acid: tt c lin kt ester u b ct t. * Thu phn bng kim: ta c acid bo dng mui, glycerophosphate v choline. Nhng choline b phn hy cho trimetylamin. Vi kim nh ch c th ct lin kt ester gia ru v acid bo. * Thu phn bng enzyme: c 4 loi enzyme lecithinase A, B, C v D tc ng ln cc lin kt ester khc nhau: B CH2O - CO - R1 A CHO - CO - R2 C OH CH2O - PO D Lecithinase A ct lin kt v tr ca lecithin cho acid bo v lisolecithin. Cephalin: Trong cu to ca cephalin X l colamine. CH2O-CO-R1 CHO-CO- R2 O CH2O-P = O O-CH2- CH2 -H3N+

44

Tng t lecithin, cephalin (X l ethanolamine) c cu tao gm hai phn a nc v ght nc, l thnh phn ca dy thn kinh v c nhiu trong no. Lisocephalin c to thnh khi ct lin kt ester v tr , cng c tnh cht ph hy hng cu nh lisolecithin. Serinphosphatid: Gi l serinphosphatid khi X l serine. Trong c th: lecithin, cephalin, serinphosphatid thng gp dng hn hp bi c s bin i tng h gia serine, choline v colamine. 2.2.2. Sphingophospholipid y l lipid phc tp, trong ru a nguyn t l sphingosine. Acid bo lin kt vi ru sphingosine bng lin kt peptid. Ty theo X ta c cc loi sphingophospholipid khc nhau
Acid bo

Sphingophospholipid quan trng nht l sphingomyelin, y X l: phosphocholine. Acid bo trong sphingomyelin l lignoceric, palmitic,

45 stearic hay nervonic. Sphingophospholipid l diaminophospholipid, khc vi phosphatid l monoaminophospholipid. Sphingophospholipid khng tan trong ethylic ether, da vo tnh cht ny tch chng ra khi hn hp lipid 2.2.3.Glycolipid Glycolipid l lipid phc tp khng cha phospho, trong thnh phn ca chng c cha hexose, thng l galactose hay cc dn xut ca galactose, i khi l glucose. Thuc nhm ny c MGDG, DGDG v sulfolipid kh ph bin trong lc lp v cc thnh phn khc ca t bo l.

6-Sulfo-6-deoxy--D-glucopyranosyldiacyglycerrol (sulfolipid)

2.2.4. Sphingolipid Cerebroside: trong phn t cerebroside ru sphingosine lin kt vi acid bo bng lin kt peptide, vi galactose (X) bng lin kt glucosidic.

46 Cc cerebroside khc nhau v thnh phn acid bo, c nhiu trong m thn kinh, hng cu, bch cu, tinh trng
Acid bo

Ganglyoside: cu to ging cerebroside nhng X l phc hp oligosaccharide

Phc hp ologosaccharide

TI LIU THAM KHO

Ti liu ting Vit
1. Phm Th Trn Chu, Trn thi ng. 1999. Ho sinh hc, NXB Gio dc, H Ni. 2. Qu Hai.2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 3. Trn Thanh Phong.2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 4. L Ngc T (ch bin), L Vn Ch, ng Th Thu, Phm Quc Thng Nguyn Th Thnh, Bi c Hi, Lu Dun, L Don Din, 2000. Ha sinh Cng nghip, Nxb KH&KT, H Ni.

Ti liu ting Anh

1. LehningerA.L. 2004. Principles of Biochemistry, 4th Edition. W.H Freeman. 2. Mead, Alfin-Slater, Howton & Popjk. 1986. Lipids: Chemistry, biochemistry and nutrion, Plenum, New York.

79

Chng 4

Nucleic acid
4.1. Thnh phn ho hc ca nucleic acid
Nucleic acid, vt cht mang thng tin di truyn ca cc h thng sng, l mt polymer c hnh thnh t cc monomer l nucleotide. Trong nucleic acid c cha cc nguyn t C, H, O, N v P. Hm lng P t 8- 10% Mi nucleotide gm 3 thnh phn kt hp vi nhau theo t l 1:1:1, bao gm: nhm phosphate, ng pentose (l ng 5 carbon) v mt base nit (nitrogen). 4.1.1 Base nit (Nitrogen) Cc base nit (nitrogen) thuc phn t nucleic acid u l dn xut ca base purine hoc pyrimidine. Cc base purine gm adenine (6-amino purine) v guanine (2-amino, 6-aminopurine), cc base nit pyrimidine gm thymine (2,6-dioxy, 5-methylpyrimidine), cytosine(2-oxy,6aminopyrimidine) v uracil (2,6 dioxypyrimidine).( Hnh 4.1.)

Hnh 4.1 Cng thc cu to ca cc base nit (nitrogen) trong nucleic acid

4.1.2 ng pentose
ng pentose trong nucleic acid gm c hai loi l ng deoxyribose v ribose. S c mt ca 2 loi ng ny l mt trong nhng c im phn bit DNA v RNA. 4.1.3 Phosphoric acid L mt acid v c - H3PO4

80

Hnh 4. 2. Cng thc cu to ca hai loi ng pentose trong nucleic acid

4.1.4. S to thnh nucleoside Nucleoside l sn phm thu phn khng hon ton ca nucleic acid. Nucleoside gm c hai thnh phn l ng pentose v mt base nit (nitrogen, thuc purine hay pyrimidine).

81

Hnh 4. 3. Cu to ho hc ca cc nucleoside v nucleotide

4.1.5. S to thnh nucleotide Nucleotide cng l sn phm thu phn khng hon ton ca nucleic acid. Nucleotide gm c ba thnh phn: ng pentose, mt base nit (nitrogen) v phosphoric acid (Hnh 4.3). 4.1.6. S to thnh nucleic acid Cc nucleotide c ni vi nhau bng lin kt phosphodiester Acide thng qua cc nhm OH v tr C3 v C5 ca ng pentose to thnh mt chui di gi l polynucleotide. Do lin kt phosphodiester c tao thnh v tr C3 v C5 nn chui polypeptide c tnh phn cc: u 5 thng c gc phosphate v u 3 thng c OH t do. Nucleic acid gm hai loi phn t c cu to rt ging nhau l DNA (desoxyribonucleic acid) v RNA (ribonucleic acid). 4.1.7. Mt s nucleotide quan trng khng tham gia cu to nucleic acid - ADP v ATP

82
ADP (adenosindiphosphate) v ATP (adenosintriphosphate) l nhng dn xut ca adenine, chng tham gia qu trnh phosphoryl ho-oxy ho. ATP c coi l ngun phosphate cao nng trong t bo (Hnh 4.4).

Hnh 4.4. Cu to ho hc ca ADP v ATP

- cAMP(AMP vng) Adenosinemonophosphate vng c hnh thnh t ATP, cAMP ch tm thy t bo ng vt v vi khun, n thng lin kt vi mng bo tng ca t bo v tham gia nhiu qu trnh chuyn ho. cAMP c th c sinh ra nh mt s hormone hot ho adenylcyclase (Hnh 4.5.) - UDP v UTP UDP (uridinediphosphate) v UTP (uridinetriphosphate) u l nhng dn xut ca uracil l nhng coenzyme quan trng trong cc phn ng trung gian chuyn ho glucose v galactose. Ngoi ra, chng cn tham gia trong vic hnh thnh nhng hp cht phosphate giu nng lng.

Hnh 4.5. Cu to ho hc ca AMP vng (cAMP)

- CDP v CTP CDP (cytidindiphosphate) v CTP (cytidinetriphosphate) l nhng dn xut ca cytidine. CTP cng l hp cht giu nng lng v co th tham gia nhiu phn ng khc nhau nh: phosphoryl ho ethanolamine

83
dn n s sinh tng hp cephaline hoc phn ng vi phosphate choline hnh thnh cytidinediphosphate-choline (CDP-Choline, hnh 4.6).

Hnh 4.6. Cu to ho hc ca CDP- cholin

4.1.8. Cc coenzyme nucleotide Hin nay ngi ta bit c mt s nucleotide tham gia cu to nn cc coenzyme quan trng nh vitamin B5 (pantothenic acid) trong coezyme A (SH-CoA), vitamin B2 (riboflavine) trong coenzyme flavin adenine inucleotide (FAD) v vitamin PP (nicotinamide) trong coenzyme nicotinamide adenine dinucleotide (NAD) v.v...Chng c phosphoryl ho khi lm chc phn nhm ngoi ca cc enzyme trong chuyn ho trung gian, (hnh 4.7)

84

FAD

Hnh 4.7. Cu to ho hc ca mt s coenzyme

4.2. Cu trc ca nucleic acid

4.2.1 DNA (Desoxyribonucleic acid) Phn t DNA l mt chui xon kp gm hai chui n. Mi chui n l mt chui nucleotide. Mi nucleotide gm ba thnh phn: nhm phosphate, ng desoxyribose v mt trong bn base v thng c k hiu bng ch ci u tin ca cc base (A-adenine, C- cytosine, Gguanine v T- thymine). Hai chui n kt hp vi nhau nh cc lin kt hydrogen hnh thnh gia cc base b sung nm trn hai chui: A b sung cho T v C b sung cho G. Mi chui n c mt trnh t nh hng vi mt u 5phosphate t do, u kia l 3 hydroxyl t do (quy c l 5 3. Hng ca hai chui n trong chui xon kp ngc nhau, nn c gi l hai chui i song. Nhng phn tch cu trc hin i cho thy

85
cu trc ca DNA khng phi lun lun tng ng vi dng c gi l B m Watson v Crick a ra. Do s tc ng ca cc hp cht c trng lng nh hoc protein dng B c th chuyn sang dng A (nn nhiu hn) hoc l dng Z (xon tri). Chng c th t gp li (DNA) hoc xon mnh, v d mt chui kp DNA c di l 20 cm c nn trong mt chromosome c kch thc l 5 m.
Lin kt hydrogen

1 nm

3,4 nm

0,34 nm (a) Cu trc ca DNA (b) Cu trc ha hc ca mt phn DNA

Hnh 4.8. Chui xon kp ca DNA

Phn t DNA trong nhim sc th ca sinh vt eukaryote dng thng, cn phn ln t bo prokaryote (vi khun) phn t DNA c dng vng. D dng no th cc phn t DNA u tn ti di dng cun cht. Trong t bo eukaryote, DNA kt hp cht ch vi cc protein l histone.

86
Trc ng-phosphate Cc base

Thymine (T)

Adenine (A)

Cytosine (C)

DNA nucleotide Phosphate ng (deoxyribose)

Guanine (G)

Hnh 4.9. Cu trc ca cc nucleotide in hnh.

DNA ca eukaryote c kch thc rt ln (v d DNA ngi c th di n 1 m) nn cu hi t ra l phn t ny phi c nn nh th no vo th tch rt hn ch ca nhn. Vic nn c thc hin nhiu mc , mc thp nht l nucleosome v mc cao nht l cu trc nhim o sc cht. Tht vy, ng knh ca chui xon DNA ch l 20 A , trong khi si nhim sc cht quan st di knh hin vi in t c ng knh o o 100 A , i khi t 300 A . iu ny chng t phn t DNA tham gia hnh o thnh nhng cu trc phc tp hn (Hnh 4.10). Si c ng knh 100 A l mt chui nhiu nucleosome. l nhng cu trc hnh thnh t mt

87
o

chui DNA qun quanh mt li gm 8 phn t histon. Si 100 A ny o c t chc thnh cu trc phc tp hn l si c ng knh 300 A . Trong nhn t bo, cc si va k trn kt hp cht ch vi nhiu protein khc nhau v c vi cc RNA to hnh nhim sc cht, mc t chc cao nht ca DNA
Chui xon kp DNA

2 nm

Dng xu chui ca nhim sc th

11 nm

Si nhim sc cht ng knh 30 nm gm cc nucleosome c ng gi

30 nm

Vng nhim sc th dng lng lo

300 nm

Vng nn cht ca nhim sc th trung k

700 nm

Nhim sc th trung k

1400 nm

Hnh 4.10. Cu trc nucleosome v nhim sc th.

Phn t DNA c sp xp trn nhim sc th lm cho chiu di ngn li hn 50.000 ln. Cc DNA eukaryote c c im khc vi DNA prokaryote. Ton b phn t DNA prokaryote u mang thng tin m ha cho cc protein trong khi DNA eukaryote bao gm nhng trnh t m ho (cc exon)

88
xen k vi nhng trnh t khng m ho (intron). Cc trnh t m ho eukaryote chm ngp trong mt khi ln DNA m cho n nay vn cha r tc dng. Ty theo mc hin din ca chng trong nhn, cc trnh t DNA c chia lm ba loi: - Cc trnh t lp li nhiu ln. V d: ng vt c v cc trnh t ny chim 10-15% genome (h gen). l nhng trnh t DNA ngn (10200 kb), khng m ho, thng tp trung nhng vng chuyn bit trn nhim sc th nh vng tm ng (trnh t CEN) hay u cc nhim sc th (trnh t TEL). Chc nng ca cc trnh t ny cha r, c th chng tham gia vo qu trnh di chuyn DNA trn thoi v sc (trnh t CEN) hoc vo qu trnh sao chp ton vn ca phn DNA nm u mt nhim sc th (trnh t TEL). - Cc trnh t c s ln lp li trung bnh. V d: genome ngi cc trnh t ny chim 25-40 %. Chng a dng hn v c kch thc ln hn (100-1.000 kb) cc trnh t lp li nhiu ln. Cc trnh t ny phn b trn ton b b gen. Chng c th l nhng trnh t khng m ha m cng c th l nhng trnh t m ha cho rRNA, tRNA v RNA 5S. - Cc trnh t duy nht: l cc gen m ha cho cc protein, c trnh t c trng cho tng gen. Mt c im ca phn t DNA c ngha rt quan trng c s dng vo phng php lai phn t. l kh nng bin tnh v hi tnh. Bin tnh l hin tng hai si n ca phn t DNA tch ri nhau khi cc lin kt hydrogen gia cc base b sung nm trn hai si b t do cc tc nhn ha hc (dung dch kim, formamide, urea) hay do tc nhn vt l (nhit). Sau , nu iu chnh nhit v nng mui thch hp, cc si n c th bt cp tr li theo nguyn tc b sung, hnh thnh phn t DNA ban u, l s hi tnh. 4.2.2 RNA (Ribonucleic acid ) Phn t RNA c cu to tng t DNA vi ba im khc bit sau: - Phn t RNA l chui n. - ng pentose ca phn t DNA l deoxyribose c thay bng ribose. - Thymine, mt trong bn loi base hnh thnh nn phn t DNA, c thay th bng uracil trong phn t RNA. Cu trc v chc nng ca RNA c s bin i r rt. V c bn RNA ch l cht mang thng tin di truyn virus, sau ngi ta chng minh rng n khng nhng ng vai tr c bn vic chuyn thng tin di truyn m cn c vai tr cu trc khi to nn phc h RNA-protein.

89
Theo mt l thuyt tin ha m i din l Manfred Eigen, RNA l cht mang thng tin di truyn, thnh vin trung gian ca s biu hin gen, thnh phn cu to v l cht xc tc. Nhm OH ru v tr th hai ca ribose cn thit cho a chc nng lm nhiu lon s to thnh chui kp, qua lm tng khng bn vng ca lin kt photphodieste. Trong t bo c ba loi RNA chnh, c cc chc nng khc nhau: - Cc RNA thng tin (mRNA) mRNA l bn sao ca nhng trnh t nht nh trn phn t DNA, c vai tr trung tm l chuyn thng tin m ha trn phn t DNA n b my gii m thnh phn t protein tng ng. Cc RNA c cu trc a dng, kch thc nh hn so vi DNA v ch cha thng tin m ha cho mt hoc vi protein v ch chim khong 2-5% tng s RNA trong t bo. Qu trnh chuyn thng tin c th hin nh sau: Phin m DNA RNA Dch m Protein

Escherichia coli, kch thc trung bnh ca mt phn t mRNA khong 1,2 kb. - RNA vn chuyn (tRNA) tRNA lm nhim v vn chuyn cc amino acid hot ha n ribosome tng hp protein t cc mRNA tng ng. C t nht mt loi tRNA cho mt loi amino acid. tRNA vn chuyn cha khong 75 nucleotide (c trng lng khong 25 kDa), l phn t RNA nh nht. Cc tRNA c cu trc dng c ba l (Hnh 7). Cu trc ny c n nh nh cc lin kt b sung hin din nhiu vng ca phn t tRNA. Hai v tr khng c lin kt b sung ng vai tr c bit quan trng i vi chc nng ca tRNA: - Trnh t anticodon gm ba nucleotide. - Trnh t CCA, c kh nng lin kt cng ha tr vi mt amino acid c trng. - RNA ribosome (rRNA) rRNA l thnh phn c bn ca ribosome, va ng vai tr xc tc v cu trc trong s tng hp protein. Ty theo h s lng rRNA c chia thnh nhiu loi: eukaryote c rRNA 28S, 18S, 5,8S v 5S, cn cc rRNA E. coli c ba loi: 23S,

90
16S v 5S. rRNA chim nhiu nht trong ba loi RNA (80% tng s RNA t bo), tip n l tRNA v mRNA ch chim 5%. Ribosome ca mi t bo u gm mt tiu n v nh v mt tiu n v ln. Mi tiu n v c mang nhiu protein v rRNA c kch thc khc nhau.
Amino acid

anticodon

.
Hnh 4.11. M hnh cu trc ca mt tRNA

- Mt s loi RNA khc. T bo sinh vt nhn chun cn cha mt s loi RNA khc, chng u c vai tr nht nh trong b my tng hp protein nh: + snRNA (small nuclear) l nhng phn t RNA nh tham gia vo vic ghp ni cc exon. + hn RNA( heterogenous nuclear) l nhng RNA khng ng nht nhn t bo. + scRNA (small cytoplasmic) l nhng RNA nh ca t bo cht.

91
Bng 4.1. Cc phn t RNA trong E. coli Loi rRNA Tng s tng i (%) 80 H s lng (S) 23 16 5 4 Khi lng phn t (kDa) 1,2 103 0,55 103 3,6 101 2,5 101 Khng ng nht S lng nucleotide 3700 1700 120 75

tRNA mRNA

15 5

Nhn chung tt c RNA trong t bo u c tng hp nh enzyme RNA polymerase. Enzyme ny i hi nhng thnh phn sau y: a) Mt khun mu, thng l DNA chui i. b) Tin cht hot ha: Bn ribonucleoside triphosphate: ATP, GTP, UTP v CTP. Sinh tng hp RNA ging DNA mt s im, th nht hng tng hp l 5 3, th hai l c ch ko di ging nhau: nhm 3-OH u cui ca chui tng hp l v tr gn kt ca nucleoside triphosphate tip theo. Th ba, s tng hp xy ra do thy phn pyrophosphate. Tuy nhin, khc vi DNA l RNA khng i hi mi (primer). Ngoi ra RNA polymerase khng c hot tnh nuclease sa cha khi cc nucleotide b gn nhm. C ba loi RNA trong t bo c tng hp trong E. coli nh mt loi RNA polymerase. ng vt c v cc RNA khc nhau c tng hp bng cc loi RNA polymerase khc nhau.

4.3. Mt s tnh cht ca nucleic acid

Dung dch nucleic acid c nht cao, c hot tnh quang hc (lm quay mt phng nh sang phn cc). Nucleic acid hp th mnh vng nh sng t ngoi c bc sng 250-280 nm, cc i hp th 260 nm. Tnh cht ny c s dng dnh lng nucleic acid v xc nh sch ca nucleic acid. Khi un nng dung dch nucleic acid nhit cao, thm acd hoc kim ion ho cc base ca n, nucleic acid b bin tnh. Phn t DNA xon kp b tho ri, hp th bc song 260 nm tng ln. S tng hp th ny gi l hin tng tng sc t (hyperchromism). Nhit lm mt mt na cu trc xon kp cu phn t DNA c gi l nhit

92
nng chy (melting temperature) vit tt l Tm. Cc DNA giu cc base G v C c nhit nng chy cao. DNA phn ng vi thuc th fucsin to thnh mu (phn ng Feulgen), phn ng ny thng s dng trong ho t bo. phn bit DNA v RNA ngi ta dng cc phn ng c trng vi thuc th orcine to thnh mu xanh lc bn, desoxyribose ca DNA phn ng vi diphenylamine to thnh mu xanh da tri bn.

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n, i Duy Ban, Nguyn Hu Chn, nh H, L c Trnh. 1980. Ho sinh hc. NXB Y hc, H Ni 2. Phm Th Trn Chu, Trn Th ng. 1999. Ho sinh hc. NXB Gio dc, H Ni. 3. H Hunh Thu Dng. 1998. Sinh hc phn t. NXB Gio dc, H Ni. 4. Phm Thnh H. 2001. Di truyn hc, (ti bn ln th 3) NXB Gio dc, H Ni.

Ti liu ting Anh

1. Lehninger A.L. , 2004. Principle of Biochemistry , 4th Ed., E-Book 2. Lodish H., 2003. Molecular Cell Biology. 5th Ed. E-book

92

Chng 5

Vitamin
Vitamin l mt nhm cht hu c c cc tnh cht l, ho hc rt khc nhau. Tc dng ca chng trn cc c th sinh vt cng rt khc nhau nhng u rt cn thit cho s sng ca sinh vt, nht l i vi ngi v ng vt. Khi thiu mt loi vitamin no s dn n nhng ri lon v hot ng sinh l bnh thng ca c th. Vitamin c tng hp ch yu thc vt v vi sinh vt. ngi v ng vt cng c th tng hp c mt s Vitamin nhng rt t nn khng tho mn nhu cu ca c th m phi tip nhn thm ngoi vo bng con ng thc n. C nhiu loi Vitamin khc nhau. Tn Vitamin c gi theo nhiu cch nh gi theo ch ci, gi theo danh php ho hc, gi theo chc nng. V d Vitamin B1 cn c tn ha hc l Thiamin, ng thi theo chc nng ca n cn c tn antinevrit. C nhiu kiu phn loi Vitamin, nhng kiu phn loi c s dng ph bin nht l da vo kh nng ho tan ca Vitamin vo cc dung mi. Ngi ta chia Vitamin ra 2 nhm: Vitamin tan trong nc v Vitamin tan trong m. Vitamin tan trong nc ch yu tham gia vo cc qu trnh lin quan ti s gii phng nng lng nh qu trnh oxi ho kh, qu trnh phn gii cc hp cht hu c... Vitamin tan trong m tham gia vo cc phn ng to nn cc cht c chc nng cu trc cc m, cc c quan.

5.1. Vitamin tan trong nc

5.1.1. Vitamin B1 (Thiamin) Vitamin B1 l loi Vitamin rt ph bin trong thin nhin, c bit trong nm men, cm go, mm la m ... Trong cm go c hm lng Vitamin B1cao nht. Vitamin B1 c tch ra dng tinh th vo nm 1912 v xc nh c cu trc ho hc ca n.

