Chng 1 HA HC PROTEIN

Cao ng Nguyn, H Trung Thng

1.1 Protein v vai tr sinh hc ca chng

1.1.1 Khi nim v protein Protein l mt nhm cc hp cht i phn t sinh hc, cng vi polysaccharide, lipid v nucleic acid, to nn cc hp phn ch yu ca c th sng. Mt cch c th, protein l cc polymer c to nn t cc trnh t xc nh cc amino acid. Protein l hp cht hu c c ngha quan trng bc nht trong c th sng. V mt s lng, protein chim khng di 50% trng lng kh ca t bo. T lu ngi ta 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. Tht vy, cc enzyme, cc khng th chng li bnh tt, cc hormon dn truyn cc tn hiu trong t bo, ... u c bn cht l protein. Ngy nay, khi hiu r vai tr to ln ca protein i vi c th sng, ngi ta cng thy r tnh cht duy vt v ngha ca nh ngha thin ti ca Engels P.: S sng l phng thc tn ti ca nhng th protein. Vi s pht trin ca khoa hc, vai tr v ngha ca protein i vi s sng ngy cng c khng nh. Cng vi nucleic acid, protein l c s vt cht ca s sng. 1.1.2 Vai tr sinh hc ca protein 1.1.2.1 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 su b; fibrolin 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. 1.1.2.2 Xc tc Hu ht tt c cc phn ng xy ra trong c th u do cc protein c bit ng vai tr xc tc. Nhng protein ny c gi l cc enzyme. Mc du 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. Tuy nhin, hu ht cc phn ng din ra trong cc c th sng u c xc tc bi cc enzyme c bn cht protein. Bi vy, nh ngha c tnh cht kinh in: enzyme l nhng protein c kh nng xc tc 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, trong s nhiu enzyme c tinh sch, kt tinh v nghin cu cu trc. 1.1.2.3 Bo v Ngoi vai tr l cht xc tc v to cu trc cho 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 1

khng th, b th v cc cytokine. Ngoi ra, mt s protein cn tham gia vo qu trnh ng mu chng mt mu cho c th. Mt s loi c th sn xut ra nhng c t c bn cht protein nh enzyme nc rn, lectin, ... c kh nng tiu dit k th bo v c th. 1.1.2.4 Vn chuyn Trong c th ng vt 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, seruloplasmin vn chuyn ng (Cu) trong mu ... Mt trong nhng protein lm nhim v vn chuyn c nhc n nhiu nht l hemoglobin. 1.1.2.5 Vn ng Nhiu protein lm nhim v vn ng co rt nh myosin v actin si c, chuyn v tr ca nhim sc th trong qu trnh phn bo, ... 1.1.2.6 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. 1.1.2.7 Dn truyn tn hiu thn kinh Nhiu loi protein tham gia vo qu trnh dn truyn tn hiu thn kinh i vi cc kch thch c hiu nh sc t th gic rodopsin vng mc mt. 1.1.2.8 iu ho Nhiu protein c kh nng iu ho qu trnh trao i cht thng qua vic tc ng ln b my thng tin di truyn nh cc hormon, cc protein c ch c hiu enzyme Mt v d in hnh l cc protein repressor vi khun c th lm ngng qu trnh sinh tng hp enzyme t cc gene tng ng. 1.1.2.9 Cung cp nng lng Protein l ngun cung cp nng lng cho c th sng. Khi thy phn protein, sn phm to thnh l cc amino acid, t tip tc to thnh hng lot cc sn phm trong c cc keto acid, aldehyd v carboxylic acid. Cc cht ny u b oxy ho dn dn to thnh CO2 v H2O ng thi gii phng ra nng lng.

1.2 Cu to ca phn t protein

1.2.1 Thnh phn nguyn t Cho n nay ngi ta thu c nhiu loi protein tinh khit dng tinh th. t lu thnh phn cc nguyn t ho hc ca cc protein phn lp c nghin cu k. Thng thng trong cu trc ca protein gm bn nguyn t chnh l C, H, O v N vi t l C 50%, H 7%, O 23% v N 16%. iu ng lu l t l N trong protein kh n nh. Li dng tnh cht ny, nh lng protein theo phng php Kjeldahl ngi ta tnh lng N ri nhn vi h s vn c quy nh ty theo i tng nghin cu. Ngoi ra trong protein cn gp mt s nguyn t khc nh S ( 0-3%), P, Fe, Zn, Cu, ...

1.2.2 Amino acid - n v cu to c bn ca protein 1.2.2.1 Cu to chung Amino acid l cht hu c m phn t cha t nht mt nhm carboxyl (-COOH) v t nht mt nhm amin (-NH2), tr prolin ch c nhm NH (thc cht l mt imino acid). Trong phn t cc amino acid tn ti trong t nhin, cc nhm -COOH v -NH 2 u gn vi carbon v tr . Hu ht cc amino acid thu nhn c khi thu phn protein u dng L-. Nh vy cc amino acid ch khc nhau mch nhnh (c k hiu l R trong hnh 1.1). 1.2.2.2 Phn loi theo quan im ho hc Hin nay ngi ta phn loi amino acid theo nhiu cch khc nhau. Mi cch phn loi u c ngha v mc ch ring. Tuy nhin, cc cch phn loi ny u da trn cu to ha hc hoc mt s tnh cht ca gc R. V d: c ngi chia cc amino acid thnh 2 nhm chnh l nhm mch thng v nhm mch vng. Trong nhm mch thng li ty theo s c mt ca s nhm carboxyl hay s nhm amin m chia ra thnh cc nhm nh, nhm amino acid trung tnh (cha mt nhm -COOH v mt nhm -NH2); nhm amino acid kim tnh (cha mt nhm -COOH v hai nhm -NH2); nhm amino acid c tnh acid (cha hai nhm -COOH v mt nhm -NH2). Trong nhm mch vng li chia ra thnh nhm vng thm hay d vng, ... C ngi li da vo tnh phn cc ca gc R t chia cc amino acid thnh 4 nhm: nhm khng phn cc hoc k nc, nhm phn cc nhng khng tch in, nhm tch in dng v nhm tch in m. y chng ti xin c gii thiu cch phn loi cc amino acid mt cch chung nht. Theo cch ny da vo gc R, cc amino acid c chia thnh 5 nhm: Nhm I: Gm 7 amino acid c gc R khng phn cc v k nc, l: glycine, alanine, proline, valine, leucine, isoleucine v methionine.

Nhm II: Gm 3 amino acid c gc R cha nhn thm, l phenylalanine, tyrosine v tryptophan.

Nhm III: Gm 5 amino acid c gc R phn cc, khng tch in, l serine, threonine, cysteine, asparagine v glutamine.

Nhm IV: Gm 3 amino acid c gc R tch in dng, l lysine, histidine v arginine.

Nhm V: Gm 2 amino acid c gc R tch in m, l aspartate v glutamate.

1.2.2.3 Phn loi theo quan im dinh dng Theo quan im dinh dng ngi ta chia 20 loi amino acid thng gp trong protein thnh 2 nhm: amino acid khng th thay th (indispensable amino acid) hay cn gi l amino acid thit yu (essential amino acid) v amino acid thay th (disapensable amino acid) hay cn gi l amino acid khng thit yu. Amino acid khng th thay th l nhng amino acid m c th ng vt khng tng hp c hoc tng hp khng p ng nhu cu sinh trng hoc sinh sn mt cch ti u. S lng amino acid khng th thay th c th l t 8-10 loi amino acid ty theo tng loi ng vt (bng 1.1). Ngc li vi cc amino acid khng th thay th, amino acid thay th l nhng amino acid m c th ng vt c th tng hp c v p ng nhu cu ca chng. B khung carbon tng hp amino acid ch yu c to ra t glucose v cc amino acid khc, cn gc amin c s dng cho tng hp amino acid ch yu c cung cp t cc amino acid khc c s lng vt qu nhu cu ca c th ng vt. Khu phn n ca vt nui hu ht u p ng s lng amino acid thay th hoc s lng nhm amin cho s tng hp cc amino acid thay th k c trong trng hp hm lng protein trong khu phn thp, do s quan tm trong dinh dng l dnh cho cc amino acid khng th thay th.
Bng 1.1: Amino acid khng th thay th i vi gia cm, ln v ngi TT Tn amino acid
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Gia cm + + + + + + + + +

Ln + + + + + + + + +

Ngi + + + + + + + +

1 Arginine (1) (2) 2 Histidine 3 Leucine 4 Isoleucine 5 Lysine 6 Methionine 7 Phenylalanine 8 Threonine 9 Tryptophan 10 Valine
(1) (2)

i vi ngi, arginine v histidine c xp vo nhm amino acid thay th; trong mt s trng hp c nhu cu ln nh giai on cn non, stress, hai loi ny c coi l amino acid khng th thay th.

i vi g con, arginine l amino acid khng thay th. Mt s amino acid khng c xp vo nhm khng th thay th hay nhm thay th m chng c xp vo nhm bn thay th (semi-dispensable) hay cn gi l bn thit yu (semi-essential). Thuc nhm ny c arginine, cysteine, tyrosine. Arginine c coi l bn thay th i vi ln v arginine c th c tng hp t glutamine. Tuy vy, s tng hp ny khng p ng nhu cu dinh dng trong giai on u ca qu trnh pht trin c th. Do khu phn cho ln sinh trng phi cha mt lng arginine nht nh. Tuy vy, giai on k t sau khi thnh thc v gii tnh v giai on mang thai, nhiu nghin cu cho thy ln c th tng hp arginine vi s lng p ng nhu cu ca chng. Tuy nhin, s tng hp arginine c th khng p ng nhu cu tit sa ca ln ni.
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Cysteine c th c tng hp t methionine. Tuy vy, cysteine v dng oxy ha ca n l cystine c th p ng khong 50% nhu cu ca tng lng amino acid cha lu hunh (methionine + cysteine). Bng cch ny, cysteine c th lm gim nhu cu i vi methionine. Tng t nh i vi nhm amino acid cha lu hunh, phenylalanine c th p ng nhu cu ca tng lng phenylalanine v tyrosine (amino acid c nhn thm) v s chuyn ha phenylalanine c th to thnh tyrosine. Tyrosine c th p ng ti thiu 50% tng nhu cu ca hai loi amino acid ny nhng n khng th l ngun duy nht v khng th thay th cho phenylalanine v n khng th chuyn c thnh phenylalanine. Bng 1.2: Cc amino acid thng gp
Tn amino acid Glycine Alanine Proline Valine Leucine Isoleucine Methionine Tyrosine Tryptophan Serine Threonine Cysteine Aspargine Glutamine Lysine Histidine Arginine Aspartate Glutamate Tn gi theo danh php ha hc Tn vit K tt hiu Gly Ala Pro Val Leu Ile Met Phe Tyr Trp Ser Thr Cys Asn Gln Lys His Arg Asp Glu G A P V L I M F Y W S T C B Q K H R D E Khi lng (Mr) 75 89 115 117 131 131 149 165 181 204 105 119 121 132 146 146 155 174 133 147

-aminoacetic -aminopropionic -pirolidincarboxylic -aminoiaovaleric -aminonoisocaproic -amino--metylvaleric -amino--metyltiobutiric -amino--hydroxyphenylpropionic -amino--idolylpropionic -amino--hydoxypropionic -amino--hydroxybutyric -amino--tiopropionic amid ca aspartate amid ca glutamate , diaminocaproic -amino--imidazolpropionic -amino--guanidinvaleric -aminosucinic -aminoglutarate

Phenylalanine -amino--phenylpropionic

Ngoi cc amino acid thng gp trn, trong phn t protein i khi cn c mt s amino acid khc khc. l nhng loi t gp. Cc amino acid ny u l dn xut ca nhng amino acid thng gp. V d, trong phn t collagen c cha 4-hydroxyproline l dn xut ca proline, 5-hydroxylysine l dn xut ca lysine, ... Mt khc, mc du khng c trong cu trc protein, nhng c hng trm loi amino acid khc c th tn ti dng t 7

do hoc lin kt vi hp cht khc trong cc m v t bo. Chng c th l tin cht hay l cc sn phm trung gian ca qu trnh chuyn ho trong c th nh citrulline, ornithine, alanine, -amino butyric acid (GABA). 1.2.2.4 Mt s tnh cht ca amino acid - Mu sc v mi v ca amino acid Cc amino acid thng khng mu, nhiu loi c v ngt kiu ng nh glycine, alanine, valine, serine, histidine, tryptophan; mt s loi c v ng nh isoleucine, arginine hoc khng c v nh leucine. Bt ngt hay cn gi l m chnh l mui ca natri ca glutamic acid (monosodium glutamate). - Tnh tan ca amino acid Cc amino acid thng d tan trong nc v kh tan trong alcohol v ether (tr proline v hydroxyproline). Chng cng d ho tan trong acid v kim long (tr tyrosine).

- Tnh hot ng quang hc ca amino acid Cc amino acid trong phn t protein u c t nht mt nguyn t carbon bt i (tr glycine). Do vy, chng u c tnh hot ng quang hc, ngha l c th lm quay mt phng ca nh sng phn cc sang phi hoc sang tri. Quay phi c k hiu bng du (+), quay tri c k hiu bng du (-). Gc quay c hiu ca amino acid ph thuc vo pH ca mi trng. Tu theo s sp xp trong cu trc phn t ca cc nhm lin kt vi carbon bt i m cc amino acid c cu trc dng D hay L (hnh 1.7) gi l ng phn lp th. Trong t nhin, tt c cc amino acid u dng L nguyn t carbon bt i . Mt cch tng qut, s ng phn lp th ca mt amino acid c tnh theo 2n (n l s carbon bt i).

Hnh 1.8: Ph hp th nh sng cc tm ca tryptophan v tyrosine

Hu ht cc amino acid hp th tia cc tm bc sng () t 220 n 280 nm. c bit cng nng 10-3M, bc sng 280 nm tryptophan hp th nh sng cc tm mnh nht, gp 4 ln kh nng hp th ca tyrosine (hnh 1.8) v phenylalanine l yu nht. Phn ln cc protein u cha tyrosine nn ngi ta s dng tnh cht ny nh lng protein. - Tnh lng tnh ca amino acid Trong phn t amino acid c nhm carboxyl (-COOH) cho nn c kh nng nhng proton (H+) th hin tnh acid, mt khc c nhm amin (-NH2) cho nn c kh nng nhn proton nn th hin tnh base. V vy cc amino acid c tnh cht lng tnh. Trong mi trng acid, amino acid dng cation (tch in dng), nu tng dn pH, amino acid ln lt nhng proton th nht chuyn qua dng lng cc (trung ho v in), nu tip tc tng pH, amino acid s nhng proton th hai chuyn thnh dng anion (tch in m) (hnh 1.9). V vy i khi ngi ta coi amino acid nh mt di-acid.

Tng ng vi phn ly H+ ca cc nhm -COOH v -NH3+ c cc tr s pK1 v pK2 (biu th phn ly ca cc nhm c 1/2). T ngi ta xc nh c pH i (pI = pH ng in) = (pK1 + pK2)/2. V d: khi ho tan glycine vo mi trng acid mnh th hu nh glycine u dng cation. Nu tng dn lng kim, thu c ng cong chun . Trn ng cong chun thy rng: glycine ln lt nhng 2 proton trc tin chuyn sang dng lng tnh v sau cng chuyn thnh dng anion.

Tng ng phn ly ca nhm -COOH c mt na c tr s pK1= 2,34 v phn ly ca -NH3+ c mt na c tr s pK2= 9,60. Nh vy ta c

Mt khc ti pK1 + 2 s phn ly H+ ca nhm -COO- glycine l 99%, ch 1% dng -COOH v pK2 -2 dng -NH3+ l 99%, ch 1% dng -NH2. Nh vy trong vng pH t pK1 + 2 n pK2 -2, phn t glycine ch yu dng lng tnh v kt qu ta c mt vng ng in. Ngoi ra cc amino acid trong gc R c thm nhm -COOH hay -NH2 s phn ly ca chng s c thm mt tr s phn ly na -pKR. - Cc phn ng ho hc ca amino acid Cc amino acid u c nhm -NH2 v -COOH lin kt vi C , v vy chng c nhng tnh cht ho hc chung. Mt khc, cc amino acid khc nhau bi gc R, v vy chng c nhng phn ng ring bit. Ngi ta chia cc phn ng ho hc ca amino acid thnh 3 nhm: + Phn ng ca gc R Do cc amino acid c cu to gc R khc nhau, cho nn ngi ta c th da vo xc nh tng amino acid ring r nh phn ng c trng ca n. V d, phn ng oxy ho kh c th c s dng nhn bit nhm SH ca cysteine, phn ng to mui nhn bit cc nhm -COOH hoc -NH2 ca glutamate hoc ca lysine, phn ng to ester nhn bit nhm -OH ca tyrosine, ... + Phn ng chung L phn ng c s tham gia ca c hai nhm -COOH v -NH2. Cc amino acid phn ng vi ninhydrin trong iu kin un nng to thnh CO2 , NH3 , aldehyd v

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ninhydrin b kh, cui cng to nn sn phm c mu xanh tm (i vi proline cho mu vng chanh). + Phn ng ring bit Cc phn ng ca nhm -COOH: Ngoi cc phn ng ca nhm -COOH thng thng nh to ester, to amid, to mui, ... th n cn c nhng phn ng c trng khc nh c th b kh thnh hp cht ru amin di s xc tc ca NaBH4.

Nhm -COOH c th to thnh phc aminoacyl-adenylate trong phn ng hot ha amino acid tng hp protein, hoc c th loi CO2 l phn ng gp rt nhiu trong qu trnh phn gii amino acid. Cc phn ng ca nhm -NH2: Nhiu phn ng ca nhm amin c dng xc nh cc ch tiu ca amino acid nh: nh lng nitrogen ca amino acid ngi ta cho phn ng vi HNO2 gii phng N2 .

nh lng amino acid ngi ta cho phn ng vi aldehyd to thnh base schiff. xc nh amino acid u N-tn cng ngi ta cho tc dng vi 2-4 dinitrofluobenzen (phn ng Sanger) hay phenyliothiocyanate (phn ng Edman). 1.2.3 Peptide 1.2.3.1 Khi nim Peptide c t hai n vi chc amino acid ni vi nhau. Chng c th c tng hp trong t nhin hoc c hnh thnh do s phn gii protein. Trong phn t peptide, cc amino acid c lin kt vi nhau thng qua lin kt peptide (hnh 1.11).

