69

3.3.2. Mt vi tnh cht ca protein 3.3.2.1. Tnh cht l-ho ca protein - Tnh tan ca protein Cc loi protein khc nhau c kh nng ho tan d dng trong mt s loi dung mi nht nh, chng hn nh albumin d tan trong nc; globulin d tan trong mui long; prolamin tan trong ethanol, glutelin ch tan trong dung dch kim hoc acid long v.v... - Tnh ngm nc ca protein Trong mi trng nc, protein kt hp vi nc trng ln tr thnh dng keo, hay ni cch khc protein trng thi hydrate ho, cc phn t nc bm vo cc nhm a nc trong phn t protein nh -NH2, -COOH..., lp o nc bao quanh phn t protein l mt trong cc yu t lm bn vng cu trc, ngn cch cc phn t protein khng cho chng dnh vo nhau thnh ta. - nht ca dung dch protein Khi protein ho tan trong dung dch, mi loi dung dch ca nhng protein khc nhau c nht khc nhau (bng 3.5). Ngi ta c th li dng tnh cht ny xc nh khi lng phn t ca protein ( nht cng cao th khi lng phn t cng cao).
Bng 3.5. nht ca mt s protein Protein Gelatin Albumin trng Gelatin Albumin trng Nng % (trong nc) 3,0 3,0 8,0 8,0 nht tng i (ca nc =1) 4,54 1,20 14,2 1,57

- Hng s in mi ca dung dch protein Khi thm cc dung mi hu c trung tnh nh ethanol, aceton vo dung dch protein trong nc th tan ca protein gim v protein s kt ta do vic gim mc hydrate ho ca cc nhm ion ho ca protein, lp o mt nc, cc phn t protein kt hp vi nhau thnh ta. Nh vy, hng s in mi ca dung mi lm ngn cn lc tnh in gia cc nhm tch in ca protein v nc. Mi lin h c c trng bi biu thc:

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L1 - l2 F= D - hng s in mi ca dung dch F- lc tnh in gia cc ion tch in L1 , l2 - in tch cc ion, r - khong cch gia cc ion - Tnh cht in li ca protein Cng nh cc amino acid, protein l cht in li lng tnh v trong phn t protein c nhiu nhm phn cc mnh (bn gc R) ca amino acid v d: nhm COOH th hai ca Asp, Glu; nhm NH2 ca Lys; nhm OH ca Ser, Thr, Tyr v.v...Trng thi tch in ca cc nhm ny ph thuc vo pH ca mi trng. mt pH no m tng in tch (+) v in tch (-) ca phn t protein bng khng, phn t protein khng di chuyn trong in trng gi l pHi (isoelectric - im ng in) ca protein. Nh vy protein cha nhiu Asp, Glu (amino acid c tnh acid mnh) th pHi trong vng acid, ngc li nhiu amino acid kim nh Lys, Arg, His th pHi trong vng kim. mi trng c pH < pHi , protein a s l mt cation, s in tch dng ln hn s in tch m. pH > pHi phn t protein th hin tnh acid, cho ion H+, do s in tch m ln hn s in tch dng, protein l mt a anion, tch in m.
Bng 3.5. Gi tr pHi ca mt s protein Protein Pepsin Albumin trng Casein Albumin huyt thanh Gelatin pHi 1,0 4,6 4,7 4,9 4,9 Protein Globulin sa Hemoglobin Ribonuclease Trypsin Cytochrom C Prolamin pHi 5,2 6,8 7,8 10,5 10,6 12,0

Dr2 Trong :

Trong mi trng c pH = pHi ca protein, protein d dng kt t li vi nhau v th ngi ta li dng tnh cht ny xc nh pHi ca protein cng nh kt ta protein. Mt khc do s sai khc nhau v pHi gia cc protein m c th iu chnh pH ca mi trng tch ring cc protein ra khi hn hp ca chng.