93

NH2 N H3C CH2 N CH3 CH2 - CH2OH S

Vitamin B1 bn trong mi trng acid, cn trong mi trng kim n rt d b phn hu khi un nng. Trong c th B1 c th tn ti trng thi t do hay dng Thiamin pyrophosphate. Thiamin pyrophosphate l dng B1 lin kt vi H3PO4 v c vai tr quan trng trong qu trnh trao i cht ca c th. Thiamin pyrophosphate l coenzyme xc tc cho qu trnh phn gii cc ceto acid nh pyruvic acid, oxaloacetic acid.... V vy khi thiu Vitamin B1 s chuyn ho cc ceto acid b ngng tr lm cho c th tch lu mt lng ln cc ceto acid lm ri lon trao i cht v gy nn cc trng thi bnh l nguy him. Vitamin B1 ho tan tt trong mi trng nc v chu nhit kh nn khng b phn hu khi nu nng. B1 c tng hp ch yu thc vt v mt s vi sinh vt. Ngi v ng vt khng tng hp c B1 m phi nhn t ngun thc n. Ngun cha nhiu Vitamin B1 l cm go, ng, la m, gan, thn, tim, no, nht l nm men. Khi thiu B1 c th pht sinh bnh beri-beri, cn gi l bnh t ph, do qu trnh trao i cht b ri lon. Nhu cu Vitamin B1 ph thuc vo iu kin ngh nghip, vo trng thi sinh l ca c th, vo la tui. Nhu cu hng ngy ca ngi ln l 1-3mg, ca tr em 0,5-2mg. 5.1.2. Vitamin B2 (Riboflavin) Vitamin B2 l dn xut ca vng Isoalloxazin, thuc nhm flavin. Trong c th B2 lin kt vi H3PO4 to nn coenzyme FMN v FAD l nhng coenzyme ca h enzyme dehydrogenase hiu kh. trng thi kh Vitamin B2 bn vi nhit v acid. Vitamin B2 c nhiu trong nm men, u, tht, sa, gan, trng. Khi thiu Vitamin B2 s tng hp cc enzyme oxi ho kh b ngng tr lm nh hng n qu trnh oxi ho kh to nng lng cho c th. ng thi khi thiu Vitamin B2 vic sn sinh ra cc t bo ca biu b rut cng

94 b nh hng gy nn s chy mu rut hay ri lon hot ng ca d dy, rut. Vitamin B2 cn gip c th khng khun tt hn. Nhu cu Vitamin B2 hng ngy ca mt ngi khong 2-3mg. CH2 - (CHOH)3-CH2OH

H3C H3C

N O Riboflavin (Vitamin B2)

NH

5.1.3. Vitamin PP (Nicotinic acid, nicotinamid) Vitamin PP l nicotinic acid v amid ca n l nicotinamid. O COOH N N Nicotinic acid Nicotinamid C NH2

Vitamin PP l thnh phn ca coenzyme NAD, NADP c trong cc enzyme thuc nhm dehydrogenase k kh. Vitamin PP gip c th chng li bnh pellagra (bnh da sn si). Khi mc bnh pellagra s dn n sng mng nhy d dy, rut, sau sng ngoi da. Vitamin PP dng nicotinic acid bn vi nhit, acid v c kim nn kh b phn hu, cn dng nicotinamid li km bn vi acid v kim.

95 Vitamin PP khng b bin i khi nu nng nn thc n gi c hm lng PP qua x l. Vitamin PP c nhiu trong gan, tht nc, tim, c bit l nm men. Nu c th thiu Vitamin PP s nh hng n cc qu trnh oxi ho kh. Vitamin PP c tc dng ngn nga bnh ngoi da, sng mng nhy rut, d dy. Hng ngy nhu cu ca mt ngi khong 15-25mg Vitamin PP. 5.1.4. Vitamin B6 (Pyridoxin) Vitamin B6 tn ti trong c th 3 dng khc nhau: Piridoxol, Pyridoxal, Pyridoxamine. Ba dng ny c th chuyn ho ln nhau
B

CH2OH HO H3C CH2OH

N Pyridoxol

Vitamin B6 l thnh phn coenzyme ca nhiu enzyme xc tc cho qu trnh chuyn ho amino acid, l thnh phn cu to ca phosphorylase... Vitamin B6 c nhiu trong nm men, trng, gan, ht ng cc, rau qu ... Nu thiu Vitamin B6 s dn n cc bnh ngoi da, bnh thn kinh nh au u, bnh rng tc, rng lng ... Hng ngy mi ngi ln cn 1,5-2,8 mg, vi tr em cn 0,5-2mg Vitamin B6. 5.1.5. Vitamin C (Ascorbic acid) Vitamin C l ascorbic acid. Trong c th Vitamin C tn ti 2 dng: dng kh l ascobic acid v dng oxy ho l dehydro ascobic.

96 CH2OH CHOH O O O O CH2OH CHOH O O

OH

OH

Ascorbic acid

Dehydro ascorbic acid

Vitamin C tham gia nhiu qu trnh sinh l quan trng trong c th: - Qu trnh hydroxyl ho do hydroxylase xc tc. - Duy tr cn bng gia cc dng ion Fe+2/Fe+3, Cu+1/Cu+2. - Vn chuyn H2 trong chui h hp ph. - Lm tng tnh khng ca c th i vi nhng iu kin khng thun li ca mi trng, cc c t ca bnh nhim trng, lm gim cc triu chng bnh l do tc dng ca phng x. Ngoi ra Vitamin C cn tham gia vo nhiu qu trnh khc c vai tr quan trng trong c th. Vitamin C c nhiu trong cc loi rau qu ti, nht l trong cc loi qu c mi nh cam, chanh, bi... Nhu cu hng ngy cn 7080mg/ngi. Nu thiu Vitamin C s dn n bnh hoi huyt, gim sc khng ca c th, b bnh chy mu rng, li hay ni quan (bnh scorbutus). 5.1.6. Vitamin B12 (Cyanocobalamin) Vitamin B12 c cu to phc tp, trong thnh phn c cha nhm CN, CO, amin. Thnh phn chnh ca Vitamin B12 l nhm porphyrin. Vitamin B12 gip cho vic to huyt cu t v hng cu. B12 tham gia cc qu trnh tng hp nucleotide nh xc tc cc phn ng metyl ho cc base Nit. Thiu B12 s gy bnh thiu mu c tnh. Ngoi cc loi Vitamin trn, trong nhm Vitamin tan trong nc cn mt s Vitamin khc nh Vitamin B5, Vitamin Bc, Vitamin H...
B B

97

5.2. Vitamin tan trong cht bo

5.2.1. Vitamin A (retinol) Vitamin A c 2 dng quan trng l A1 v A2. Vitamin A c hnh thnh t .caroten l tin Vitamin A. T .caroten to thnh 2 phn t Vitamin A.
CH3 CH3 CH3 CH3

CH2OH

CH3

Vitamin A1

Vitamin A c nhiu trong du c, lng trng. Trong thc vt c nhiu tin Vitamin A (.caroten) nht l trong c c rt, qu c chua, qu gc,... qu u . Vitamin A c vai tr quan trng trong c ch tip nhn nh sng ca mt, tham gia vo qu trnh trao i protein, lipid, saccharide. Thiu Vitamin A s b bnh qung g, kh mt, chm ln, st cn, gim kh nng khng ca c th i vi cc bnh nhim trng. Nhu cu Vitamin A hng ngy i vi ngi ln 1-2mg, tr em di 1 tui 0,5-1mg. 5.2.2. Vitamin D Trong c th tn ti nhiu loi Vitamin D, trong quan trng nht l dng D2 v D3. Cc Vitamin D l dn xut ca cc sterol. Trong c th Vitamin D c to ra t tin Vitamin D c sn di da nh nh sng mt tri c tia t ngoi. Thiu hoc tha Vitamin D u nh hng n nng photpho v canxi trong mu. Thiu Vitamin D tr em d b bnh ci xng, ngi ln b bnh long xng. Vitamin D c nhiu trong du c, m b, lng trng. Tin Vitamin D c sn trong m ng vt. Hng ngy mi ngi cn khong 10-20mg, tr em di 30 thng cn nhiu hn: 20-40mg.

98 5.2.3. Vitamin E (Tocopherol) Vitamin E c nhiu dng khc nhau. l cc dng , , , ... tocopherol. Cc dng khc nhau ny c phn bit bi s lng v v tr ca cc nhm metyl gn vo vng thm ca phn t. Trong cc loi Vitamin E, dng -tocopherol c hot tnh cao nht:
CH3 O H3C HO CH3 CH3 CH3 CH3 CH3

(CH2)3 - CH - (CH2)3 - CH - (CH2)3- CH - CH3

- Tocopherol

Vitamin E c nhiu cc loi rau xanh, nht l x lch, ht ng cc, du thc vt, gan b, lng trng, mm ht ho tho ... Vitamin E c tc dng nh cht chng oxi ho nn c tc dng bo v cc cht d b oxi ho trong t bo. Vitamin E cn c vai tr quan trng trong sinh sn. Nhu cu Vitamin E hng ngy khong 20mg cho mt ngi ln. 5.2.4. Vitamin K C nhiu loi Vitamin K, vi cng thc tng qut l O CH3 R O Vitamin K cn cho qu trnh sinh tng hp cc yu t lm ng mu (prothrombin) cho nn Vitamin K l Vitamin chng chy mu, thiu Vitamin K tc ng mu gim, mu kh ng.

99 Vitamin K c nhiu trong c linh lng, bp ci, rau m, c chua, u, ng cc, lng trng, tht b ... Thng ngi kho mnh, vi khun ng rut c kh nng cung cp Vitamin K cho nhu cu ca c th, ch cn b sung thm khong 0,2-0,3mg/ngy/ngi. 5.2.5. Vitamin F Vitamin F l cc acid bo khng no nh linoleic acid, linolenic acid, arachidonic acid ... Vitamin F c tc dng nui da, tiu m. Thiu Vitamin F ng vt chm ln, vim da, rng lng, hoi t ui. Vitamin F c nhiu trong cc loi du thc vt. 5.2.6. Vitamin Q (Ubiquinon) Vitamin Q ln u tin c tch ra t m ng vt vo nm 1955. Cu trc v chc nng ca Vitamin Q gn tng t nh Vitamin K v F. Vitamin Q tham gia vo cc qu trnh oxi ho-kh ca c th vi chc nng thnh vin ca chui vn chuyn in t ca ty th. Vitamin Q c trong nhiu i tng nh vi sinh vt, thc vt, ng vt .... 5.2.7. Vitamin P (Rutin) Vitamin P l nhm Vitamin c cu trc t flavon. Hin nay ngi ta pht hin khong 150 loi cht flavonoid khc nhau c tc dng nh Vitamin P, trong c 10 cht c nghin cu k. Thiu Vitamin P xy ra hin tng tng tnh thm ca mao qun, chy mu bt thng, mi mt, suy nhc c th. Vitamin P c tc dng lm gim tnh thm ca thnh mao qun. Vitamin P cn c th tham gia vo qu trnh oxi ho kh ca c th nh chc nng ca Vitamin C. Nhu cu Vitamin P khong bng 50% nhu cu Vitamin C.

100

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT, H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc, H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc, H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 Sinh ha hc hin i theo s . Nxb Y hc, H Ni.

Ti liu ting Anh

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.

101

Chng 6

Enzyme
Enzyme l protein c kh nng xc tc c hiu cho cc phn ng ha hc. Chng thc y mt phn ng xy ra m khng c mt trong sn phm cui cng. Enzyme c trong nhiu i tng sinh hc nh thc vt, ng vt v mi trng nui cy vi sinh vt.Hin nay ngi ta thu c nhiu loi ch phm enzyme khc nhau v s dng rng ri trong nhiu lnh vc nh y hc , nng nghip, cng nghip

6.1. Bn cht ha hc ca enzyme

Ngoi tr mt nhm nh RNA c tnh xc tc, tt c enzyme u l protein. Tnh cht xc tc ph thuc vo cu to ca protein. Nu mt enzyme b bin tnh hay phn tch thnh nhng tiu n v th hot tnh xc tc thng b mt i, tng t khi bn thn protein enzyme b phn ct thnh nhng amino acid. V vy cu trc bc 1, 2, 3, 4 ca protein enzyme l cn thit cho hot tnh xc tc ca chng. Enzyme, cng nh nhng protein khc, c trng lng phn t khong 12.000 n hn 1000.000.Mt s enzyme cu to gm ton nhng phn t L amino acid lin kt vi nhau to thnh, gi l enzyme mt thnh phn. a s enzyme l nhng protein phc tp gi l enzyme hai thnh phn. Phn khng phi protein gi l nhm ngoi hay coenzyme. Mt coenzyme khi kt hp vi cc apoenzyme khc nhau (phn protein) th xc tc cho qu trnh chuyn ha cc cht khc nhau nhng chng ging nhau v kiu phn ng. Mt s enzyme cn ion kim loi cho hot ng nh: Cu2+ Cytochrome oxidase 2+ 3+ Cytochrome oxidase, catalase, peroxidase Fe hoc Fe + K Pyruvate kinase 2+ Mg Hexokinase, glucose 6-phosphatase, pyruvate kinase Mn2+ Arginase, ribonucleotide reductase Mo Dinitrogenase Ni2+ Urease Se Glutathione peroxidase 2+ Zn Carbonic anhydrase , alcohol dehydrogenase, cc carboxypeptidase A v B

102 Mt s coenzyme v chc nng vn chuyn nhm tng ng ca chng nh sau: Biocytin Coenzyme A 5- Deoxyadenosylcobalamin (coenzyme B12) Flavin adenine dinucleotide Lipoate Nicotinamide adenine dinucleotide Pyridoxal phosphate Tetrahydrofolate Thiamine pyrophosphate CO2 Nhm Acyl Nguyn t H v nhm alkyl in t in t v nhm acyl Ion Hydride (:H-) Nhm Amino Nhm 1 Carbon Aldehyde

6.2. C ch tc dng
Nhng quan im hin nay nhm gii thch c ch tc dng ca enzyme u cho rng khi enzyme (E) tong tc vi c cht (S) s lm gim nng lng hot ha cc phn ng ha sinh. Mun lm gim nng lng hot ha cc phn ng enzyme cn tri qua nhiu giai on trung gian v to thnh phc cht nht nh gia E v S. Khi kt hp vi phn t enzyme, do kt qu ca s cc ha, s chuyn dch ca cc electron v s bin dng ca cc mi lin kt tham gia trc tip vo phn ng s lm thay i ng nng v th nng nn phn t c cht tr nn hot ng v d dng tham gia phn ng. Vic to thnh phc hp E-S giai on u xy ra rt nhanh v rt khng bn. Do sau mt thi gian di mi c chng minh bng thc nghim. Bng chng r rng nht v s tn ti ca phc hp E-S l thnh cng ca hai nh ha sinh Nht Bn K. Iaglu v T. Ozava l tch c phc E-S trong phn ng kh amin bng cch oxy ha (loi tr nhm amine) mt amino acid dy D do oxydase xc tc. Nhn chung ta c th hnh dung c ch tc dng ca enzyme ln c cht to sn phm bng phng trnh tng qut nh sau: E+S ES P+E Giai on 1: E kt hp vi S to thnh E-S. Giai on ny xy ra rt nhanh, nh cc lin kt khng bn nh lin kt hydro, tng tc tnh

103 din, tng tc Van der Waals Mi loi lin kt i hi nhng iu kin khc nhau v chu nh hng khc nhau khi c nc. Giai on 2: Sau khi to phc, c cht c nhng bin i nht nh v mt in t, cu hnh lm c cht tr nn hot ng hn, phn ng c d dng to thnh sn phm P. Trong nhiu phn ng do enzyme xc tc c 2 hay nhiu lai c cht, v d hexokinase xc tc phn ng: ATP + glucose hexokinase ADP + glucose 6 phosphate C ch enzyme xc tc cho phn ng 2 c cht c th nh sau: a/ C ch to phc 3 thnh phn S2

b/ C ch khng to phc 3 thnh phn

y l trng hp c cht th 2(S2) ch kt hp vo enzyme ( trng thi E) sau khi P1 c to thnh.

6.3. Trung tm hot ng (TTH) ca enzyme

T kt qu nghin cu v bn cht ho hc, v cu trc trung tm hot ng , c ch tc ng, v trung tm hot ng chng ta c th c mt s nhn xt chung v trung tm hot ng nh sau: - L b phn dng lin kt vi c cht. - Ch chim t l rt b so vi th tch ton b ca enzyme. - Gm cc nhm chc ca amino acid ngoi ra c th c c cc ion kim loi v cc nhm chc ca cc coenzyme.

104

i vi E mt thnh phn: TTH ch gm nhng nhm chc ca cc amino acid nh nhm hydroxy ca serin, carboxy ca glutamic, vng imidazol Cc nhm chc ca cc amino acid c th xa nhau trong chui polypeptide nhng nh cu trc khng gian nn n gn nhau v mt khng gian. i vi E hai thnh phn: TTH cng nh trn, cc nhm chc ca cc amino acid tham gia to thnh TTH lin kt vi nhau bng cc lin kt hydro. Ngoi ra trong TTH ca loi ny cn c s tham gia ca coenzyme v c th c ion kim loi. Theo Fisher TTH c cu trc c nh, khi kt hp vi c cht to phc E-S ta c th hnh dung ging nh cha kha v kha. Ngy nay ngi ta chng minh c rng: TTH ca enzyme ch c cu to hon chnh khi c s tng tc vi c cht (thuyt tip xc cm ng ca Koshland).

6.4. Tnh c hiu ca enzyme

Ngi ta chia tnh c hiu ra lm 3 kiu: + c hiu phn ng + c hiu c cht + c hiu khng gian a/ c hiu phn ng: l biu hin ca mt enzyme ch thng xuyn xc tc cho mt kiu phn ng nht nh, v d vn chuyn hydro t cht cho (ru bc nht hay ru bc hai) n cht nhn (NAD+ hay NADP+) hay chuyn nhm amin t mt amino acid n mt ceto acid. Cc phn ng loi th nht do dehydrogenase xc tc, cn phn ng loi th hai do aminotransferase xc tc.

105 b/ c hiu c cht: Tu mc ngi ta chia thnh: c hiu tng i v c hiu tuyt i + c hiu tuyt i: Enzyme ch tc dng ln mt c cht nht nh, mt v d c tnh cht kinh in v chuyn ho tuyt i l urease, enzyme ch phn gii ure: Hng trm th nghim trn cc dn xut ca ure u cho thy chng khng b phn gii di tc ng ca urease. Thc ra ngi ta pht hin kh nng phn gii c cht hydroxyure nhng vi tc b hn khong 120 ln. + c hiu nhm tuyt i: Cc enzyme ny ch tc dng ln nhng cht c cng mt kiu cu trc phn t, mt lin kt v c nhng yu cu xc nh i vi nhm nguyn t i vi nhm nguyn t gn lin kt chu tc dng. v d : maltase ch phn gii lin kt glucosidic c to thnh t glucoside ca glucose vi -OH ca monose khc. + c hiu nhm tng i: Cc enzyme khng c nhng yu cu i vi nhm chc gn lin kt chu tc dng. v d lipase thu phn lipid. c/ c hiu khng gian: Cc enzyme ch xc tc cho mt dng ng phn no nh dng L hay dng D, dng cis hay trans m thi.

6.5. Cc yu t nh hng n tc ca phn ng enzyme

6.5.1. nh hng ca nng enzyme Trong iu kin d tha c cht, ngha l [S] >>>[E] th tc phn ng ph thuc vo [S], v= K[E] c dng y = ax. Nh ngi ta o [E] bng cch o vn tc phn ng do enzyme xc tc. C nhiu trng hp trong mi trng c cha cht km hm hay hot ho th vn tc phn ng do enzyme xc tc khng ph thuc tuyn tnh vi [E] . v

[E]
Hnh 6.1: S ph thuc ca vn tc phn ng vo [E]

6.5.2 nh hng ca nng c cht [S] Ta kho st trng hp n gin nht : ch mt c cht.

106

E v S

(1) Gi v1 l vn tc ca phn ng to thnh phc cht ES. Gi v-1 l vn tc ca phn ng phn ly phc cht ES to thnh Gi v2 l vn tc ca phn ng to thnh E v P (sn phm). v1 = k1[E][S] v-1 = k-1[ES] v2 = k2[ES] Khi h thng t trng thi cn bng ta c: k-1[ES]+k2[ES] = k1[E][S] (k-1+k2)[ES] = k+1[E][S] (2) Gi E0 l nng ban u: [E0]=[E]+[ES]=>[E]=[E0]-[ES] (3) Thay tr s [E] t (3) vo (2) ta c: (k-1+k2)[ES] = k1([E0]-[ES]) [S] k1 [E0] [S] [ES] = -------------k-1+ k2+k1[S] Nu t Km= k-1+k2/ k1 (Km: gi l hng s Michaelis Menten). Ta c : [ES] = [E0][S]/ Km+[S] Mt khc vn tc phn ng to thnh sn phm P l: V = k2[ES] Thay [ES] bng gi tr trn ta thu c: k2[E0] [S] v = ----------------- (4) Km + [S]

107 Qua y ta thy nng enzyme cng cao th vn tc phn ng enzyme cng ln. Vn tc t cc i khi ton b enzyme lin kt vi c cht, ngha l: Vmax= k2[E0] Thay vo phng trnh (4) ta c: [S] v = Vmax ---------(5) Km+ [S] Phng trnh (5) gi l phng trnh Michaelis Menten. Km gi l hng s Michaelis Menten c trng cho mi enzyme Km c trng cho i lc ca enzyme vi c cht, Km c tr s cng nh th i lc ca enzyme vi c cht cng ln, ngha l vn tc ca phn ng do enzyme xc tc cng ln.

[S]
Hnh 6.2. Bin thin vn tc phn ng theo nng c cht.

Khi tng [S] th v phn ng tng, tng [S] n mt gi tr no th v t n gi tr vmax v s khng tng na nu ta vn tip tc tng [S]. Khi Km=[S] th v =1/2 Vmax Nm 1934. Lineweaver v Burk, trn c s ca phng trnh (5) nghch o bin thnh dng ng thng y=ax+b, n c ngha ln i vi vic nghin cu km hm enzyme.

108

1/v

1/Vmax -1/Km 1/[S]

Hnh 6.3: S ph thuc ca tc phn ng vo nng c cht theo Lineweaver-Burk

6.5.3. nh hng ca cht km hm (inhibitor) L cht c tc dng lm gim hot hay lm enzyme khng cn kh nng xc tc bin c cht thnh sn phm. Km hm enzyme c th thc hin bng nhiu cch khc nhau (thun nghch hay khng thun nghch). Thun nghch c: Cch 1: Km hm cnh tranh (competitive inhibition) Trong trng hp km hm cnh tranh l c cht v cht km hm u tc dung ln trung tm hot ng ca enzyme, Cht km hm chon ch ca c cht enzyme.

Hnh 6.4. Kiu km hm cnh tranh

Khi c cht d tha, nng cht km hm thp th c th loi b tc dng ca cht km hm, cn nng c cht thp v nng cht km hm cao th li c tc dng km hm hon ton.

1/v= (Km/Vmax) 1/S +1/Vmax

= 1+[I]/KI

109 1/v

[I]

1/Vmax

Khng c cht km hm

1/[S]
Hnh 6. 5. S ph thuc ca tc phn ng vo nng c cht theo Lineweaver-Burk khi c km hm cnh tranh

Ngi ta thy km hm nh vy phn ln xy ra gia cht km hm v c cht c s tng ng v mt ho hc. v d: malic acid c cu trc gn ging vi succinic acid nn km hm cnh tranh enzyme succinatedehydrogenase, l enzyme xc tc cho s bin i succinic acid thnh acid fumaric acid. Trng hp c bit ca km hm cnh tranh l km hm bng sn phm. Trng hp ny xy ra khi mt sn phm phn ng tc dng tr li enzyme v chon v tr hot ng phn t enzyme. ng thng c cht km hm th c xin ln hn v ct trc tung mt im l 1/Vmax Cch 2: Km hm phi cnh tranh (uncompetitive inhibition) c trng ca kiu km hm ny l cht km hm ch lin kt vi phc hp ES, m khng lin kt vi enzyme t do.