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Lin kt peptide c bn cao bi cu trc ca n c 4 e/, 2e/ thuc v lin kt C=O cn 2e/ thuc v b i e/ t do ca nguyn t N. Lin kt gia C-N l lin kt phc tp. N c th chuyn t dng n dng lai (trung gian) l mt phn ghp i ca lin kt (hnh 1.12). Ngi ta cho rng t l ca lin kt kp ny l khong 30% i vi lin kt C-N v 70% i vi lin kt gia C v O. u ca mt chui peptide l amino acid c nhm -amin (-NH2) t do c gi l u N-tn cng v u kia c nhm -carboxyl (-COOH) t do c gi l u C tn cng. Lin kt peptide to nn b khung chnh ca chui peptide hoc polypeptide, cn cc gc R to nn mch bn ca chui.

1.2.3.2 Mt s peptide c hot tnh sinh hc quan trng Trong t nhin tn ti nhiu dng peptide c chc phn quan trng lin quan n hot ng sng ca c th nh l cc hormone, cc cht khng sinh ... Bn cnh cng c nhng peptide c chc phn cha r rng. Mt s peptide l sn phm thy phn ang cn dang d ca protein. Sau y s gii thiu mt s peptide quan trng, c nhiu ngha i vi hot ng sng ca sinh vt. - Cc hormone sinh trng (GH) Hormon sinh trng ca ngi (HGH-human growth hormone) cn c tn gi STH (somatotropin hormone) l mt chui polypeptide bao gm 191 amino acid. Trong cu trc c hai cu disulfur c to thnh gia amino acid 53-165 v gia amino acid 182-189. HGH c cu to rt ging vi hormone lactogen ca nhau thai (85% trnh t amino acid ging nhau) v gn ging prolactin ca ngi (32% trnh t amino acid ging nhau). Hormone sinh trng c tc dng kch thch s tng trng ni chung, kch thch s to sn hn l to xng. N cng l mt hormone chuyn ho. Hormone sinh trng kch thch s tng hp protein t nhng amino acid c vn chuyn d dng vo trong t bo nh HGH, v l hormone gy tng ng huyt, sinh i tho ng, ng thi kch thch s phn gii lipid m bo nhu cu v nng lng cho c th, gy tng acid bo t do trong huyt tng. S thiu ht HGH nu xy ra trc tui dy th s dn n chng ngi ln, s d tha HGH nu xy ra trc tui dy th s dn n chng ngi khng l, nu xy ra sau tui dy th s dn n chng ngi b to cc (pht trin chiu dy ca u, xng v mt).

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- Insulin T 1953, Sanger (2 gii thng Nobel v ha hc vo nm 1958 v 1980) nghin cu, tinh ch v xc nh hon ton cu trc ca phn t insulin. Phn t insulin bao gm 51 amino acid, c cu trc gm 2 chui polypeptide, vi khi lng phn t 5.700. Chui A c 21 amino acid, chui B c 30 amino acid. Hai chui c ni vi nhau bng 2 cu disulfur. Trong chui A cng hnh thnh 1 cu disulfur gia amino acid th 6 v amino acid th 11. Phn c hiu (c trng ca mt loi) ch tp trung vo cc amino acid th 89-10, 12-14 ca chui A v c bit l amino acid th 30 ca chui B (hnh 1.13). Ngi ta 13

cng xc nh c cu trc ba chiu ca insulin v thy rng cu trc phn t insulin c gi vng bi nhiu lin kt mui, lin kt hydro v cu ni disulfur gia chui A v chui B. Insulin c tc dng r nht trong tt c cc hormon ca tuyn ty, c bit i vi qu trnh chuyn ho carbohydrate. N c tc dng h ng huyt. Insulin cn kch thch qu trnh tng hp v c ch qu trnh phn gii glycogen c, gan v m m. c bit insulin tng cng tng hp acid bo, protein v kch thch s ng phn. Tc dng quan trng nht ca insulin l kch thch s xm nhp glucose, mt s ng khc, amino acid trong t bo c v m. Do vy insulin lm gim lng glucose trong mu. Ngoi ra insulin cng lm gim s tng hp mi glucose do lm gim nng enzyme nh pyruvate carboxylase v fructose 1-6 diphosphatase. - Oxytocin, vasopressin v vasotocin Oxytocin l mt peptide c 9 amino acid. ng vt c v, oxytocin ch khc s thay i ca 2 amino acid l: amino acid v tr th ba l isoleucine v amino acid v tr th tm l leucine (bng 1.3). Vasopressin ca loi ch nhi c cu trc trung gian gia vasopressin v oxytocin ca ng vt c v: amino acid th ba l isoleucine v amino acid th tm l arginine. Vasopressin l mt peptide c cu trc gm 9 amino acid. Phn ln ng vt c v amino acid th 8 ca vasopressin l arginine (arg-vasopressin), tr ln v h m, amino acid th 8 l lysine (lys-vasopressin). ng vt c xng sng nhng khng c v, amino acid th 3 l isoleucine; vasopressin c tn l arginin-vasotocin (bng 1.3). Oxytocin c tc dng trn c trn ca t cung v tuyn v, gy co t cung khi sinh con v kch thch s tit sa khi cho con b. Vasopressin c tc dng chng li niu, tng cng ti hp thu nc thn, ng thi lm co mch, do c tc dng tng huyt p.

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1.2.4 Cc bc cu trc protein

V mt cu trc ngi ta phn bit protein gm bn bc: bc 1, bc 2, bc 3 v bc 4 (hnh. 1.14).

1.2.4.1 Cu trc bc 1

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Cu trc bc 1 biu th trnh t cc gc amino acid trong chui polypeptide, cu trc ny c gi vng bng lin kt peptide (lin kt cng ha tr). Cu trc bc 1 l phin bn ca m di truyn, vic xc nh c cu trc bc 1 l c s tng hp nhn to protein bng phng php ho hc hoc bng bin php cng ngh sinh hc. 1.2.4.2 Cu trc bc 2 Biu th s xon ca chui polypeptide, l tng tc khng gian gia cc gc amino acid gn nhau trong mch polypeptide. Ni cch khc, l dng khng gian cc b ca tng phn trong mch polypeptide. Cu trc ny c lm bn nh cc lin kt hydro c to thnh gia cc lin kt peptide gn nhau. 1.2.4.3 Cu trc bc 3 Biu th s xon v cun khc ca chui polypeptide thnh khi, c trng cho protein dng cu, l tng tc khng gian gia cc gc amino acid xa nhau trong mch polypeptide. Trong nhiu protein dng cu c cha cc gc cysteine to nn lin kt disulfur gia cc gc Cys xa nhau trong mch polypeptide lm cho mch b cun li. Ngoi ra, cu trc bc 3 cn c gi vng bng cc loi lin kt khc nh lin kt k nc, lc Van der Waals, lin kt hydro, lin kt tnh in gia cc gc amino acid. 1.2.4.4 Cu trc bc 4 Biu th s kt hp ca cc chui c cu trc bc 3 trong phn t protein. Nhng phn t protein c cu trc t 2 hay nhiu chui protein hnh cu, tng tc vi nhau sp xp trong khng gian to nn cu trc bc 4. Mi mt chui polypeptide c gi l mt tiu n v, chng gn vi nhau nh cc lin kt hydro, tng tc Van der Waals gia cc nhm phn b trn b mt ca cc tiu n v lm bn cu trc bc 4.

1.3 Mt s tnh cht ca protein 1.3.1 Tnh cht l hc

- Phn t lng C nhiu phng php xc nh phn t lng ca protein nh phng php ha hc, thm thu, tc lng, Protein c phn t lng ln > 10.000. Khi nim protein v polypeptide thnh thong c hiu ging nhau, tuy vy polypeptide c trng lng phn t < 10.000. - c tnh keo Khi ha tan, protein to thnh cc dung dch keo v cho hin tng Tindal. Xung quanh mi phn t protein c bao quanh bng mt lp dy c cc phn t dung mi, nu dung mi l nc th s to thnh tng hydrate. - p sut thm thu Protein ha tan trong nc to nn dung dch keo c p sut thm thu gi l p sut keo nh hn rt nhiu so vi p sut thm thu ca dung dch tht. p sut keo ng vai tr quan trng trong vic vn chuyn nc cng cc cht dinh dng v cn b qua thnh mch. 16

- S khuych tn Trong dung dch, protein khuych tn chm. Do chng c kch thc ln cho nn protein khng i qua c cc mng bn thm nh mng t bo, mng cellophan, Nu cho dung dch protein vo mng bn thm ri nhng vo cc nc th cc phn t c kch thc nh nh NaCl i qua mng bn thm mt cch d dng, cn protein vn nm li trong ti. y l s thm tch v mng bn thm c gi l mng thm tch. Ngi ta ng dng phng php ny loi mui khi dung dch protein. - nht Khi ha tan, protein to thnh dung dch keo c nht kh cao v ln hn nht ca nc. Cc loi protein khc nhau th c nht khc nhau. - S hp ph Protein c th hp ph cc cht kh, lng hoc rn ln trn b mt ca chng. Nh tnh cht ny m nhiu protein ng vai tr l cc cht vn chuyn trong c th. C ch tc dng ca mt s loi thuc hoc ca cc cht ha hc cng c gii thch da trn tnh cht hp ph ca protein.

1.3.2 Tnh lng tnh

Phn t protein gm nhiu amino acid kt hp li vi nhau thnh mt hoc nhiu chui polypeptide. Trn mi chui polypeptid u c mt nhm -NH2 ca amino acid Ntn v mt nhm -COOH ca amino acid C-tn. Ngoi ra, trong phn t protein cn c th c mt s nhm -NH2 t do ca cc amino acid kim tnh v mt s nhm -COOH ca cc amino acid acid tnh. Trong dung dch cc nhm -NH2 v -COOH ny c th in li to thnh -NH+3 v -COO-. Do , protein cng c tnh lng tnh nh amino acid v trong dung dch chng cng c th tn ti di 3 dng ion: anion, cation v ion lng cc vi cc nng khc nhau ty theo pH ca mi trng. Mi loi protein cng c im ng in (pI) tng ng v t l ca mi dng ion trong dung dch ph thuc vo pH ca dung dch ha tan protein. Nu pH ca mi trng = pI th protein tn ti dng ion lng cc nhiu nht v protein s khng di chuyn trong in trng. Nu pH ca mi trng > pI th anion chim nhiu nht v protein s di chuyn v pha cc dng ca in trng. Nu pH < pI th dng cation chim nhiu nht v protein s di chuyn trong in trng v cc m. Tnh cht ny c ng dng tch protein bng k thut in di, ngoi ra cn c ng dng chit xut v tinh ch protein bng phng php sc k trao i ion.

1.3.3 c tnh ha tan

trng thi ha tan, phn t protein c hydrate ha, lp o nc bao quanh l mt trong cc yu t lm bn vng dung dch protein, lm cho cc phn t protein ngn cch v khng dnh vo nhau. C nhiu yu t nh hng n tnh tan ca protein. - nh hng ca pH tan ca protein thp nht pH = pI ca n, tan ca protein tng ln khi pH nm xa pI v khi pH = pI th phn t protein khng tch in cho nn chng khng c lc y tnh in v d b ng kt. Khi pH pI th cc phn t protein tch in cng du v y nhau cho nn khng khng b ng kt do tan tng ln. - nh hng ca nng mui Mui trung tnh nng thp lm tng tan ca nhiu loi protein. Khi tng nng mui ti mt gii hn nht nh th tan ca protein gim xung v nu nng

17

mui tip tc tng ln th protein c th ng kt hon ton. Tnh cht ny c ng dng tch cc loi protein theo phng php dim tch. - nh hng ca dung mi Khi cho cc dung mi nh ethanol, aceton, vo dung dch protein th tan ca protein b gim v c th dn n ng kt do gim lp v hydrate ha. - nh hng ca nhit Trong khong nhit 0 - 40oC, tan ca a s protein tng khi nhit tng. khong nhit 45 - 70oC a s protein mt tnh bn vng v c th b bin tnh (tr protein ca mt s sinh vt chu nhit nh vi khun chu nhit).

1.3.4 S ng kt v bin tnh

Phn t protein ch th hin c c tnh sinh hc ca n trong mt khong gii hn nht nh ca mi trng. Nu vt qu gii hn s lm cho protein b mt tnh cht ban u, mt tnh cht sinh hc, l s bin tnh ca protein (gim tnh tan, tnh cht sinh hc khng cn na). S bin tnh khng lm t cc lin kt peptid m ch lm t cc lin kt yu nh lin kt hydro, lin kt mui, V vy, cu trc khng gian ca protein b o ln, cc nhm k nc quay ra ngoi, cc nhm a nc quay vo trong, s hydrate ha b gim, cc chui polypeptid d kt hp li vi nhau, do tan gim v protein d b ng kt. trong ng tiu ha ca ng vt, s bin tnh protein gip cho qu trnh tiu ha protein din ra d dng hn. C hai loi bin tnh: - Bin tnh thun nghch Cu trc khng gian ca phn t protein b bin i tm thi v c th tr li trng thi ban u nu loi tr nguyn nhn gy bin tnh. Mt s yu t gy bin tnh thun nghch nh dung mi hu c trung tnh (ether, ethanol, aceton, ), mt s mui trung tnh ((NH4)2SO4). - Bin tnh khng thun nghch Cu trc khng gian ca phn t protein b ph v vnh vin, khng ti lp d iu kin no. Cc yu t gy bin tnh khng thun nghch nh acid v c m c (HCl, H2SO4, HNO3, ), acid hu c (CCl3-COOH), nhit cao,

1.3.5 Tnh cht ha hc

- Protein c th b thy phn bi enzyme hoc bi mt s yu t khc nh acid hoc base c (nng 6N) nhit cao v trong nhiu gi to thnh cc sn phm d dang hoc sn phm cui cng l cc amino acid. - Protein cho phn ng biurea (c trng ca lin kt peptid). - Cc gc amino acid trong phn t protein cng cho cc phn ng c trng ca amino acid t do tng ng. - Phn t protein c nhm -NH2 u N-tn cng v -COOH u C-tn cng, ngoi ra chng cn c th c thm mt s nhm -NH2 v COOH ca cc gc amino acid nm gia chui polypeptid. Do vy, protein c th cho hoc nhn proton (H+) th hin tnh acid hoc base. Nh c im ny cho nn protein l mt h m gp phn duy tr pH ca c th ng vt tng i n nh.

1.3.6 Tnh c hiu sinh hc

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Chc nng sinh hc ca protein rt a dng, tuy vy trong nhiu trng hp khng phi ton b phn t protein u c gi tr nh nhau i vi hot tnh sinh hc ca n m c b phn lm nhim v gi khung khng gian, c b phn trc tip quyt nh hot tnh sinh hc. S thay i cc lin kt, cc nhm ha hc hoc cc amino acid nht nh trong phn t protein, c bit l b phn quyt nh trc tip hot tnh sinh hc ca n u dn ti s thay i hot tnh sinh hc ca protein. Mt s v d v tnh c hiu sinh hc ca protein nh tnh c hiu kiu phn ng v c hiu c cht ca enzyme, hin tng chong khi truyn mu, hin tng khng dung ha khi v da hoc ghp m,

1.4 Phn loi protein 1.4.1 Phn loi theo hnh dng
1.4.1.1 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 bc cao v d collagen gn v m xng, elastin m lin kt n hi, -keratin tc v da, ... 1.4.1.2 Protein dng cu C dng gn nh hnh cu hoc hnh 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 hormone, cc protein vn chuyn nh albumin huyt thanh, hemoglobin ... 1.4.1.3 Protein dng trung gian Mt s protein c dng trung gian, va c c im ca protein si, va c c im ca protein cu. V d, myosin (yu t cu trc v chc nng quan trng 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.

1.4.2 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 hoc nhiu chui polypeptide c cu trc phc tp. Cn c s c hay vng mt ca mt s thnh phn c bn cht khng phi protein m ngi ta chia protein thnh: 1.4.2.1 Protein n gin Protein n gin l nhng phn t protein m phn t ca n gm ton amino acid. V d, mt s enzyme ca ty 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), ... Da vo kh nng ha tan trong nc hoc trong dung dch, ngi ta c th chia cc protein n gin ra mt s nhm nh nh:

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- 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%. - Glutelin: 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. 1.4.2.2 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 ghp (nhm ngoi). Ty 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 t ch bn cht nhm ngoi: - Lipoprotein: nhm ngoi l lipid. - Nucleoprotein: nhm ngoi l nucleic acid. - Glycoprotein: nhm ngoi l carbohydrate v dn xut ca chng. - Phosphoprotein: nhm ngoi l phosphoric acid. - Cromoprotein: nhm ngoi l hp cht c mu. Ty theo tnh cht ca tng nhm ngoi m c nhng mu sc khc nhau nh mu ( hemoglobin), mu vng ( flavoprotein), ... Nu nhm ngoi l kim loi th protein s c tn gi chung l metaloprotein.

Ti liu tham kho

Ti liu ting Vit Trn Th n, i Duy Ban, Nguyn Hu Chn, nh H, L c Trnh, 1980. Ho sinh hc. NXB Y hc. Phm Th Trn Chu, Trn thi ng, 1999. Ho sinh hc. NXB Gio dc, H Ni. Nguyn Ln Dng, Phm Vn Ty, Nguyn nh Quyn, 1999. Vi sinh vt hc. NXB Gio dc. L c Trnh, 1998. Hormon. NXB Y hc, H ni. Nguyn Vit Tu, 1980. Phng php nghin cu ho hc cy thuc. NXB Y hc, Tp. H Ch Minh. Ti liu ting Anh Copeland, R. A., 2000. Enzymes; a practical introduction to structure mechanism and data analysis, 2nd ed. Willey-VCH. A John Willey & Sons, INC., Pub.

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Dennison, C., 2002. A guide to protein isolation. Kluwer Academic Publishers, New York, Boston, Dordrecht, Lodon, Moscow. Hans, U. B., 1974. Methods of enzymatic analysis. Second English Edition Acdemic Press, Inc., New York, San Francisco, London, Vol., 4. Liebler, D. C, 2002. Introduction to proteomics. Humana Press Inc. Totuwa, New Jersey. Lodish, H., 2003. Molecular Cell Biology. 5th ed. E-Book. Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA. NRC, 1998. Nutrient requirements of swine. Tenth revised edition, National Academy Press, Washington D. C., USA. Walker, J. M., 1996. The protein protocols, handbook, 2nded. Humana Press Inc, Totuwa, New Jersey. Ti liu ting c Arbeitsgemeinschaft fuer Wirkstoffe in der Tierernaehrung e.V. (AWT), 1998. Aminosaeren in der Tierernaehrung. Agrimedia Buchedition, Verlag Alfred Strothe Deutscher Fachverlag, Frankfurt am Main.