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- S kt ta bng mui ca dung dch protein Mui trung tnh c nh hng r ti ho tan ca protein hnh cu: vi nng thp chng lm ho tan nhiu protein. Tc dng khng ph thuc vo bn cht ca mui trung tnh, m ph thuc vo nng mui v s in tch ca mi ion trong dung dch, tc l ph thuc vo lc ion ca dung dch ( = 1/2 C1 Z1, 2 trong l k hiu ca tng, C1 l nng ca mi ion, Z1 l in tch ca mi ion). Cc mui c ion ho tr 2 (MgCl2, MgSO4...) lm tng ng k tan ca protein hn cc mui c ion ho tr 1 (NaCl, NH4Cl, KCl...). Khi tng ng k nng mui trung tnh th tan ca protein bt u gim v nng mui rt cao, protein c th b kt ta hon ton. Cc protein khc nhau b kt ta nhng nng mui trung tnh khc nhau. Ngi ta s dng tnh cht ny chit xut v tch ring protein khi hn hp. l phng php dim tch (kt ta protein bng mui). Th d dng mui amonium sulfate 50% bo ho kt ta globulin v dung dch amonium sulfate bo ho kt ta albumin t huyt thanh. - Biu hin quang hc ca protein Cng nh nhiu cht ho hc khc, protein c kh nng hp th v bc x nh sng di dng lng t h. V vy c th o cng hp th ca protein trong dung dch hay cn gi l mt quang thng k hiu bng ch OD (Optical Density). Da trn tnh cht ngi ta sn xut ra cc loi my quang ph hp th phn tch protein. Nhn chung protein u c kh nng hp th nh sng trong vng kh kin (t 350nm- 800nm) v vng t ngoi (t 320nm xung ti 180nm). Trong vng nh sng kh kin protein kt hp vi thuc th hp th mnh nht vng nh sng 750nm (nh lng protein theo Lowry). i vi vng t ngoi dung dch protein c kh nng hp th nh sng t ngoi hai vng bc sng khc nhau: 180nm-220nm v 250nm - 300nm. bc sng t 180nm-220nm l vng hp th ca lin kt peptide trong protein, cc i hp th 190nm. Do lin kt peptide c nhiu trong phn t protein nn hp th kh cao, cho php nh lng tt c cc loi protein vi nng thp. Tuy nhin vng hp th ny ca cc lin kt peptide trong protein c th b dch v pha c bc sng di hn khi c mt s tp cht ln trong dung dch protein. Mt khc chnh cc tp cht ny cng hp th nh sng t ngoi vng bc sng 180nm-220nm. V th trong thc t thng o hp th ca dung dch protein bc sng 220nm-240nm. bc sng t 250nm-300nm l vng hp th cc amino acid thm (Phe, Tyr, Trp) c trongphn t protein hp th cc i 280nm. C th s dng phng php o hp th ca dung dch protein bc sng 280nm

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nh tnh v nh lng cc protein c cha cc amino acid thm. Hm lng cc amino acid thm trong cc protein khc nhau thay i kh nhiu, do dung dch ca cc protein khc nhau c nng ging nhau c th khc nhau v hp th bc sng 280nm. Ngoi ra c nhiu cht khc trong dung dch cng c nh hng n hp th protein. V vy, cc phng php o hp th vng nh sng t ngoi thng c dung nh lng protein c tinh sch hoc xc nh protein trong cc phn on nhn c khi sc k tch cc protein qua ct. - Kt ta thun nghch v khng thun nghch protein Khi protein b kt ta n thun bng dung dch mui trung tnh c nng khc nhau hoc bng alcohol, acetone nhit thp th protein vn gi nguyn c mi tnh cht ca n k c tnh cht sinh hc v c th ho tan tr li gi l kt ta thun nghch. Cc yu t kt ta thun nghch c dng thu nhn ch phm protein. Trong qu trnh kt ta thun nghch mui trung tnh va lm trung ho in va loi b lp v hydrate ho ca protein, cn dung mi hu c vn ho nc s ph hy lp v hydrate nhanh chng. Trong ch phm protein nhn c cn ln cc cht dng kt ta, cn s dng phng php thch hp loi b cc cht ny. V d c th dng phng php thm tch loi b mui. Ngc li kt ta khng thun nghch l protein sau khi b kt ta khng th phc hi li trng thi ban u. S kt ta ny thng c s dng loi b protein ra khi dung dch, lm ngng phn ng ca enzyme. Mt trong nhng yu t gy kt ta khng thun nghch n gin nht l un si dung dch protein (s ni k hn trong phn bin tnh protein sau). - Cc phn ng ho hc ca protein Cng nh cc amino acid v peptide, protein c cc phn ng ho hc tng t, l: phn ng ca cc nhm -COOH, -NH2, gc R v phn ng to mu c trng ca lin kt peptide nh phn ng biure (xem chng 2 v 3. y xin c gii thiu thm mt s phn ng mu c trng khc, c ngha quan trng trong pht hin protein v cc gc amio acid trong chui polypeptide. + Phn ng vi thuc th Folin-Ciocateau Thuc th Folin-Ciocateau c cha phosphomolipdic acid v phosphovolframic acid cc cht ny lm tng nhy ca phn ng biure, mt khc phn ng vi gc Tyr v Trp trong phn t protein. Cc gc amino acid ny tham gia trong qu trnh to phc cht mu xanh da tri. + Cc phn ng mu c trng khc ca protein