110

Hnh 6.6. Kiu km hm phi cnh tranh

1/v=(Km/Vmax)1/[S] + /Vmax
1/v [I]

-1/Km

khng c cht km hm

1/[S]
Hnh 6.7. S ph thuc ca tc phn ng vo nng c cht theo Lineweaver-Burk khi c km hm phi cnh tranh

111

Cch 3: Km hm hn tp( mixed inhibition )

Hnh 6.8. Kiu km hm hn tp

Trong , cht km hm khng nhng lin kt vi enzmye t do m cn lin kt vi c phc hp ES to thnh phc hp EIS khng to c sn phm P. Tng t nh trn ta c phng trnh :

1/v= (Km/Vmax)1/[S] +/vmax

1/v [I]

1/Vmax

khng c cht km hm

1/[S]
Hnh 6.9. S ph thuc ca tc phn ng vo nng c cht theo Lineweaver-Burk khi c km hm hn tp

112 Cc gi tr , c nh ngha nh trn. Trng hp = gi l km hm khng cnh tranh (noncompetitive inhibition). Mt trng hp km hm cn gp na l km hm enzyme bng nng cao ca c cht gi l km hm c cht nh km hm urease khi nng ure cao, ngoi ra cn c cc enzyme khc nh lactatdehydrogenase, carboxypeptidase, lipase, pyrophotphatase, photphofructokinase (i vi ATP). Nguyn nhn ca nhng hin tng ny cn cha c bit r. c th l: + Tn ti nhiu trung tm lin kt vi c cht bng cc i lc khc nhau. Khi nng c cht thp th enzyme c th ch lin kt vi mt phn t c cht, cn khi nng c cht cao n lin kt vi nhiu c cht dn n hnh thnh phc hp ES khng hot ng. + C cht cng c th c lin kt nh nhng v tr c bit ca enzyme. l mt nhm enzyme quan trng (enzyme d lp th) bn cnh trung tm xc tc cn c trung tm iu chnh. + C cht c th lin kt vi mt cht hot ho v bng cch ny n tch khi E. + C cht c th chon ch (ngn cn) mt cofactor ( ng yu t ) hay mt coenzyme. + C cht c th nh hng n ion lc ca mi trng v qua lm mt i tnh chuyn ho ca enzyme. 6.5.4. nh hng ca cht hot ha (activator) L cht lm tng kh nng xc tc nhm chuyn ha c cht thnh sn phm. Thng thng l nhng cation kim loi hay nhng hp cht hu c nh cc vitamin tan trong nc. V d: Mg++ hot ha cc enzyme m c cht c phosphoryl ha nh pyrophosphatase (c cht l pyrophosphate), adenosinetriphosphatase (c cht l ATP). Cc cation kim loi c th c tnh c hiu, tnh i khng v tc dng cn tu thuc vo nng . 6.5.5. nh hng cu nhit Ta c th tng vn tc ca mt phn ng ha hc bng cch tng nhit mi trng, hin tng ny tun theo quy lut Vant-Hoff. iu ny c ngha khi tng nhit ln 100C th tc phn ng tng ln khang 2 ln. i vi phn ng do enzyme xc tc cng c th p dng c quy lut ny nhng ch trong mt phm vi nht nh,v bn cht enzyme l protein.Khi ta tng nhit ln trn 40-500C xy ra qu trnh ph hu cht

113 xc tc. Sau nhit ti u tc phn ng do enzyme xc tc s gim. Nh tn ti nhit ti u ngi ta phn bit phn ng ho sinh vi cc phn ng v c thng thng. Mi enzyme c mt nhit ti u khc nhau, phn ln ph thuc ngun cung cp enzyme, thng thng trong khong t 40-600C , cng c enzyme c nhit ti u rt cao nh cc enzyme ca nhng chng a nhit. Cc chng vi sinh vt a nhit, c bit cc vi khun chu nhit c cha enzyme chu nhit cao. Hat

nhit
Hnh 6.10. nh hng ca nhit ln hat enzyme

6.5.6. nh hng ca pH S phn li khc nhau ca mt phn t protein cc gi tr pH khc nhau lm thay i tnh cht ca trung tm lin kt vi c cht v tnh cht hot ng ca phn t enzyme.iu ny dn n gi tr xc tc khc nhau ph thuc vo gi tr pH. Nh bit mi enzyme c mt pH ti u,mi enzyme c ng biu din nh hng pH ln vn tc phn ng do chng xc tc. ng biu din c dng nh hnh sau: Hat

Hnh 6.10. nh hng ca pH ln hat enzyme

114 nh hng ca gi tr pH n tc dng enzyme c th do cc c s sau: a/ Enzyme c s thay i khng thun nghch phm vi pH cc hp. b/ hai sn ca pH ti u c th xy ra s phn ly nhm prosthetic hay coenzyme. c/ Lm thay i mc ion ho hay phn ly c cht. d/ Lm thay i mc ion ho nhm chc nht nh trn phn t enzyme dn n lm thay i i lc lin kt ca enzyme vi c cht v thay i hot tnh cc i. Nh xc nh Vmax v Km ph thuc gi tr pH cho php nhn nh li bn cht ca cc nhm tham gia vo lin kt c cht v qu trnh t xc tc. 6.5.7. Cc yu t khc + nh sng: C nh hng khc nhau n tng loi enzyme, cc bc sng khc nhau c nh hng khc nhau, thng nh sng trng c tc ng mnh nht, nh sng c tc ng yu nht. nh sng vng t ngoi cng c th gy nn nhng bt li, enzyme trng thi dung dch bn hn khi c kt tinh dng tinh th, nng enzyme trong dung dch cng thp th cng km bn, tc ng ca tia t ngoi s tng ln khi nhit cao. V d di tc ng ca tia t ngoi nhit cao enzyme amylase nhanh chng mt hot tnh. + S chiu in: in chiu vi cng cng cao th tc ng ph hu cng mnh. Tc ng s mnh hn i vi dch enzyme c nng thp. C th do to thnh nhng gc t do, t tn cng vo phn ng enzyme. + Sng siu m: Tc ng rt khc nhau i vi tng loai enzyme, c enzyme b mt hot tnh, c enzyme li khng chu nh hng. Nhn xt chung: bn ph thuc vo trang thi tn ti ca enzyme, cng tinh khit th enzyme cng km bn, dch cng long th bn cng km, tc ng ca mt s ion kim loi trong dch vi nng khong 10-3M nh Ca++ lm tng tnh bn. Enzyme allosteric ( Enzyme d lp th, d khng gian) Cho n nay, ngi ta m t enzyme m hat tnh enzyme ph thuc nng c cht khng c dng hyperbol m c dng sigmoid l enzyme allosteric (Hnh 6.11 ):

115

Hnh 6.11. Bin thin vn tc phn ng theo nng c cht

i vi enzyme ny, khi nng c cht thp th tc phn ng tng chm, sau tip tc tng nng th tc nhanh chng t gi tr cc i. Nh ta bit, enzyme tun theo ng hc Michaelis-Menten th 1 hay nhiu c cht cng ch lin kt vo 1 v tr trn phn t enzyme, iu ny s dn n enzyme bo ha c cht. Cn enzyme c ng cong tc sigmoid ch xut hin khi enzyme l mt oligomer, nn c th lin kt vi nhiu phn t c cht. iu ny c ngha trn enzyme allosteric c nhiu trung tm lin kt, mi monomer c 1 trung tm lin kt. Ngi ta cho rng, trong trng hp ny c tnh hp tc gia cc v tr lin kt c cht trong phn t enzyme oligomer. Cc enzyme oligomer ny c Monod gi l allosteric enzyme d lp th (allosteric enzyme). ng cong tc sigmoid c th b cht iu ha (modulator) y v pha tri hay phi. Cht iu ha dng tc lm tng i lc ca enzyme allosteric vi c cht, ngc li l cht iu ha m. Cc cht iu ha c th lm nh hng khc nhau n cc thng s ng hc, lm thay i gi tr ring l mt trong hai gi tr Km hay Vmax.

Hnh 6.12 Minh ha khi c modulator

116

6.6. Cch gi tn v phn loi enzyme

Nh ta bit mi enzyme xc tc cho mi kiu phn ng ho hc duy nht (nh oxy ho mt kiu c cht nht nh, thu phn mt kiu lin kt nht nh,vn chuyn mt nhm cht nht nh t mt cht cho n mt cht nhn c a ch, trong c c vic bin i ch mt c cht duy nht), mt khc cn c mt kiu phn ng ho sinh nht nh c th c xc tc bng cc enzyme khc nhau. Da vo tnh c hiu phn ng ca enzyme, nm 1961 tiu ban enzyme hc quc t trnh by mt bo co, trong c ngh nhng nguyn tc nh tn v phn loi enzyme. Ngi ta chia enzyme ra lm 6 lp: 1. Oxydoreductase: cc enzyme xc tc cho cc phn ng oxi ho-kh. 2. Transferase: cc enzyme xc tc cho cc phn ng chuyn v. 3. Hydrolase: cc enzyme xc tc cho cc phn ng thy phn. 4. Lyase: cc enzyme xc tc cho cc phn ng phn ct khng cn nc. 5. Isomerase: cc enzyme xc tc cho cc phn ng ng phn ho. 6. Ligase (synthetase): cc enzyme xc tc cho cc phn ng tng hp c s dng lin kt giu nng lng ca ATP .v.v. Mi lp chia thnh nhiu t (di lp), mi t chia thnh nhiu nhm (siu lp). Tn enzyme thng c gi: Tn c cht c hiu - loi phn ng xc tc cng thm tip v ng ase. ng trc tn enzyme thng c 4 con s: s th nht ch lp, s th hai ch t, s th ba ch nhm, s th t ch s hng enzyme trong nhm. V d: (2.6.1.1) L.aspartate: -cetoglutarate aminotransferase. Enzyme ny xc tc cho phn ng chuyn nhm amine t L.aspartate n -cetoglutarate. L.aspartate +-cetoglutarate oxaloacetate + glutamate

6.7. Cc coenzyme quan trng

Nicotinamide adenine dinucleotide

117

Flavine adenine dinucleotide

Lipoic acid

Coenzyme A

Biotin

118

Thiamin diphosphate

(Nhm - amino protein, Schiff base)

Pyridoxal phosphate

Tetrahydrofolic acid

119

TI LIU THAM KHO

Ti liu ting Vit
1. Phm Th Trn Chu, Trn thi ng. 1999. Ho sinh hc, NXB Gio dc, H Ni. 2. Qu Hai. 2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 3. Trn Thanh Phong. 2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 4. L Ngc T (ch bin), L Vn Ch, ng Th Thu, Phm Quc Thng Nguyn Th Thnh, Bi c Hi, Lu Dun, L Don Din, 2000. Ha sinh Cng nghip, Nxb KH&KT, H Ni.

Ti liu ting Anh

1. Bergmeyer H. U. 1968. Methods of enzymatic analysis, translated from the third German edition, Acanamic, New York. 2. Copeland R. A. 2000. Enzymes, copyright by Wiley-VCH, Inc. 3. Lehninger A. L. 2004. Principles of Biochemistry, 4th Edition. W.H Freeman. 4. Mikkelsen S. R. 2004. Bioanalytical chemistry, copyright by John Wiley & Sons, Inc.

120

Chng 7

Hormone
7.1. C ch tc dng ca hormone
Hormone l nhng cht hu c c to thnh trong c th c tc dng iu ho cc hot ng sng trong c th. Lng hormone trong c th thng rt thp. Hormone c c thc vt v ng vt. ng vt hormone c sn xut ti cc tuyn ni tit v tc ng n cc m khc ni n c to ra. Hormone t tuyn ni tit c tit trc tip vo mu v c mu vn chuyn n cc m chu tc dng. Hormone c tnh c hiu. Hormone c tc dng iu ho cc qu trnh sinh l, ho sinh trong c th m khng tham gia trc tip vo cc phn ng ca c th. Hormone c tc ng n tc sinh tng hp protein, enzyme, nh hng n tc xc tc ca enzyme; thay i tnh thm ca mng t bo, qua iu ho hot ng sng xy ra trong t bo. Mt s hormone tc ng n c th thng qua cht trung gian AMP vng. AMP vng l cht truyn tin th 2, cn hormone l cht truyn tin th nht. Theo c ch ny tc dng ca hormone ln t bo ch xy ra qua nhiu giai on kh phc tp. - Trong mng nguyn sinh cht ca t bo c cha cht nhn hormone, cht ny s kt hp c hiu vi hormone. - S kt hp kch thch lm tng hot ca adenylatcyclase xc tc cho phn ng chuyn ho ATP thnh AMP vng. - Adenylatcyclase xc tc cho phn ng chuyn ho ATP thnh AMP vng. - AMP vng lm thay i vn tc ca cc qu trnh xy ra trong t bo lin quan n hot ng ca hormone. - Nh vy tc dng ca hormone theo c ch ny phi thng qua AMP vng m khng tc ng trc tip vo t bo. - Qu trnh hot ho adenylatcyclase bi phc hormone-cht nhn c thc hin qua cht trung gian l protein G. Phn t protein ny c kh nng kt hp vi GDP hay GTP. Dng phc protein G-GTP c tc dng hot ho adenylatcyclase, cn protein G-GDP khng c tc dng ny. Nh vy mun chuyn sang dng hot ng phi c s tham gia ca GTP, nu l protein G-GDP cn c s thay th GDP bng GTP nh phc

121 hormone-cht nhn xc tc. Dng thng tin c truyn t cht nhn hormone n protein G ri n adenylatcyclase. - Protein G khng ch c vai tr trung gian mang thng tin t cht nhn hormone n adenylatcyclase m cn c hot tnh ca GTPase, l kh nng thu phn GTP. Nh kh nng nn n xc tc cho qu trnh chuyn phc proteinG-GTP hot ng thnh dng proteinG-GDP khng hot ng do thu phn GTP trong phc proteinGTP thnh GDP to nn phc proteinG-GDP. Bng c ch protein G c vai tr quan trng trong qu trnh hot ho hay phn hot ho adenylatcyclase. Khi lng hormone gim adenylatcyclase tr thnh dng khng hot ng. - Nhiu hormone c c ch tc ng thng qua vai tr trung gian ca AMP vng. Nh vy AMP vng tham gia vo nhiu qu trnh khc nhau trong c ch tc ng ca hormone. l do AMP vng c tc dng hot ho proteinkinase l enzyme xc tc qu trnh photphoryl ho nhiu loi protein khc nhau. Thng cc protein enzyme dng phosphoryl ho l dng c hot tnh sinh hc. - Cc hormone tc dng theo c ch qua AMP vng, tn hiu c khuych i ln nhiu ln, do vy nng cc hormone trong mu rt thp, ch khong 10-10M, nhng ch cn hot ho c mt phn t adenylatcyclase c th to ra c nhiu phn t AMP vng nn nng AMP vng trong t bo ch cao hn nhiu lng hormone trong mu. Tc dng hot ho proteinkinase nh AMP vng li lm cho tn hiu c khuych tn tip tc v nhiu phn t protein c hot ho nh proteinkinase. iu gii thch c ti sao nng hormone trong mu rt thp m tc dng kch thch ca n li rt mnh. - Mt c ch tc ng th hai ca hormone l khng qua AMP vng.Insulin l hormone tc ng n t bo ch khng qua bc trung gian l lm tng lng AMP vng. Insulin lin kt cht ch vi cht nhn c hiu ca n trn mng nguyn sinh cht ca t bo ch. Tng tc gia Insulin v cht nhn bo m cho tc ng ca Insulin c th hin nhanh chng. Insulin cn c tc dng phosphoryl ho protein tham gia vo c ch kch thch qu trnh trao i glycogen. - C ch tc dng ca cc hormone thc vt hon ton khc hormone ng vt. Cc hormone thc vt tc ng ln hot tnh cc enzyme bng cch lin kt vi enzyme to phc hot ng. Khi lin kt vi hormone hot tnh ca enzyme c tng ln. - Hormone thc vt cn lm thay i tnh cht ca mng cellulose, mng nguyn sinh qua tc ng kch thch qu trnh sinh trng ca t bo.

122 - Mt c ch tc ng quan trng na ca hormone thc vt l thay i tnh cht ca nguyn sinh cht ca t bo, t nh hng n cc hot ng sinh l, trao i cht ca t bo

7.2. Cc hormone quan trng

7.2.1. Hormone ng vt - Hormone ng vt c nhiu loi vi cu to v chc nng rt khc nhau. Da vo cu to ho hc c th chia hormone ng vt thnh 3 nhm: - Hormone steroid l dn xut ca cholesterol. - Hormone l dn xut ca amino acid. - Hormone l peptide hay protein. 7.2.1.1. Hormone l steroid - y l nhm hormone c s lng ln, c vai tr quan trng v a dng. Ngi ta chia steroid thnh 5 nhm nh vi nhiu loi khc nhau:
T T 1 Nhm Progestagen i din Progesterol Ni to thnh -Th vng -V thng thn Vai tr Hormone dng thai gip trng pht trin - Kch thch tng glycogen v tch glycogen gan. - Kch thch phn protein, lipid. - Chng vim, nc mui. hp lu gii tch

Glucocorticoid 2

Cortisol

V thng thn

3 4

Mineral corticoid

Andosterol

V thng thn Tinh hon

- Tng hp th Na+, Cl- Tng tch nc. - Bi tit K+ Pht trin cc c im ca nam gii.

Androgen

Testosterol

Estrogen

Estron

- Pht trin cc c im n gii. Bung trng - Pht trin nim mc d con.

123 7.2.1.2. Hormone l dn xut amino acid n nay ngi ta bit mt s hormone l dn xut amino acid nh adrenaline, noradrenaline, thyroxine... - Adrenaline v noradrenaline l cc hormone do tuyn thng thn to ra. Cc hormone ny c tc dng kch thch s phn gii glycogen, lm gim s tng hp glycogen nn lm tng hm lng glucose trong mu. . OH

HO

CHOH - CH2 - NH - CH3 Adrenaline OH CHOH - CH2 - NH2 Noradrenaline

HO

Thiroxine l hormone do tuyn gip sn xut c tc dng tng cng qu trnh trao i cht, gip cho c th pht trin bnh thng. Nu thiu thyroxine gy nn trng thi thiu nng tuyn gip lm cho c th ln, km pht trin, n n. Ngc li nu tha thyroxine cng gy bnh l u nng tuyn gip lm cho ngi cao qu kh, khng cn i. I HO I O I Thyroxine (Tetraiodothyronine) I CH2 - CH NH2 COOH

124 y l nhm hormone c vai tr quan trng trong qu trnh iu ho trao i cht trong c th, c bit l iu ho lng ng trong mu. Mt s hormone l peptide:
STT 1 2 3 4 5 6 7 8 9 10 Hormone Tyrocalcitonin Insulin Glucagon Oxytoxin (HGF) Vasopressin (ADH) Melanotropin (MSH) Somatotropin (STH) Corticotropin (ACTH) Thyreotropin (TSH) Kch nang t (FSH) Ni to ra Tuyn ty Tuyn ty Tuyn yn Tuyn yn Tuyn yn Tuyn yn Tuyn yn Tuyn yn Tuyn yn Vai tr Gim lng ng trong mu Tng lng ng trong mu Gy co d con, kch thch Tng p, chng bi tit Kch thch tng sc t da Kch thch tng trng, tng TC Kch thch tuyn trn thn Kch thch tuyn gip Kch thch to estradiol

Tuyn gip Gim hm lng Ca++ trong mu

Sau y s cp n mt s hormone trong nhm ny: - Insulin: Insulin c tit t t bo beta ca o Langhan ca tuyn ty khi lng ng trong mu cao. Insulin kch thch cc qu trnh tng hp, km hm cc qu trnh phn gii glycogen gan, m m. Insulin cn kch thch s phn gii glucose. Nh insulin lm gim lng ng trong mu, do chng li bnh i tho ng. Insulin c khi lng phn t l 5800. Cu to insulin gm 2 chui polypeptide: chui A c 21 amino acid, chui B c 30 amino acid. Hai chui lin kt vi nhau bng 2 lin kt disunfit. Tin cht ca insulin l proinsulin v preproinsulin.T preproinsulin bin i thnh proinsulin, sau insulin c to nn t proinsulin. - Glucagon l hormone peptide, c tc dng ngc vi insulin. Khi lng ng trong mu gim qa mc cho php th tuyn tu sn sinh ra glucagon c tc dng lm tng lng ng trong mu nh km hm qu trnh tng hp glycogen. Glucagon c khi lng phn t 3.500, bao gm 29 gc amino acid to chui polypeptide mch thng.

125 7.2.2. Hormone thc vt Hormone thc vt l cc cht c vai tr quan trng trong qu trnh sinh trng, pht trin ca thc vt. C nhiu loi hormone khc nhau trong c th thc vt. Cc loi hormone ny khc nhau v bn cht ho hc, v vai tr i vi thc vt. C th chia hormone thc vt thnh 5 nhm: - Auxin. - Gibberellin. - Cytokinin. - Absisic acid. - Ethylen. 7.2.2.1. Auxin Auxin l nhm hormone quan trng, ph bin nht thc vt. C nhiu loi auxin khc nhau vi cu trc ho hc khc nhau. Loi auxin quan trng nht l -indol-acetic acid (IAA), ngoi ra mt s auxin khc cng kh ph bin l napthalen-acetic acid (NAA), phenyl-acetic acid (PAA) ...
CH2-COOH CH2-COOH

CH2-COOH
N H

IAA

NAA

PAA

Auxin c vai tr nhiu mt i vi thc vt: - Kch thch s sinh trng t bo, t kch thch s sinh trng cc c quan v ton c th. - C vai tr quyt nh hin tng u th nh. - C vai tr quyt nh cc c ng sinh trng nh hng sng, hng trng lc. - Kch thch qu trnh ny mm, rt ngn thi k ng ca ht, c. - c ch s rng l, kch thch s to qu.

126 - Kch thch cc hot ng sinh l, cc qu trnh trao i cht v nng lng ca c th. 7.2.2.2. Gibberellin Gibberellin l nhm hormone quan trng th hai thc vt. Gibberellin c cc nh khoa hc Nht pht hin ln u tin loi nm gy bnh la von (Gibberellin fujcoroi). C nhiu loi Gibberellin khc nhau, n nay tm thy hn 70 loi Gibberellin c mt thc vt, vi sinh vt. Ngi ta t tn cc Gibberellin theo th t thi gian pht hin GA1. GA2 .... GAn, trong quan trng nht c th k n l GA3. Cc Gibberellin u l dn xut ca vng gibban.

HO

C=O

OH COOH CH2

CH3

Cu to GA3 Gibberellin c vai tr quan trng trong qu trnh sinh trng, pht trin ca thc vt: - Kch thch s sinh trng ca t bo, qua kch thch s sinh trng ca cc c quan v c th. - Kch thch qu trnh ny mm, ph trng thi ng ca ht, c. - Kch thch s ra hoa ca cy ngy di. - Kch thch cc hot ng sinh l, cc qu trnh trao i cht v nng lng ca c th. 7.2.2.3. Cytokinin Cytokinin l cc dn xut ca base Adenine. C nhiu loi cytokinin khc nhau, quan trng nht l kinetin v zeatin.