Chng 2

HA HC NUCLEIC ACID
Cao ng Nguyn, H Trung Thng

2.1 Thnh phn cu to Nucleic acid, vt cht mang thng tin di truyn cua cac h thng sng, la mt polymer c hinh thanh t cac monomer la nucleotide. Mi nucleotide gm 3 thanh phn: nhom phosphate, ng pentose (l ng 5 carbon) va mt base nit. 2.1.1 Base nit Cc base nit c trong phn t nucleic acid u l nhng dn xut ca base purine hoc pyrimidine. Cc base purine gm adenine (6-amino purine) v guanine (2-amino, 6aminopurine), cc base nit pyrimidine gm thymine (2,6-dioxy, 5-methylpyrimidine), cytosine (2-oxy, 6-aminopyrimidine) v uracil (2, 6 dioxypyrimidine) (hnh 2.1).

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2.1.2 ng pentose ng pentose trong nucleic acid gm c hai loi l ng deoxyribose v ribose (hnh 2.2). S c mt ca 2 loi ng ny l mt trong nhng c im phn bit DNA v RNA. 2.1.3 Phosphoric acid L mt acid v c - H3PO4 2.2 Lin kt to thnh nucleic acid 2.2.1 S to thnh nucleoside Nucleoside l sn phm thy phn khng hon ton ca nucleic acid. Nucleoside gm c hai thnh phn l ng pentose v mt base nit (thuc purine hay pyrimidine).

2.2.2 S to thnh nucleotide 22

Nucleotide cng l sn phm thy phn khng hon ton ca nucleic acid. Nucleotide gm c ba thnh phn: ng pentose, base nit v phosphoric acid (hnh 2.3).

2.2.3 S to thnh nucleic acid Cac nucleotide c ni vi nhau bng lin kt phosphodiester 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 to thnh v tr C3 v C5 cho 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 co cu tao rt ging nhau la DNA (desoxyribonucleic acid) va RNA (ribonucleic acid).

2.3 Mt s nucleotide quan trng khng tham gia cu to nucleic acid 2.3.1 ADP v ATP
ADP (adenosine diphosphate) v ATP (adenosine triphosphate) u 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.

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2.3.2 cAMP (AMP vng)

Adenosine monophosphate 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. Ngoi cAMP, mt s nucleotide khc cng tham gia vo cc qu trnh chuyn ha trong c th, v d cGMP v ppGpp.

2.3.3 UDP v UTP UDP (uridine diphosphate) v UTP (uridine triphosphate) u l nhng dn xut ca uracil. Chng u l nhng cht quan trng trong cc phn ng trung gian chuyn ho glucose v galactose. Ngoi ra, chng cn tham gia trong qu trnh hnh thnh nhng hp cht phosphate giu nng lng. 2.3.4 CDP v CTP CDP (cytidine diphosphate) v CTP (cytidine triphosphate) l nhng dn xut ca cytidine. CTP cng l hp cht giu nng lng v c th tham gia nhiu phn ng khc nhau nh: phosphoryl ho ethanolamine dn n s sinh tng hp cephaline hoc phn ng vi phosphate choline hnh thnh cytidine diphosphate-choline (CDP-choline).

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2.3.5 Cc coenzyme nucleotide Hin nay ngi ta bit c mt s nucleotide tham gia cu to nn mt s coenzyme quan trng nh coenzyme A (CoA hoc CoA-SH), flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD+), nicotinamide adenine dinucleotide phosphate (NADP+), ... to thnh cc coenzyme ny cn c cc vitamin nh vitamin B5 (pantothenic acid) trong cu trc ca CoA, vitamin B2 (riboflavine) trong cu trc ca FAD, vitamin PP (nicotinic acid) trong cu trc ca NAD+ v NADP+.

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2.4 Phn loi cu hnh v cu trc ca nucleic acid 2.4.1 Deoxyribonucleic acid (DNA) Phn t DNA la mt chui xon kep gm hai si n. Mi si n la mt chui polynucleotide. Mi nucleotide gm ba thanh phn: nhom phosphate, ng desoxyribose va mt trong bn base v thng c k hiu bng ch ci u tin ca cc base (A: adenine, C: cytosine, G: guanine va T: thymine). Hai si n kt hp vi nhau nh cac lin kt hydrogen hinh thanh gia cac base b sung nm trn hai si: A b sung cho T va C b sung cho G. Mi si n co mt trinh t inh hng vi mt u 5phosphate t do, u kia la 3 hydroxyl t do (quy c la 5 3). Hng cua hai si n trong chui xon kep ngc nhau, cho nn c goi la hai si i song. Nhng phn tich cu truc hin ai a cho thy cu truc cua DNA khng phai lun lun tng ng vi dang c goi la B ma Watson va Crick a a ra. Do s tac ng cua cac hp cht co trong lng nho hoc protein, dang B co th chuyn sang dang A (nen nhiu hn) hoc la dang Z (xon trai). Chung co th t gp lai (DNA) hoc xon manh, vi du mt si kep DNA co dai la 20 cm c nen trong mt chromosome co kich thc la 5 m.

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Phn t DNA trong nhim sc th cua sinh vt eukaryote dang thng, con phn ln t bao prokaryote (vi khun) phn t DNA co dang vong. Du dang nao thi cac phn t DNA u tn tai di dang cun cht. Trong t bao eukaryote, DNA kt hp cht che vi cc protein la histone. DNA eukaryote co kich thc rt ln (vi du DNA ngi co th dai n 1 m) cho nn cu hoi t ra la phn t nay phai c nen nh th nao vao th tich rt han ch cua nhn. Vic nen c thc hin nhiu mc , mc thp nht la nucleosome va mc cao nht la cu truc nhim sc cht.

27

28

Tht vy, ng kinh cua chui xon DNA chi la 20 , trong khi si nhim sc cht quan sat di kinh hin vi in t co ng kinh 100 , i khi at 300 . iu nay chng to phn t DNA tham gia trong vic hinh thanh nhng cu truc phc tap hn (hinh 2.6). Si co ng kinh 100 la mt chui nhiu nucleosome. o la nhng cu truc hinh thanh t nay c t chc thanh mt si DNA qun quanh mt loi gm 8 phn t histon. Si 100

cu truc phc tap hn la si co ng kinh 300 . Trong nhn t bao, cac si va k trn kt hp cht che vi nhiu protein khac nhau va ca vi cac RNA tao hanh nhim sc cht, mc t chc cao nht cua DNA. Phn t DNA c sp xp trn nhim sc th lm cho chiu di ngn li hn 50.000 ln. Cac DNA eukaryote co c im khac vi DNA prokaryote. Toan b phn t DNA prokaryote u mang thng tin ma ha cho cac protein trong khi o DNA eukaryote bao gm nhng trinh t ma hoa (cac exon) xen ke vi nhng trinh t khng ma hoa (intron). Cac trinh t ma hoa eukaryote chim ngp trong mt khi ln DNA ma cho n nay vn cha ro tac dung. Ty theo mc hin din cua chung trong nhn, cac trinh t DNA c chia lam ba loai: - Cac trinh t lp lai nhiu ln. V du: ng vt co vu cac trinh t nay chim 1015% genome (h gene). o la nhng trinh t DNA ngn (10-200 kb), khng ma hoa, thng tp trung nhng vung chuyn bit trn nhim sc th nh vung tm ng (trinh t CEN) hay u cac nhim sc th (trinh t TEL). Chc nng cua cac trinh t nay cha ro, co th chung tham gia vao qua trinh di chuyn DNA trn thoi v sc (trinh t CEN) hoc vao qua trinh sao chep toan ven cua phn DNA nm u mut nhim sc th (trinh t TEL).

29

30

- Cac trinh t co s ln lp lai trung binh. Vi du: genome ngi, cac trinh t nay chim 25-40 %. Chung a dang hn va co kich thc ln hn (100-1.000 kb) cac trinh t lp lai nhiu ln. Cac trinh t nay phn b trn toan b b gen. Chung co th la nhng trinh t khng ma ha ma cung co th la nhng trinh t ma ha cho rRNA, tRNA va RNA 5S. - Cac trinh t duy nht: la cac gen ma ha cho cac protein, co trinh t c trng cho tng gen. Mt c im cua phn t DNA co y nghia rt quan trong c s dung vao phng phap lai phn t. o la kha nng bin tinh va hi tinh. Bin tinh la hin tng hai si n cua phn t DNA tach ri nhau khi cac lin kt hydro gia cac base b sung nm trn hai si bi t do cac tac nhn ha hoc (dung dich kim, formamide, urea) hay do tac nhn vt ly (nhit). Sau o, nu iu chinh nhit va nng mui thich hp, cac si n co th bt cp tr lai theo nguyn tc b sung, hinh thanh phn t DNA ban u, o la s hi tinh.

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2.4.2 Ribonucleic acid Phn t RNA co cu tao tng t DNA vi ba im khac bit sau: - Phn t RNA la chui n. - ng pentose cua phn t RNA la ribose thay vi deoxyribose. - Thymine, mt trong bn loai base hinh thanh nn phn t DNA, c thay th bng uracil trong phn t RNA. Cu truc va chc nng cua RNA co s bin i ro rt. V c ban RNA chi la cht mang thng tin di truyn virus, sau o ngi ta chng minh rng chng khng nhng ong vai tro c ban vic chuyn thng tin di truyn ma con co vai tro cu truc khi tao nn phc h RNA-protein. Theo mt ly thuyt tin ha ma ai din la Manfred Eigen, RNA la cht mang thng tin di truyn, thanh vin trung gian cua s biu hin gene, thanh phn cu tao va la cht xuc tac. Nhom OH ru vi tri th hai cua ribose cn thit cho a chc nng lam nhiu loan s tao thanh si kep, qua o lam tng khng bn vng cua lin kt phosphodiester. Trong t bao co ba loai RNA chinh, co cac chc nng khac nhau: 2.4.2.1 Cac RNA thng tin (mRNA)

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mRNA la ban sao cua nhng trinh t nht inh trn phn t DNA, co vai tro trung tm la chuyn thng tin ma ha trn phn t DNA n b may giai ma thanh phn t protein tng ng. Cac RNA co cu truc a dang, kich thc nho hn so vi DNA vi chi cha thng tin ma ha cho mt hoc vai protein va chi chim khoang 2-5% tng s RNA trong t bao. Escherichia coli, kich thc trung binh cua mt phn t mRNA khoang 1,2 kb. 2.4.2.2 RNA vn chuyn (tRNA) tRNA lam nhim vu vn chuyn cac amino acid hoat ha n ribosome tng hp protein t cac mRNA tng ng. Co it nht mt loai tRNA cho mt loai amino acid. tRNA vn chuyn cha khoang 75 nucleotide (co trong lng khoang 25 kDa), la phn t RNA nho nht. Cac tRNA co cu truc dang co ba l (hnh 2.7). Cu truc nay c n inh nh cac lin kt b sung hin din nhiu vung cua phn t tRNA. Hai vi tri khng co lin kt b sung ong vai tro c bit quan trong i vi chc nng cua tRNA: - Trinh t anticodon gm ba nucleotide. - Trinh t CCA, co kha nng lin kt cng ha tri vi mt amino acid c trng. 2.4.2.3 RNA ribosome (rRNA) rRNA la thanh phn c ban cua ribosome, va ong vai tro xuc tac va cu truc trong s tng hp protein. Ty theo h s lng, rRNA c chia thanh nhiu loai: eukaryote co rRNA 28S, 18S, 5,8S va 5S, con cac rRNA E. coli co ba loai: 23S, 16S va 5S. rRNA chim nhiu nht trong ba loai RNA (80% tng s RNA t bao), tip n la tRNA va mRNA chi chim 5%. Ribosome cua moi t bao u gm mt tiu n vi nho va mt tiu n vi ln. Mi tiu n vi co mang nhiu protein va rRNA co kich thc khac nhau.

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34

2.4.2.4 Mt s loi RNA khc T bao sinh vt nhn chun con 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 nho tham gia vao ghep ni cac exon. - hn RNA( heterogenous nuclear) l nhng RNA khng ng nht nhn t bo. - scRNA (small cytoplasmic) l nhng RNA nh t bo cht. Nhn chung tt ca RNA trong t bao u c tng hp nh enzyme RNA polymerase. Enzyme nay oi hoi nhng thanh phn sau y: - Mt khun mu, thng la DNA si i. - Tin cht hoat hoa: Bn ribonucleoside triphosphate: ATP, GTP, UTP va CTP. Sinh tng hp RNA ging DNA mt s im, th nht hng tng hp la 5 3, th hai la c ch keo dai ging nhau: nhom 3-OH u cui cua chui tng hp la vi tri gn kt cua nucleoside triphosphate tip theo. Th ba, s tng hp xay ra do thy phn pyrophosphate. Tuy nhin, khac vi DNA la RNA khng oi hoi mi (primer). Ngoai ra RNA polymerase khng co hoat tinh nuclease sa cha khi cac nucleotide bi gn nhm. Ca ba loai RNA trong t bao c tng hp trong E. coli nh mt loai RNA polymerase. ng vt co vu, cac RNA khac nhau c tng hp bng cac loai RNA polymerase khac nhau. Ti liu tham kho Ti liu ting Vit Trn Th n, i Duy Ban, Nguyn Hu Chn, nh H, L c Trnh, 1980. Ho sinh hc. NXB Y hc, H Ni. Phm Th Trn chu, Trn Th ng, 1999. Ho sinh hc. NXB Gio dc, H Ni. H Hunh Thu Dng, 1998. Sinh hc phn t. NXB Gio dc, H Ni. Phm Thnh H, 2001. Di truyn hc, (ti bn ln th 3). NXB Gio dc, thnh ph H Ch Minh. Ti liu ting Anh Lodish, H., 2003. Molecular Cell Biology. 5th Ed. E-book.

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Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA.

Chng 3 HA HC CARBOHYDRATE
L Vn An, H Trung Thng
Carbohydrate l nhng phn t sinh hc c s lng ln nht trn tri t. Mt s loi carbohydrate nh tinh bt v ng l ngun thc n ch yu ca hu ht mi ni trn th gii v qu trnh oxy ha carbohydrate l con ng khai thc nng lng trung tm ca hu ht cc t bo khng quang hp. thnh t bo vi khun, thnh t bo thc vt v t bo ca m lin kt ng vt, cc carbohydrate khng ha tan ng vai tr l yu t cu trc hoc bo v. Mt s carbohydrate ng vai tr l cht nhn bi trn cc khp xng hoc tham gia vo c ch nhn bit hoc kt dnh gia cc t bo. Mt s carbohydrate phc tp khc lin kt cht ch vi protein hoc lipid hot ng nh mt tn hiu v c gi l glycoconjugate. Nhiu loi carbohydrate c cng thc cu to l (CH2O)n, mt s loi carbohydrate cn cha nit, phosphor, lu hunh. Da vo kch thc phn t ngi ta chia carbohydrate thnh ba nhm: monosaccharide, oligosaccharide v polysaccharide.

3.1. Monosaccharide 3.1.1 nh ngha

Monosaccharide hay cn gi l ng n l nhng carbohydrate n gin nht, c ti thiu 2 nhm hydroxyl (-OH) v c th tn ti di mt trong hai dng aldehyde v ketone. Ty theo s lng carbon m cc monosaccharide c th c gi l triose (3 carbon), tetrose (4 carbon), pentose (5 carbon), hexose (6 carbon),

Monosaccharide c th tn ti dng mch thng hoc mch vng. Khi dng mch thng, mt nguyn t carbon c lin kt i vi mt nguyn t oxy to thnh nhm carbonyl (-C=O) cn mi mt nguyn t carbon cn li ca phn t monosaccharide c mt nhm hydroxyl (-OH). Nu nhm carbonyl nm cui cng ca mch carbon th s hnh thnh nhm aldehyde (-CHO), trong trng hp ny monosaccharide l mt aldose. Nu nhm carbonyl nm bt c mt v tr no khc trn chui mch th s to thnh nhm ketone, trong trng hp ny monosaccharide l mt ketose (hnh 3.1).

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3.1.2 ng phn lp th
Tt c cc monosaccharide tr dihydroxyacetone u c t nht mt nguyn t carbon bt i xng, do c th to thnh cc dng ng phn lp th dng D hoc dng L (hnh 3.2). Aldose n gin nht l glyceraldehyde c mt nguyn t carbon bt i xng, do c hai dng ng phn lp th. Monosaccharide c n nguyn t carbon bt i xng s c 2n ng phn lp th.

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3.1.3 Cu trc dng vng

n gin, cu to ca cc aldose v ketose c th hin di dng mch thng. Tuy vy, trong thc t tt c cc monosaccharide c s nguyn t carbon 5 tn ti ch yu di dng mch vng trong nhm carbonyl thc hin lin kt cng ha tr vi mt nguyn t oxy ca mt nhm hydroxyl trong phn t monosaccharide. S hnh thnh cu trc dng vng l kt qu ca phn ng gia alcohol vi aldehyde hoc ketone to thnh dn xut gi l hemiacetal hoc hemiketal (hnh 3.4). Cc cht ny c thm mt nguyn t carbon bt i xng, do c hai ng phn lp th c k hiu l v (hnh 3.5 v 3.6).

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3.1.4 Cc dn xut ca hexose

Ngoi cc dng hexose n gin nh glucose, galactose v mannose, trong c th sinh vt cn c nhiu dn xut khc ca ng (hnh 3.7). Cc dn xut ny c hnh thnh do nhm hydroxyl ca cc monosaccharide b thay th bng mt nhm khc hoc mt nguyn t carbon b oxy ha thnh nhm carboxyl, v d glucosamine, galactosamine, mannosamine. N-acetylglucosamine l mt phn ca nhiu loi polymer cu trc bao gm c thnh t bo vi khun. Thnh t bo vi khun cn c thm mt dn xut ca glucosamine l N-acetylmuramic acid. S thay th nhm hydroxyl v tr C6 ca Lgalactose v L-mannose bng mt nguyn t hydro s to thnh L-fucose v L-rhamnose, cc loi ng kh oxy ny c tm thy trong polysaccharide ca thc vt v trong cu to ca cc oligosaccharide ca cc glycoprotein v glycolipid. S oxy ha carbon carbonyl (aldehyde) ca glucose n carboxyl s to thnh gluconic acid, cn ca cc aldose khc s to thnh aldonic acid. S oxy ha carbon tn cng ca glucose, galactose, mannose s to thnh cc uronic acid tng ng: glucuronic, galacturonic, mannuronic acid. Ngoi ra, mt dn xut ca N-acetylmannosamine l N-acetylneuraminic acid (cn gi l sialic acid) l mt thnh phn ca nhiu loi glycoprotein v glycolipid trong m bo ng vt.