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Nhng phn ng ny c c l do s c mt ca cc nhm nh chc ha hc xc nh trong phn t protein. C th s dng chng pht hin amino acid, protein trong dung dch. Phn ng xanthproteic: cc gc amino acid Tyr, Trp, Phe trong protein tc dng vi HNO3 c to thnh mu vng v sau khi thm kim s chuyn thnh da cam. Phn ng Pauli: cc gc Tyr, His trong protein tc dng vi diazobenzosulfonic acid to thnh mu anh o. Phn ng Millon: gc Tyr tc dng vi thu ngn nitrate trong HNO3 c to thnh kt ta mu nu t. Phn ng Saccaguichi: gc Arg tc dng vi dung dch kim ca -naphtolv hypobromitecho mu anh o. Phn ng Adamkievich: gc Trp tc dng vi glyoxylic acid v H2SO4 c to thnh vng tm mt phn cch. 3.3.2.2. Tnh cht v chc nng sinh hc ca protein Protein l hp cht hu c c ngha quan trng bc nht trong c th sng, v mt s lng, n chim khng di 50% trng lng kh ca t bo. T lu, bit rng protein tham gia mi hot ng sng trong c th sinh vt, ngoi vai tr l thnh phn chnh trong cu trc ca t bo v m, protein cn c nhiu chc nng phong ph khc quyt nh nhng c im c bn ca s sng nh s truyn t thng tin di truyn, s chuyn ho cc cht do cc enzyme, cc khng th chng li bnh tt, cc hormon dn truyn cc tn hiu trong t bo v.v... u c bn cht l cc protein. Trong c th protein c mt s vai tr sau y: - Chc nng to hnh Ngoi cc protein lm nhim v cu trc nh v virus, mng t bo, cn gp nhng protein thng c dng si nh: sclerotin c trong lp v ngoi ca su b; fibroin ca t tm, nhn; collagen, elastin ca m lin kt, m xng. Collagen m bo cho bn v tnh mm do ca m lin kt. - Chc nng xc tc Hu ht cc phn ng xy ra trong c th u do cc protein c bit ng vai tr xc tc, nhng protein c gi l cc enzyme. Mc d gn y ngi ta pht hin c mt loi RNA c kh nng xc tc qu trnh chuyn ho tin RNA thng tin (pre-mRNA) thnh RNA thng tin (mRNA), ngha l enzyme khng nht thit phi l protein. Nhng nh ngha c tnh cht kinh in: enzyme l nhng protein c kh nng xc tc