127

HN - CH2

O
N

HN - CH2- CH = CH

CH2OH CH3

N N H N N N H

Kinetin

Zeatin

Xitokinin tham gia v nhiu hot ng sng quan trng ca thc vt: - Kch thch s phn bo qua kch thch s sinh trng ca t bo. - Lm chm qu trnh ho gi ca t bo, m. - Gip cho thc vt chng li cc stress ca mi trng c hiu qu. - L thnh phn cu to ca nucleic acid (trong mt s loi RNA) nn c vai tr trong qu trnh trao i nucleic acid v protein. - Kch thch cc hot ng sinh l, cc qu trnh trao i cht v nng lng ca c th. 7.2.2.4. Absisic acid Acid absisic (ABA) l nhm cht c ch sinh trng c tc dng ngc li 3 nhm cht trn. Absisic acid l dn xut ca triterpen. CH3 CH3
OH COOH O

CH3

CH3 ABA

128 Tc dng ch yu ca ABA l c ch qu trnh sinh trng ca t bo, gy hin tng rng l, rng qu. ABA ko di thi gian ng ca ht, c. Do c ch s sinh trng ca thc vt nn ABA phi hp vi nhm cht kch thch sinh trng iu ho qu trnh sinh trng ca thc vt xy ra cn i. 7.2.2.5. Ethylen Ethylen (CH2 = CH2) l nhm hormone thc vt c tc dng gn ging ABA nn thuc nhm cht c ch sinh trng. Etylen thc y qu trnh chn ca qu, qu trnh rng l. Khc vi hormone ng vt, hormone thc vt c tng hp trong cc phn khc nhau ca cy m khng c cc tuyn tit chuyn bit. Cc hormone thc vt c tng hp cc vng khc nhau ca cy. Auxin, gibberellin ch yu c tng hp ti cc phn non ca cy, nht l vng sinh trng nh nh sinh trng, tng tng Sau khi tng hp Auxin, gibberellin c vn chuyn trong cc m dn hay qua h thng t bo sng a n cc vng tc dng. Hormone thc vt cng khng c t bo ch chuyn bit nh ng vt m tc ng ln ton c th. Cytokinetin c tng hp mnh phn r non, cn absisic acid , ethylen l c tng hp nhiu cc phn gi ca cy. Sau khi tng hp cc hormone ny cng c vn chuyn n cc vng khc nhau trong c th thc hin cc chc nng ca chng.

129

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT. H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc. H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc. H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 2. Sinh ha hc hin i theo s . Nxb Y hc. H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.

130

Chng 8

Khi nim v s trao i cht v trao i nng lng

Trao i cht v trao i nng lng l bn cht ca hot ng sng ca mi c th sinh vt, l biu hin tn ti s sng. S trao i cht ca c th lun gn lin vi s trao i v chuyn ha nng lng. Chnh v vy, trao i cht v trao i nng lng l hai mt ca mt qu trnh lin quan cht ch vi nhau.

8.1. Khi nim chung v s trao i cht

C th sng tn ti, pht trin trong mi trng v khng ngng lin h mt thit vi mi trng . N hp th cc cht khc nhau t mi trng ngoi, lm bin i cc cht v mt mt to nn cc yu t cu to ca bn thn c th sng, mt khc li thi vo mi trng ngoi cc sn phm phn gii ca chnh c th cng nh cc sn phm hnh thnh trong qu trnh sng ca c th. Qu trnh thc hin c l do cc bin i ha hc lin tc xy ra trong c th. Ngi ta gi ton b cc bin i ha hc l s trao i cht. S trao i cht bao gm nhiu khu chuyn ha trung gian. Cc qu trnh ny xy ra phc tp trong tng m, tng t bo bao gm 2 qu trnh c bn l ng ha (tng hp) v d ha (phn gii) to nn chu k trao i cht lin tc gia cht nguyn sinh v cht nhn vo. Qu trnh ng ha l s hp th cc cht mi t mi trng bn ngoi, bin i chng thnh sinh cht ca mnh; bin i cc cht n gin thnh cht phc tp hn, s tch ly nng lng cao hn. y l qu trnh bin i cc cht khng c hiu (cc cht hu c ca thc n nh glucid, lipid, protein) t cc ngun khc nhau (thc vt, ng vt, vi sinh vt) thnh cc cht hu c khc (glucid, lipid, protein) c hiu ca c th. c im ca qu trnh ny l thu nng lng. Nng lng cn thit cung cp cho cc phn ng tng hp trn ch yu dng lin kt cao nng ca ATP. Qu trnh d ha l qu trnh ngc li ca qu trnh ng ha, l s bin i cc cht phc tp thnh cc cht n gin v gii phng nng lng cn thit cho hot ng sng. Nh vy y l qu trnh phn gii cc cht d tr, cc cht c trng ca c th thnh cc sn phm phn t nh khng c trng v cui cng thnh nhng cht thi (CO2, H2O,

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NH3...) thi ra mi trng. Nng lng c tch tr trong ATP v c s dng cho nhiu phn ng thu nng lng khc. Hai qu trnh ng ha v d ha xy ra lin tc lin quan vi nhau v khng tch ri nhau. Qu trnh ng ha l qu trnh i hi nng lng cho nn ng thi phi xy ra qu trnh d ha cung cp nng lng cho qu trnh ng ha. Do s trao i cht v trao i nng lng l hai mt ca mt vn . Ty theo kiu trao i cht, ngi ta chia sinh vt ra thnh hai nhm: nhm sinh vt t dng v nhm sinh vt d dng. Nhm sinh vt t dng bao gm tt c cc sinh vt t tng hp cht dinh dng cn thit cho chng. tn ti v pht trin, nhm ny ch cn H2O, CO2, mui v c v ngun nng lng. C hai hnh thc t dng. l hnh thc t dng quang hp v hnh thc t dng ha hp. Hnh thc u th hin cy xanh v vi khun ta, vi khun lu hunh vn dng quang nng tng hp cht hu c. Hnh thc sau c th hin mt s vi khun nhn nng lng trong qu trnh oxy ha cc cht v c. Nhm sinh vt d dng bao gm cc sinh vt khng c kh nng t tng hp cht dinh dng t cc cht v c m phi sng nh vo cc cht dinh dng ca nhm sinh vt t dng tng hp nn. Nh vy, qu trnh trao i cht ca th gii sinh vt lin quan cht ch vi nhau, to nn chu k trao i cht chung. nh sng Glucid, lipid, protein O2

Sinh vt t dng

Sinh vt d dng

CO2, H2O, mui cha Nitrogen Ngoi cch chia trn, cng theo kiu trao i cht, ngi ta chia sinh vt thnh hai nhm ln: nhm hiu kh (aerob) v nhm k kh (anaerob).

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Nhm hiu kh l kiu trao i cht m cc qu trnh oxy ha c s tham gia ca oxy kh quyn. Nhm k kh l kiu trao i cht m cc qu trnh oxy ha khng c s tham gia ca oxy kh quyn. a s cc sinh vt thuc nhm hiu kh. Nhm k kh ch l mt phn nh ca nhm sinh vt d dng bc thp. Tuy vy, gia cc c th hiu kh v k kh khng c ranh gii r rng. Sinh vt hiu kh biu hin r rng nht nh ngi chng hn cng c thc hin mt phn cc qu trnh trao i cht theo con ng k kh (v d nh m c) Qu trnh chuyn ha trong c th sng mang tnh thng nht v ring bit. Cc con ng chuyn ha ln trong mi c th ng vt, thc vt n bo, a bo u theo nhng giai on tng t nhau. Tuy vy, nu i su vo tng m, c quan, c th tng loi th li c nhng nt ring bit. Cc phn ng ha hc trong c th xy ra lin tc pH trung tnh, 370C, di tc dng xc tc ca enzyme. ng vt, cc qu trnh chuyn ha c iu khin bi h thng thn kinh

8.2. Khi nim chung v trao i nng lng v nng lng sinh hc
Trao i cht lun gn lin vi trao i nng lng. i vi c th ngi, ng vt v phn ln vi sinh vt th ngun nng lng duy nht l nng lng ha hc ca cc cht trong thc n. Trong c th, cc cht dinh dng ch yu v quan trng l glucid, lipid v protein u b oxy ha. Lipid v glucid i vo c th b t chy s sinh ra CO2, H2O v NH3, cht ny tc dng vi CO2 chuyn thnh carbamid (ure). Cc qu trnh oxy ha kh sinh hc thuc cc phn ng d ha c ngha rt quan trng. Chng khng nhng ch l ngun nng lng quan trng dng thc hin cc phn ng tng hp khc nhau m cn l ngun cung cp cc hp cht trung gian dng lm nguyn liu cho cc phn ng tng hp v ng vai tr ht sc quan trng trong vic lin hp cc qu trnh trao i cht. tn ti v pht trin, c th cn phi c cung cp lin tc nng lng. Trong hot ng sng ca mnh, c th bin i nng lng t dng ny sang dng khc v s bin i nng lng trong c th sng cng tun theo cc quy lut vt l nh s bin i nng lng gii v c.

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So snh v nng lng sinh hc v nng lng k thut ta thy c nhng c im sau: th nht, c th khng s dng nhit nng thnh cng c ch c; th hai, s gii phng nng lng trong c th l dn dn, tng bc; th ba, s gii phng nng lng i km theo s phosphoryl ha ngha l nng lng gii phng c c nh li lin kt este vi phosphoric acid trong phn t ATP vn c gi l lin kt cao nng. T dng nng lng trung gian ny (ATP) m c th d tr v s dng nng lng vo cc hot ng sng; th t, c th khng s dng c nng lng t do ca tt c cc loi phn ng pht nhit m ngun nng lng duy nht c th s dng l ca cc qu trnh oxy ha. 8.2.1. S bin i nng lng t do S thay i v i lng ca nng lng t do l mt ch tiu quan trng nht ca hiu ng nng lng tc l h s ca tc dng hu hiu ca phn ng. C th nh ngha nng lng t do l lng nng lng m mt nhit nht nh no c th bin thnh cng. T bo c th to ra v duy tr c cu trc trt t v phc tp ca mnh nh chng lin tc tip nhn nng lng t do t mi trng dng quang nng hoc ha nng v bin ha n thnh cc dng nng lng sinh hc phc v cho cc qu trnh hot ng sng. S bin ha, tch ly v s dng nng lng sinh hc xy ra song song vi s chuyn ha vt cht v tun th cc nguyn tc ca nhit ng hc. Nhng bin i nng lng t do ca h thng phn ng c k hiu bng G c gi tr l Kcal/mol. i lng ca G l hiu s gia lng nng lng t do ca trng thi cui (sau phn ng) G2 v nng lng t do ca trng thi u (trc phn ng) G1. Nu G<0 (c gi tr m), phn ng ta nhit, c th xy ra mt cch t pht. V d cc phn ng thy phn u thuc loi phn ng ny. Nu G = 0, h thng trng thi cn bng. Nu G>0 (c gi tr dng), phn ng thu nhit, mun thc hin phn ng cn phi cung cp nng lng. Cc phn ng thu nhit ch c th c thc hin cng vi cc phn ng ta nhit, ngha l vic tng nng lng t do ch c th c c do cc phn ng lin hp khc tin hnh vi vic gim nng lng t do. Cc qu trnh c bn gn lin vi hot ng sng ca c th, nhiu kiu lm vic ca t bo, cc phn ng tng hp u l nhng phn ng thu nhit lun lun lin hp vi cc phn ng ta nhit. G c tnh theo cng thc:

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G=

G 0 + RT lnK

trong G0 l s bin i nng lng t do tiu chun ca phn ng 250C khi nng ca tt c cc cht phn ng l 1 mol v p sut l 101,3 KPa (1atm), R l hng s kh, T l nhit tuyt i, K l hng s cn bng ca phn ng bng [C]c. [D]d /[A]a[B]b tc l nng ca cc cht tham gia phn ng A + B C + D; a, b, c, d l s lng phn t A, B, C, D tham gia phn ng. Trong h thng sinh hc, khi tnh gi tr G0 cn ch n pH, nng H+ l 1 mol, pH=0. Trng thi ion ha ca nhiu hp cht sinh hc b bin i khi pH thay i. V vy, thun tin cho vic tnh ton, xem trng thi chun ca pH l 7 v k hiu s thay i nng lng t do chun pH 7,0 l G0'. 8.2.2. Lin kt cao nng v vai tr ca ATP Cc lin kt ha hc gia cc nguyn t u l nhng tc nhn mang ch yu ca nng lng t do trong cc cht hu c. V vy, trong vic bin to ca cc lin kt ha hc trong phn t, mc nng lng t do ca hp cht s thay i. Xt v mt nng lng trong cc hp cht hu c c hai loi lin kt: Lin kt thng v lin kt cao nng (lin kt giu nng lng). Lin kt thng l lin kt m khi phn gii hoc to thnh n c s bin i nng lng vo khong 3 Kcal trn mt phn t gam (V d nh lin kt este); cn i vi lin kt cao nng s bin i ny ln hn nhiu t 7 12 kcal/mol. Trong cc hot ng sng ca c th sinh vt, cc qu trnh tng hp cc cht phn t ln t cc cht n gin, vn chuyn tch cc cc cht qua mng t bo, qu trnh vn ng v.v. lun i hi nng lng t do. Trong h thng sng cn c cc cht, cc h thng nhn nng lng t do t cc qu trnh ny chuyn n cho cc qu trnh khc. ATP l cht ph bin gi vai tr ny, l cht c vai tr trung tm trong trao i nng lng t bo v c th sng, l cht lin kt hoc c th ni l mt xch gia h thng s dng nng lng v h thng sn sinh ra nng lng. Trong phn t ATP c 3 gc phosphate, 1 gc kt hp vi gc ribose qua lin kt este, 2 lin kt gia 3 gc phosphate l lin kt anhydric. l cc lin kt cao nng c k hiu bng du ~ . ATP ( Adenosine Tri Phosphate) c biu th mt cch khi qut nh sau: Adenosine - P ~ P ~ P (trong P l cc gc phosphoric acid ). Khi ct t cc lin kt cao nng ny, s gii phng s nng lng ln gp hn 2 ln so vi lin kt este: ATP + H2O ADP + H3PO4 G0 = -7 Kcal/mol ( P )

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ATP + H2O

AMP + H4P2O7 ( P ~ P )

G0 = - 8,5 Kcal/mol

Nu tip tc thy phn lin kt este ca AMP to thnh adenosine v phosphate v c, nng lng t do c gii phng ca phn ng ny thp hn nhiu. S chuyn ha tng h gia ATP v ADP c vai tr c bit quan trng trong qu trnh trao i nng lng ca h thng sng. Trong a s trng hp thng thy phosphore hoc sulphure tham gia to thnh lin kt cao nng (Bng 8.1).
Bng 8.1. Mt s dng lin kt cao nng thng gp G0 ( Kcal/mol) -7 -7 - 10 1,3Diphosphoglyceric acid Aminoacyl-AMP Phosphoenol Pyruvic acid Creatin phosphate Arginin phosphate - 10,5 -7 - 12,8

Dng lin kt

Kiu lin kt - Anhydrid phosphate (pyro phosphate)

C trong cc cht ATP, GTP....... ADP, GDP.....

P
O~

~O ~P

- Acyl phosphate

O RCO~ P

- Enol phosphate RCO~ P CH2 - Amid phosphate N~ P ( phosphoguanidin) R C NH ~ P NH - Thioeste C~S O RC~SR

Acetyl coenzyme A Acyl coenzyme A

- 8,8

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8.3. Qu trnh oxy ha kh sinh hc

C th nh ngha qu trnh oxy ha kh l qu trnh trao i in t. S oxy ha l s tch mt hay nhiu in t, ngc li s kh oxy l s thu in t. Tt c cc cht tham gia vo qu trnh oxy ha kh c th sng u c kh nng nhng hoc thu in t. chnh l kh nng oxy ha kh. Song song vi s oxy ha c s kh oxy v in t c chuyn t cht b oxy ha sang cht b kh:
- 2e

V d: 2Fe

2+

+ Cl2

2Fe 3+ + 2Cl-

i lng c trng cho kh nng oxy ha kh ca mi cht gi l th nng oxy ha kh. C th tnh c th nng oxy ha kh theo cng thc sau: E'n= E'o + RT
nF

ln

[dng oxy ha] [ dng kh]

(1)

Trong : En l th nng oxy ha kh ca mt cht nht nh trong nhng iu kin nht nh. E0 l th nng oxy ha kh cc iu kin tiu chun ( nng ca hai dng bng nhau) R l hng s kh, T l nhit tuyt i, F l tr s Faraday do Bng 8.2 trnh by E0, hiu in th oxy ha kh Go ca mi h. E0 v nng lng t

Th nng oxy ha kh cn dng tnh nng lng t do (Go) c gii phng ra trong qa trnh oxy ha kh theo phng trnh: Go = -nF.E'o (2) (Cc k hiu c gii thch cng thc tnh th nng oxy ha kh v lin quan n bng 8.2 trn) * Tin trnh ca s oxy ha sinh hc: S phn gii cht dinh dng v gii phng nng lng ca t bo (s d ha) c th c chia thnh 3 giai on c bn: giai on u: cc hp cht cao phn t b thy phn thnh cc cht n gin c phn t nh hn: cc glucid (tinh bt, glucogen v.v...) thnh cc

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monosaccharid (glucose), cc protein thnh cc amino acid, cc lipid thnh cc acid bo. giai on th hai: bin nhng cht n gin thnh nhng cht 2 carbon l acetyl CoA (CH3 - COSCoA) (thiu). Acetyl CoA c coi l sn phm thoi ha ca cc cht glucid, lipid v protein. N c hnh thnh do s -oxy ha acid bo, do s oxy ha ca khong mt na s -amino acid cng nh do s oxy ha hiu kh glucose.
Bng 8.2. Th nng oxy ha tiu chun ca mt s h thng Eo (volt) pH7, 30oC -0,42 -0,32 -0,10 +0,04 +0,23 +0,26 +0,29 +0,55 +0,81 +0,22 +0,14 +0,19 +0,03 +0,03 +0,26 +0,26 +1,13 -10,1 -6,4 -8,7 -1,4 -1,4 -12,0 -12,0 -52,0 3 1 1 1 E0 (volt) Go (kcal/pH7, 30oC) Phosphoryl ha ADP ATP

H thng oxy ha kh

in cc hydro 2H+/ H2 NAD+/ NADH + H+ FAD/ FADH2 Cytochrome b Fe3+/ Fe2+ Cytochrome c1 Fe3+/ Fe2+ Cytochrome c Fe3+/ Fe2+ Cytochrome a Fe3+/ Fe2+ Cytochrome a3 Fe3+/ Fe2+ in cc oxy 1/2 O2 / O2-

giai on th ba: Acetyl CoA c hnh thnh giai on th hai s b oxy ha hon ton trong chu trnh Szent-Gyrgyi-Krebs (chu trnh citrat) hnh thnh CO2, H2O v gii phng nng lng. Phn ln nng lng c gii phng ra giai on th ba ny (khong 2/3) Trong giai on th hai v th ba khong 30-40% nng lng ha hc c bin thnh nhit, hn 60% nng lng ny c s dng tng hp cc hp cht cao nng.

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Trong chu trnh citrat, cc hydrogen tch ra s c oxy ha qua chui h hp to nn nng lng v H2O. Nng lng gii phng c tch tr cc phn t ATP. Ton b qu trnh c th c minh ha bng s trn hnh 8.3.
Thc n glucid lipid

protein Amino acid

glucose

Acid bo Acetyl CoA

CO2

Chu trnh citrat ATP 1/2O2

H2NADFADCoQCytbCytc1CytcCytaCyta3 2H+ ADP+P

Hnh 8.3. Tin trnh oxy ha sinh hc

O H2O

8.4. Chui h hp t bo v s phosphoryl ha oxy ha

8.4.1. Chui h hp t bo Chui h hp t bo l mt h thng cc enzyme xc tc vn chuyn H+ v eletron t c cht n phn t oxygen to H2O. Trong t bo, oxygen l cht oxy ha vn nng, cn cc phn t hu c khc nhau ng vai tr cht cho in t. y, in t v ion hydrogen ca phn t c cht khng chuyn trc

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tip cho oxygen khng kh m c chuyn dn qua mt chui phc tp nhiu mt xch, bao gm cc h enzyme oxy ha kh, c th nng oxy ha kh nm trong khong gia th nng oxy ha kh ca c cht v ca oxygen. Cc h enzyme ny c sp t theo mt trt t tng dn th nng oxy ha kh to thnh mt chui, gi l chui h hp hay chui vn chuyn in t ca t bo. Vai tr ca chui h hp l oxy ha tng bc hydrogen ca c cht n H2O. C ch hot ng ca chui h hp t bo c th tm lc nh sau: Cht cho nguyn t hydrogen l NADH + H+ hoc trong mt s trng hp l FADH2. Nguyn t hydrogen s c chuyn ti h coenzyme Q (CoQ) thng qua h trung gian flavoprotein cha st v lu hunh. Tip theo hai in t ca nguyn t hydrogen c tch ra v i vo h thng vn chuyn in t theo trnh t cc cytochrome b-c1-a-cytochromeoxydase (a3), cui cng in t c chuyn cho oxygen. Nguyn t oxygen b kh ( trng thi ion ha) s kt hp vi 2H+ (proton) to ra phn t nc. Qu trnh chuyn hydrogen v in t trong chui h hp c th phn thnh 4 giai on: - Giai on 1: Thng thng hydrogen c tch t c cht bi dehydrogenase c coenzyme NAD+(hoc NADP +). Hydrogen ca c cht gn vo NAD+, c cht t dng kh chuyn thnh dng oxy ha v NAD+ t dng oxy ha bin sang dng kh. Mi c cht c mt dehydrogenase c hiu tng ng: AH2 + NAD+ A + NADH +H+ (Trong AH2 v A l c cht dng kh v dng oxy ha) NADH khng th t oxy ha bi oxygen c, tc l khng th trc tip chuyn hydrogen cho oxygen m phi chuyn sang cho dehydrogenase khc c coenzyme l FMN hoc FAD. - Giai on 2: NADH (hoc NADPH) b oxy ha bi dehydrogenase. Enzyme ny l mt flavoprotein c coenzyme l FMN hoc FAD. Hai eletron c chuyn t NADH + H+ ti FMN (hoc FAD) cho FMNH2 (hoc FADH2): NADH + H+ + FMN NAD+ + FMNH2 NADH dehydrogenase cng cha st, cht ny c l gi vai tr vn chuyn eletron. st khng tham gia vo mt nhm hem no. NADH dehydrogenase l mt protein cha st khng thuc hem.