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3.1.5 Mt s monosaccharide quan trng

3.1.5.1 Triose: D-glyceraldehyde (aldotriose) v dihydroxyacetone (ketotriose) l nhng sn phm chuyn ha trung gian ca carbohydrate. 3.1.5.2 Pentose: Ribose v deoxyribose (b kh oxy v tr C2) l hai pentose quan trng tham gia cu to nucleic acid. 3.1.5.3 Hexose: l loi ng gp nhiu nht trong c th sinh vt so vi cc monosaccharide khc. - D-glucose: l loi monosaccharide ph bin nht trong t nhin, l thnh phn cu to nhiu loi oligosaccharide v polysaccharide nh saccharose, tinh bt, glycogen, cellulose. Glucose c mt khp cc dch v m bo trong c th ng vt, l c cht chuyn ha ch yu ca carbohydrate trong c th ngi v ng vt. - D-fructose: l mt hexose quan trng, c nhiu trong qu chn. Cng vi glucose, fructose l sn phm trung gian ca nhiu con ng trao i cht quan trng nh ng phn, chu trnh pentose, Trong c th, fructose thng tn ti di dng -Dfructofuranose.

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- D-galactose: l thnh phn cu to ca lactose (ng sa).

3.2 Oligosaccharide 3.2.1 nh ngha

Oligosaccharide l carbohydrate c t 2 n 20 gc monosaccharide, cc gc monosaccharide lin kt vi nhau bng lin kt glycoside. Oligosaccharide ph bin nht l disaccharide c hai gc monosaccharide m in hnh l sucrose (saccharose hay ng ma).

3.2.2 Mt s oligosaccharide ph bin

3.2.2.1 Maltose: c nhiu trong mm la, ko mch nha, ... Maltose t tn ti dng t do m ch yu c to thnh khi thy phn tinh bt bi enzyme amylase v khi ht ny mm. Maltose c cu to t hai phn t D-glucose ni vi nhau bng lin kt glycoside gia C1 ca mt gc glucose vi C4 ca mt gc glucose cn li. Trong cu to ca maltose cn c mt nhm -OH ti v tr C1 trng thi t do, do maltose l mt ng kh. 3.2.2.2 Lactose: cn gi l ng sa (c trong sa ca ng vt v ngi), l disaccharide c to thnh t mt phn t D-galactose v mt phn t D-glucose. Lactose l mt disaccharide c tnh kh do mt nhm -OH v tr C1 ca gc ng glucose cn trng thi t do. 3.2.2.3 Saccharose: cn c gi l sucrose hay ng ma, c nhiu trong c ci ng, thn cy ma v nhiu loi c qu khc. Saccharose khng c to ra trong c th ng vt. Khi thy phn saccharose bng acid hoc enzyme saccharase s thu c -Dglucopyranose v -D-fructofuranose. Hai phn t ng n ny lin kt vi nhau bng lin kt 1,2-glycoside (hnh 3.8). Khc vi maltose v lactose, saccharose khng c tnh kh. Saccharose l mt sn phm trung gian quan trng ca quang hp, trong c th thc vt, saccharose l dng vn chuyn ng t l v cc b phn khc ca c th thc vt.

3.3 Polysaccharide 3.3.1 Khi nim

Trong t nhin hu ht carbohydrate tn ti dng polysaccharide. Polysaccharide l carbohydrate cha trn 20 gc monosaccharide, mt s polysaccharide cha ti hng trm 41

hoc hng ngn gc monosaccharide. Polysaccharide cn gi l glycan, chng khc nhau v n v cu to, di ca mch v phn nhnh. Polysaccharide khng c v ngt, khng c kh nng to thnh dung dch tht khi tan trong nc, nu tan trong nc chng ch to ra dung dch keo. Trong c th sinh vt, polysaccharide ch yu gi vai tr cu trc hoc d tr. Khc vi protein, polysaccharide khng c phn t lng c trng. S khc nhau ny l do s khc nhau trong c ch sinh tng hp hai loi polymer ny. Protein c tng hp da trn khun mu (mRNA) vi trnh t v di xc nh. Sinh tng hp polysaccharide khng theo khun mu, chng trnh sinh tng hp polysaccharide do bn cht ca enzyme xc tc v khng c im kt thc c trng trong qu trnh tng hp.

3.3.2 Phn loi polysaccharide

Polysaccharide c chia thnh hai nhm: polysaccharide thun (homopolysaccharide) v polysaccharide tp (heteropolyssaccharide). Ngoi ra, theo quan im dinh dng ng vt, ngi ta cn chia polysaccharide thnh hai nhm: tinh bt v polysaccharide phi tinh bt (nonstarch polysaccharide: NSP).

3.3.2.1 Polysaccharide thun Polysaccharide thun ch cha mt loi n v cu to duy nht (hnh 3.9). Mt s polysaccharide thun ng vai tr d tr cc gc ng n cung cp nhin liu cho c th. Tinh bt v glycogen l nhng polysaccharide thun thuc nhm ny. Mt s polysaccharide thun khc nh cellulose v chitin ng vai tr l yu t cu trc ca thnh t bo thc vt v v cng ca nhiu loi ng vt nh cn trng, gip xc.

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- Tinh bt: c to ra t thc vt, l ngun nng lng d tr quan trng ca thc vt v l thc n quan trng ca ng vt v ngi. Tinh bt c nhiu ht, c, qu. Trong cc m thc vt, tinh bt thng tn ti dng ht c hnh dng kch thc v cu to khc nhau ty theo ngun gc. Cc ht tinh bt khng tan trong nc lnh. Khi un nng v khuy u trong nc, tinh bt n ra v khuych tn thnh dung dch keo gi l h tinh bt, h tinh bt tc dng vi iod cho mu xanh tm.

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Tinh bt c hai thnh phn l amylose (chim khong 20%) v amylopectin (chim khong 80%) v khng c tnh kh. Hai thnh phn ny c mt s c im cu to v tnh cht khc nhau, tuy vy chng u c cu to t cc n phn l -D-glucose. Amylose tan c trong nc, cho mu xanh vi iod, c cu to mch thng do cc n v -Dglucose lin kt vi nhau bng lin kt ( 1 4) glycoside (hnh 3.10 v 3.12). Khi tan trong nc, mch amylose c th b xon li. Amylopectin khng tan trong nc, to thnh tnh keo ca h tinh bt v cho mu tm vi iod. Amylopectin c to thnh t cc n v -D-glucose ni vi nhau bng lin kt ( 1 4) v ti im phn nhnh l ( 1 6). T 24 n 30 gc glucose li c mt im phn nhnh. Tinh bt b thy phn bi acid v c nhit cao hoc bi enzyme trong ng tiu ha to thnh cc sn phm trung gian c tn gi chung l dextrin cho mu khc nhau vi iod, sau to thnh maltose v cui cng l glucose: h tinh bt (mu xanh tm vi iod) Amylodextrin (mu tm vi iod) Erythrodextrin (tm nht n nu vi iod) Acrodextrin (mu nu vi iod) Maltose (khng cho mu vi iod) Glucose (khng cho mu vi iod).

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- Glycogen: l polysaccharide d tr ca cc t bo ng vt. Cng ging nh amylopectin, glycogen l mt polymer c to thnh do cc n v glucose ni vi nhau bng lin kt ( 1 4) v lin kt ( 1 6) ti im phn nhnh. Tuy vy, s lng nhnh trong phn t glycogen ln hn nhiu so vi s lng nhnh trong phn t amylopectin, c 8 n 10 gc glucose li c mt im phn nhnh. Glycogen c nhiu trong gan v c vn ca ng vt. Trong m gan, glycogen c th chim n 7% trong lng ca gan ti v ht glycogen c kch thc ln do s to thnh cc b glycogen t nhng ht c kch thc nh hn. Cui mi nhnh c mt u khng c tnh kh, do s lng u khng c tnh kh ca glycogen bng s lng nhnh c trong phn t. Khi glycogen c s dng nh mt ngun nng lng, cc n v glucose c tch dn ra khi phn t glycogen t u khng c tnh kh. Enzyme phn gii glycogen ch c th xc tc t u khng c tnh kh, nhng c th thc hin vic xc tc ng thi trn nhiu nhnh khc nhau, do m bo tc phn gii nhanh glycogen thnh monosaccharide, cc thng tin chi tit v s phn gii glycogen s c trnh by trong chng chuyn ha carbohydrate. Vic d tr glucose di dng glycogen c ngha quan trng i vi tnh thm thu cng nh i vi kha cnh nng lng. Ngi ta tnh ton rng t bo gan d tr glycogen vi s lng tng ng nng glucose 0,4M. Glycogen l dng khng ha tan v ch ng gp mt phn rt nh n kh nng thm thu (osmolarity) ca t bo cht (ch khong 0,01 M). Nu nng glucose trong t bo cht l 0,4M th kh nng thm thu s tng cao n mc e da s sng ca t bo, s dn n s di chuyn ca nc t ngoi vo t bo cht v t bo c th b v. Ngoi ra, vi nng glucose ni bo l 0,4M trong lc nng glucose ngoi bo ch khong 5mM, s thay i nng lng t do trong qu trnh thu nhn glucose ngc gradient nng qu ln l rt ln v qu trnh thu nhn glucose t ngoi vo t bo c th b ngn tr.

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- Dextran: l polysaccharide ca vi khun v nm men c to thnh t cc n v D-glucose ni vi nhau bng lin kt ( 1 6). Tt c cc loi dextran u c mch nhnh lin kt ( 1 3), mt s dextran c mch nhnh lin kt ( 1 2) v ( 1 4).

- Cellulose: l cht c cu trc dng si v khng tan trong nc, c tm thy trong thnh t bo thc vt. Cellulose l thnh phn chnh ca g, bng cha cellulose gn nh nguyn cht. Cng ging nh amylose, cellulose l polysaccharide thun khng phn nhnh, c cu to t 10.000 n 15.000 n v D-glucose. Tuy vy c s khc nhau rt quan trng: cellulose c to thnh t cc n v -D-glucose trong khi amylose, amylopectin v glycogen c to thnh t cc n v -D-glucose. Cc n v glucose trong cellulose ni vi nhau bng lin kt ( 1 4) glycoside, ngc li vi amylose, amylopectin v glycogen l lin kt ( 1 4) glycoside. S khc nhau ny dn n s khc nhau rt ln v mt cu trc v tnh cht vt l gia cellulose v amylose. Nhiu loi ng vt khng th s dng cellulose lm ngun nhin liu v khng c enzyme thy phn lin kt (1 4) glycoside. ng vt nhai li v mt s loi ng vt khc nh th, nga c kh nng s dng cellulose lm ngun nhin liu chnh do chng c h vi sinh vt phong ph trong ng tiu ha, nhng vi sinh vt ny tit ra enzyme phn gii cellulose. Phng php Weende (phn tch gn ng / approximate analysis) dn n thnh phn x th khi phn tch thnh phn dinh dng ca thc n c ngun gc thc vt. Thnh phn ch yu ca x th l cellulose, pentosane v lignin (dn xut ca phenylpropan). - Chitin: l mt polysaccharide thun c cu to t cc n v Nacetylglucosamine ni vi nhau bng lin kt (1 4) glycoside. S khc nhau duy nht v mt ha hc gia chitin v cellulose l s thay th nhm hydroxyl v tr C2 bng mt nhm c acetyl ha (CH3-CO-NH-) (hnh 3.17). Chitin c dng si ging nh cellulose v ng vt cng khng tiu ha c chitin. Chitin l thnh phn c bn ca lp v cng ca nhiu loi sinh vt, l polysaccharide ph bin trong t nhin ch sau cellulose.

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- Inulin: l mt polysaccharide d tr ca thc vt, c cu to t cc n v fructose. Phn t lng ca inulin thp v n ch c khong 30 gc fructose, do polysaccharide ny d dng ha tan trong nc. - Glucane: c cu to tng t cellulose, chng c to thnh t cc n v glucose. Tuy vy, ngoi lin kt (1 4) cn c lin kt (1 3) glycoside vi t l gia hai loi lin kt ny l t 3-4/1. 3.3.2.2 Polysaccharide tp Polysaccharide tp l nhng i phn t c cu trc phc tp, c to ra t nhiu loi monosaccharide khc nhau, dn xut ca cc monosaccharide hay cn c thm nhng cht khc. Polysaccharide c nhiu trong t nhin v c nhiu vai tr sinh hc quan trng. m bo ng vt, nhiu loi polysaccharide tp c trong khong gia cc t bo, chng to thnh matrix gia cc t bo, t lm cho cc t bo c th lin kt c vi nhau, gip t bo gi c hnh dng, bo v v tr gip cho t bo, m, v c quan trong nhiu hot ng khc nhau. - Hemicellulose: Hemicellulose l mt nhm cht gm nhiu loi polysaccharide ca thc vt, cng vi cellulose to nn thnh t bo thc vt, c nhiu trong cc b phn ca thc vt nh rm r, b ng, tru, v cng ca ht, cc b phn ha g ca thc vt. V mt ha hc, hemicellulose c to thnh t polysaccharide ca xylose (xylan), arabinose (araban), mannose (mannan), galactose (galactan). - Pentosane: Pentosane l polysaccharide c cu to t arabinose v xylose, l mt b phn cu to ca hemicellulose. Hm lng pentosane c th chim n 20% vt cht kh ca c. - Pectine: Pectine l polysaccharide ca thc vt, c cu to t galacturonic acid. Mt s nhm carboxyl ca cc gc galacturonic acid c ester ha vi methanol. Pectine c nhiu trong c, qu, m mm nh l cy.

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- Polysaccharide tp ca thnh t bo vi khun (peptidoglycan) v to (agar): Thnh t bo vi khun cha polysaccharide tp, polysaccharide ny c to thnh do s thay i lun phin gia cc gc N-acetylglucosamine (GlcNAc) v N-acetylmuramic acid (Mur2Ac), cc gc ny ni vi nhau bng lin kt (1 4) (hnh 3.18). Cc phn t ny nm cnh nhau v c gn vi nhau bng lin kt ngang (cross-link) thng qua mt on peptide mch ngn, cu trc chnh xc ca on peptide ny ph thuc vo tng loi vi khun. Cc on peptide ny gip cho cc chui polysaccharide to thnh mt lp v chc bao xung quanh t bo v ngn chn s trng n dn n s tan v t bo do s ht nc t ngoi mi trng vo t bo. Lysozyme tiu dit vi khun bng cch thy phn lin kt (1 4) gia N-acetylglucosamine v N-acetylmuramic acid. Mt v mt s loi virus ca vi khun cng c kh nng tit lysozyme. Penicillin v cc khng sinh lin quan khc tiu dit vi khun bng cch ngn cn s tng hp on peptide mch ngn lm nhim v ni cc chui polysaccharide, do lm cho thnh t bo vi khun b yu v c th b tan v. Mt s loi to v rau cu c thnh t bo cha agar, l mt polysaccharide tp. Agar c hai thnh phn: mt thnh phn khng phn nhnh l agarose v mt thnh phn phn nhnh l agaropectin (hnh 3.19).

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- Polysaccharide tp ca matrix ngoi bo: Khong gia cc t bo trong cc m bo ca ng vt a bo tn ti mt cht c qunh tng t gel gi l matrix ngoi bo. Matrix ngoi bo lin kt cc t bo vi nhau, to ra cc l cho s khuych tn ca cc cht dinh dng v oxy i n tt c cc t bo. Matrix ngoi bo c to ra do s ci vo nhau ca heteropolysaccharide v cc protein hnh si nh collagen, elastin, fibronectin v laminin. Cc polysaccharide tp ny l cc glycosaminoglycan (mucopolysaccharide). Cc glycosaminoglycan c cu to t cc n v disaccharide, mi disaccharide c mt monosaccharide l N-acetylglucosamine hoc l N-acetylgalactosamine, trong hu ht cc trng hp mt monosaccharide cn li l uronic acid m thng thng l D-glucuronic hoc L-iduronic acid. Trong mt s glycosaminoglycan, mt hoc nhiu nhm hydroxyl ca ng amin c ester ha vi sulfate. Glycosaminoglycan c gn vo cc phn t protein ngoi bo to thnh proteoglycan (hnh 3.20). C th k mt s glycosaminoglycan quan trng nh sau:

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+ Hyaluronic acid: c cu to t N-acetylglucosamine v D-glucuronic acid. Hyaluronic acid to thnh dung dch trong v rt nht c tc dng lm trn cc khp xng v lm cho thy tinh dch ca mt ng vt c xng sng c nh thch. Hyaluronic acid cn l thnh phn quan trng ca matrix ngoi bo ca sn v gn, lm tng tnh co gin v chc ca cc t bo ny v hyaluronic acid tng tc mnh vi cc thnh phn khc trong matrix ngoi bo. Hyaluronidase l enzyme c tit ra bi mt s vi khun gy bnh, enzyme ny c th thy phn lin kt glycoside ca acid hyaluronic lm cho cc m bo d b vi khun xm nhp. nhiu loi sinh vt, mt enzyme tng t c trong tinh dch c th xc tc cho qu trnh thy phn glycosaminoglycan bao xung quanh t bo trng gip tinh trng xm nhp vo bn trong t bo cht. + Chondroitin sulfate: l mt glycosaminoglycan c nhiu trong t chc sn, t chc lin kt (gn, da, van tim v thnh ng mch).

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+ Heparin: l cht chng ng mu do t bo ln (mast cell, mt loi t bo bach cu) tit ra v c a vo mu. Heparin chng ng mu bng cch gn vo phn t protein antithrombin. S gn kt ny ca heparin to ra s gn kt ca antithrombin vo thrombin v c ch enzyme protease ny. + Keratan sulfate: l glycosaminoglycan khng c uronic acid. Keratan sulfate c trong gic mc, sn, xng, sng, tc, mng, vut. - Glycoprotein v glycolipid: + Glycoprotein: c mt hoc nhiu chui oligosaccharide lin kt cng ha tr vi mt phn t protein (hnh 3.21). Glycoprotein c tm thy trn b mt ngoi ca mng nguyn sinh cht, trong matrix ngoi bo v trong mu. trong t bo, glycoprotein c trong mt s bo quan nh phc h golgi, cc ht thi tit v lysosome. T l oligosaccharide ca glycoprotein t hn rt nhiu so vi cc chui glycosaminoglycan trong proteoglycan. + Glycolipid: l lipid mng trong cc phn t oligosaccharide nm u a nc (hnh 3.22). Ging nh glycoprotein, oligosaccharide hot ng nh l v tr nhn bit ca cc protein lin kt vi carbohydrate.