74
c hiu cho cc phn ng ho hc, l cht xc tc sinh hc vn c ngha c bit quan trng. Hin nay ngi ta bit c khong 3.500 enzyme khc nhau, nhiu enzyme c tinh sch, kt tinh v nghin cu cu trc. - Chc nng bo v Ngoi vai tr l thnh phn chnh trong xc tc, cu trc ca t bo v m, protein cn c chc nng chng li bnh tt bo v c th. l cc protein tham gia vo h thng min dch, c bit nhiu loi protein thc hin cc chc nng ring bit to nn hiu qu min dch c hiu v khng c hiu. Cc protein min dch c nhc n nhiu hn c l cc khng th, b th v cc cytokine. Ngoi ra protein cn tham gia vo qu trnh ng mu chng mt mu cho c th hoc mt s loi c th sn xut ra nhng c t c bn cht l protein nh enzyme nc rn, lectin v.v...,c kh nng tiu dit k th bo v c th. - Chc nng vn chuyn Trong c th c nhng protein lm nhim v vn chuyn nh hemoglobin, mioglobin, hemocyanin vn chuyn O2, CO2 v H+ i khp cc m, cc c quan trong c th. Ngoi ra cn c nhiu protein khc nh lipoprotein vn chuyn lipid, ceruloplasmin vn chuyn ng (Cu) trong mu v.v...Mt trong nhng protein lm nhim v vn chuyn c nhc n nhiu nht l hemoglobin. - Chc nng vn ng. Nhiu protein lm nhim v vn ng co rt nh myosin, actin si c, chuyn v tr ca nhim sc th trong qu trnh phn bo v.v... -Chc nng d tr v dinh dng Cc protein lm nhim v d tr nh casein ca sa, ovalbumin ca trng, feritin ca lch (d tr st) v.v...Protein d tr ny chnh l ngun cung cp dinh dng quan trng cho cc t chc m, phi pht trin. - Chc nng dn truyn tn hiu thn kinh Nhiu loi protein tham gia vo vic dn truyn tn hiu thn kinh i vi cc kch thch c hiu nh: sc t th gic rodopsin mng li mt. - Chc nng iu ho Cc protein lm nhim v iu ho qu trnh trao i cht thng qua tc ng ln b my thng tin di truyn nh cc hormon, cc protein c ch c hiu enzyme u c chc nng iu ho nhiu qu trnh trao i cht, chng hn cc protein repressor vi khun c th lm ngng qu trnh sinh tng hp enzyme t cc gen tng ng.

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- Chc nng cung cp nng lng Protein cng l ngun cung cp nng lng quan trng cho mi hot ng sng trong c th. Trong c th cc protein b phn gii thnh cc amino acid, t tip tc to thnh hng lot cc sn phm, trong c cc ceto acid, aldehyde v carboxylic acid. Cc cht ny u b oxy ho dn dn to thnh CO2 v H2O ng thi gii phng ra nng lng. 3.3.3. Phn loi protein Protein l hp cht a dng v thnh phn cu trc, chc nng; khi lng phn t ca chng cng rt khc nhau (bng 3.6). Do s phn loi protein c th da vo nhng mc ch khc nhau. C hai cch phn loi thng thng nht: 3.3.3.1. Phn loi theo hnh dng - Protein dng si C hnh dng di, thng l hnh si, chiu di ca phn t protein si ln hn ng knh ca n hng trm ln. Protein si tng i bn vng, khng tan trong nc v dung dch mui long, cc chui polypeptide ca protein si nm dc theo mt trc thnh nhng si di. Protein si l yu t cu trc c bn ca m lin kt ng vt cao cp, th d collagen gn v m xng,elastin m lin kt n hi, -keratin tc da v.v... - Protein dng cu C dng gn nh hnh cu hoc hinh bu dc, chiu di ca phn t protein cu c th ln hn ung knh ca n t 3 n 10 ln. Protein cu khng bn vng bng protein si, a s tan trong dung dch nc v d khuych tn, thng c chc nng hot ng sng ca t bo nh cc enzyme, cc hormon, cc protein vn chuyn nh albumin huyt thanh, hemoglobin v.v... - Protein dng trung gian Mt s protein dng trung gian, va c c im ca protein si, va c c im ca protein cu. Th d myosin (yu t cu trc v chc nng quan trong ca c) c cu trc hnh que di l c im ca protein si, nhng li tan trong dung dch mui l c im ca protein cu. Ngoi ra, trong nhm trung gian ny cn c cht tin thn ca fibrin l fibrinogen. 3.3.3.2. Phn loi theo thnh phn ho hc Protein gm hng trm, hng ngn, thm ch hng vn amino acid ni vi nhau bng lin kt peptide to nn mt hay nhiu chui polypeptide c cu trc rt phc tp.