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- Giai on 3: H+ v eletron c chuyn t FMNH2 ti coenzyme Q l mt dn xut quinone, cn c gi l ubiquinon (UQ). Coenzyme Q l mt cht tc dng chuyn vn kh linh hot eletron gia flavoprotein v h thng cytochrome. Ubiquinon c th nhn 1 hoc 2e- v to ra semiquinone (UQH-) hoc ubiquinol (UQH2). c tnh ny cho php n lm cu ni vn chuyn e- t cht cho 2e- sang cht nhn 1e-. Ngoi ra, v phc UQ nh v k nc, nn n d dng di chuyn trong lp lipid i ca mng ty th lm con thoi vn chuyn e- gia cc phc vn chuyn e- cng knh khc trong mng ty th. - Giai on 4: Cc enzyme vn chuyn eletron t CoQH2 n oxygen. l h thng cytochrome, n gi vai tr trung tm trong h hp t bo. Mi cytochrome l mt protein enzyme vn chuyn electron c cha nhm ngoi hem. cc phn t cytochrome, nguyn t st lin tc i t trng thi st hai (Fe2+) - dng kh ti trng thi st ba (Fe3+) - dng oxy ha trong qu trnh chuyn vn eletron. Nhm hem chuyn vn mt eletron; ngc li vi NADH, flavin v coenzyme Q l nhng cht chuyn vn hai electron. C 5 cytochrome gia CoQ v O2 trong chui chuyn vn electron. Th nng Oxy ha kh ca chng tng theo th t: cytb, cytc1, cytc, cyta, cyta3. Cu trc v tnh cht ca cc Cytochrome ny khc nhau. Nhm ph ca Cytochrome b, c1, c l protoporphyrin c st, thng gi l hem. Cytochrome a v a3 l nhng thnh phn cui ca chui h hp t bo, chng dng mt phc cht gi l Cytochrome oxydase. Electron c chuyn ti phn Cytochrome a ca phc cht, ri ti Cytochrome a3 c cha ng (Cu+) dng kh trong qu trnh vn chuyn electron, c l n tham gia xc tc vn chuyn electron t hem A ca Cytochrome a3 ti oxygen. Qu trnh vn chuyn electron qua h thng Cytochrome c tm lc nh sau: 2e2 cytb Fe2+ 2 cytc1 Fe2+ 2 cytc Fe2+ 2 cyta Fe2+ 2 cyta3 Fe3+ + 2 cytb Fe3+ + 2 cytc1 Fe3+ + 2 cytc Fe3+ + 2 cyta Fe3+ + 2 cyta3 Fe3+ + 1/2 O2 2 cytb Fe2+ 2 cytb Fe3+ 2 cytc1 Fe3+ 2 cytc Fe3+ 2 cyta Fe3+ 2 cyta3 Fe3+ + 2 cytc1 Fe2+ + 2 cytc Fe2+ + 2 cyta Fe2+ + 2 cyta3 Fe2+ + 1/2 O2-

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Ton b chui h hp t bo t c cht dng kh AH2 ti oxygen phn t qua NAD, flavoprotein, coenzyme Q, h thng Cytochrome c trnh by hnh 8.4. AH2 NAD ATP FADH2 (FMNH2) CoQ 2Fe2+ ATP 2Fe3+ 2Fe2+ 2Fe3+ ATP 2Fe2+
1/2O2

A NADH + H+ ADP + P FAD (FMN) CoQH2 2cytb 2Fe3+ ADP + P 2cytc1 2Fe2+ 2cytc 2cyta 2Fe3+ 2Fe2+ ADP + P 2cyta3 2Fe3+ H2O
1/2O2-

2H+

Hnh 8.4. Chui h hp t bo

Kt qu ca chui h hp t bo thng thng l H2O, nhng vn c trng hp to thnh gc superoxyd (O2) v hydrogenperoxyd (H2O2). y l cc cht c i vi t bo v chng tn cng cc acid bo khng no cu to lipid mng t bo gy s bin cht ca cu trc mng. Theo cc s liu thc nghim th v tr to thnh O2 chnh l vng CoQ - cytochrome b do qu trnh t oxy ha ca cibi-semiquinone. Nh vy, thng xuyn c s r r 1 in t trong ty th v ty th s dng khong 1 - 2% s lng electron vn chuyn n cytochrome oxydase to thnh O2.

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Superoxyd dismutase cha Mn (Mn.SOD) c mt trong matrix ch chuyn c khong 80% O2 do s r r in t thnh H2O2. 20% O2 to thnh c chuyn vo cytoplasme, y superoxyd dismutase ca cytoplasme (SOD) cng hp tc vi cc h thng bo v khc s phn hy tip. C th biu th cc qu trnh trn nh sau: O2 + e- O2 (gc superoxyd ) 2O2 + 2H+ 2H2O2
C-ase SOD

H2O2 + O2

2H2O + O2

SOD v C-ase l cc enzyme chng oxy ha (antioxydant enzymes), bo v t bo chng li cc gc t do c hi. Nh vy, qu trnh vn chuyn hydrogen n oxygen to ra H2O, thc cht l mt qu trnh trao i electron (cho v nhn) mt cch lin tc. Bn cht ca n l mt qu trnh oxy ha kh. V vy, ngi ta gi h hp t bo l oxy ha kh sinh hc. Mt iu cn lu thm l: chui h hp t bo trnh by l chui h hp t bo bnh thng, nhng trong mt s trng hp, chui c th ko di hoc ngn hn ph thuc vo th nng oxy ha kh ca c cht. Quan nim hin i v h hp t bo cn b sung thm nhiu chi tit ca qu trnh h hp t bo kinh in nh trnh by. Nhng dng di chuyn in t v hydrogen cn ph thuc vo trng thi c cht n cc phc hp khc nhau. 8.4.2. S phosphoryl ha oxy ha Qu trnh tng hp ATP l qu trnh phosphoryl ha: ADP + H3PO4 ATP y l qu trnh cn nng lng. Nh chng ta bit, mi lin kt cao nng trong ATP cha nng lng t do l 7Kcal/mol nn tng hp c ATP t ADP theo phn ng trn cn cung cp nng lng tng ng 7Kcal/mol. Ngun nng lng cung cp cho qu trnh phosphoryl ha rt khc nhau. S phosphoryl ha quang ha l qu trnh tng hp ATP lc lp th nh nng lng nh sng xy ra trong quang hp. S phosphoryl ha oxy ha l qu trnh tng hp ATP ty th nh nng lng thi ra trong cc phn ng oxy ha kh. Theo quan nim hin nay, s phosphoryl ha oxy ha l qu trnh hnh thnh ATP bng cch chuyn electron v proton trong chui h hp t bo. S to thnh ATP trong chui h hp t bo c th hin hnh 8.4. Theo

143

phng trnh (2) cn c s chnh lch th nng oxy ha kh gia cc cht tham gia trong chui h hp t bo vo khong 0,152 volt to thnh mt phn t ATP
Eo =
'

G nF

7 = 0,152volt 2.23,06

Trong chui h hp c 3 im tng hp gia s h hp vi s phosphoryl ha: 1) gia NADH vi flavoprotein; 2) gia cytochrome b v c1; 3) gia cytochrome a v cytochrome oxydase (hnh 8.4.). iu c ngha l proton v electron oc chuyn t NADH + H+ ti oxygen to c 3 im phosphoryl ha, cn proton v electron c chuyn trong chui h hp t bo t FADH2 ch c 2 im phosphoryl ha. Mi tng quan P/O (t s P/O) l s phn t phosphate v c oc chuyn thnh dng hu c i vi s tiu th mt nguyn t oxygen. T s ny biu th s tng quan gia qu trnh phosphoryl ha v s oxy ha kh t bo, c gi l ch s . Nh vy c th ni rng s phosphoryl ha oxy ha qua h thng vn chuyn in t ca chui enzyme h hp l con ng ch yu i vi cc sinh vt hiu kh nhm khai thc nng lng ca cc hp cht hu c mt cch hu hiu nht phc v cho cc hot ng sng ca mnh.

144

TI LIU THAM KHO

Ti liu ting Vit
1. Nguyn Hu Chn, 1983. Enzyme v xc tc Sinh hc. Nxb Y hc, H Ni. 2. Nguyn Hu Chn, Nguyn Th H, Nguyn Nghim Lut, Hong Bch Ngc, V Th Phng, 2001. Ha sinh. Nxb Y hc, H Ni. 3. Phm Th Trn Chu, Trn Th ng, 2000. Ha sinh hc. Nxb Gio dc, H Ni. 4. L Don Din, 1975. Ha sinh thc vt. Nxb Nng nghip, H Ni 5. Nguyn Tin Thng, Nguyn nh Huyn, 1998. Gio trnh sinh ha hin i. Nxb Gio dc, H Ni 6. Nguyn Xun Thng, o Kim Chi, Phm Quang Tng, Nguyn Vn ng, 2004. Ha sinh hc. Nxb Y hc, H Ni. 7. L Ngc T, La Vn Ch, Phm Trn Chu, Nguyn Ln Dng, 1982. Enzyme vi sinh vt. Nxb KH&KT, H Ni. 8. L Ngc T (ch bin), L Vn Ch, ng Th Thu, Phm Quc Thng Nguyn Th Thnh, Bi c Hi, Lu Dun, L Don Din, 2000. Ha sinh Cng nghip, Nxb KH&KT, H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Fehr J. - Verekei A., 1985. Szabad Gyk Reakcik Jelntsge az orvostudomnyban. Medicina Knyv Kiad Budapest. 3. Karlson. P., 1972. Biokmia. Medicina Knyv Kiad Bydapest. 4. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman, 2004. 5. Stryer L., 1981. Biochemistry. W.H.Freeman and company. San Francisco.

145

Chng 9

S trao i saccharide
9.1. S phn gii saccharide
9.1.1. S phn gii polysaccharide v disaccharide Ngoi bin php dng acid phn gii th polysaccharide v disaccharide cn c th b phn gii bi s thy phn hay bi qu trnh phosphoryl- phn (phosphorolysis). S thy phn nh phn gii tinh bt thnh glucose, maltose hay dextrin ty thuc vo tnh cht ca enzyme: -amylase ch ct lin kt D-glucosidic-1,4 c kh nng ct khong gia, -amylase cng ch ct lin kt 1,4 nhng c kh nng ct bt u t u khng kh, -amylase c bit c tng hp t vi sinh vt c kh nng ct lin kt 1,4 v enzyme loi tr (kh) s phn nhnh (debranching enzyme, c hat tnh glucosidase) ct dy ni 1,6 trong amylopectin v glycogen. Cc polysaccharide b thy phn bi cac enzyme tng ng khc nh cellulose l cellulase, pectin l pectinase,... Vi cc disaccharide s b phn gii thnh cc monose nh cc enzyme tng ng nh sucrose bi sucrase to thnh glucose v fructose, maltose bi maltase to thnh 2 phn t glucose... Qu trnh phosphoryl- phn (phosphorolysis) l qu trnh to glucose-1-P nh enzyme phosphorylase (glycogen phosphorylase hay phosphorylase tinh bt) vi s hin din ca ion phosphate. Phosphorylphn khc vi s thy phn lin kt glucosidic l nng lng gii phng c dng cho s to lin kt ester trong glucose-1-P (Hnh 9.1.) Enzyme phosphorylase c coenzyme: Pyridoxal phosphate, nhm phosphate tn cng nh cht xc tc acid, tn cng lin kt glucosidic bng Pi . Phosphorylase tn cng vo u khng kh ca glycogen (hay amylopectin) n khi cch ch phn nhnh 4 n v glucose th ngng li. Chng s hat ng tr li sau khi enzyme loi tr (kh) s phn nhnh (debranching enzyme) thc hin chc nng transferase v glucosidase. (Hnh 9.2.) Cc disaccharide cng c th b phosphoryl-phn (phosphorolysis) bi enzyme tng ng to ra mt dn xut phosphate ca monose ng thi gii phng monose th hai. V d maltose phosphorylase chuyn ho maltose thnh glucose-1-P v glucose.

146

u khng c tnh kh

Chui glycogen (glucose)n u khng c tnh kh

Hnh 9.1. S phosphoryl-phn to glucose-1-phosphate

9.1.2. S oxy ho monosaccharide Di tc ng ca h thng nhiu enzyme khc nhau c trong ty th, cc monosaccharide b oxy ha to ra CO2, H2O, cc hp cht cao nng v cc sinh cht trung gian khc cn cho cc qu trnh ha sinh xy ra trong c th. Sn phm to thnh ph thuc vo iu kin mi trng:

147 u khng kh lin kt 1-6

Enzyme loi tr s phn nhnh

Enzyme loi tr s phn nhnh

Hnh 9.2: S phn gii glycogen bng glycogen phosphorylase

9.1.2.1. Qu trnh phn gii k kh( glycolysis) Qu trnh ny cn c gi l qu trnh Embden-Meyerhof-Parnas, y l qu trnh chuyn ha hexose thnh pyruvate trong iu kin khng c oxy, c th khi qut s chuyn ha qua hai giai an gm nhiu phn ng trn hnh 9.3. Phn ng 1: Glucose c phosphoryl ha C6 cho sn phm glucose-6-P, ngun phosphate l ATP.

148

Trong iu kin t bo y l phn ng mt chiu, c xc tc bi enzyme hexokinase. Kinase l tn chung c dng cho cc enzyme xc tc chuyn gc phosphate t ATP cho cc cht nhn, lp ph ca transferase . Hexokinase khng nhng xc tc s phosphoryl ha glucose m cn xc tc s phosphoryl ha cc hexose khc nh fructose, manose. Hexokinase, cng nh cc kinase khc cn Mg2+ cho hot tnh ca n v c cht tht ca enzyme khng phi l ATP4- m l ATP2Hexokinase ph bin tt c cc loi t bo. T bo gan trng thnh c cha hexokinase gi l hexokinase D hay glucokinase c hiu cho glucose, khc vi cc dng khc v ng hc v tnh cht iu ha. Phn ng 2: Chuyn ha glucose-6-P thnh fructose-6-P Enzyme phosphohexose isomerase xc tc s chuyn ha ng phn glucose-6-P thnh fructose-6-P, bin mt aldose thnh mt ketose.

149

Phn ng mi th 1

Phn ng mi th 2 S tch ca ng phosphate 6C thnh ng phosphate 3C

S oxy ha v s phosphoryl ha

Phn ng th 1 to ATP

Phn ng th 2 to ATP (s phosphoryl ha mc c cht

Hnh 9.3. Qu trnh ng phn (glycolysis)

150 Phn ng 3: Phosphoryl ha fructose-6-P thnh fructose1,6 biphosphate

Trong iu kin ca t bo phn ng do PFK-1 xc tc l phn ng mt chiu. vi sinh vt, sinh vt n bo(protista) v hu ht hay tt c thc vt u c phosphofructokinase dng P~P, khng dng ATP lm ngun cung cp phosphate to fructose1,6 biphosphate Mg2+ Fructose-6-P + PPi Fructose1,6 biphosphate + Pi

Phn ng 4: Phn ct Fructose 1,6 biphosphate Fructose1,6 biphosphate b phn ct thnh triose phosphate :3phosphate glyceraldehyde v dihydroxy acetonphosphate. Aldolase ca m ng vt c xng khng cn cation ha tr 2, nhng nhiu aldolase ca vi sinh vt cn Zn2+ cho hat ng ca chng.

151

Glycogen, tinh bt, disaccharide, hexose i vo pha chun b (preparatory phase) c th hin r hnh 9.4. Phn ng 5: Chuyn ha ni phn t triose phosphate Ch mt trong hai triose phosphate l aldose: 3-P glyceraldehyde tham gia tip vo qu trnh ng phn. Nhng dihydroxyaceton-P c th c chuyn ha thnh 3-P glyceraldehyde nh triose phosphate isomerase.

152

Glycogen; tinh bt

Hnh 9.4: Mi lin quan gia qu trnh ng phn v mt s saccharide

Phn ng 6: Oxy ha 3-P glyceraldehyde thnh 1,3 biphosphoglycerate Xc tc cho phn ng ny l enzyme 3-P glyceraldehyde dehydrogenase, c coenzyme NAD+, trong trung tm hot ng c nhm -SH C ch phn ng c nghin cu y :

153 Sau khi to phc hp E-S v NADH+H+, l phc khng bn nn khi c mt phosphate v c n s to thnh 1,3 biphosphoglycerate v gii phng enzyme trng thi t do.

Hnh 9.5: C ch tc ng ca glyceraldehyde 3 phosphate dehydrogenase

Phn ng 7: Trong phn ng ny gc phosphate cao nng ca 1,3 biphosphoglycerate chuyn cho ADP to ATP ( oxy ha phosphoryl ha mc c cht) v 3P glycerate

154

Phn ng 8: Chuyn ha 3P glycerate thnh 2P glycerate (chuyn gc P ni phn t) nh enzyme phosphoglycerate mutase cn Mg2+ cho hot ng ca n. y l phn ng thun nghch:

155 C ch:

Phn ng 9: 2P glycerate b loi nc to thnh phosphoenolpyruvate, l phn ng thun nghch c xc tc bi enzyme enolase.

Phn ng 10: Chuyn nhm phosphate t phosphoenolpyruvate n ADP, phn ng c xc tc bi pyruvat kinase, to ATP v pyruvate. Pyruvat kinase b km hm bi ATP, khi nng ATP cao th n gy km hm d khng gian. ng vt c xng sng pyruvat kinase c t nht 3 isozyme, hi khc nhau trong phn b cc m v trong vic p ng i vi nhng cht iu ha (modulator) .

156

T pyruvate, tu thuc mi c th, iu kin mi trng c th chuyn ha thnh cc sn phm khc nhau (k kh) 2 Pyruvate (k kh) 2 Lactate (ln men lactate)

2 Ethanol + 2CO2 (hiu kh) (ln men ru nm men ) 2 Acetyl-CoA

Chu trnh citric acid

4CO2 + 4H2O (ng vt, thc vt v nhiu t bo vi sinh vt trong iu kin hiu kh). T pyruvate c th c 3 kh nng phn gii nh trn, ngoi ra n cn l ngun tng hp mt s cht khc m ta khng cp y. Trong iu kin k kh, pyruvate c th ln men to lactic acid: Di tc dng ca lactate dehydrogenase, pyruvate b kh thnh lactic acid. Phn ng ny xy ra trong m c ng vt s to thnh L-lactic acid,

157 cn trong qu trnh ln men do vi sinh vt gy ra (ln men sa chua, mui da, c ) s to thnh D-lactic acid.

Ln men ru: Nm men v mt s vi khun khc c th chuyn ha pyruvate thnh ethanol v CO2. Qu trnh tri qua 2 bc

Trong bc 1, pyruvate b kh cacboxyl-ha vn c xc tc bi enzyme pyruvate decarboxylase, enzyme ny cn Mg2+ v c coenzyme l TPP. Bc 2, acetaldehyde b kh thnh ethanol vi NADH+H+ c to ra t s oxy ha kh 3 P glyceraldehyde. 9.1.2.2. Qu trnh phn gii ho kh glucose. Chu trnh Krebs C th chia qu trnh ny ra lm 4 giai on chnh: - Phn gii glucose thnh pyruvate (xem qu trnh ng phn). - Chuyn ha pyruvate thnh acetyl- CoA. - Oxy ha acetyl- CoA thng qua chu trnh Krebs (chu trnh citric acid). - Oxy ha cc coenzyme kh qua chui h hp(xem phn khi nim v s trao i cht). - Chuyn ha pyruvate thnh acetyl-CoA(hiu kh)

158

- Oxy ha acetyl-CoA qua chu trnh Krebs: Do trong chu trnh c mt cc sn phm trung gian l cc di- v tricarboxylic nn chu trnh Krebs cn c tn l chu trnh tricarboxylic, hay chu trnh citric acid. Chu trnh Krebs bao gm 8 phn ng sau (Hnh 9.6). Phn ng 1: L phn ng trng hp acetyl-CoA v oxaloacetate to thnh citrate. Nng lng cn cho s trng hp do s phn gii lin kt cao nng trong acetyl-CoA cung cp.

Phn ng 2: Citrate b bin i thnh isocitrate, l qu trnh thun nghch c xc tc bi enzyme aconitase. Cis-aconitate thng khng tch khi enzyme, t bo thng to isocitrate v isocitrate s c chuyn ha tip theo trong chu trnh, d cn bng pH= 7,4, nhit 25oC ch c it hn 10% isocitrate. Isocitrate c nhm H-C-OH, m ch 2 nguyn t hydro v tr ny mi d dng tch khi c cht kt hp vi coenzyme NAD+ hoc NADP+.

159

Phn ng 3:

Kt qu ca s oxy ha di tc dng xc tc ca enzyme isocitrate dehydrogenase l 2 nguyn t hydro c chuyn cho NAD(P)+ v 1 nguyn t C c tch ra khi c cht di dng CO2. Phn ng 4: Sn phm ketoglutarate va b oxy ha va b kh carboyl ha di tc dng xc tc ca phc enzyme -ketoglutarate dehydrogenase. Ging nh phn ng 3, NADH+H+, CO2 v succinyl CoA c to thnh.

Phc -ketoglutarate dehygrogenase

160 Phn ng 5:

Nng lng trong lin kt cao nng ca succinyl CoA c dng to ATP thng qua GTP. y l chng phn ng duy nht ca chu trnh Krebs xy ra s tch ly nng lng trong ATP. Phn ng 6:

y c s km hm cnh tranh enzyme gia succinate v malonate. Coenzyme kh FADH2 qua chui h hp to ATP. Phn ng 7: L phn ng hydrate ha fumarate to malate di tc dng ca enzyme fumarase.

Trng thi chuyn tip Carbanion

Fumarase c tnh c hiu rt cao, xc tc s hydrate ha ni i ca fumarate (dng trans) m khng tc ng ln maleate( ng phn dng cis ca fumarate).

161 Phn ng 8: Malate to ra phn ng 7 s tip tc b oxy ha cho ra oxaloacetate, enzyme xc tc cho phn ng ny l malate dehydrogenase. Nh vy 1 vng chu trnh khp kn, oxaloacetate c to ra y khc vi oxaloacetate m u ca phn ng 1 v thnh phn carbon, oxaloacetate mi c b sung 2 carbon t acetyl-CoA. Oxaloacetate m u ca phn ng 1 c 2 carbon tham gia to CO2 phn ng 3 v 4.

ngha ca qu trnh ng phn v chu trnh Krebs 1/ L cc ng hng phn gii to ra cc sn phm trung gian to thnh cc c cht khc nhau cn cho s sng. 2/ To cc coenzyme kh v ATP. Vic to ra nng lng, s dng nng lng v coenzyme kh qua qu trnh ng phn (glycolyis) v chu trnh Krebs c tm tt nh sau:
Glucoseglucose 6-phosphate Fructose 6-phosphate fructose 1,6-bisphosphate 2 Glyceraldehyde 3-phosphate 2 1,3-bisphosphoglycerate 2 1,3-Bisphosphoglycerate 2 3-phosphoglycerate 2 Phosphoenolpyruvate 2 pyruvate 2 Pyruvate 2acety-CoA 2 Isocitrate 2 -ketoglutarate 2 -Ketoglutarate 2 succinyl-CoA 2 Succinyl-CoA 2 succinate (hoc 2 GTP) 2 Succinate 2 fumarate 2 Malate 2 oxaloacetate -1 ATP -1 ATP 2 NADH 2 ATP 2 ATP 2 NADH 2 NADH 2 NADH 2 ATP 2 FADH2 2 NADH

162

Chu trnh citric acid

Hnh 9.6. S tng qut ca chu trnh citric acid

thc vt v mt s vi khun cn c ng hng khc trong vic chuyn ha acetyl-CoA. Cc phn ng ca s chuyn ha ny to nn chu trnh gi l chu trnh glyoxylate. Gia chu trnh ny v chu trnh Krebs c nhng giai an ging nhau (hnh 9.7).

163

Chu trnh Glyoxylate

Hnh 9.7. Tm tt chu trnh glyoxylate

9.1.2.3. Chu trnh pentose phosphate L s phn gii trc tip glucose 6 Phosphate khng qua qu trnh ng phn, gm 2 giai on oxy ha v ti to hexose phosphate. Pentose phosphate (hexose monophosphate) gy ra s oxy ha v s kh carboxyl ha C1 ca glucose 6 Phosphate, kh NADP+ thnh NADPH v pentose phosphate. NADPH cn cho cc phn ng sinh tng hp v pentose phosphate cn cho s tng hp nucleotid v nucleic acid. Pha th nht ca pentose phosphate l qa trnh oxy ha glucose 6 Phosphate to ribulose 5 phosphate v kh NADP+ thnh NADPH. Pha th hai (nonoxidative) chuyn ha pentose phosphate thnh glucose 6 Phosphate v bt u chu trnh tr li.

164

Hnh 9.8. S pha th nht ca chu trnh pentose phosphate

Trong pha th hai, cc phn ng c xc tc bi transaldolase v transketolase.