Ti liu tham kho

51

Ti liu ting Vit Trnh Vn Bo, Phan Th Hoan, 2002. Cht sng. Trong: Cc nguyn l sinh hc; Bin son bi Trnh Vn Bo (ch bin), Phan Th Hoan, Trn Th Thanh Hng, Trn Th Lin, Trn c Phn, Phm c Phng, Nguyn Vn Rc, Nguyn Th Trang; trang 11-29. NXB Y hc, H Ni. Phm Thi Trn Chu, Trn Th ng, 1999. Ha sinh hc. NXB Gio dc, trang 62-76. o Kim Chi, 2004. Glucid. Trong: Ha sinh hc; bin son bi Nguyn Xun Thng (ch bin), o Kim Chi, Phm Quang Tng, Nguyn Vn ng; trang 10-28. NXB Y hc, H Ni. Nguyn Th Ho, 2005. Ha hc glucid. Trong: Ha sinh y hc; bin son bi nh H (ch bin), ng Th Hoi An, Nguyn Th Ho, nh H, Phm Th Mai, Trn Thanh Lan Phng, Th Thanh Thy, L Xun Trng; trang 17-34. NXB Y hc, Tp. H Ch Minh. Phan Hi Nam, 2000. Ha sinh glucid. Trong: Ha sinh y hc; bin son bi Hong Quang (ch bin), Nguyn nh , Trng Th Minh c, Bch Vng Hi, Phan Hi Nam, on Trng Ph; trang 87-112. NXB Qun i nhn dn, H Ni. L Th Kim Thu, 2002. Ha sinh lm sng. NXB Y hc, H Ni. Ti liu Ting c Bergmann, H. und Flachowsky, G., 1995. Naturwissenschaftliche Grundlagen und globales Futterpotential. In: Nutztierernaehrung (Abel Hj., Flachowsky, G., Jeroch, H., Molnar S., Hrsg.), pp. 39-50. Gustav Fischer Verlag, Jena-Stuttgart, Gerrmany. Engelhardt W. v., Breves G. (Hrsg.), 1999. Physiologie der Haustiere. Enke Verlag, Stuttgart, Germany. Jeroch H., Drochner W., Simon, O., 1999. Ernaehrung landwirt-schaftlicher Nutztiere. Verlag Eugen Ulmer Stuttgart, Germany. Kirchgessner, M., 1997. Tiernaehrung, 10. neubearbeitete Auflage, DLG-Verlag, Frankfurt am Main, Germany. Loeffler G., 1998. Kohlenhydrate. In: Biochemie und Pathobiochemie, 6. Auflage (Loeffler G., Petrides P. E., Hrsg.), pp. 121-132. Springer Verlag, Berlin, Germany. Weissbach F., 1993. Wertbestimmende Bestandteile der Futtermittel. In: Futtermittelkunde (Jeroch H., Flachowsky G., Weissbach F., Hrsg.), pp. 18-32. Gustav Fischer Verlag Jena-Stuttgart, Germany. Ti liu ting Anh McDonald P., Edwards R. A., Greenhalgh J. F. D., Morgan C. A., 1995. Animal Nutrition, fifth edition. Longman Scientific & Technical, New York - USA, pp. 9-27. Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA.

Chng 4

HA HC LIPID
L Vn An, H Trung Thng

52

Lipid l mt nhm cht hu c a dng v mt ha hc v ph bin trong t nhin. c im quan trng ca lipid l khng tan hoc rt t tan trong nc (l dung mi phn cc) nhng li d tan trong cc dung mi hu c. Trong nhng iu kin nht nh, lipid tan trong nc to thnh dng nh tng (l loi dung dch c kch thc ht phn tn tng i ln: > 10-5 cm).

4.1 Thnh phn cu to ca lipid

Trong phn ln cc cht lipid c cha hai thnh phn chnh l alcol v acid bo. Alcol v acid bo c ni vi nhau bng lin kt ester (alcol-acid bo) hoc bng lin kt amid (aminoalcol-acid bo). Ngoi ra, lipid cn c th kt hp vi carbohydrate (to thnh glycolipid) c vai tr cu trc (thnh phn cu to mng, ) hoc kt hp vi protein (to thnh lipoprotein) gi vai tr quan trng trong vic ha tan v vn chuyn lipid trong mu, gip hp thu vitamin tan trong lipid,

4.1.1 Acid bo
4.1.1.1 c im chung v danh php Acid bo l nhng acid hu c monocarboxyl c cng thc chung l R-COOH. Cc acid bo c chui hydrocarbon bin ng t 4 n 36 carbon (C4 n C36). Cc acid bo gp trong thin nhin thng c s carbon chn, phn ln c t 14 n 22 carbon, thng gp nht l cc acid bo c 16, 18 v 20 carbon. Tuy nhin trong thin nhin cng c th gp cc acid bo mch ngn (trong sa) hoc cc acid bo c s carbon l (11, 13, 15, 17 hoc 19 carbon, v d undecylenic acid c 11 carbon do tuyn nhn da u tit ra tui dy th c kh nng tr nm tc). C nhng acid bo mch thng v no (khng c lin kt i), tuy vy cng c nhng acid bo khng no (c mt hoc nhiu lin kt i). Mt s acid bo c vng 3 nguyn t carbon, nhm hydroxyl hoc nhm methyl. Cc acid bo c th c gi theo tn thng thng hoc tn h thng, v d acid bo c cng thc C15H31COOH c tn thng thng l palmitic acid v tn h thng l hexadecanoic acid. Cch nh s carbon ca acid bo nh sau:

4.1.1.2 Phn loi acid bo theo mch carbon Acid bo c bn c phn loi theo chiu di v s lng lin kt i, gc R (mch thng, mch nhnh, mch vng). - Acid bo mch ngn v mch trung bnh

53

Cc acid bo thuc nhm ny c t 4-14 carbon. Tt c cc acid bo thuc nhm ny u l acid bo no, khng tham gia cu trc mng sinh hc, chc nng ca cc acid bo nhm ny l to nng lng.
Bng 4.1: Cu to, c im v danh php ca cc acid bo mch ngn v acid bo mch trung bnh Khung carbon 4:0 6:0 8:0 10:0 12:0 14:0 Cu to ha hc CH3(CH2)2COOH CH3(CH2)4COOH CH3(CH2)6COOH CH3(CH2)8COOH CH3(CH2)10COOH Tn h thng n-Butanoic acid n-Hexanoic acid n-Octanoic acid n-Decanoic acid n-Dodecanoic acid Tn thng gi im nng chy (oC) Butyric acid Caproic acid Caprylic acid Capric acid Lauric acid - 7,9 - 3,9 16,3 31,3 44,2 53,9

CH3(CH2)12COOH n-Tetradecanoic acid Myristic acid

- Acid bo mch di
Bng 4.2: Cu to, c im v danh php ca mt s acid bo mch di
Khung carbon 16:0 18:0 20:0 16:1 ( ) 18:1 (9) 18:2 (9,12) 18:3 (9,12,15) 20:4 (5,8,11,14)
9

Cu to ha hc CH3(CH2)14COOH CH3(CH2)16COOH CH3(CH2)18COOH CH3(CH2)5CH= CH(CH2)7COOH CH3(CH2)7CH= CH(CH2)7COOH CH3(CH2)4CH=CHCH2CH= CH(CH2)7COOH CH3CH2CH=CHCH2CH= CHCH2CH=CH(CH2)7COOH CH3(CH2)4CH=CHCH2CH= CHCH2CH=CHCH2CH= CH(CH2)3COOH

Tn h thng n-Hexadecanoic acid n-Octadecanoic acid n-Eicosanoic acid

Tn thng gi im nng (acid + ) chy (oC) Palmitic acid Stearic acid Arachidic acid 63,1 69,6 76,5 1-0,5 13,4 1-5 - 11 - 49,5

cis-9-Hexadecenoic acid Palmitoleic acid cis-9-Octadecenoic acid cis-,cis-9,12Octadecadienoic acid cis-,cis-,cis-9,12,15 Octadecatrienoic acid cis-,cis-,cis-,cis-5,8,11,14 Eicosatetraenoic acid Oleic acid Linoleic acid Linolenic acid Arachidonic acid

Acid bo mch di l nhng acid bo c t 16 carbon tr ln, thng gp nht l cc acid bo c 16, 18 v 20 carbon, c th no (bo ha) hoc khng no (cha bo ha). Acid bo mch di c nhiu vai tr sinh hc quan trng nh vai tr to nng lng, d tr nng lng (l thnh phn ca triglycerid), lipid mng, vt liu cho cc qu trnh sinh tng hp mt s hp cht c vai tr sinh hc quan trng khc nh prostaglandin, leukotrien, . Trong s cc loi acid bo ny c nhng acid bo c th khng tng hp c m phi c cung cp t thc n, cc acid bo ny c gi l acid bo thit yu (linoleic, linolenic v arachidonic acid). Cc loi du thc vt nh du u nnh, du m, du lc 54

cha nhiu acid bo thit yu. Trong t bo, cc acid bo t do tn ti dng mui, do tn acid bo c th c gi theo tn mui ca n v d palmitate, palmitoleate, stearate. - Acid bo mch rt di Cc acid bo nhm ny c t 22 nguyn t carbon tr ln. Do cu trc mch carbon rt di v k nc cho nn cc acid bo nhm ny rt kh tan. Cc acid bo mch rt di tham gia cu to mng sinh hc, v d docosahexaenoic acid (DHA) c trong phospholipid ca mt s m nh vng mc.
Bng 4.3: Cu to, c im v danh php ca mt s acid bo mch rt di
Khung carbon Cu to ha hc Tn h thng Tn thng gi im nng chy (oC) 80 86,0 87,7 < - 78

22:0 24:0 26:0 22:4 (

4,8,12,15,19

CH3(CH2)20COOH CH3(CH2)22COOH CH3(CH2)24COOH C22H34O2 )

n-Docosanoic acid n-Tetracosanoic acid n-Hexacosanoic acid Docosapentaenoic acid

Behenic acid Lignoceric acid Cerotic acid Clupanodonic acid

Bng 4.4: Cc h acid bo khng no H acid bo khng no 7 hay n-7 9 hay n-9 6 hay n-6 3 hay n-3 Khung carbon 16:1 (9) 18:1(9) 18:2(9,12) 18:3(9,12,15) Tn thng gi Palmitoleic acid Oleic acid Linoleic acid Linolenic acid Cu to ha hc CH3(CH2)5CH= CH(CH2)7COOH CH3(CH2)7CH= CH(CH2)7COOH CH3(CH2)4CH=CHCH2CH= CH(CH2)7COOH CH3CH2CH=CHCH2CH= CHCH2CH=CH(CH2)7COOH

- Acid bo khng no Trong t nhin thng gp cc acid bo khng no c t 18-20 carbon. Theo s lng lin kt i, ngi ta chia cc acid bo khng no thnh hai loi: acid bo khng no mt lin kt i (monoethylenic/ monounsaturated) v acid bo khng no nhiu lin kt i (polyethylenic/ polyunsaturated). V tr cc lin kt i sp xp theo quy lut: a s cc acid bo c mt lin kt i th v tr lin kt i gia carbon s 9 v s 10 ( 9), cc lin kt i khc thng v tr 12 v 15. Hu ht cc acid bo khng no trong t nhin u dng ng phn cis v c im nng chy thp hn acid bo no (cng s carbon). Mt s acid bo khng no nh elaidic acid, -eleostearic acid, vacenic acid, . c lin kt i dng trans.

55

Bng 4.5: Mt s acid bo khng no thng gp

S carbon v v H tr lin kt i Acid bo 1 lin kt i 16:1 (9) 18:1 (9) 7 Palmitic acid cis-9-Hexadecenoic acid C hu ht cc cht bo 9 Oleic acid cis-9-Octadecenoic acid L thnh phn acid bo chung ca hu ht cc cht bo trans-9-Octadecenoic acid cis-13-Docosenoic acid Thng gp trong cerebrosid Thng gp dng b kh Tn thng thng Tn h thng Thng gp

18:1 (9) 22:1 (13) 24:1 ( )

15

9 9

Elaidic acid Erucic acid

9 Nervonic acid cis-15-Tetracosenoic acid

Acid bo 2 lin kt i 18:2 (9,12) 6 Linoleic acid cis-9,12C nhiu trong bp, u Octadecadienoic acid nnh,

Acid bo 3 lin kt i 18:3 (6,9,12) 18:3 (9,12,15) 6 3 -Linolenic acid -Linolenic acid cis-6,9,12octadecatrienoic acid cis-9,12,15octadecatrienoic acid C nhiu thc vt, trng, Thng gp cng vi acid linoleic

Acid bo 4 lin kt i 20:4 (5,8,11,14) 6 Arachidonic acid cis-5,8,11,14eicosatetraenoic acid L thnh phn quan trng ca phospholipid ng vt, c nhiu trong du lc

Acid bo 5 lin kt i 20:5 (5,8,11,14,17) 3 Timnodonic acid cis-5,8,11,14,17eicosapentaenoic (EPA) C nhiu trong du c v trong trng

22:5 (7,10,13,16,19) 3 Clupanodonic cis-7,10,13,16,19C nhiu trong du c, acid docosapentaenoic acid phospholipid no (DPA) Acid bo 6 lin kt i 22:6 (4,7,10,13,16,19) 3 Cervonic acid cis-4,7,10,13,16,19- C nhiu trong du c, docosahexaenoic acid phospholipid no (DHA)

ng phn cis c nhit nng chy thp hn ng phn dng trans, v d oleic acid l dng cis nng chy nhit 13,4oC cn elaidic acid l dng trans th nng chy nhit 43,7oC. Ngoi ra, ng phn dng cis km bn vng hn ng phn dng trans. ng vt, acid bo khng no c th c ti 22 carbon v 6 lin kt i, cc lin kt i ng k tip cch nhau 3 carbon. Trong lc thc vt cc acid bo khng no c khng qu 20 carbon v 4 lin kt i v cc lin kt i ng k tip c th ch cch nhau 2 carbon (v 56

d: eleostearic acid: 18:3 (9,11,13)). Trong h acid bo 3, acid bo EPA (20:5)v DHA (22:6) gi vai tr c bit quan trng trong qu trnh pht trin ca t bo no thi k u ca qu trnh sinh trng. Trong cc nhm acid bo khng no c nhm eicosanoic acid (c to thnh t arachidonic acid: 20:4) gi vai tr quan trng, l tin cht ca nhiu cht c hot tnh sinh hc quan trng nh prostaglandin, thromboxan v leukotrien. - Mt s loi acid bo khc Ngoi cc loi acid bo thng gp ni trn, trong t nhin cn c nhng acid bo khc nh acid bo c nhm -OH, acid bo mch nhnh, acid bo c mch vng, + ng vt c v, trong nhiu glycolipid c acid bo c nhm OH, v d: cerebronic acid (CH3-(CH2)21-CH(OH)-COOH), l mt dn xut ca lignoceric acid, c trong cerebrosid. + Prostaglandin c nhiu loi t bo c bit c nhiu trong tinh dch l dn xut t acid bo mch vng prostanoic.

4.1.2 Alcol ca lipid

Alcol ca lipid c chia thnh nhiu nhm khc nhau: glycerol, cc alcol bc cao, aminoalcol v sterol. Ngoi ra, trong t nhin cn c alcol khng no, v d phytol (l mt thnh phn ca chlorophyl v lycophyl) l mt dihydroxylalcol khng no c nhiu lin kt i l cht mu ta trong c chua.

- Glycerol l mt alcol a chc c trong thnh phn cu to ca glycerid v phosphatid. - Cc alcol cao phn t thng tham gia vo thnh phn ca cc cht sp. - Aminoalcol tham gia vo thnh phn cu to ca cerebrosid v mt s phosphatid. Aminoalcol thng gp l : + Sphingosin (thnh phn cu to ca sphingolipid): CH3-(CH2)12-CH=CH-CH(OH)-CH(NH2)-CH2-OH + Cerebrin (c nhiu trong nm men, ht ng): CH3-(CH2)13-CH(OH)-CH(OH)-CH(NH2)-CH2-OH - Sterol: tiu biu cho sterol l cholesterol trong m bo ng vt, sterol khi ester ha vi acid bo to thnh sterid.

4.2 Phn loi lipid 4.2.1 Lipid thun

4.2.1.1 Triacylglycerol Triacylglycerol l loi lipid n gin nht v cn c gi l triglycerid, m hoc m trung tnh. Triacylglycerol c to thnh do ba phn t acid bo kt hp vi glycerol

57

bng cc lin kt ester. Cc acid bo ny c th cng loi tt c ba v tr v trong trng hp ny c gi l triacylglycerol n gin. Tuy vy hu ht cc triacylglycerol trong t nhin l dng hn hp ca nhiu loi acid bo, chng c hai hoc ba loi acid bo (hnh 4.3). V nhm hydroxyl phn cc ca glycerol v nhm carboxyl phn cc ca acid bo nm trong lin kt ester, cho nn triacylglycerol l nhng phn t khng phn cc, k nc v khng tan trong nc. T trng ca lipid nh hn t trng ca nc. ng vt, triacylglycerol c nhiu trong t bo m, cn nhiu loi thc vt triacylglycerol c d tr di dng du v l cht d tr nng lng ng thi l tin cht ca qu trnh ny mm.

Hu ht cc loi lipid trong t nhin nh du thc vt, m sa v m ng vt u l hn hp ca triacylglycerol n gin v triacyglycerol hn hp. Chng c nhiu loi acid bo khc nhau v di mch carbon cng nh mc bo ha. Cc loi du thc vt nh du ng, du u nnh, du lc, du m (vng) u cha nhiu triacylglycerol vn c cc acid 58

bo khng no, do chng tn ti th lng nhit phng. Triacylglycerol ch cha cc acid bo no nh tristearin l thnh phn chnh ca m b tn ti dng c nhit phng. Nu thc n giu lipid tip xc vi oxy khng kh lu ngy s b i. Mi v v kh chu l do s phn ct lin kt i ca acid bo khng no, s oxy ha ny s to thnh aldehyde v carboxylic acid c mch carbon ngn hn v d bay hi hn. 4.2.1.2 Sterol v cc hp cht steroid

Steroid v dn xut ca chng l mt nhm ln trong t nhin c c im chung l khng thy phn c v c cha nhn steran hay cn gi l nhn steroid (cyclopentanoperhydrophenanthren) (hnh 4.4). Cc steroid quan trng ng vt bao gm cc sterol v cc dn xut nh acid mt, mui mt, vitamin D, cc loi hormone steroid. Sterol l lipid cu trc, tham gia cu to mng ca hu ht cc t bo eukaryote. Cholesterol l mt sterol tiu biu ca m bo ng vt. c im cu to ca cholesterol l c mt u phn cc (nhm hydroxyl C-3) v phn hydrocarbon cn li khng phn cc. Phn khng phn cc ny bao gm nhn steroid v chui hydrocarbon v tr C-17.