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Cn c s c hay vng mt ca mt s thnh phn c bn cht khng phi protein m ngi ta chia protein thnh hai nhm:
Bng: 3.6 Khi lng (MW) v cu trc phn t ca mt s protein Protein Glucagon Insulin Ribonuclease (ty b) Lysozyme (lng trng trng) Myoglobin (tim nga) Chymotripsin (ty b) Hemoglobin (ngi) Albumin (huyt thanh ngi) Hexokinase (men bia) Tryptophan-synthetase (E.coli) -globulin (nga) Glycogen-phosphorylase (c th) Glutamate-dehydrogenase (gan b) Synthetase ca acid bo (men bia) Virus khm thuc l Khi lng (Dalton) 3482 5733 12.640 13.930 16.890 22.600 64.500 68.500 96.000 117.000 149.000 495.000 1.000.000 2.300.000 40.000.000 S gc amino acid 29 51 124 129 153 241 574 550 800 975 1.250 4.100 8.300 20.000 336.500 S chui polypeptide 1 2 1 1 1 3 4 1 4 4 4 4 40 21 2.130

- Protein n gin Protein n gin l nhng protein m phn t ca chng gm ton amino acid. Th d mt s enzyme ca tu b nh ribonuclease gm ton amino acid ni vi nhau thnh mt chui polypeptide duy nht (c 124 gc amino acid, khi lng phn t 12.640), chymotrypsin gm ton amino acid ni vi nhau thnh ba chui polypeptide (c 241 gc amino acid, khi lng phn t 22.600) v.v...Da theo kh nng ho tan trong nc hoc trong dung dch ngi ta c th chia cc protein n gin ra mt s nhm nh nh: - Albumin: tan trong nc, b kt ta nng mui (NH4)2SO4 kh cao (70-100%). - Globulin: khng tan hoc tan t trong nc, tan trong dung dch long ca mt s mui trung tnh nh NaCl, KCl, Na2SO4..., v b kt ta nng mui (NH4)2SO4 bn bo ho. - Prolamin: khng tan trong nc hoc dung dch mui long, tan trong ethanol, isopropanol 70-80%.

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- Glutein: ch tan trong dung dch kim hoc acid long. - Histon: l protein c tnh kim d tan trong nc, khng tan trong dung dch amoniac long. - Protein phc tp Protein phc tp l nhng protein m phn t ca chng ngoi cc amino acid nh protein n gin cn c thm thnh phn khc c bn cht khng phi l protein cn gi l nhm thm (nhm ngoi). Tu thuc vo bn cht ca nhm ngoi m ngi ta chia cc protein phc tp ra cc nhm nh v thng gi tn cc protein bt u bng tip u ng ch bn cht nhm ngoi: - Lipoprotein: nhm ngoi l lipid. - Nucleoprotein: nhm ngoi l nucleic acid. - Glucoprotein: nhm ngoi l carbohydrate v dn xut ca n. - Phosphoprotein: nhm ngoi l phosphoric acid. - Chromoprotein: nhm ngoi l hp cht c mu. Tu theo tnh cht ca tng nhm ngoi m c nhng mu sc khc nhau nh ( hemoglobin), vng ( flavoprotein)...

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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. Nguyn Ln Dng, Phm Vn Ty, Nguyn nh Quyn. 1999. Vi sinh vt hc. NXB Gio dc,H Ni. 4. L c Trnh. 1998. Hormon. NXB Y hc. H ni

Ti liu ting Anh

1. Dennison C. 2002. A Guide To Protein Isolation. Kluwer Academic Publishers. New York, Boston, Dordrecht, Lodon, Moscow. 2. Fersht A., 1998, Structure and Mechanism in Protein Science, W. H. Freeman, 3rd Rev Edit. 3. Lehninger A. L. 2004. Principle of Biochemistry, 4th Edition. W.H Freeman, 2004 4. Liebler D. C. 2002. Introduction to proteomics. Humana Press Inc. Totuwa, New Jersey. 5. Lodish H, 2003. Molecular Cell Biology. 5th ed, W.H Freeman, 2003 6. Walker J. M. 1996. The Protein Protocols Hand book. 2nd ed. Humana Press Inc. Totuwa, New Jersey.

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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

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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).

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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

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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

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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)

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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

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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.

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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.

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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