165

Hnh 9.9. S pha th hai ca chu trnh pentose phosphate

166

9.2. S tng hp saccharide

9.2.1. S tng hp saccharide n gin. Qu trnh quang hp Cy xanh c th hp th CO2, kh n thnh saccharide. y l qu trnh cn c s tham gia ca nh sng v dip lc . Ta c th khi qut qu trnh quang hp bng phn ng sau: nh sng 6 CO2 + 6 H2O Chlorophyll Qu trnh quang hp gm hai giai on v c chc nng ring: Giai on 1: Xy ra qu trnh quang phn ly nc ng thi gii phng oxy phn t: 2H2O nh sng Cng chlorophill v h thng chuyn in t, ATP s c tng hp t ADP v H3PO4. V vy ngi ta cn gi qu trnh ny l s phosphoryl ha quang hp hay quang phosphoryl ha. Theo Arnon, hnh nh in t b tch ra khi clorophyll a khi c kch hat bi photon n s i theo hai con ng khc nhau: Con ng quang phosphoryl ha vng xy ra h quang ha I, in t xut pht t P700 qua h thng chuyn in t ri tr li P700. Con ng ny ch cho php tng hp ATP. Con ng quang phosphoryl ha khng vng xy ra khi c s tham gia ca h quang ho I v II, Con ng ny cho php tng hp ATP v NADPH2. Khi mt in t, chlorophyll ca h I tip tc nhn in t h II qua cc khu chuyn trung gian . in t ca phn t sc t h II c b sung t H2O. Nh vy con ng i ca in t trong qu trnh ny khng khp kn v c gi l qu trnh quang phosphoryl ha khng vng (hnh9.10). Giai on 2: kh CO2 thnh saccharide nh NADPH v ATP c tng hp giai on 1. Ty theo c ch, ngi ta phn bit: 4 H+ +O2 C6H12O6 + 6 O2

167

H quang ha I

H quang ha II

nh sng Phc cyt b

nh sng Phc Mn tch H2O

Hnh 9.10 : S tm tt qu trnh vn chuyn in t quang phosphoryl ho khng vng

Chu trnh Calvin ( chu trnh C3): Chu trnh c nh CO2 ny do M. Calvin v cng s tm ra nm 1951 v c gi l chu trnh Calvin hay chu trnh C3. u tin phn t CO2 kt hp vi ribulose 1,5 biphosphate to 2 phn t 3-phosphoglycerate. Hai phn t 3-phosphoglycerate c

168 phosphoryl ha nh enzyme 3-phosphoglycerate kinase xc tc to thnh 1,3-biphosphoglycerat. Cht ny b kh di tc dng ca glyceraldehyde 3-phosphate dehydrogenase chuyn thnh glyceraldehyde 3-phosphate (hnh 9.11, 9.12).

Hnh 9.11. S kt hp CO2 vo ribulose1,5-biphosphate

169

Tinh bt

Hnh 9.12 : S tng hp ng v tinh bt t bo thc vt

Nh tc dng ng phn ha ca triose phosphate isomerase: glyceraldehyde 3-phosphate to thnh dihydroxyacetone phosphate. Sau c s kt hp gia 2 phn t glyceraldehyde 3- phosphate v dihydroxyacetone phosphate bng phn ng chuyn aldose to thnh fructose 1,6 bi phosphate. Fructose 1, 6 biphosphate mt di 1 phosphate to thnh fructose 6 phosphate, n chnh l nguyn liu to thnh cc hexose khc nh glucose 6 phosphate. T cc dng ng ny tng hp cc oligosaccharide v polysaccharide khc (hnh 9.12). Mt phn fructose 6 phosphate chuyn ha thnh ribulose 1,5biphosphate, ng thi khp kn chu trnh (hnh 9.13).

170

Hnh 9.13: S to thnh phn t khi u chu trnh Calvin - ribulose 1,5-biphosphate Ghi ch: Cc enzyme xc tc cho chui phn ng: (1) Transaldolase 2)Fructose 1,6 -biphosphatase , 3)Transketolase , 4)Transaldolase , 5)Sepdoheptulose 1,7-biphosphatase , 6)Transketolase , 7)Ribose 5-phosphate isomerase , 8)Ribulose 5-phosphate epimerase , 9)Ribulose 5-phosphate kinase.

Chu trnh C3 l chu trnh c bn nht ca th gii thc vt xy ra trong tt c thc vt, d l thc vt bc cao hay bc thp, d thc vt C3, C4 hay thc vt CAM.

171

Chu trnh Hatch-Slack (chu trnh C4) :

Nm 1966, hai nh khoa hc l Hatch v Slack nghin cu v pht hin ra ngoi chu trnh Calvin, mt s thc vt nhit i nh la min, ng, ma, c g... c qu trnh ng ho CO2 theo con ng khc. thc vt ny sn phm quang hp u tin ca quang hp l oxalo acetic acid, mt phn t c 4 carbon, ch khng phi l 3-phosphoglycerate . Chu trnh c nh CO2 nh vy gi l chu trnh C4 hay chu trnh Hatch-Slack v cc thc vt c nh CO2 theo con ng ny gi l thc vt C4. C s chuyn ho trong vic thc hin chc nng quang hp ca cy C4: mt loi lc lp chuyn trch c nh CO2 vi hiu qu cao nht, cn mt loi lc lp chuyn kh CO2 thnh cc cht hu c cho cy. V vy m hot ng quang hp ca cy C4 c hiu qu hn cc nhm thc vt khc. Kt qu l nng sut sinh hc ca cy C4 thng rt cao.
T bo Mesophyll ATP Pyruvic acid AMP PEPA CO2 NADPH2 Oxalo acetic acid NADP Malic acid

Pyruvic acid CO2 NADPH2 NADP

Malic acid

Chu trnh Calvin

Tinh bt

C6

T bo b mch

Hnh 9.12. Chu trnh Hatch-Slack

172 Con ng c nh CO2 thc vt CAM(Crassulaceae Acidetabolism) y l con ng quang hp thch nghi vi iu kin kh hn ca thc vt mng nc. Nh con ng quang hp ny m kh nng chu hn ca chng rt cao, hn hn cc thc vt chu hn khc.Do quang hp trong iu kin qu kh khn nn cng quang hp ca cc thc vt nhm ny thng thp, nng sut sinh hc khng cao v sinh trng chm hn cc thc vt khc. Qu trnh c nh CO2: qu trnh c nh CO2 c thc hin vo ban m. Ban m khi nhit khng kh gim xung th kh khng m ra thot hi nc v CO2 s xm nhp vo l qua kh khng m. Qu trnh tng hp monosaccharide (qu trnh kh CO2): qu trnh ny din ra vo ban ngy khi c nh sng hot ho h thng quang ho v kh khng ng li. Malic acid b kh carboxyl ho gii phng CO2 cung cp cho chu trnh C3. 9.2.2. Tng hp oligosaccharide S sinh tng hp oligosaccharide bng phn ng chuyn gc glucosyl, di tc dng ca enzyme : glucosyl transferase, v d:
sucrose glucosyl

Glucose 1 phosphate + fructose

transferase

sucrose + H3PO4

Ngoi ra dng UDP-glucose cng d dng chuyn glucose cho fructose to thnh sucrose. Cc dn xut UDP ca ng l nhng cht cho gc glucosyl rt hot ng (Hnh 9.13.) Tng hp polysacharide cng xy ra bng con ng chuyn gc glucosyl nh tng hp oligosaccharide . Cht cho gc glucosyl cn c th l oligosaccharide nh maltose, sucrose S chuyn gc khng ch ti C4 m ti c C6 to mch nhnh.

173

Hnh 9.13. S to thnh Sucrose t UDP - glucose v Fructose 6 - phosphate

174

TI LIU THAM KHO

Ti liu ting Vit
1. Phm Th Trn Chu, Trn thi ng. 1999. Ho sinh hc, NXB Gio dc, H ni. 2. Qu Hai. 2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu. 3. V Mai Hng. 2004. Gio trnh Sinh l thc vt, Ti liu lu hnh ni b Trng HKH Hu. 4. Trn Thanh Phong. 2004. Gio trnh Ha sinh i cng, Ti liu lu hnh ni b Trng HKH Hu.

Ti liu ting Anh

1. Halliwell, R. 1984.Chloroplast Metabolism: the structure and function of Chloroplast in green leaf cells, Clarendon, Oxford. 2. Lehninger A. L.. 2004. Principles of Biochemistry, 4th Edition. W.H Freeman.

175

Chng 10

Trao i Lipid
10.1. S phn gii lipid
10.1.1. Phn gii glycerid Glycerid d dng b thy phn do s xc tc ca cc loi lipase.
H2COOCR1 HCOOCR2 + 3H2O H2COOCR3 lipase H2COH HCOH H2COH + R1COOH R2COOH R3COOH

ng vt s thy phn glycerid xy ra nhanh chng nh s tc ng ca mui acid mt lm nh tng ha glycerid nn d b thy phn. 10.1.2. S oxi ha acid bo Acid bo b phn gii bng nhiu con ng: - oxi ha. - oxi ha. - oxi ha. Trong con ng ph bin v quan trng nht l .oxi ha. 10.1.2.1. .oxi ha acid bo S phn gii acid bo bng cch ct dn tng cp C, tc l ti v tr C ca chui carbon. Cc acid bo c mch carbon chn v cc acid bo c mch carbon l c c ch .oxi ha khc nhau giai on cui. Acid bo no v acid bo khng no c s khc nhau giai on sau. * i vi acid bo no c mch C chn: Qu trnh .oxi ha xy ra qua nhiu phn ng phc tp:

176

R - CH2 CH2 CH2 COOH A CoA-SH 1 ATP

(Acid bo)

AMP +(P-P)

R.CH2 CH2 CH2 CO SCoA (Acyl-CoA) FAD 2 FADH2 R-CH2 CH = CH CO SCoA (Enoyl-CoA) H2O 3 R-CH2 CHOH CH2 CO SCoA (-Hydroxy-acyl-CoA) NAD 4 NADH2 R-CH2 CO CH2 CO SCoA (-Ceto-acyl-CoA) CoASH 5 CH3CO SCoA (Acetyl-CoA)

R - CH3- CO SCoA (Acyl-CoA)

177 Cc enzyme tham gia xc tc cc phn ng trn l: 1. Acyl-CoA-Synthetase. 2. Acyl-CoA-Dehydrogenase. 3. Enoyl-CoA-Hydratase. 4. Hydroxy-acyl-Thiolase. Qua mt chu k phn ct, phn t acid bo ngn bt i 2 carbon, kt qu cui cng ca cc chu k phn ct .oxi ha ca acid bo l cc phn t acetyl-CoA . Nu phn t acid bo c n nguyn t C th s to ra n/2 phn t acetyl-CoA. Cc phn t acetyl-CoA tip tc b phn gii qua chu trnh Krebs to CO2 v H2O. V mt nng lng, qu trnh .oxi ha to nn ngun nng lng ln cung cp cho cc hat ng sng ca t bo. Mi ln phn ct bt 2C s to nn 1 NADH2, 1FADH2, qua chui h hp s tng hp c 5 ATP. ng thi mi phn t Acetyl-CoA b phn gii thng qua chu trnh Krebs s to ra c 12ATP. T ngi ta tnh c tng s ATP c to ra do s phn gii phn t acid bo no, mch cacbon chn c n nguyn t C l: n n 5 2 1 1 + 2 . 12 ATP * i vi acid bo no c mch C l i vi cc acid bo no c mch C l, qu trnh phn gii theo phng thc .oxi ha xy ra ging vi acid bo no c mch carbon chn ni trn nhng sau ln phn ct cui cng khng phi to ra 2 phn t Acetyl-CoA m cho ta 1 phn t Acetyl-CoA v 1 phn t propionyl-CoA. T propionyl-CoA li tip tc bin i thm mt chu k .oxi ha na to ra 1 phn t CO2 v 1 phn t Acetyl-CoA.

178

CH3 CH2 CO SCoA FAD

FADH2 CH2 = CH COSCoA H2O

CH2OH CH2 COSCoA

CoA-SH CH2OH CH2 COOH NAD NADH2 CHO CH2 COOH CoASH NAD NADH2 CH3CO SCoA Cc enzyme xc tc ging nh chu trnh trc * i vi acid bo khng no.

CO2

179 Vi acid bo khng no, qu trnh phn gii xy ra ty v tr ni i. - Nu v tr ni i ng vo v tr th qu trnh xy ra ging nh i vi acid bo no nhng khng xy ra phn ng 2. - Nu v tr ni i v tr khc th trc khi phn gii, acid bo khng no b kh thnh acid bo no tng ng ri tip tc phn gii theo con ng .oxi ha. Acid bo khng no FADH2 FAD Acid bo no

10.1.2.2. .oxi ha Phng thc .oxi ha l s phn gii acid bo xy ra ti v tr C, mi ln phn gii mch C b ct ngn i 1 nguyn t C v to ra CO2.
3 2 1

R CH2 CH2 - COOH H2O2

CO2
3 2

R CH2 CHO NAD

NADH2
3 2

R CH2 COOH

180 10.1.3. Phn gii glycerin Sau khi gii phng khi lipid n gin, glycerin tip tc c bin i bng nhiu cch to nn cc sn phm khc nhau
CH2OH CHOH CH2OH Glycerin CH2OH CHOH CH2O P Glycerol-P CHO CHOH CH2OP Aldehyde phospho glycerol (ALPG)

ATP

ADP

NAD

NADH2

T ALPG bin i thnh pyruvic acid nh trong qu trnh ng phn, sau pyruvic acid b phn gii tip thng qua chu trnh Krebs to CO2 v H2O. Nh vy s phn gii glycerin xy ra qua qu trnh ng phn v qua chu trnh Krebs to sn phm cui cng l CO2 v H2O. Nng lng gii phng trong qu trnh phn gii ny c dng tng hp ATP cung cp cho cc hat ng sng ca t bo. 10.1.4. Phn gii glycero phospho lipide Cc glycero phospholipide b phn gii qua nhiu giai on to ra nhiu sn phm trung gian tham gia vo nhiu qu trnh trao i cht. Qu trnh phn gii ny do nhiu enzyme thy phn xc tc. - Phospholipase A1 thu phn lin kt gia glycerin v acid bo th nht. - Phospholipase A2 thy phn lin kt gia glycerin v acid bo th hai. - Phospholipasse B thy phn c hai loi lin kt trn. - Phospholipase C thy phn lin kt gia glycerin v H3PO4. - Phospholipase D thy phn lin kt gia H3PO4 vi choline, ethanolamine hay serine. Phi hp tc dng ca tt c cc loi enzyme ni trn, phn t glycero phospholipid s b phn gii thnh glycerin, acid bo, H3PO4 v choline (hay ethanolamine, serine)

181

A1

R1CO
A2

O CH2 O CH H2 C O C O P OH D CH2 CH2 N (CH3)3

R2CO

10.2. Tng hp lipid

10.2.1. Tng hp acid bo Acid bo c tng hp t acetyl-CoA. S tng hp acid bo no v khng no giai on u ging nhau. Trc ht acid bo no c tng hp sau hnh thnh acid bo khng no bng cch oxi ha cc acid bo tng ng. Qu trnh tng hp acid bo t acetyl-CoA xy ra ngc vi qu trnh .oxi ha. T cc acetyl-CoA c ni dn li vi nhau thnh chui trung bnh ri dn n vic to thnh Stearic acid (c 18C) l loi acid bo no ch yu ca cc m. T Stearic acid c th tip tc ko di thm chui carbon to nn cc acid bo c mch C di hn. Trc ht t acetyl-CoA v CO2 kt hp vi nhau to nn malonyl-CoA. Qu trnh ny c xc tc bi acetyl-CoA-carboxylase.
H2O CH3COCoA COOH CH2 CO SCoA CO2 ATP ADP

Acetyl-CoA

Malonyl - CoA

tin hnh phn ng ngng t gia acetyl-CoA vi malonylCoA cn c s tham gia ca mt loi protein c vai tr vn chuyn nhm acyl, l protein vn chuyn gc acyl-ACP.
COOH CH2 CO ~ SCoA COOH CH2 CO ~ SACP ACPSH CoASH

Malonyl - CoA

Malonyl - ACP

182 Tip theo acetyl-SACP v malonyl-SACP ngng t vi nhau vi s xc tc ca enzyme acyl-synthetase. Khi cc phn t acetyl-ACP v malonyl-ACP tc dng vi enzyme acyl-synthetase s xy ra s chuyn cc gc acetyl v malonyl t nhm SH ca ACP sang nhm SH ca enzyme ng thi CO2 c gii phng.
COOH CH2 CO ~ SACP + COOH CO ~ SACP CH3CO CH2 CO ~ SACP + CO2 + ACP - SH

T CH3COCH2CO ~ SACP (aceto acetyl-ACP) b kh thnh .hydroxy-butyryl-ACP.

CH3CO CH2 CO ~ SACP NADH2 NAD CH3CHOHCH2CO ~ SACP

.hydroxy butyryl - ACP

Phn ng tip theo l .hydroxy butyryl-ACP b kh nc to nn crotonyl-ACP.

CH3CHOHCH2CO ~ SACP CH3CH = CHCO ~ SACP

(Crotonyl-ACP)

Crotonyl-ACP b kh to nn butyryl-ACP
CH3CH = CHCO ~ SACP NADH2 NAD CH3CH2CH2CO ~ SACP

Butyryl-ACP

T butyryl-ACP tip tc mt chu k mi ngng t vi malonylACP cho ta phnt c 6 nguyn t C. Qu trnh c tip din nh vy cho n khi to ra s C cn thit ca acid bo, sau Acyl-ACP ny s bin i tr li thnh Acyl-CoA v cui cng to ra acid bo no bng cch ct b CoA-SH.

183 Nh vy acid bo c mch cacbon chn vi n nguyn t C th qu n trnh s din ra 1 chu k. 2 Nu acid bo c mch C l th trong cc ln ni di mch C ni trn, ln u tin khng phi phn ng xy ra t 2 Acetyl CoA m xy ra t Acetyl-CoA v propionyl-CoA to ra acyl-CoA c 5 nguyn t C. T c mi chu k li ni thm 1 phn t Acetyl-CoA lm cho phn t acid bo di thm 2 nguyn t cacbon cui cng to nn phn t acid bo c s nguyn t cacbon l. Cc acid bo khng no c to ra t cc acid bo no tng ng bng cch b oxy ha bi FAD. Acid bo no
FAD FADH2

Acid bo khng no

10.2.2. Tng hp glycerin Glycerin c tng hp bng nhiu con ng. Con ng ph bin l t cc sn phm trung gian ca qu trnh trao i glucose l AlPG v PDA bin i thnh AlPG
NADH2 NAD

Glycero-P

Glycerin + H3PO4

PDA
NADH2 NAD

Glycero-P

Glycerin + H3PO4

10.2.3. Tng hp glyceride T cc acid bo v glycerin c tng hp s to thnh glyceride theo cc phn ng sau y:

184
R1-CO-SCoA Glycerin Glycero-P
ATP ADP Phosphatidic acid + 2CoA-SH

R2-CO-SCoA

R3-CO-SCoA Photphatidic acid H3PO4 Diglyceride Triglyceride

10.2.4. Tng hp glycero phospholipid T photphatidic acid s to nn cc loi phospho lipid khc nhau

185

Photphatic acid

CTP CDP-diacyl-glyceride Serine Glycero - P

Phosphatidyl serine CO2

Phosphatidyl inosid

Phosphatidyl Glycero-P

H3PO4

Phosphatidyl ethanolamine 3CH3 Phosphatidyl - choline

Phosphatidyl Glycerin

10.2.5. Tng hp sterid Sterid c to nn bi sterol v acid bo. Nguyn liu tng hp sterol l acetyl-CoA. Qu trnh sinh tng hp sterol c th chia lm 3 giai on vi nhiu phn ng rt phc tp. - Giai on chuyn acetyl-CoA thnh mevalonic acid. - Giai on tng hp squalen. - Giai on chuyn squalen thnh cholesterol.

186

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT. H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc. H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc. H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 Sinh ha hc hin i theo s . Nxb Y hc. H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.

187

Chng 11

Trao i Protein
11.1. S phn gii protein v amino acid
11.1.1. Phn gii protein Thy phn l con ng phn gii protein ph bin thc vt v ng vt. Qu trnh thy phn protein xy ra ti lysosome, ni cha nhiu enzyme thy phn protein l protease. Qu trnh thy phn xy ra qua 2 giai on - Nh peptid-peptido hydrolase, protein b thy phn thnh cc on peptid ngn. - Nh peptid-hydrolase thy phn tip cc peptid thnh amino acid. Kt qu chung l
protease

Protein
+(n-1)H2O

n.amino acid

ng vt c v, s phn gii protein u tin do tc ng ca pepsin. T bo nim mc d dy tit ra pepsinogen. Nh pepsin v HCl ca dch d dy, pepsinogen bin i thnh pepsin hat ng v pepsin hat ng s thy phn protein thnh amino acid. 11.1.2. Phn gii amino acid C nhiu con ng phn gii amino acid. 11.1.2.1. Kh amine Bng nhiu con ng khc nhau, cc amino acid b kh nhm amine to ra cc sn phm tng ng. - Kh amin bng cc enzyme kh. Nh enzyme kh xc tc, amino acid b kh thnh acid tng ng v gii phng NH3. Dehydrogenase R CH COOH NH2 amin bng con ng oxi ha. - Kh NADH+H+ NAD+ R CH2 COOH + NH3

188 Nh amino acid oxydase, amino acid b oxi ha to ceto acid tng ng v NH3 R CH COOH NH2 + O2 oxydase

R CO COOH + NH3

- Kh amine bng con ng thy phn. Nh tc dng ca enzyme thy phn hydrolase, amino acid b thy phn to oxiacid tng ng v NH3
R CH COOH NH2 + H2O

hydrolase
R CHOHCOOH + NH3

Ngoi cc con ng ra, aspartic acid cn b kh amin bng con ng kh ni phn t nh enzyme dezaminase xc tc
COOH CH2 CH COOH

Dezaminase
COOH CH = CH - COOH + NH3

NH2 Sn phm ca con ng kh amine cc amino acid l cc loi acid tng ng v NH3. 11.1.2.2. Kh carboxyl S loi carboxyl ca amino acid l cch phn gii amino acid rt ph bin nh decarboxylase xc tc R CH COOH NH2 Sn phm to ra l cc amine, l cc cht c hat tnh sinh hc cao c vai tr trong qu trnh trao i cht, cc hot ng sinh l ca c th nh histamine. 11.1.2.3. Chuyn v amine Bng con ng chuyn v nhm amine sang cho mt cetoacid, amino acid bin i thnh ceto acid tng ng, phn ng nh enzyme vn chuyn nhm amin xc tc amino transferase Decarboxylase R CH2 NH2 + CO2

189

R1 CH COOH NH2

R2 C COOH O

Amino Transferase

R1 C COOH O

R2 CH COOH NH2

Phn ng ny thc hin 2 chc nng: va phn gii 1 amino acid thnh ceto acid, ng thi tng hp mi amino acid khc t ceto acid tng ng. Tr threonine v lysine, tt c cc amino acid cn li u c th tham gia vn chuyn nhm amine bin i thnh cc ceto acid tng ng, v d:
CH3 CH COOH NH2 CH3 CO COOH + COOH CH3 CH COOH NH2 + COOH CH2 CO COOH

11.1.2.4. S bin i cc sn phm ca qu trnh phn gii amino acid Cc ng hng phn gii amino acid trnh by trn to ra nhiu loi sn phm. Cc sn phm ny tip tc c bin i to sn phm cui cng. - Cc cht hu c tip tc bin i bng cch oxy ha nh qu trnh phn gii acid bo to acetyl-CoA, t tham gia vo chu trnh Krebs phn gii tip. - Cc amine c bin i thnh cc acid tng ng sau tip tc bin i nh cc acid khc R CH2 NH2 + O2 R CH = NH + H2O R CHO + O2 R CH = NH + H2O R CHO + NH3 R COOH

NH3 tip tc bin i bng nhiu con ng gii c cho c th v NH3 tch ly nhiu s gy c. + NH3 c dng lm nguyn liu tng hp tr li amino acid bng con ng amine ha, amide ha (s trnh by trong phn tng hp amino acid).