59

Mt s sterol tng t cholesterol c tm thy trong cc t bo eukaryote khc, v d stigmasterol thc vt, ergosterol nm. Vi khun khng th tng hp sterol, tuy vy mt s vi khun c th kt hp sterol t mi trng ngoi vo mng ca chng. Ngoi vai tr tham gia cu to mng, sterol cn ng vai tr l tin cht tng hp nn nhiu cht c vai tr sinh hc quan trng khc, v d: cc steroid hormone l nhng tn hiu sinh hc iu khin s biu hin gene, cc acid mt l cc dn xut phn cc ca cholesterol hot 60

ng nh l nhng cht ty (detergent) trong rut v nh tng ha m ca thc n, t lm cho chng c th tip cn c vi enzyme lipase ca ng tiu ha. 4.2.1.3 Sp (serid) Sp l ester ca acid bo cao phn t no hoc khng no (C14-C36) v alcol cao phn t (C16-C30). im nng chy ca sp thng cao hn im nng chy ca triacylglycerol v thng nm trong khong 60-100oC. Vai tr sinh hc ca sp a dng v lin quan n tnh cht k nc v rn ca sp. Tuyn da ca ng vt c xng sng tit ra cht sp c tc dng bo v tc v da, gi cho da mm, trn v khng thm nc. Chim v gia cm c bit l nhng loi thy cm tit ra cht sp t tuyn phao cu c tc dng chng thm nc. Sp khng ch c ng vt m cn c thc vt. L ca nhiu loi thc vt nhit i c bao ph bi mt lp sp mng c tc dng chng thot hi nc qu mc ng thi bo v trc s tn cng ca cn trng

4.2.2 Lipid phc tp

4.2.2.1 Glycerophospholipid Glycerophospholipid cn c gi l phosphoglyceride l lipid mng. Cu trc ca glycerophospholipid gm c 2 acid bo lin kt vi glycerol bng cu ni ester vi carbon s 1 v s 2 ca glycerol v mt nhm phn cc hoc tch in c gn vo glycerol bng lin kt phosphodiester vi nguyn t carbon s 3. Hp cht glycerophosspholipid n

61

gin nht l phosphatidic acid. T hp cht ny s c nhiu dn xut khc nhau nh phosphatidylcholine, phosphatidylethanolamine (hnh 4.8, hnh 4.9, bng 4.6).

Nhiu loi acid bo c th tham gia cu to glycerophospholipid do c nhiu loi phn t glycerophospholipid khc nhau. Ni chung, glycerophospholipid cha mt acid bo no c 16 hoc 18 carbon v tr s 1 v mt acid bo khng no c 18 hoc 20 carbon v tr s 2. M bo ng vt cha nhiu ether lipid (hnh 4.10). Trong cu to ca ether lipid mt trong hai chui acyl c gn vo glycerol bng lin kt ether ch khng phi bng lin kt ester. Chui c lin kt ether ny c th no hoc cha mt lin kt i v tr carbon s 1 v carbon s 2 (v d trong plasmalogen).

62

Chc nng ca cc ether lipid hin cha bit ht, c th mt trong nhng chc nng ca n l khng li s xc tc ca phospholipase nhm ct t lin kt ester ca acid bo trong lipid mng. Vai tr ca mt loi ether lipid c lm sng t, ether lipid ny l yu t hot ha tiu cu (platelet-activating factor). Yu t hot ha tiu cu l mt tn hiu mc phn t quan trng, n c gii phng t bch cu basophil (bch cu a kim, t bo ). Yu t ny kch thch s t li ca tiu cu v gii phng serotonin (l cht lm co mch qun / vasoconstrictor) t tiu cu. Ngoi ra, yu t ny cn c nhiu nh hng i vi nhiu m bo khc nhau nh gan, c trn, tim, t cung v phi v ng thi ng mt vai tr quan trng trong phn ng vim v d ng.

4.2.2.2 Sphingolipid Sphingolipid l mt nhm lipid mng ln, chng c mt u phn cc v hai ui khng phn cc, tuy vy chng khng ging vi glycerophospholipid v galactolipid v trong cu trc ca sphingolipid khng c glycerol (hnh 4.11). Alcol ca sphingolipid l sphingosine, l mt aminoalcol mch di. Khi ch c mt acid bo gn vi sphingosine v tr carbon s 2 s to thnh hp cht tng ng c tn gi l ceramid. Sphingolipid c chia thnh 3 nhm nh khc nhau: sphingomyelin (1), glycolipid trung tnh (khng tch in) (2) v ganglioside (3). Sphingomyelin cha phosphocholine hoc phosphoethanolamine, do n cn c xp vo nhm phospholipid. Sphingomyelin c trong mng nguyn sinh ca t bo ng vt, c bit mng myelin ca t bo thn kinh l mng bao xung quanh v c tc dng cch in cho phn axon ca t bo thn kinh.

63

Glycosphingolipid c nhiu mt ngoi ca mng nguyn sinh cht. Thnh phn cu to ca glycosphingolipid c mt hoc nhiu gc ng lin kt trc tip vi nhm -OH v tr carbon s 1 ca ceramide v khng cha phosphate. Trong cu trc ca cerebroside ch c mt gc ng n galactose hoc glucose. Cerebroside c mng nguyn sinh ca t bo, tuy vy cerebroside c gc ng galactose c trng cho nhng m bo trung tnh; ngc li, cerebroside c gc ng l glucose c trng cho nhng m bo khng trung tnh. Globoside l cc glycosphingolipid trung tnh (khng tch in) c hai hoc nhiu gc ng m cc gc ng ny thng l D-glucose, D-galactose, hoc N-acetyl-Dgalactosamine. Cerebroside v globoside cn c gi l glycolipid trung tnh v chng khng tch in pH = 7. Ganglioside l sphingolipid phc tp nht, trong thnh phn cu to cha oligosaccharide. Cu trc ca oligosaccharide c mt hoc nhiu gc N-acetylneuraminic acid, acid ny thng c gi l sialic acid. S c mt ca sialic acid lm cho ganglioside tch in m pH = 7.

64

4.2.2.3 Galactolipid v sulfolipid

Nhm lipid ny c thc vt. Galactolipid c nhiu lc lp v trong thnh phn cu to ca galactolipid c mt hoc hai gc galactose lin kt trc tip vi 1,2diacylglycerol. i vi sulfolipid, mt gc glucose c sulfonate ha lin kt trc tip vi mt diacylglycerol bng lin kt glycoside. Ngoi phng php phn loi da theo thnh phn cu to nh trn, lipid cn c phn loi theo chc nng. Theo cch ny, lipid c chia thnh lipid d tr (cn gi l lipid trung tnh) v lipid mng (lipid phn cc). Triacylglycerol l lipid d tr. Phospholipid (glycerophospholipid, sphingolipid) v glycolipid (sphingolipid, galactolipid) u l nhng lipid mng.

4.3 Khi nim v lipoprotein

Cholesterol, cholesteryl ester cng nh triacylglycerol v phospholipid u khng tan trong nc, do i t m bo ny sang m bo khc, chng phi c vn chuyn trong mu bng cch kt hp vi protein to thnh cc lipoprotein (LP) gi l lipoprotein huyt tng. Phn protein ca cc LP lm nhim v vn chuyn lipid c gi l apolipoprotein (cn gi l apoprotein). b mt ca cc LP, apolipoprotein c tc dng n nh cu trc ca cc LP, gip cho cc phn t LP phn tn, vn chuyn c trong mu. Ngoi ra, apolipoprotein cn c nhng chc nng khc ty theo cu trc v t trng ca tng loi lipoprotein.
Bng 4.8: Cc loi lipoprotein huyt tng ch yu ca ngi
Lipoprotein Thnh phn (% trng lng)

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T trng (g/ml) Chylomicron VLDL LDL HDL < 1,006 0,95-1,006 1,006-1,063 1,063-1,210

Protein 2 10 23 55

Phospholipid Cholesterol Cholesteryl Triacylglycerol t do ester 9 18 20 24 1 7 8 2 3 12 37 15 85 50 10 4

Cu to ca LP c hai lp: lp li trung tm v lp v b mt. Lp li gm cc lipid k nc (v d: triglycerid), lp o b mt l cc phn t lipid phn cc (v d: phospholipid) v cc phn t protein. Phospholipid l phn lipid phn cc nht ch yu nm gia phn lipid khng phn cc v phn protein gi vai tr l cht gn kt hai nhm cht ny. S lin kt gia apolipoprotein vi lipid to thnh nhiu loi ht lipoprotein, chng khc nhau v t trng bin ng t thp nht (chylomicron) ti cc lipoprotein c t trng cao. - Chylomicron: c kch thc ln nht (750-1000nm), do t trng nh nht. Chylomicron c tng hp li ni nguyn sinh trn ca t bo nim mc rut non sau c vn chuyn v gan ri nhp vo dng mu nh h thng mch bch huyt. Chylomicron ch cha mt t l protein rt thp (< 5%) v thnh phn lipid ch yu l triacylglycerol. Apolipoprotein ca chylomicron bao gm apoB-48, apoE v apoC-II. ApoC-II hot ha lipoprotein lipase trong mao mch ca m m, tim, c vn v tuyn v thi k tit sa. Vi s xc tc ca enzyme ny, acid bo s c gii phng vo cc m bo ny. Nh vy, vai tr ca chylomicron l vn chuyn acid bo ti cc m bo phc v nhu cu nhin liu hoc s dng cho mc ch d tr. Sau khi i qua mao mch, hu ht triacylglycerol c gi li nhng m bo ny, phn cn li ca chylomicron (mt t triacylglycerol, cholesterol, apoE, apoB-48) s di chuyn theo dng mu v gan. Receptor ca t bo gan s lin kt vi apoE ca phn cn li ca chylomicron v n s c a vo t bo gan bng con ng endocytosis (ni thc bo). trong t bo gan, phn cn li ca chylomicron s gii phng cholesterol v cholesterol s c phn gii trong lysosome.

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- Lipoprotein c t trng rt thp (VLDL): c kch thc nh hn v nhiu protein hn chylomicron. VLDP vn chuyn triacylglycerol t gan ti cc m ngoi bin. Ngoi triacylglycerol, VLDL cn cha mt t cholesterol, cholesteryl ester, v cc apolipoprotein nh apoB-100, apoC-I, apoC-II, apoC-III v apoE. Cc LP ny c vn chuyn trong mu t gan n m c v m m. Khi n cc m ny, apoC-II s hot ha lipoprotein lipase, do acid bo t do s c gii phng. T bo m s s dng cc acid bo ny tng hp mi triacylglycerol v d tr di dng cc ht m trong t bo, cn t bo c s s dng cc acid bo cho mc ch khai thc nng lng. - Lipoprotein c t trng thp (LDL): khi VLDL mt dn thnh phn triacylglycerol, VLDL s tr thnh VLDL-remnant (phn cn li ca VLDL hay cn gi l IDL: Intermediate density lipoprotein). Nu tip tc mt triacylglycerol, IDL s tr thnh LDL vi c im rt giu cholesterol v cholesteryl ester v apoB-100 l apolipoprotein chnh, LDL vn chuyn cholesterol n cc m bo ngoi gan. Cc m bo ny c receptor nhn bit apoB-100, do s thc y hp thu cholesterol v cholesteryl ester. - Lipoprotein c t trng cao (HDL): c kch thc nh nht, c ngun gc t gan v rut non. Thnh phn cu to ca HDL giu protein (khong 50% l protein), rt t cholesterol v khng c cholesteryl ester. HDL cha apoA-I, apoC-I, apoC-II v nhiu loi apolipoprotein khc nh enzyme lecithin-cholesterol acyl transferase (LCAT).

4.4 Vai tr ca lipid i vi c th ng vt

Cu to ha hc ca lipid a dng, do chc nng ca lipid cng rt a dng.

4.4.1 Kin to c th

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Lipid l thnh phn quan trng cu to nn mng sinh hc v to thnh hng ro bao xung quanh t bo v cc b phn ca t bo. Ty theo tng loi m bo khc nhau m c thnh phn v t l cc loi lipid khc nhau. Cc loi lipid tham gia cu to mng sinh hc bao gm glycerophospholipid, sphingomyelin v sterol (cholesterol). Ty theo tng m bo v loi mng, hm lng lipid c th thay i rt khc nhau; v d mng myelin cha 79% l lipid, trong khi mng trong ca ti th ch c 24% lipid. Lipid mng chim ti 510% vt cht kh ca hu ht cc loi t bo. Lipid d tr chim n trn 80% trng lng ca mt t bo m. Chi tit v chc nng kin to c th ca lipid c m t trong chng 11: Mng sinh hc v c ch vn chuyn xuyn mng.

4.4.2 D tr nng lng

M v du thc vt l dng d tr nng lng ch yu nhiu loi sinh vt. Trong c th ng vt, triacylglycerol (cn gi l lipid trung tnh) ng vai tr l cht d tr nng lng v c d tr trong cc t bo m (adipocyte) cc m m. T bo m cha enzyme lipase, enzyme ny xc tc qu trnh thy phn triacylglycerol d tr, do acid bo c gii phng v c vn chuyn n cc khu vc cn c cung cp nhin liu. Khi so snh d tr nhin liu di dng triacylglycerol v dng polysaccharide nh glycogen v tinh bt ngi ta thy d tr nhin liu di dng triacylglycerol c hai li th c ngha. Th nht, vi trng lng nh nhau, khi oxy ha triacylglycerol s cung cp nng lng nhiu hn rt nhiu (hn gp i) so vi khi oxy ha carbohydrate (bng 4.9). Th hai, v triacylglycerol c tnh k nc do vy khng b hydrate ha, sinh vt s dng m nh l mt ngun nhin liu d tr khng phi mang theo mt lng nc hydrate ha. i vi polysaccharide l cht d tr, c 01 g polysaccharide th c 02 g nc hydrate ha. Tuy vy, carbohydrate nh glucose v glycogen c u im l cung cp nng lng nhanh, tc thi cho nhu cu ca c th, mt trong nhng l do l chng ha tan trong nc.
Bng 4.9: Gi tr nng lng khi t chy mt s cht Cht Protein (trung bnh) Casein Du lc M ln M sa Tinh bt Glycogen Nng lng tng s (kJ/g) 23,8 24,5 39,7 38,8 38,9 17,3 17,5 Glucose Saccharose Lactose Acetic acid Propionic acid Methan Cht Cellulose thc n Nng lng tng s (kJ/g) 17,8 15,6 16,5 16,4 14,6 20,8 55,2

4.4.3 Dung mi ha tan vitamin

Lipid l dung mi ha tan cc vitamin A, D, E v K.

4.4.4 Gi nhit cho c th

i vi nhng ng vt sng vng nhit thp, lp m di da c tc dng gi nhit cho c th. Vai tr ny c bit quan trng i vi ng vt sng vng cc (hi cu, hi m, chim cnh ct) v cc ng vt ng ng (gu). 68

4.4.5 Bo v c hc
Lp m di da ca ng vt c tc dng bo v c th ng vt trc cc tc ng c hc.

4.4.6 Cung cp nc ni sinh

i vi cc ng vt ng ng, ng vt di c, ng vt sng nhng vng kh hn nh trn sa mc, lipid cn l ngun cung cp nc v mt lng nc ln c to ra khi oxy ha m.

4.4.7 Mt s vai tr quan trng khc

4.4.7.1 Vai tr ca cc hormone steroid Cc hormone steroid vn chuyn thng tin gia cc t bo. Steroid l dn xut b oxy ha ca sterol, chng c nhn steroid nhng khng c chui alkyl gn vo vng D ca cholesterol v phn cc hn so vi cholesterol. Testosterone, estradiol, cortisol, aldosterone u l nhng hormone steroid. Testosterone l hormone sinh dc c c sn xut tinh hon. Estradiol l hormone sinh dc ci c sn xut bung trng v nhau thai. Cortisol v aldosterone l cc hormone c tng hp min v tuyn thng thn c tc dng iu ha chuyn ha glucose v o thi mui. Prednisolone v prednisone u l nhng steroid tng hp c s dng trong nhiu ng dng y hc khc nhau nh: yu t khng vim; yu t c ch gii phng archidonate do s xc tc ca phospholipase A2 , t c ch tng hp leukotrien, prostaglandin v thromboxane; thuc iu tr hen suyn v vim khp. Cc hormone ny di chuyn theo dng mu (da vo protein vn chuyn) t ni n c sn xut ra n cc m bo mc tiu. Sau khi vo t bo mc tiu, hormone s gn vo receptor c tnh c hiu cao trong nhn, t to ra s thay i s biu hin gene v s chuyn ha. 4.4.7.2 Vai tr ca phosphatidylinositol v cc dn xut ca sphingosine Phosphatidylinositol v cc dn xut phosphoryl ha ca n c nh hng nhiu mc khc nhau iu khin cu trc v s trao i cht ca t bo. Phosphatidylinositol 4,5-bisphosphate nm mt trong ca mng nguyn sinh cht ng vai tr l im kt ni cho cc protein thuc h thng cytoskelet. N cn ng vai tr l cht d tr ca cc phn t thng tin, cc phn t thng tin ny c gii phng trong t bo khi t bo p ng li cc tn hiu ngoi bo tng tc vi cc receptor nm mt ngoi ca mng nguyn sinh cht. 4.4.7.3 Vai tr ca cc sphingolipid C nhiu loi sphingolipid khc nhau c pht hin trong mng t bo, c bit l trong mng nguyn sinh ca t bo thn kinh. Nhiu loi sphingolipid c tm thy ti v tr nhn bit trn b mt ca t bo, tuy vy ch mt s t loi sphingolipid c xc nh r vai tr ca chng trong c th. Sphingolipid mng cng c th ng vai tr l cht thng tin ni bo. Ceramide v sphingomyelin l cht iu ha i vi enzyme protein kinase. Ceramide hoc cc dn xut ca n c lin quan n cc qu trnh: iu ha phn chia t bo, bit ha t bo, s di tr ca t bo, s cht ca t bo theo chng trnh. ngi, mt s loi sphingolipid quyt nh nhm mu. 4.4.7.4 Vai tr ca cc cht eicosanoid Cc cht eicosanoid mang thng tin n cc t bo bn cnh. Cc cht eicosanoid l nhng hormone ch c nh hng n t bo bn cnh v tr tng hp ra cc hormone ny m khng c vn chuyn trong mu n cc m bo hoc c quan khc. Tt c cc 69