190 + NH3 b bin i thnh ure qua chu trnh ornithine thi ra ngoi qua con ng nc tiu ng vt. Chu trnh ornithine chia lm 3 giai on 1) Tng hp carbamyl-phosphate CO2 + NH3 + ATP O=C NH2 O~P Phn ng c xc tc bi enzyme carbamyl phosphate synthetase. 2) Tng hp arginine. T carbanyl-phosphate v ornithine s to thnh citrullin bng mt phn ng ngng t. Sau citrullin kt hp vi aspactic acid nh arginino-succinic-synthetase to arginino-succinic acid. Tip theo arginino-succinic acid b phn gii thnh arginine v fumaric acid nh arginino-succinate-ligase. 3) Arginine b phn gii nh arginase to ornithine v ure. Ure c thi ra ngoi cn ornithine tip tc tham gia vo chu trnh mi. Trn y l nhng ng hng chung ca s phn gii amino acid. Tuy nhin mi amino acid u c con ng phn gii ring. Cc amino acid bin i theo cc ng hng trn u dn n vic to nn cc sn phm tham gia vo chu trnh Krebs phn gii thnh CO2 v H2O. + ADP

11.2. Tng hp amino acid

11.2.1. Amine ha Mt s acid bo khng no v ceto acid c th amine ha to nn amino acid tng ng
COOH CH = CH COOH + NH3
COOH CH CH2 COOH NH2

Fumaric acid
COOH CH2 CH2 CO COOH + NH3

aspartic acid
COOH CH2 CH2 CH COOH

-cetoglutaric acid
COOH CH2 CO COOH + NH3

glutamic acid

NH2

COOH CH2 CH COOH

oxaloacetic acid

aspartic acid

NH2

191 V nguyn tc, mi amino acid u c th c tng hp bng con ng ny t cc acid tng ng. Nhng trong t bo ch c 2 enzyme l glutamate dehydrogenase v pyruvate dehydrogenase c hot mnh thc hin xc tc loi phn ng trn, cn cc enzyme khc khng c kh nng xc tc cho nn trong thc t ch c glutamic acid v alanin l 2 amino acid c tng hp bng con ng ny. 11.2.2. Amide ha T 2 loi amino acid l aspactic acid v glutamic acid do c 2 nhm carboxyl nn c th c amide ha to amino acid mi, dng amide ca aspactic acid v glutamic acid l asparagine v glutamine
COOH CH2 CH COOH NH2 O = C CH2 CH COOH NH2

+ NH3

Asparagine

NH2

Aspartic acid
COOH CH2 CH2 CH COOH NH2 + NH3 O = C CH2 CH2 CH COOH NH2

Glutamine

NH2

Glutamic acid 11.2.3. Tng hp amino acid nh ATP Qu trnh tng hp amino acid nh ATP xy ra qua 2 giai on - NH3 + ATP AMP ~ NH2 + P - P y l phn ng hat ha nhm NH2 nh ATP. AMP ~ NH2 thc hin phn ng chuyn v amine cho ceto acid to amino acid tng ng AMP ~ NH2 + R C COOH O AMP + R CH COOH NH2

Thc cht ng hng ny cng l hnh thc amine ha cc ceto acid nhng khng s dng cc dehydrogenase m s dng ATP.

192 11.2.4. Chuyn v amine Nh trnh by trn (Mc 11.1.2.3) amino acid c th b phn gii bng con ng chuyn v amine ng thi vi vic tng hp mt amino acid khc. Nh qu trnh ny m thnh phn cc amino acid lun c i mi ph hp vi nhu cu ca c th trong qu trnh sng. 11.2.5. Oxim ha mt s vi sinh vt v thc vt c kh nng c nh Nit t do qu trnh c nh m. Qua qu trnh c nh N2, NH2OH c hnh thnh lm nguyn liu cho qu trnh tng hp amino acid theo cch oxim ha. Ngoi ra mt s vi sinh vt v thc vt cn c qu trnh kh nitrat (NO3-) thnh ammoniac (NH3). Trong qu trnh bin i theo ng hng ny NH2OH c to thnh trc khi to NH3. NH2OH lm nguyn liu tng hp amino acid bng cch oxim ha. Qu trnh oxim ha xy ra qua 2 giai on - Cc ceto acid kt hp vi NH2OH to nn oxim tng ng R C COOH O Ceto acid + NH2OH Oximase R C COOH NOH Oxim + H2O

- Cc oxim b kh to amino acid tng ng Oxim dehydrogenase R C COOH NOH

NADH+H+ NAD+

R CH COOH NH2 + H2O

vi sinh vt v thc vt, y l con ng tng hp amino acid quan trng.

11.3. Tng hp protein

Qu trnh tng hp protein l vn quan trng ca sinh hc phn t. Qu trnh xy ra phc tp vi s tham gia ca nhiu thnh phn.

193 11.3.1. Cc thnh phn tham gia tng hp protein 11.3.1.1. Nucleic acid Tham gia vo qu trnh tng hp protein c cc loi nucleic acid vi cc chc nng khc nhau - DNA: mang thng tin v cu trc phn t protein theo dng m ha. Mi protein c m ha trn 1 on DNA, l gen. - RNAm: lm nhim v truyn thng tin v cu trc phn t protein t gen sang chui polypeptide. - RNAt: lm nhim v vn chuyn cc amino acid t cc vng trong t bo n ribosome tin hnh tng hp chui polypeptide ti . ng thi nhn bit v tr b ba m ha amino acid trn RNAm t amino acid vo ng v tr ca n trn chui polypeptide. - RNAr: cng vi protein, RNAr cu to nn ribosome, ni thc hin qu trnh tng hp chui polypeptide. 11.3.1.2. Cc enzyme Tham gia xc tc qu trnh tng hp protein, c nhiu loi enzyme - Aminoacyl-adenilat-synthetase l enzyme xc tc qu trnh hat ha amino acid, phn ng gn amino acid vo RNAt. - Transpeptidase: xc tc phn ng to lin kt peptide ni cc amino acid li thnh chui polypeptide v chuyn dch chui polypeptide trong ribosome t v tr P sang v tr A. - Translocase: l enzyme xc tc qu trnh di chuyn ca ribosome trn RNAm. Ngoi cc enzyme chnh ny cn c enzyme ct amino acid m u ra khi chui polypeptide, enzyme xc tc s to cc cu trc khng gian ca protein 11.3.1.3. Nng lng Qu trnh tng hp protein cn nng lng. Nng lng cung cp cho qu trnh ny l ATP v GTP. - ATP cung cp nng lng cho giai on hat ha amino acid. - GTP cung cp nng lng cho giai on tng hp chui polypeptide ribosome. 11.3.1.4. Nguyn liu Nguyn liu tng hp protein l cc amino acid. Trong s cc amino acid c loi amino acid m u l methionine Eucariote v formyl methionine Procariote.

194 11.3.1.5. Ribosome Ribosome l ni tin hnh tng hp chui polypeptide. Thnh phn ribosome gm protein v RNAr. Cu trc ribosome gm 2 tiu th: tiu th ln v tiu th b. Trong ribosome c 2 vng hat ng: vng A l ni tip nhn cc amino acid mi cn vng P l ni to nn chui polypeptide. tiu th b cha mt loi RNAr, trn phn t RNAr ny c 1 on c thnh phn cc nucleotide tng ng b sung vi on khng m ha trn RNAm. Nh khi bt u qu trnh tng hp, RNAm n gn vo ribosome v t ng b ba m u ca n vo v tr P nh s lin kt gia on khng m ha trn RNAm vi on b sung trn RNAr. 11.3.1.6. Cc yu t tham gia tng hp protein * Yu t m u. l nhng phn t protein vi chc nng tham gia vo vic kch thch s m u trong qu trnh tng hp chui poplypeptide.
Procariote Yu t IF-1 IF-2 Chc nng Kch thch hat ng ca IF2, IF3 Lm d dng qu trnh kt hp f.Met-RNAt vi tiu th b 30S Gn vi tiu th b 30S, ngn khng kt hp vi tiu th ln 50S Yu t eIF-1 eIF-2 Eucariote Chc nng Gn vi RNAm Lm d dng s kt hp Met-RNAt vi tiu th b 40S Kt hp vi tiu th b 40S Kt hp vi m ca RNAm Kt hp vi RNAm Tch ri cc yu t khi u khi 40S v kt hp vi 60S Tch ribosome thnh 2 tiu th. 80S

IF-3

eIF-3

CBP-1 eIF-4a eIF-5

eIF-6

* Yu t ko di Tham gia vo giai on ko di c cc yu t:

195 - EF-Tu gip cho RNAtAa n gn vo v tr A ca ribosome. - EF-Ts gip s gii phng GDP khi phc EF-Tu-GDP. - EF-G xc tc s di chuyn ca ribosome trn RNAm theo chiu 5-3. 11.3.2. Tng hp chui polypeptide ti ribosome 11.3.2.1. Giai on hat ha amino acid tham gia vo qu trnh tng hp protein cc amino acid phi c hat ha v gn vo RNAt. Qu trnh ny xy ra hai phn ng, c xc tc bi enzyme aminoacyl-adenylat-synthetase [AMP ~ amino acid] E + P-P Trong phn ng th nht ny amino acid kt hp vi ATP to ra amino acid-AMP v gii phng pyrophosphat (P-P). Aminoacid-AMP khng trng thi t do m gn vi enzyme to phc linh ng Enzyme Amino acid + ATP [AMP ~ Amino acid] E + P-P RNAt mang amino acid s di chuyn n ribosome thc hin qu trnh tng hp chui polypeptide . 11.3.2.2. Giai on m u Tham gia vo giai on m u c cc yu t m u. procariote yu t m u l IF-1, IF-2, IF-3, cn Eucariote yu t m u l eIF-1, eIF-2, eIF-3. Nng lng cung cp cho giai on m u l GTP. c bit thc hin giai on tng hp nn amino acid m u cn c RNAt mang amino acid m u. procariote amino acid m u l mt loi methionine b bin i thnh dng formyl methionine. H2N CH COOH (CH2)2 S CH3
Methionine

CHO NH CH COOH
Formyl ha

(CH2)2 S
Formyl Methionine

CH3

Tham gia vn chuyn formyl methionine v methionine l 2 loi RNAt c cng b ba i m l UAC tng ng b sung vi m m u

196 AUG trn RNAm. Nh vy, RNAt mang formyl methionine vo m u qu trnh tng hp chui polypeptide, cn vic vn chuyn methionine a vo thnh phn chui polypeptide ch xy ra khi trn RNAm c b ba m ha methionine AUG. Eucariote amino acid m u l methionine nn RNAt vn chuyn methionine va lm nhim v m u nu gii m cho b ba m u AUG, va lm nhim v a methionine vo tham gia thnh phn chui polypeptide nu gii m cho b ba AUG nm cc v tr khc v tr m u. Giai on m u c thc hin bi s tch ribosome thnh 2 tiu n v ( procariote l 50S v 30S, cn Eucariote tng ng l 60S v 40S). Tip theo tiu n v b lin kt vi yu t m u IF3 to phc I (IF3 -30S). ng thi RNAt mang amino acid m u (f.Met hay Met) gn vi GTP, yu t m u IF2 to phc th II l (RNAtGTP-IF2). Tip theo phc I v phc II kt hp li vi nhau ng thi RNAm n gn vo tiu th b ca t hp trn. on khng m ha trn RNAm gn b sung vi mt on trn RNAr ca tiu th b nh t b ba m u ca RNAm vo ng v tr P ca tiu th ln khi tiu th b n gn vo phc trn. Cui cng t hp (30S-IF3-RNAt-GTP-IF2-RNAm) gn vo tiu th ln, khi phc li ribosome v gii phng cc yu t m u, GDP v H3PO4. Kt qu ca giai on ny l to nn phc m u, trong RNAt amino acid m u gn vo m m u ca RNAm nm v tr P ca ribosome

197

Ribosome 80S khng hot ng

RNAm

S ca giai on m u chui polypeptide Eucariota 11.3.2.3. Giai on ko di chui polypeptide Sau khi phc m u c to nn, qu trnh tng hp chui polypeptide bt u tin hnh. Tham gia giai on ko di chui c cc yu t ko di (EF procariote, eEF Eucariote) GTP cung cp nng lng, cc enzyme, cc amino acid-RNAt v phc h m u. Qu trnh ko di chui polypeptide xy ra theo trt t cc b ba trn RNAm k t sau b ba m u, theo chiu 5-3. ng vi cc b ba cc RNAt tng ng mang cc amino acid ca n trong phc hp aminoacidRNAt n gn ng v tr bng cch nhn bit gia b ba m ha ca RNAm vi b ba i m ca RNAt theo nguyn l b sung. Bng cch t ng v tr cc amino acid trn chui polypeptide. M u giai on ko di chui, amino acid-RNAt mang amino acid u tin n gn vo v tr A ca ribosome ang b trng nh to lin kt b sung gia b ba m ha trn RNAm vi b ba i m ca RNAt. Sau khi phc hp amino acid-RNAt gn vo v tr A ca ribosome, amino acid m u v tr P c tch khi RNAt ca n v chuyn sang v tr A lin kt vi amino acid bng lin kt peptid. Qu trnh c xc tc bi peptidyl Transferase. Nh vy v tr P ch cn RNAt khng mang amino acid cn v tr A c RNAt mang 2 amino acid. Bc

198 tip theo l nh locaferase xc tc ribosome trt trn RNAm theo chiu 5-3 mt on 3 nucleotide. Kt qu l t hp RNAt mang 2 amino acid chuyn sang v tr P cn v tr A ca ribosome li b trng nh phc h m u v kt thc vic ni di thm 1 amino acid vo chui polypeptide. Cc amino acid tip theo vo ni di chui cng c tin hnh qua cc bc nh vi amino acid th nht trn. Th t cc b ba trn RNAm quy nh trnh t cc amino acid tng ng vo tham gia qu trnh ni di chui polypeptide. Nh vy trt t cc b ba trn RNAm quyt nh trt t cc amino acid trn chui polypeptide.
RNAtAla + GTP+ EF

Trans peptidase

Trans locase RNAt m u

S giai on ko di chui 11.3.2.4. Giai on kt thc s tng hp chui polypeptide Qu trnh ko di chui s ngng khi gp tn hiu kt thc. Tn hiu kt thc l b ba kt thc. Khi b ba kt thc ca RNAm (trn mt RNAm c 1 trong 3 b ba UAG, UGA v UAA) nm vo v tr A ca ribosome. S xut hin 1 trong 3 b ba trn, qu trnh tng hp chui polypeptide

199 c kt thc do cc b ba ny khng m ha amino acid nn qu trnh ko di chui b ngt qung, chui polypeptide c tng hp b tch khi RNAt cui cng m khng c RNAt tip gn vo nn c gii phng ra khi ribosome v kt thc qu trnh tng hp. Tham gia vo qu trnh kt thc c yu t gii phng RF lm nhim v nhn bit m kt thc v gii phng chui polypeptide ra khi ribosome. 11.3.3. Hon thin phn t protein Chui polypeptide c tng hp ti ribosome phi qua nhiu bin i mi tr thnh phn t protein hon thin. Trc ht methionine u chui b ct b nh peptidase xc tc. Sau t cc nhm chc trn cc amino acid ca chui hnh thnh cc lin kt ni phn t to nn protein c cc mc cu trc khc nhau. Trc ht t chui polypeptide lin kt hydro c hnh thnh t cc nhm CO v NH ca cc amino acid to nn cu trc bc II ca protein. T protein bc II cc lin kt disulfua, lin kt ion, lin kt k nc to ra lm cho phn t protein c cu trc bc II cun xon to nn phn t protein bc III. T mt s phn t protein bc III cng chc nng c th lin kt vi nhau bng lin kt hydrogen, tng tc Van der Waals to nn protein c cu trc bc IV. Cc phn t protein c hon thin s c a n cc ni s dng thc hin chc nng ca chng trong t bo. 11.3.4. iu ha tng hp protein Qu trnh tng hp protein xy ra trong t bo c iu ha ph hp vi nhu cu ca c th. Khi c th cn loi protein no th qu trnh iu ha t iu chnh cho qu trnh tng hp protein xy ra, ngc li khi khng cn mt loi protein no na th qu trnh tng hp protein b c ch. Cng trnh c gi tr u tin v c ch iu ha tng hp protein do J.Monod v Jacob (1956) xut. Cc tc gi ny a ra thuyt Operon gii thch c ch iu ha tng hp protein. Theo thuyt operon phn t DNA cha nhiu loi gen: - Gen cu trc-cistron,structural gene (S): gen ny m ha phn t protein. Mi operon c th c nhiu gen cu trc m ha cho mt nhm protein c chc nng lin quan nhau nh cc enzyme xc tc mt chui phn ng. - Gen tc ng operator (O). - Gen khi ng promotor (P). Bn cnh mi operon c gen iu ha Regulator (R) vai tr iu ha hot ng ca operon, quyt nh s ng hay m ca operon.

200 Trt t cc gen trong operon nh sau R P O S1 S2 S3 S4

Mi operon chu trch nhim iu ha s tng hp mt nhm protein-enzyme cng tham gia xc tc mt chui phn ng, trong mi protein-enzyme do mt gen cu trc m ha. C nhiu hnh thc iu ha tng hp protein theo operon nh iu ha m tnh, iu ha dng tnh. Trong mi loi trn li c nhiu hnh thc khc nhau nh iu ha bng cch cm ng, iu ha bng cch c ch. 11.3.4.1. iu ha m tnh iu ha m tnh l c ch iu ha m khi khng c phc h c ch bm vo operon th operon m v tng hp protein xy ra. C 2 hnh thc iu ha theo dng ny l iu ha cm ng m tnh v iu ha c ch m tnh. * iu ha cm ng m tnh. in hnh ca dng iu ha ny l hot ng iu ha ca operon lac. Bn cht ca dng iu ha ny l khi trong mi trng c cht cm ng th operon m , qu trnh tng hp protein xy ra. Cht cm ng y l c cht ca cc enzyme do operon kim sot, l lactose. Khi trong mi trng c lactose th operon m v qu trnh tng hp nn cc enzyme tham gia phn gii lactose xy ra. Ngc li khi trong mi trng khng c lactose th operon ng v qu trnh tng hp protein khng xy ra. C ch qu trnh iu ha xy ra nh sau: operon lac c gen khi ng, gen tc ng v 3 gen cu trc Z.Y.A trong - Gen Z: m ha enzyme .galactosidase. - Gen Y: m ha enzyme permease . - Gen A: m ha enzyme transacetylase. Ba enzyme trn tham gia qu trnh phn gii lactose. Gen iu ha tng hp protein c ch. Cht c ch ny nu trng thi t do s c i lc vi operon v bm vo operon lm cho operon b ng. Cn khi c cht cm ng do cht cm ng c i lc vi cht c ch mnh hn operon nn lin kt vi cht c ch to phc khng hat ng. Phc ny khng bm c vo operon nn operon m. Nh vy trong t bo c lactose lm cht cm ng lin kt vi cht c ch do gen iu ha tng hp ra lm cho cht c ch khng bm vo operon, gen khi ng hat ng kch hot RNA polymerase. RNA

201 polymerase s xc tc cho qu trnh sao m cc gen cu trc thnh cc RNAm, t cc RNAm s tng hp ra 3 loi enzyme trn. Ba enzyme xc tc cho qu trnh phn gii lactose. Khi lactose b phn gii ht, khng cn cht cm ng lin kt vi cht c ch nn cht c ch bm vo operon, c ch hot ng ca RNA polymerase, RNA polymerase khng hat ng sao m c nn qu trnh tng hp protein cng khng xy ra.

RNAm

RNAm

* iu ha c ch m tnh. in hnh ca dng iu ha ny l hat ng iu ha ca operon Tryp. Bn cht ca dng iu ha ny l khi trong mi trng c cht ng c ch th operon ng, qu trnh tng hp protein khng xy ra. Cn khi khng c cht ng c ch th operon m, qu trnh tng hp protein xy ra. Cht ng c ch y l sn phm ca cc enzyme do operon kim sot, l Tryptophan. Khi trong mi trng khng c Tryptophan, operon m, qu trnh tng hp cc enzyme tham gia tng hp Tryptophan xy ra. Ngc li, khi trong mi trng c Tryptophan, operon ng, qu trnh tng hp cc enzyme khng xy ra. C ch iu ha xy ra nh sau. Operon Tryp c 5 gen cu trc m ha cho 5 enzyme tham gia qu trnh tng hp Triptophan trong t bo: Tryp E, Tryp D, Tryp C, Tryp B v Tryp A. Gen iu ha tng hp protein c ch. Khi trong t bo c Tryptophan ng vai tr cht ng c ch th cht c ch kt hp vi cht ng c ch to phc hat ng. Phc ny c i lc vi operon nn bm vo operon lm cho operon khng hot ng, RNA-polymerase khng sao

202 m nn khng c cc RNAm cho qu trnh tng hp protein, tng hp protein-enzyme khng xy ra. Khi trong t bo khng c Triptophan, cht c ch do gen iu ha tng hp ra khng c i lc vi operon nn khng bm vo operon, RNApolymerase khng b c ch nn xc tc cho qu trnh sao m t cc gen cu trc to ra cc RNAm tng ng. T cc RNAm tin hnh qu trnh tng hp protein to nn cc protein-enzyme.

RNAm

RNAm

11.3.4.2. iu ha dng tnh iu ha dng tnh l c ch iu ha tng hp protein m khi c phc h c ch bm vo operon th operon m, qu trnh tng hp protein xy ra. Khi khng c phc h c ch bm vo operon th operon ng, qu trnh tng hp protein dng li. C bn ca qu trnh ny l c c ch iu ha ngc hnh thc iu ha m tnh phn tch trn.

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TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT. H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc. H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc. H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 Sinh ha hc hin i theo s . Nxb Y hc. H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.

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Chng 12

Trao i nucleic acid

12.1. S phn gii nucleic acid
12.1.1. Thy phn nucleic acid S thy phn nucleic acid thnh mononucleotide c xc tc bi cc enzmie thy phn tng ng. DNA nh desoxyribonuclease xc tc bin i thnh cc desoxyribonucleotide cn RNA do cc ribonuclease xc tc s b phn gii thnh cc ribonucleotide. 12.1.2. Phn gii mononucleotide Mononucleotide b phn gii bi tc dng ca cc phosphatase hoc nucleotidase to nn cc nucleoside v H3PO4. Cc nucleoside li tip tc b thy phn bi cc nucleosidase to base nit v pentose. Cc sn phm ca qu trnh phn gii trn tip tc bin i - H3PO4 tham gia vo cc qu trnh trao i saccharide hay cc qu trnh trao i cht khc. - Base Nit tip tc b phn gii to cc sn phm tham gia vo qu trnh trao i cht ca t bo. 12.1.3. Phn gii base purine Adenine v guanine bin i thnh xanthine, t xanthine qua mt s phn ng tip theo to sn phm cui cng l ure v glyoxylic acid.
(1)

Adenine

Xanthin
(3)

(2)

Guanine

Allantoic acid
(4)

Ure +glyoxylic acid Phn ng 1 v 2 do enzime desaminase xc tc, phn ng 3 do xanthineoxydase v phn ng 4 do allantoicase xc tc.