cht eicosanoid u c tng hp t arachidonic acid (20:4 (5,8,11,14)). Ngi ta bit l chng c lin n cc qu trnh sau: sinh sn, vim, st, au do tn thng hoc bnh l, ng mu v iu ha huyt p, sn sinh acid ca d dy v nhiu qu trnh quan trng khc. C ba loi eicosanoid: prostaglandin, thromboxane v leukotrien. - Prostaglandin (PG): c chia thnh 2 nhm: PGE l PG tan trong ether v PGF l PG tan trong m phosphate, t cc nhm ny li c chia thnh nhiu loi khc nhau nh PGE1, PGE2, PG ln u tin c tm thy tuyn tin lit (prostate gland), c trong nhiu loi m ca ng vt c xng sng. nh hng ca PG nhiu loi m bo thng qua vic iu khin qu trnh tng hp cht thng tin ni bo 3',5'-AMP vng (cAMP). cAMP nh hng n hot ng ca nhiu loi hormone khc nhau, do vai tr sinh hc ca PG rt a dng v phc tp: kch thch co c trn t cung, nh hng n dng mu i n mt s m bo, chu k thc-ng, phn ng ca mt s m bo i vi hormone nh hormone epinephrine v glucagon, lm gia tng thn nhit (to ra st), gy nn d ng v au. - Thromboxane: c to ra bi tiu cu, c tc dng ch yu trong qu trnh ng mu. - Leukotrien: ln u tin c tm thy bch cu (leukocyte). Leukotrien D4 lm co c kh qun, do khi leukotrien D4 c to thnh qu nhiu s dn n kh th. Triu chng ny gp trong nhiu trng hp nh hen suyn, hoc phn ng d ng khi b ong t, penicillin hoc cc tc nhn khc. 4.4.7.5 Vai tr ca sterol Ngoi cc vai tr nh c m t cc phn nu trn, cholesterol cn l nguyn liu tng hp nn acid mt. Acid mt c vai tr quan trng trong qu trnh nh tng ha lipid ca thc n v vn chuyn acid bo t xoang rut vo t bo nim mc rut non (xem thm chng chuyn ha lipid). Vitamin D3 cn c gi l cholecalciferol c to thnh trong da t 7-dehydrocholesterol thng qua mt phn ng quang ha di tc dng ca nh sng mt tri. Bn thn vitamin D3 khng c hot tnh sinh hc m n s c chuyn ha trong gan v thn to thnh 1,25-dihydroxycholecalciferol. Hp cht 1,25dihydroxycholecalciferol l mt hormone iu ha vic hp thu calcium trong rut non v iu ha mc calcium trong thn v xng. Ging nh cc hormone steroid khc, 1,25dihydroxycholecalciferol iu ha s biu hin gene, v d tng hp mt protein c kh nng gn vi calcium trong rut non. Ngoi vitamin D3, vitamin D2 (ergocalciferol) c to thnh t ergosterol ca t bo nm men bng cch chiu tia UV (tia t ngoi: ultraviolet). Cu trc ca vitamin D2 tng t vitamin D3, chng ch c s sai khc nh chui hydrocarbon gn vo vng D ca nhn steroid. C hai loi vitamin D2 v D3 u c vai tr sinh hc ging nhau.

Ti liu tham kho

Ti liu ting Vit Trnh Vn Bo, Phan Th Hoan, 2002. Cht sng. Trong: Cc nguyn l sinh hc; Bin son bi Trnh Vn Bo (ch bin), Phan Th Hoan, Trn Th Thanh Hng, Trn Th Lin, Trn c Phn, Phm c Phng, Nguyn Vn Rc, Nguyn Th Trang; trang 11-29. NXB Y hc, H Ni. Phm Th Trn Chu, Trn Th ng, 1999. Ha sinh hc. NXB Gio dc, trang 62-76.

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o Kim Chi, 2004. Lipid. Trong: Ha sinh hc; bin son bi Nguyn Xun Thng (ch bin), o Kim Chi, Phm Quang Tng, Nguyn Vn ng; trang 29-50. NXB Y hc, H Ni. Trn Thanh Lan Phng, 2005. Ha hc lipid. Trong: Ha sinh y hc; bin son bi nh H (ch bin), ng Th Hoi An, Nguyn Th Ho, Phm Th Mai, Trn Thanh Lan Phng, Th Thanh Thy, L Xun Trng; trang 35-61. NXB Y hc, tp. H Ch Minh. Nguyn nh , 2000. Ha sinh lipid. Trong: Ha sinh y hc; bin son bi Hong Quang (ch bin), Nguyn nh , Trng Th Minh c, Bch Vng Hi, Phan Hi Nam, on Trng Ph; trang 113-146. NXB Qun i nhn dn, H Ni. L Th Kim Thu, 2002. Ha sinh lm sng. NXB Y hc, H Ni. Ti liu ting c Jeroch H., Drochner W., Simon, O., 1999. Ernaehrung landwirt-schaftlicher Nutztiere. Verlag Eugen Ulmer Stuttgart, Germany. Kirchgessner M., 1997. Tiernaehrung, 10. neubearbeitete Auflage, DLG-Verlag, Frankfurt am Main, Germany. Loeffler G., 1998. Lipide. In: Biochemie und Pathobiochemie, 6. Auflage (Loeffler G., Petrides P. E., Hrsg.), pp. 135-147. Springer Verlag, Berlin, Germany. Weissbach F., 1993. Wertbestimmende Bestandteile der Futtermittel. In: Futtermittelkunde (Jeroch H., Flachowsky G., Weissbach F., Hrsg.), pp. 18-32. Gustav Fischer Verlag Jena-Stuttgart, Germany. Ti liu ting Anh McDonald P., Edwards R. A., Greenhalgh J. F. D., Morgan C. A., 1995. Animal Nutrition, fifth edition. Longman Scientific & Technical, New York - USA, pp. 9-27. Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA.

Chng 5

VITAMIN
Qu Hai, H Trung Thng

5.1 Khi nim chung 5.1.1 nh ngha, cch gi tn v phn loi vitamin
Vitamin l mt nhm cht hu c c phn t tng i nh v c cc tnh cht l, ha 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. Danh t vitamin bt ngun t ch "vita" c ngha l s sng v ch "amine" ni ln s cn thit ca amine cho s sng.

71

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 cho 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 ha hc, gi theo chc nng, tc l theo tn bnh xy ra khi thiu mt vitamin no v thm tip u ng "anti". 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-ha 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. 5.1.2. Vai tr ca vitamin trong c th ng vt i vi cc ng vt khc nhau, nhu cu vitamin cng khc nhau. Khng phi tt c cc ng vt bc cao u cn thit tt c cc vitamin nh nhau. Chnh v vy, ty theo chc nng sinh l ca tng loi vitamin, chng c vai tr, tc dng khc nhau n s sinh trng pht trin ca ng vt. Cc s liu thc nghim cho bit, vic b sung vitamin B1 c th lm tng nhanh tc sinh trng v pht trin ca ln con. Khi thiu vitamin B2, kh nng ng ha carbohydrate v protein ca gia sc b hn ch, gia sc chm ln, cc trng thi sinh l chuyn bin xu i. i vi ln, thiu vitamin B6 s gy ra hin tng thiu mu, ph hu s trao i st bnh thng. i vi gia cm, thiu vitamin B6 s gy ra hin tng lit cc mc khc nhau. ng vt nhai li khng i hi vic cung cp vitamin B12. Kh nng tng hp vitamin B12 ca khu h vi khun sng trong d c v trong rut ca chng l rt ln. Ln v gia cm cn ly thm vitamin B12 trong thc n. Thiu vitamin B12 s lm ln con chm ln, lm thng tn n s pht trin bnh thng ca phi g, lm g con thiu mu, chm ln, gim kh nng khng i vi bnh tt. Thiu folic acid ln s km n, gim hng cu, gim hemoglobin; gia cm cng gy ra trng thi thiu mu. Thiu pantothenic acid gia sc s mc bnh vim da, mt mu lng, rng lng, thng tn h thn kinh, h tiu ha, ... Biu hin s thiu pantothenic acid g con l lng gin v ri, mt nng lc iu ho s c ng, ... Ln trng thnh c th t tng hp lng biotin cn thit i vi chng, Thiu biotin s dn n vim da, st cn c gia sc v gia cm. Phn ln ng vt t nhiu u c kh nng t tng hp c vitamin C, nhng nu c cung cp lng vitamin ny th s c nh hng tch cc i vi s sinh trng v pht trin ca chng. Thiu vitamin A s lm cho ln con chm ln, chm pht trin, mt sng, qung g; lm g vt gim kh nng trng, gim t l n trng, ... Thiu vitamin D s lm cho ln con chm ln, khp xng phng, chn co git, phi g pht trin khng bnh thng... Vic b sung ch phm vitamin D vo thc n nui ln, nui g khng nhng phng tr c bnh ci xng, m cn nh hng r rt i vi tc sinh trng v pht trin ca chng. Vitamin E c vai tr quan trng i vi b my di truyn ca ng vt. Th nghim cho thy thiu vitamin E s nh hng n kh nng sinh sn chut, ngn cn s to phi, d sy thai. Thiu vitamin K gia sc, gia cm s b chy mu ( di da, hc bng, c), do dn n 72

thiu mu. Vitamin K c bit cn thit i vi gia cm. Vitamin E cn thit i vi s sinh trng bnh thng ca ng vt, n tham gia tch cc vo qu trnh ng ha lipid, nng cao tnh bn vng v tnh n hi ca thnh mch mu. 5.2 Vitamin tan trong nc 5.2.1 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 B 1 cao nht. Vitamin B1 c tch ra dng tinh th vo nm 1912 v ngi ta xc nh c cu trc ha hc ca n (hnh 5.1).

Vitamin B1 bn trong mi trng acid, cn trong mi trng kim n rt d b phn hy 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 keto acid nh pyruvic acid, oxaloacetic acid, .... V vy khi thiu vitamin B1, s chuyn ha cc keto acid b ngng tr lm cho c th tch ly mt lng ln cc keto acid lm ri lon s trao i cht v gy nn cc trng thi bnh l nguy him. Vitamin B1 ha tan nhiu trong mi trng nc v chu nhit kh cho nn khng b phn hy 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. 5.2.2 Vitamin B2 (riboflavin) Vitamin B2 l dn xut ca vng isoalloxazin, thuc nhm flavin. Trong c th, vitamin 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.

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5.2.3 Vitamin PP (nicotinic acid, nicotinamid) Vitamin PP l nicotinic acid v amid ca n l nicotinamid. Vitamin PP l thnh phn ca coenzyme NAD+, NADP+ c trong cc enzyme thuc nhm dehydrogenase k kh. N cn c tn l niacin, niacinamid hoc vitamin B5. Vitamin PP gip c th chng li bnh pellagra. 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 cho nn kh b phn hu, cn dng nicotinamid li km bn vi acid v kim. Vitamin PP khng b bin i khi nu nng cho 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 ha kh. Vitamin PP c tc dng ngn nga bnh ngoi da, sng mng nhy rut, d dy.

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5.2.4 Vitamin B6 (pyridoxin) Vitamin B6 tn ti trong c th 3 dng khc nhau: pyridoxol (cn c tn l pyridoxine), pyridoxal, pyridoxamine. Ba dng ny c th chuyn ha ln nhau. Vitamin B6 l thnh phn coenzyme ca nhiu enzyme xc tc cho qu trnh chuyn ha 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, ... 5.2.5. Vitamin C (ascorbic acid) Vitamin C l ascorbic acid. Trong c th vitamin C tn ti 2 dng: dng kh l ascorbic acid v dng oxy ha l dehydro ascorbic acid. Vitamin C tham gia nhiu qu trnh sinh l quan trng trong c th: - Qu trnh hydroxyl ha 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 70-80mg/ngi. Nu thiu vitamin C s dn n bnh hoi huyt, gim sc khng ca c th, b bnh chy mu rng, chy mu li hay ni quan (bnh scorbutus). 5.2.6 Vitamin B12 (cyanocobalamin) Vitamin B12 c cu to phc tp, trong thnh phn c cha nhm CN, Co, amine. 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 ha cc base nit (nitrogen). Thiu B12 s gy bnh thiu mu c tnh.

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5.2.7 Pantothenic acid (vitamin B3) Vitamin B3 rt ph bin cc i tng sinh vt khc nhau, do c tn l pantothenic acid (danh t pantothen theo ting Latin l khp ni). N bao gm hai phn l panthoic acid v -alanin. Vitamin B3 l tin cht ca coenzyme A, thiu n lm vim da. Pantothenic acid c nhiu nm men, gan, cc phn xanh ca thc vt v c tng hp bi cc vi khun ng rut.

5.2.8 Biotin (vitamin H)

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y l mt loi monocarboxylic acid c cu trc vng, trong vng A l vng imidazol v vng B l vng thiophen. Biotin l thnh phn cu to ca cc enzyme xc tc cho cc phn ng carboxyl ha. Thiu biotin s xut hin cc triu chng nh sng da, rng tc, ...

5.2.9 Folic acid (vitamin Bc) Folic acid bao gm c mt nhm cht tng t nhau v cu trc ha hc v tnh cht sinh hc ty thuc vo ngun cha vitamin . Folic acid mang tn l vitamin Bc (c l ch vit tt ca t chicken, ting Anh c ngha l g con) v n cng cn thit cho s pht trin ca g con. Folic acid bao gm ba gc lin kt vi nhau l gc pterin, gc paraaminobenzoic acid v gc glutamic acid. Thiu vitamin ny cng s b thiu mu. T folic acid d dng chuyn thnh tetrahydrofolic acid (CoF, FH4) l coenzyme ca cc enzyme xc tc cho phn ng chuyn v cc nhm cha mt carbon (nhng khng phi l CO2).

5.3 Vitamin tan trong cht bo 5.3.1 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 s to thnh 2 phn t vitamin A khi thy phn nh enzyme carotenase xc tc.

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Vitamin A c nhiu trong du c v 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. 5.3.2 Vitamin D Trong c th tn ti nhiu loi vitamin D, trong quan trng nht l dng D 2 v D3. Cc vitamin D u l dn xut ca cc sterol. Trong c th, vitamin D c to ra t tin vitamin D c sn di da nh tia t ngoi ca nh sng mt tri.

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Thiu hoc tha vitamin D u c nh hng n nng phosphorus v calcium 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. 5.3.3 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 methyl gn vo vng thm ca phn t. Trong cc dng vitamin E, dng -tocopherol c hot tnh cao nht. Vitamin E c nhiu cc loi rau xanh, nht l x lch, ht ng cc, du thc vt, gan b, lng trng, mm ht ha tho, ... Vitamin E c tc dng nh cht chng oxy ha cho nn c tc dng bo v cc cht d b oxy ha trong t bo. Vitamin E cn c vai tr quan trng trong sinh sn.

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5.3.4 Vitamin K C nhiu loi vitamin K, chng u l nhng dn xut ca naphtoquinon. Hai dng vitamin K1 v K2 u c mch bn R di, cc dng tng hp (vitamin K3) c cu trc phn t n gin hn. 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 b gim, mu kh ng. 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 0,20,3mg/ngy/ngi. Ngoi cc loi vitamin ni trn, trong hai nhm vitamin tan trong nc v trong m cn mt s vitamin khc nh vitamin P (Rutin), vitamin F, vitamin Q (Ubiquinon).

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Ti liu tham kho Phm Th Trn Chu, Trn Th ng, 1999. Ha sinh hc. NXB Gio dc, H Ni. Nguyn Th Ho, 2005. Vitamin. Trong: Ha sinh y hc; bin son bi nh H (ch bin), ng Th Hoi An, Nguyn Th Ho, Phm Th Mai, Trn Thanh Lan Phng, Th Thanh Thy, L Xun Trng; trang 35-61. NXB Y hc, Tp. H Ch Minh.

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Loeffler G., 1998. Lipide. In: Biochemie und Pathobiochemie, 6. Auflage (Loeffler G., Petrides P. E., Hrsg.), pp. 135-147. Springer Verlag, Berlin, Germany. McDonald P., Edwards R. A., Greenhalgh J. F. D., Morgan C. A., 1995. Animal Nutrition, fifth edition. Longman Scientific & Technical, New York - USA, pp. 9-27. Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA.

Chng 6

ENZYME V S XC TC SINH HC
Qu Hai

6.1 Enzyme - ng lc ca qu trnh sng 6.1.1 Khi nim v enzyme

Trong cc t bo ca c th sng lun lun xy ra qu trnh trao i cht. S trao i cht ngng th s sng khng cn tn ti. Qu trnh trao i ca mt cht l tp hp cc quy lut ca rt nhiu phn ng ho hc khc nhau. Cc phn ng ho hc phc tp ny c lin quan cht ch vi nhau v iu chnh ln nhau. Enzyme l cc hp cht c bn cht protein xc tc cho cc phn ng ho hc . Chng c kh nng xc tc c hiu cc phn ng ha hc nht nh v m bo cho cc phn ng xy ra theo mt chiu hng nht nh vi tc nhp nhng trong c th sng. Chng c trong tt c cc loi t bo ca c th sng. Enzyme cn c gi l cht xc tc sinh hc (biocatalysator) nhm phn bit vi cc cht xc tc ho hc.

6.1.2 Vi nt v qu trnh pht hin v nghin cu enzyme

Enzyme hc c coi nh ct sng ca ho sinh hc cho nn phn ln cc nghin cu ho sinh t trc n nay u lin quan nhiu n enzyme. Trc th k 17 ngi ta bit s dng cc qu trnh enzyme trong i sng, tuy nhin ch c tnh cht kinh nghim thc t v thng qua hot ng ca vi sinh vt. Sau th k 17 ngi ta khi qut hin tng ln men nh l hin tng ph bin trong s sng v enzyme l yu t gy nn s chuyn ho cc cht trong qu trnh ln men. u th k 19 c nhng nghin cu ghi nhn s c mt ca enzyme trong qu trnh tiu ho nc bt, d dy v rut. T cc khi nim enzyme c t chc (enzyme khng th tch khi t bo c) v enzyme khng c t chc (enzyme c th thc hin tnh xc tc ca chng ngoi c th) ca Pasteur (1856) n kt qu thc nghim ca Buchner (1897) l c mt qu trnh nhm chng minh cho s tham gia ca cc enzyme vn c trong t bo nm men trong vic chuyn ho ng thnh ru. Nm 1926, Sumner kt tinh c enzyme urease ln u tin. Sau Northrop (1930) kt tinh c pepsin, Northrop v Kunitz (1931) kt tinh c trypsin cho php xc nh c mt cch dt khot bn cht ca enzyme l protein v t nn mng cho nhng nghin cu c bn v enzyme.