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12.1.4. Phn gii base pyrimidine Cc base pyrimidine b phn gii to nn sn phm cui cng l NH3, CO2, .amino isobutyric acid v alanine.
Cytosine H2O Thymine NH3 Uracil Dihydro Uracil NADPH2 NADP Dihydro Thymine H2O H2O + alanine CO2 + NH3

NH3+CO2+ .aminoisobutyric acid

NADPH2 NADP

CO2, NH3 to ra trong cc qu trnh bin i trn c thi ra ngoi, cn alanine v .aminoisobutyric acid tip tc bin i nh cc amino acid khc.

12.2. Sinh tng hp nucleotide purine

Gc purine c to ra t nhiu thnh phn khc nhau: CO2, aspartic acid, glycine, formate, glutamine.
CO2

aspartic acid formate

glycine formate

N-H

glutamine

Trong qu trnh tng hp khung purine s xy ra ng thi c qu trnh tng hp nucleotide. Tm tt kt qu qu trnh nh sau: Riboso5P + 2.glutamine + glycine + CO2 + 2 formate + aspactic acid + H2O inosinic acid T inosinic acid s to nn GMP v AMP - Inozinic acid + aspactic acid + GTP AMP + fumaric acid GDP + Pv - Inozinic acid + NAD + ATP + NH3 GMP + NADH2 + AMP + Pv

206

Ngoi ra, cc nucleotide purine cn c th c tng hp trc tip t base purine v phosphoriboso-pyrophosphate (PRPP) Adenine + PRPP AMP + P-P Guanine + PRPP GMP + P-P

12.3. Sinh tng hp nucleotide pyrimidine

Khung pyrimidine c to ra t NH3, CO2 v aspartic acid
N

NH3 CO2

Asparic acid
N

Qu trnh tng hp nucleotide pyrimidine xy ra qua cc giai on sau: CO2 + NH3 + ATP carbamyl-P Carbamyl-P + Aspactic acid Orotic acid Orotic acid + Riboso5P UMP CMP TMP. Cc nucleotide pyrimidine cn c tng hp trc tip t base nit pyrimidine vi PRPP Uracil + PRPP UMP + P-P Thymine + PRPP TMP + P-P Cytosine + PRPP CMP + P-P

12.4. Tng hp DNA

Qu trnh tng hp DNA, hay cn gi l s ti bn, c ngha rt quan trng trong i sng c th lin quan n c ch di truyn. y l mt qu trnh phc tp c s tham gia ca nhiu yu t v xy ra nhiu hnh thc. C th chia qu trnh ti bn DNA thnh 3 kiu - Ti bn bo th. L qu trnh tng hp DNA t 1 phn t DNA gc to ra 2 phn t DNA con, trong c 1 phn t chnh l phn t DNA gc cn 1 phn t c tng hp mi hon ton.

207

- Ti bn gin on. L qu trnh tng hp DNA t 1 phn t DNA gc to ra 2 phn t DNA con c cc on mi tng hp v cc on c ca DNA gc xen k. Hai hnh thc ti bn trn t ph bin. - Ti bn bn bo th. y l hnh thc tng hp DNA t 1 phn t DNA gc to ra 2 phn t DNA con, trong mi phn t DNA con mt chui ly t DNA gc v mt chui mi tng hp. Hnh thc ny c Meselson v Stahl pht hin nm 1958 bng thc nghim nui cy E.coli. Trc ht E.coli c nui cy trong mi trng ch cha 15N (Nit nng) nn DNA c tng hp nn ch cha 15N s to nn phn t DNA c t trng cao hn DNA thng. Sau chuyn E.coli vo mi trng cha 14 N (Nit thng) v theo di, phn tch cc th h DNA mi c to ra bng phng php li tm phn on vi CSCl. Qua kt qu phn tch li tm cho thy th h th nht 100% phn t DNA dng lai, mt chui cha 15N v 1 chui cha 14N. th h th 2 c 50% dng lai v xut hin 50% dng 14N. iu chng t c ch ti bn DNA l dng bn bo th. 12.4.1. Cc yu t tham gia ti bn DNA - DNA khun. tng hp phn t DNA mi cn c phn t DNA lm khun. DNA va lm chc nng khun va tham gia trong sn phm ca qu trnh tng hp. - Nguyn liu. tng hp DNA mi cn c cc nguyn liu. Nguyn liu l cc desoxy Ribonucleotide Triphosphate (dATP, dGTP, dCTP, dTNP) dTMP va lm nguyn liu va cung cp nng lng cho qu trnh tng hp DNA mi. - Enzyme. Tham gia xc tc qu trnh ti bn DNA c nhiu loi enzyme + DNA-polymerase, + Topoisomerase, + Helicase, + DNA-ligase. - Protein. C nhiu loi protein tham gia vo qu trnh ti bn DNA vi chc nng h tr, kch thch + Protein bm si n SBS, + Protein DnaA, + Protein DnaB, + Protein DnaG.

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12.4.2. C ch ti bn DNA procariote 12.4.2.1. Giai on m u - Protein DnaB lm nhim v m xon DNA bng cch phn hy cc lin kt hydrogen gia 2 chui, tch 2 chui ra to nn chc ti bn. - Protein SBS n gn vo chc ti bn. - Primase xc tc s to RNA mi b sung vo chui khun 3-5. 12.4.2.2. Giai on ko di * Tng hp chui sm - Trn chui khun 3-5 sau khi to on RNA mi, cc nucleotide tip tc n gn vo u 3-OH ca chui theo nguyn tc b sung vi chui lm khun nh enzyme DNA-polymerace III xc tc. - Chui sm c tng hp lin tc, tho xon n u cc nucleotide t do trong mi trng t bo tng ng b sung vi cc nucleotide trn chui khun ln lt n gn vo u 3-OH bng cch to lin kt phosphodiester vi nucleotide cui cng u 3. ng thi pirophosphate c tch ra. * Tng hp chui mun Trn chui khun 3-5 ca DNA khun, chiu tho xon v chiu tng hp ngc nhau nn qu trnh tng hp khng din ra lin tc m to ra cc on okazaki ngc chiu vi chiu pht trin ca chc ti bn. Mi on okazaki c RNA mi ring c tng hp nh primase. Mi c tng hp b sung vi chui khun 5-3 v ngc chiu tho xon. Qu trnh tho xon xy ra c mt on khong 300 nucleotide mi tng hp RNA mi theo chiu ngc li. Xc tc cho qu trnh tng hp chui mun l phc hp protein c tn l primosom. Primosom di chuyn trn chui khun 5-3 v tin hnh tng hp on RNA mi nh primase sau tng hp tip on DNA b sung vo chui khun nh DNA-polymerase to nn on Okazaki. Sau khi on Okazaki hon chnh, RNA mi c tch ra nh DNApolymerase I sau thay vo v tr on RNA mi l on DNA tng ng. Sau cng nh DNA-ligase ni 2 on Okazaki li vi nhau. 12.4.2.3. Giai on kt thc Qu trnh ko di c tip din cho n khi ht phn t DNA khun. Kt qu t 1 phn t DNA khun to ra 2 phn t DNA mi, trong mi phn t DNA mi c 1 chui mi c tng hp t cc nucleotide trong mi trng, cn mt chui l ca DNA khun.

209

12.4.3. Ti bn DNA Eucariote Eucariote qu trnh ti bn DNA c bn ging procariote nhng cng c mt s c trng ring. - Trn mt phn t DNA khun qu trnh ti bn xy ra ng thi nhiu im. - Vn tc ti bn chm hn procariote + procariote vn tc 500 nucleotide/S. + Eucariote vn tc 50 nucleotide/S. - Mt s enzyme khc procariote + DNA polymerase , + DNA polymerase , + DNA polymerase , + DNA polymerase .

12.5. Tng hp RNA (sao m)

12.5.1. Cc yu t tham gia tng hp RNA 12.5.1.1. Khun tng hp RNA cn c khun. Khun c th l DNA, cng c th l RNA. phn ln cc sinh vt RNA c tng hp t DNA, do DNA lm khun. Phn t DNA lm khun ch s dng 1 on, tng ng 1 gen tng hp nn 1 phn t RNA. Nh vy t 1 phn t DNA c th tng hp ra nhiu RNA. ng thi trn 2 chui ca DNA, ch s dng chui 3-5 lm khun. 12.5.1.2. Nguyn liu Cng nh tng hp DNA, trong qu trnh tng hp RNA cn cc Ribonucleotide-Triphosphat lm nguyn liu v ngun nng lng. 12.5.1.3. Cc enzim v protein * RNA-polymerase. C 2 loi RNA-polymerase, mt loi xc tc qu trnh tng hp RNA t DNA mt loi xc tc qu trnh tng hp RNA t RNA. procariote RNA-polymerase c cu to phc tp. Phn t RNApolymerase gm 5 tiu n v , , , ,

210

Tiu n v

S lng 2 1 1 1 1

M 40.000 155.000 165.000 95.000 95.000

Chc nng Nhn bit v tr m u To lin kt P-diester Gn DNA khun Cha r Nhn bit chui lm khun v im m u

* Yu t Rho (): Rho l mt loi protein tham gia vo qu trnh kt thc tng hp RNA. 12.5.2. C ch sao m 12.5.2.1. Giai on m u Bc vo giai on m u RNA-polymerase tch yu t ra khi enzyme. Li enzyme tin hnh m xon DNA. Yu t nhn bit bit chui lm khun v im m u nh cc tn hiu trn promotor. Hai chui DNA tch ra 1 on 30 nucleotide to nn vng sao m. Chui n ca DNA (chui 3-5) nhn 1 nucleotide gn b sung vo nucleotide m u trn DNA. Tip theo nucleotide th 2 n gn vi nucleotide u bng lin kt phosphodiester v to lin kt b sung vi nucleotide trn chui khun. Sau khi lin kt phosphodiester u tin ny c to ra, yu t tch khi vng sao m v kt thc giai on m u. 12.5.2.2. Giai on ko di chui Nh li enzyme cc nucleotide trong mi trng n ko di chui theo nguyn tc b sung vi cc nucleotide trn chui khun DNA. Qu trnh ko di chui xy ra rt phc tp gm nhiu phn ng lin hon to ra s n nh ca vng m xon. Qu trnh xy ra theo trnh t sau: - Tho xon trn DNA khun u 3 chui khun. - Ko di thm 1 nucleotide trn chui RNA. - Tho xon kp lai DNA-RNA u 5. - ng xon trn DNA khun u 5.

211

Qu trnh c din ra theo chu k nh li enzyme xc tc cho n khi gp tn hiu kt thc. 12.5.2.3. Giai on kt thc C 2 kiu kt thc: kt thc nh yu t Rho v kt thc khng nh yu t Rho. - Kt thc nh yu t Rho: trn b mt ca mt s v tr kt thc c loi protein Rho. Rho di chuyn trn RNA mi c tng hp v i ti vng sao m, Rho lm nhim v tch xon lai DNA-RNA, gii phng RNA v kt thc qu trnh sao m. - Kt thc khng cn yu t Rho: Trn RNA c 1 on c cu trc ngc chiu (palindrome) khi sao m to ra vng palindrome, vng ny s to lin kt kp hnh thnh cu trc ci kp tc nn lm ngng qu trnh sao m. 12.5.3. Qu trnh trng thnh ca RNA Phn t RNA c sao t DNA l proRNA. T proRNA phi qua qu trnh bin i phc tp mi to RNAm. 12.5.3.1. Gn m vo u 5 ProRNA cha c m nn trc ht cn gn thm m vo u 5 ca Pro-RNA. M c tng hp sn trong nhn. M c gn vo u 5 bng lin kt anhydric acid vi nhm Triphosphate ca ProRNA ch khng gn vo u 3 nh qu trnh ko di chui. 12.5.3.2. Gn ui vo u 3 Cng nh m, ui ca RNAm khng m ha trong gen m c tng hp ring trong nhn. ProRNA cha c ui. ui c ni vo vi ProRNA u 3 nh polyA-polymerase. 12.5.3.3. Ct b cc on Intron trn proRNA. Trn Pro-RNA c cc on khng m ha amin acid (Intron I) cho nn to ra RNAm cn ct b cc on I v ni cc on m ha (ExonE) li. tn hiu di truyn c truyn t chnh xc, s ct ni cn c chnh xc cao v ch cn ct sai 1 nucleotide s lm thay i ton b cc m di truyn pha sau v tr ct. Gia cc on E v I c cc trnh t nucleotide c trng ging nhau mi pro-RNA. - u 3 ca E pha trc lun l AG, - u 5ca E pha sau lun l G,

212

- u 5 ca I lun l GU, - u 3 ca I lun l G. Trong Intron c mt on c vai tr quan trng trong c ch ct ni ca pro-RNA. l v tr tch nhnh. Qua v tr ny, di tc ng ca enzyme ct. Cc Intron b ct b ra v cc Intron ni li vi nhau. Kt qu ca qu trnh bin i trn to nn phn t RNAm trng thnh tham gia vo qu trnh dch m.

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT. H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc. H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc. H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 Sinh ha hc hin i theo s . Nxb Y hc. H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.

213

Chng 13

Mi lin quan gia cc qu trnh trao i cht

Trong c th mi qu trnh bin i cc cht khng xy ra ring r m c mi quan h qua li rt cht ch. Mi quan h th hin c trong qu trnh bin i ca mt nhm cht ln trong mi quan h ca qu trnh bin i cc nhm cht khc nhau. Trong cng mt nhm cht mi quan h th hin qua qu trnh ng ha v d ha. Mi quan h tng h ca s ng ha v d ha th rhin c hai qu trnh xy ra u c cc cht trung gian chung. V d phosphoglyceric aldehyde va l sn phm trung gian trong qu trnh phn gii (d ha) va l sn phm trung gian ca qu trnh tng hp (ng ha) saccharose. Gia cc nhm cht mi quan h din ra phc tp hn, nhiu hnh thc hn. Trc ht mi quan h c th hin qua cc cht trung gian. Mt cht l sn phm phn gii ca nhm cht ny li l nguyn liu tng hp cho nhm cht khc. V d Acetyl-CoA l sn phm ca qu trnh phn gii glucose ng thi n l nguyn liu tng hp acid bo. Mi lin quan tng h gia cc qu trnh trao i saccharide, lipid, protein, nucleic acid c ngha quan trng trong s sng ca sinh vt. V d vic chuyn ha tinh bt thnh ng sau to cc cht bo trong nhng thng ma ng thc vt cng nh ng vt, c ngha quan trng trong vic tng kh nng chu rt ca chng. Vic chuyn bin cht bo thnh ng khi ht ny mm s m bo thc n cho phi. Trong qu trnh hat ng sng ca c th, saccharide l cht d tr quan trng; khi c th cn s phn gii thnh phosphoglyceric acid ri thnh pyruvic acid lm nguyn liu cho s tng hp amino acid, acid bo v cc cht khc. Cht bo d tr cng c vai tr tng t, khi phn gii cht bo to nn acetyl-CoA t li tng hp amino acid, saccharide Khi trong c th ht ngun d tr saccharide v cht bo th protein s b phn gii to ra cc sn phm t c th tng hp lipid, saccharide.

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Qu trnh chuyn ha trn c vai tr quan trng c i vi sinh vt t dng ln sinh vt d dng. sinh vt t dng ngun cht hu c s cp do quang hp to ra l glucid, trc ht l glucose. T ngun saccharide , nh s chuyn ho tng h mi to ra c cc cht hu c m c th cn n nh lipid, protein, nucleic acid. sinh vt d dng s dng nhng cht hu c c sn trong thc n to nn cc cht c trng cho c th. T cc cht protein, lipid c trong thc n qua qu trnh bin i s kin to nn nhng cht c trng cho c th. Do thnh phn thc n khng th c y i vi nhu cu ca c th cho nn trong qu trnh ng ha vic chuyn ha nhm cht ny thnh cht hu c khc rt cn thit cho c th.

13.1. Mi lin quan gia qu trnh trao i saccharide v trao i lipid

Hai qu trnh trao i lipid v saccharide c mi quan h cht ch vi nhau. Saccharide d dng bin i thnh lipid v ngc li thng qua cc cht trung gian l AlPG, PDA v Acetyl-CoA T glucose qua qu trnh ng phn s to nn pyruvic acid. T Pyruvic acid b oxy ha s to nn acetyl-CoA. Acetyl-CoA l nguyn liu tng hp acid bo. ng thi trong qu trnh ng phn cn to ra AlPG, t AlPG bin i thnh glycero-P , t to nn glycerin. Nh vy t sn phm phn gii ca saccharide to nn nguyn liu c bn tng hp lipid l glycerin v acid bo. Ngc li qua s phn gii lipid s to nn cc cht trung gian l acetyl-CoA, glycerin. T acetyl-CoA, qua chu trnh ornithine s tng hp tr li saccharide. T glycerin to nn glycero-P v t tng hp li saccharide. thc vt, vi khun, nm mc, chu trnh glyoxylic l con ng ni trc tip qu trnh trao i lipid vi qu trnh trao i saccharide. Qua chu trnh ny acid bo sau khi phn gii thnh acetyl-CoA s bin i thnh oxalo acetic acid, t tng hp nn glucose. Ngc li t glucose s to acetyl-CoA v t tng hp tr li lipid.

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Mi lin quan gia s trao i saccharide v trao i lipid Saccharide

AlPG

Pyruvic acid

Acetyl-CoA

Chu trnh glyoxylic

Glycero-P

Glycerin Acid bo Acetyl-CoA

Lipid

13.2. Mi lin quan gia s trao i saccharide v trao i protein

Pyruvic acid l mt xch ch yu ni lin qu trnh trao i protein v trao i saccharide. Glucose qua qu trnh ng phn to ra pyruvic acid. Pyruvic acid l nguyn liu tng hp mt s amino acid trong h alanine: alanine, leucine, valine Trong qu trnh tng hp protein cn nng lng ATP, m ATP l sn phm quan trng ca qu trnh trao i saccharide.

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S mi lin quan gia trao i saccharide v trao i protein Saccharide

Ribuloso5P

Erylozo4P Erytroso4P
Shikimic acid

APEP Ser
Gly, Cys

APG
Pyruvic acid

His

Phe, Tyr, Trip

Ala, val, leu

Asp

Glu

Protein

Tre, Ila, Arg Met, Lys

Arg, Pro

13.3. Mi lin quan gia trao i saccharide v trao i nucleic acid

Trong qu trnh phn gii saccharide theo con ng pentosophosphate, Ribozo5-phosphate c to nn. T Riboso-5phosphate hnh thnh nn phospho-ribosyl-pyrophosphate (PRPP) l nguyn liu tng hp nn cc nucleotide purine v nucleotide pyrimidine. Ngc li trong qu trnh phn gii nucleic acid Riboso-5P c to thnh. T Riboso 5P s hnh thnh cc monosaccharide khc. Kiu lin quan th 2 gia qu trnh trao i nucleic acid v trao i saccharide l s lin quan cht ch gia qu trnh sinh tng hp cc nucleotide diphosphate v nucleotide triphosphate vi mc phn gii saccharide trong t bo v qu trnh phn gii ny gn lin vi qu trnh phosphoryl ho oxy ho. S phn gii saccharide to nng lng tng hp cc nucleotide diphosphate v nucleotide triphosphate. Qua qu trnh tng hp saccharide li cn s tham gia ca cc sn phm to ra trong qu trnh trao i nucleic acid, nh UTP tham gia vo qu trnh tng hp polysaccharide.

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13.4. Mi lin quan gia s trao i protein v trao i lipid

Trao i protein v trao i lipid c mi lin quan cht ch thng qua cc cht trung gian. S phn gii lipid to nn glycerin v acid bo v mt s cht khc nh serine, choline, sphingosine, H3PO4 Trc ht acid bo b phn gii to ra acetyl-CoA lm nguyn liu tng hp nn nhiu loi amino acid. Glycerin trong qu trnh phn gii to ra phosphoglyceric acid, t lm nguyn liu tng hp nn nhiu amino acid. Nhng mi quan h ny xy ra tng t nh mi lin quan gia saccharide vi protein phn tch trn. Ngc li, khi phn gii protein cng to nn cc hp cht trung gian, t tng hp nn lipid. Cc amino acid do thoi ho protein to ra, b kh amine s to nn cc acid nh pyruvic acid, oxalo acetic acid, -cetoglutaric acid. Trong s cc acid va nu th pyruvic acid c vai tr quan trng trong qu trnh tng hp lipid. T pyruvic acid, acetyl-CoA c to ra, acetyl-CoA l nguyn liu tng hp nn cc acid bo ng thi t pyruvic acid cng c th to ra glycerophosphate v t to thnh glycerin. Glycerin v acid bo l nguyn liu tng hp lipid. Trong qu trnh phn gii lipid s to thnh nn mt lng ln ATP l ngun nng lng cho qu trnh trao i protein. Ngc li protein vi chc nng enzyme c vai tr quyt nh i vi cc phn ng xy ra trong trao i lipid cng nh cc cht khc v khng c enzyme th khng c cc phn ng ho sinh xy ra tc l khng c trao i cht.

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S mi lin quan gia trao i lipid v trao i protein Lipid

Glycerin Glycero-P AlP

Nhng cht khc

Acid bo Acetyl-CoA Oxalo acetic acid AsP Tre, Met, Ile, Cys -Ceto glutaric acid Glu

Pyruvic acid APEP Ser Gly, Xyl Phe Protein Ala, val, leu

Fru.1.6d Rib5P His

Arg, Pro

13.5. Mi lin quan gia trao i protein v trao i nucleic acid

Gia qu trnh trao i protein v trao i nucleic acid c mi quan h c bit quan trng m biu hin r nht l trong c ch truyn t thng tin di truyn. DNA lm khun sao m thnh RNAm t tng hp nn protein. Cu trc phn t protein c m ho trong DNA. Qu trnh trao i nucleic acid li ph thuc vo s c mt ca cc phn t protein-enzyme. ng thi mt s amino acid l nguyn liu cho qu trnh tng hp nucleic acid nh aspartic acid l nguyn liu tng hp nucleotide pyrimidine, aspartic acid, glutamin, glycine l nguyn liu tng hp nucleotide purine.

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V mi quan h m nhiu nh nghin cu cho rng s tng hp protein l s cp cn s tng hp nucleic acid l qu trnh th cp lm nhim v tham gia vo qu trnh tng hp protein. Trong qu trnh phn gii protein vi s tham gia ca ATP to nn cc nucleotide-peptid. Cc phn t ny lai to nn nhng protein mi lm i mi thnh phn cc protein trong c th. ng thi s tng hp protein cng cn cc nucleotide triphosphat lm ngun nng lng (ATP, GTP).

13.5. Mi lin quan gia trao i lipid v trao i nucleic acid

Gia lipid v nucleic acid c t mi lin quan trc tip, ch yu l lin quan gin tip qua s trao i saccharide v protein. Tuy nhin cng c mi lin quan gia hai qu trnh trao i cht ny. S .oxi ho acid bo l ngun duy tr y cho s tng hp cc nucleotide diphosphat v nucleotide triphosphat qua qu trnh phosphoryl ho. Ngoi ra trong qu trnh tng hp lipid nh tng hp phospholipid c s tham gia ca CTP.

TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n (ch bin). 1979. Ha sinh i cng (tp I, II). NxB KH&KT. H Ni. 2. Phm Th Trn Chu, Trn Th ng. 2000. Ha sinh hc. Nxb Gio dc. H Ni. 3. Nguyn B Lc. 1997. Ha sinh. Nxb Gio dc. H Ni

Ti liu dch
1. Musil J.G., Kurz .K., Novakava .O. 1982 Sinh ha hc hin i theo s . Nxb Y hc. H Ni.

Ti liu ting nc ngoi

1. Farkas G. 1984. Nvnyi anyagcserelettan. Akadmiai Kiad Budapest. 2. Lehninger A. L., 2004. Principle of Biochemistry, 4th Edition. W.H Freeman.