6.1.3 Enzyme v cng ngh sinh hc

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Nhng thnh tu nghin cu c bn v enzyme l c s pht trin cc nghin cu ng dng enzyme trong thc t. Hin c trn 30 ngnh sn xut khc nhau s dng ti cc ch phm ca enzyme. Con s ny ngy cng tng v s lng enzyme ngy cng ln. So vi cc cht ha hc, enzyme c nhng tnh cht u vit hn hn nh: phn ng xc tc c th din ra ngay nhng iu kin bnh thng v p sut, pH v nhit . Chng c th tc ng nng rt thp. Phn ng do enzyme xc tc l nhng phn ng c tnh c hiu rt cao, t sinh ra cc sn phm ph. V vy, chng to iu kin rt thun li cho vic tin hnh cc qu trnh sn xut cng nghip. Tuy nhin, ta c th iu chnh d dng cc chuyn ha ca enzyme. N c th c s dng trong nhiu lnh vc. Enzyme thng c s dng theo 2 cch: - Khng tch enzyme khi nguyn liu m ch to iu kin cho mt s enzyme sn c trong nguyn liu chng chuyn ha cc cht c cng trong nguyn liu y theo hng mong mun. - Tch enzyme khi nguyn liu dng ch phm s dng khi cn thit. Do c th tn dng cc ngun nguyn liu giu enzyme tch enzyme, dng ch phm enzyme ny ch bin cc nguyn liu khc nhau hoc s dng vo nhng mc ch khc nhau. Vic s dng enzyme theo cch ny ngy cng pht trin dn n vic hnh thnh ngnh cng nghip enzyme nhiu nc, hng nm sn xut hng trm tn ch phm enzyme phc v cho cc ngnh sn xut khc nhau v cho y hc. Nh vy, enzyme - mt trong nhng tc nhn sinh hc mc di t bo, tham gia mt cch tch cc vo cc qu trnh thuc lnh vc cng ngh sinh hc. T cng ngh enzyme tr thnh mt nhnh quan trng ca cng ngh sinh hc.

6.2 Tn gi v phn loi enzyme 6.2.1 Cch gi tn enzyme

Trong thi gian u khi enzyme hc cha pht trin, ngi ta thng gi tn enzyme mt cch ty tin, ty theo tc gi. V d nh cc tn pepsin, trypsin, chymotrypsin hin nay vn c dng gi l tn thng dng. Sau , ngi ta thng gi tn enzyme bng cch ly tn c cht c hiu ca enzyme cng thm tip v ng ase. V d, urease l enzyme tc dng ln urea, protease l enzyme tc dng ln protein, lipase l enzyme tc dng vo lipid, amylase l enzyme tc dng ln tinh bt. i vi cc nhm enzyme cng xc tc mt loi phn ng, ngi ta ly tn ca phn ng enzyme thm tip v ng ase, v d nhng enzyme xc tc s oxy ho c gi l oxydase, nhng enzyme kh hydrogen c gi l dehydrogenase, ... Tn gi y , chnh xc theo quy c quc t - tn gi h thng ca enzyme c gi theo tn c cht c hiu ca chng cng vi tn ca kiu phn ng m chng xc tc, cng thm tip v ng ase, v d enzyme xc tc cho s thy phn urea (carbamid) c tn h thng l carbamid-amidohydrolase (tn thng dng l urease).

6.2.2 Phn loi enzyme

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Mc ch ca vic phn loi enzyme l nhn mnh mt cch chnh xc v tng qut, mi quan h v nhng iu ging nhau ca mt loi enzyme. Tiu ban v enzyme (Enzyme Commission: EC) c thnh lp bi Hi Ho sinh quc t (International Union of Biochemistry: IUB) a ra cch phn loi thng nht da trn cc loi phn ng v c ch phn ng. Theo cch phn loi ny th enzyme c chia ra lm su lp ln nh s t 1 n 6. Cc s th t ny l c nh cho mi lp. Su lp enzyme theo phn loi quc t gm c: - Oxydoreductase: Cc enzyme xc tc cho phn ng oxy ho - kh. Trong nhm ny c tt c cc enzyme c cc tn thng thng bit nh dehydrogenase, oxydase, cytochromreductase v peroxydase. Trong cc phn ng do chng xc tc xy ra s vn chuyn hydrogen, s chuyn electron, s oxy ho bi oxy phn t, bi hydrogen peroxide hoc bi cc cht oxy ho khc. - Transferase: Cc enzyme xc tc cho phn ng chuyn v. Cc transferase do bn cht ca nhng gc m chng vn chuyn c th tham gia vo cc qu trnh trao i cht rt khc nhau. Trong lp transferase bn cnh transaminase v methyltransferase cn c cc kinase khc nhau (xc tc ch yu cho s vn chuyn ca gc phosphate t hp cht cao nng ti cht khc), mt phn ln cc enzyme trc kia gi l mutase v mt phn vo loi synthetase, v d cc enzyme tng hp DNA v RNA. - Hydrolase: cc enzyme xc tc cho phn ng thu phn. Trong lp ny c cc enzyme phn gii ester (v d lipid), glycoside, amide, peptide, protein. - Lyase: cc enzyme xc tc cho phn ng phn ct khng cn nc, loi nc to thnh ni i hoc kt hp phn t nc vo ni i. Thuc vo lp ny c cc enzyme c gi l hydratase, aldolase, decarboxylase cng nh mt s desaminase. - Isomerase: Cc enzyme xc tc cho phn ng ng phn ho. Tnh cho n cng th chng xc tc cho nhng phn ng chuyn cc nhm khc nhau bn trong phn t. Trong lp ny khng nhng c nhng enzyme chuyn ho cc ng phn hnh hc v ng phn quang hc (nh alaninracemase) m c cc enzyme xc tc cho cc phn ng v d s chuyn ho aldose thnh cetose (glucose phosphate isomerase, trc kia gi l phosphohexoisomerase) hoc bin i v tr ca lin kt ester bn trong phn t (v d phosphoglucomutase) - Ligase: Cc enzyme xc tc cho phn ng tng hp c s dng lin kt giu nng lng ATP, ... y cn ch thm l cc enzyme phn ct c phn loi vi tn lyase. Nu cn bng chuyn dch v pha tng hp th enzyme cng c th c gi l synthase. Ngc li chng ta gi cc enzyme xc tc cho phn ng kt hp 2 phn t c s tham gia ca ATP hoc nucleotide triphosphate tng t hoc c s dng ca mi lin kt giu nng lng l synthetase. Tn gi theo h thng phn loi ca lp ny l ligase trnh s nhm ln vi tn synthase ni trn.

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Mi lp li c chia thnh nhiu lp ph v phn lp ph, ri sau th t ca enzyme trong phn lp ph. Nh vy, mi enzyme trong h thng c phn loi v t tn theo m 4 ch s biu th phn ng xc tc: con s u ch lp, s th hai ch lp ph, s th ba ch phn lp ph, s th t ch r s bc th t ca enzyme. V d, enzyme xc tc cho phn ng: V d: Ethanol + NAD+ acetaldehyde + NADH + H+ c tn gi l alcohol dehydrogenase (ADH) C tn quc t theo kha phn loi l: Alcohol: NAD oxydoreductase, EC 1.1.1.1 Trong , m s 1 u tin biu th tn lp enzyme l oxydoreductase (lp 1); m s 1 th hai biu th lp ph 1: tc dng ln nhm CH - OH ca cc cht cho; m s 1 th ba biu th phn lp ph 1: cht nhn l NAD hay NADP v m s 1 cui cng ch s th t ca enzyme. Nh vy, trong cch gi h thng ca enzyme ADH trn c tn ca c cht v ca coenzyme cng nh tn ca qu trnh chuyn ho ha hc c xc tc vi tn cng ase. Sau tn ca enzyme l s ca n theo danh sch cc enzyme do tiu ban v enzyme ra.

6.3 Cu trc ca enzyme

Enzyme c kh nng v hiu lc xc tc rt ln, c tnh c hiu rt cao. m bo cho chc nng ca enzyme l cht xc tc sinh hc, cu trc ca enzyme phi rt tinh vi v phc tp.

6.3.1 Bn cht ho hc ca enzyme

T gn mt th k trc y, cc nh khoa hc x vo vic xc nh bn cht ho hc ca enzyme. Cho n nay, c th ni rng, ngoi mt nhm nh phn t RNA c hot tnh xc tc, tuyt i a s enzyme c bn cht protein v s th hin hot tnh xc tc ph thuc vo cu trc bc 1, 2, 3 v 4 ca phn t protein v trng thi t nhin ca n. Thc t l bn cht ho hc ca enzyme ch c xc nh ng n t sau khi kt tinh c enzyme. Enzyme u tin nhn c dng tinh th l urease ca u tng (Sumner, 1926), tip theo l pepsin v trypsin (Northrop v Kunitz, 1930, 1931). Sau nhng tc gi khc cng kt tinh c mt s enzyme khc v c bng chng xc nhn cc tinh th protein nhn c chnh l cc enzyme. Kt qu nghin cu tnh cht ho l ca enzyme cho thy enzyme c tt c cc thuc tnh ho hc ca cc cht protein v hnh dng phn t: a s enzyme c dng hnh cu (dng ht). T l gia trc di v trc ngn ca phn t vo khong 1 - 2 hoc 4 - 6.

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V khi lng phn t: cc enzyme c khi lng phn t ln, thay i rt rng t 1.2000 dalton n 1.000.000 dalton hoc ln hn. V d ribonuclease c khi lng phn t l 12.700, glutamate dehydrogenase c khi lng phn t l 1.000.000. a s enzyme c khi lng phn t t 20.000 n 90.000 hoc vi trm nghn. Do kch thc phn t ln, cc enzyme khng i qua c mng bn thm. Enzyme tan trong nc, khi tan to thnh dung dch keo; chng cng tan trong dung dch mui long, glycerin v cc dung mi hu c phn cc khc. Enzyme khng bn v d dng b bin tnh di tc dng ca nhit cao. Enzyme b bin tnh th mt kh nng xc tc. Mc gim hoot tnh ca enzyme tng ng vi mc bin tnh ca protein trong ch phm. Kim, acid mnh, kim loi nng cng lm cho enzyme bin tnh. Cng nh protein, enzyme cng c tnh cht lng tnh.

6.3.2 Thnh phn cu to ca enzyme

Cng nh protein, enzyme c th l protein n gin hoc protein phc tp. Trn c s , ngi ta thng phn enzyme thnh hai nhm: enzyme mt thnh phn (enzyme mt cu t) v enzyme hai thnh phn (enzyme hai cu t). Trng hp enzyme l mt protein n gin c gi l enzyme mt thnh phn. Trng hp enzyme l mt protein phc tp ngha l ngoi protein n gin cn c mt nhm ngoi no khng phi protein c gi l enzyme hai thnh phn. Phn protein ca enzyme hai thnh phn c gi l apoprotein hay apoenzyme, cn phn khng phi protein c gi l nhm ngoi hoc coenzyme. Phn khng phi protein thng l nhng cht hu c c hiu c th gn cht vo phn protein hoc ch lin kt lng lo vi protein v c th tch khi phn protein khi cho thm tch qua mng. Coenzyme l phn khng phi protein ca enzyme trong trng hp khi chng d tch khi phn apoenzyme khi cho thm tch qua mng bn thm v c th tn ti c lp. Phn khng phi protein ca enzyme c gi l nhm ngoi hay nhm prosthetic, khi n lin kt cht ch vi phn protein ca enzyme bng lin kt ng ho tr. Mt phc hp hon chnh gm c apoenzyme v coenzyme c gi l holoenzyme. Mt coenzyme khi kt hp vi cc apoenzyme to thnh cc holoenzyme khc nhau xc tc cho qu trnh chuyn ho cc cht khc nhau nhng ging nhau v kiu phn ng. Coenzyme trc tip tham gia phn ng xc tc, gi vai tr quyt nh kiu phn ng m enzyme xc tc v lm tng bn ca apoenzyme i vi cc yu t gy bin tnh. Cn apoenzyme c tc dng nng cao hot tnh xc tc ca coenzyme v quyt nh tnh c hiu ca enzyme. Cc coenzyme thng l cc dn xut ca cc vitamin ha tan trong nc. Cn ch l s phn bit coenzyme v nhm ngoi ch l tng i, v kh c th c mt tiu chun tht rnh mch phn bit lin kt cht ch v lin kt khng cht ch, nht l trong nhng nm gn y, ngi ta chng minh c rng, nhiu coenzyme cng kt hp vo apoenzyme ca chng bng lin kt ng ha tr. Do , ngy nay ngi ta t ch n s phn bit coenzyme v nhm ngoi. Ngoi ra, trong thnh phn cu to, rt nhiu enzyme c cha kim loi. Thuc loi enzyme hai thnh phn gm c hu ht cc enzyme ca cc lp 1, 2, 4, 5, 6. Cc enzyme thy phn (lp 3) thng l enzyme mt thnh phn c cha ion kim loi hoc i hi ion kim loi lm cofactor.

6.3.3 Trung tm hot ng ca enzyme

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Ton b cu trc khng gian ca phn t enzyme c vai tr quan trng i vi hot tnh xc tc ca enzyme. Tuy nhin, hot ng ca enzyme lin h trc tip vi mt phn xc nh trong phn t enzyme. Trung tm hot ng ca enzyme l phn ca phn t enzyme trc tip kt hp vi c cht, tham gia trc tip trong vic to thnh v chuyn ha phc cht trung gian gia enzyme v c cht to thnh sn phm phn ng. Trung tm hot ng bao gm nhiu nhm chc nng khc nhau ca amino acid, phn t nc lin kt v nhiu khi c c cofactor (coenzyme) hu c v v c. cc enzyme mt thnh phn, trung tm hot ng thng bao gm mt t hp cc nhm chc nng ca amino acid khng tham gia to thnh trc chnh ca si polypeptide. V d nhm - SH ca cysteine, - OH ca serine, threonine v tyrosine, - NH2 ca lysine, -COOH ca glutamic acid, aspartic acid, vng imidazol ca histidine, indol ca tryptophan, nhm guanidin ca arginine. Cc nhm ny c th xa nhau trong mch polypeptide nhng li gn nhau trong khng gian, c nh hng xc nh trong khng gian cch nhau nhng khong cch nht nh sao cho chng c th tng tc vi nhau trong qu trnh xc tc. V d: trung tm hot ng ca -chymotrypsin bao gm nhm hydroxyl ca Ser 195, imidazol ca His-57 v nhm carboxyl ca Asp-102. Cc gc ny kh xa nhau trong chui polypetid nhng gia cc nhm chc nng ca chng ch cch nhau t 2,8 - 3,0. Trung tm hot ng ca cc enzyme hai thnh phn thng bao gm nhm ngoi (vitamin, ion kim loi, ...) v cc nhm chc nng ca cc amino acid phn apoenzyme. S tng ng v cu hnh khng gian gia trung tm hot ng v c cht c hnh thnh trong qu trnh enzyme tip xc vi c cht. Theo quan nim ca Fisher th trung tm hot ng ca enzyme c hnh thnh sn vi mt cu to nht nh ch cho php c cht c cu to tng ng kt hp vo. Do c th v s tng ng nh kha vi cha kha (hnh 6.1a).

Gi thuyt ny tuy cng gii thch c mt s hin tng nhng khng gii thch tha ng c nhiu kt qu thu c trong thc nghim. V vy, Koshland a ra mt gi thuyt khc hp dn v t nh hn. Theo thuyt ny th c im ca vng trung tm

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hot ng l rt mm do v linh hot, cc nhm chc nng ca trung tm hot ng ca enzyme t do cha t th sn sng hot ng, khi tip xc vi c cht, cc nhm chc nng trong phn trung tm hot ng ca phn t enzyme thay i v tr trong khng gian, to thnh hnh th khp vi hnh th ca c cht (hnh 6.1.b). Cng v vy, ngi ta gi m hnh ny l m hnh tip xc cm ng hoc khp cm ng. Gia c cht v trung tm hot ng to thnh nhiu tng tc yu, do c th d dng b ct t trong qu trnh phn ng gii phng enzyme v sn phm phn ng. Trung tm hot ng ca cc enzyme c cu trc bc 4 c th nm trn mt phn di n v (subunit) hoc bao gm cc nhm chc nng thuc cc phn di n v khc nhau.

6.3.4 Cc dng phn t ca enzyme

Trong cu trc phn t ca enzyme, tnh cht tinh vi v phc tp khng ch gii hn phm vi tng phn t, t thnh phn cu to v cc bc cu trc cho n cu to ca trung tm hot ng cng vi vai tr ca cc nhm chc nng m cn th hin tnh a dng ca cc phn t enzyme. Tnh a dng ca nhiu enzyme khc nhau c pht hin cc c th sng khc nhau t ngi, ng vt, thc vt n vi sinh vt. Ngi ta thy rng c nhng enzyme xc tc cng mt phn ng ha hc v c cng tnh c hiu c cht nhng c ngun gc khc nhau cho nn th hin nhiu tnh cht khc nhau. Aldolase c ngun gc t nm men c nhiu tnh cht khc vi aldolase ca m ng vt; pepsin, trypsin, chymotrypsin, xanthin - oxydase v lysozyme cng c nhng dng phn t khc nhau. Nhiu enzyme tng t nhau thu c t cng mt loi m nhng ca nhng loi khc nhau cng c nhng tnh cht khc bit nhau: -amylase ca dch nc bt v dch ty ca ngi th ging nhau, nhng chng khc vi -amylase thu c t ty ln v ha tan, v pH thch hp v mt s tnh cht khc. Nhng enzyme c ngun gc t nhng m khc nhau ca cng mt loi, tuy xc tc cng mt loi phn ng ha hc, nhng khc nhau rt r rt v tnh c hiu c cht nh trng hp ca nhng cholinesterase. Nhng enzyme xc tc nhng phn ng chuyn ha ging nhau trong cc t bo ca nhiu m khc nhau c tnh cht c hiu c quan, v d lactate dehydrogenase ca c tim v c xng khc nhau r rt v tc di chuyn in di v nhiu tnh cht khc. Ngay trong mt m hay mt c quan, cng tn ti nhng dng phn t khc nhau: trong c tim t nht cng c hai dng phn t ca lactate dehydrogenase c tc di chuyn in di khc nhau, t nm men c th tch ra c bn dng phn t ca phosphoglyceraldehyde dehydrogenase. Trong mt t bo, mt enzyme no cng c th c nhng dng phn t khc nhau tn ti trong cc b phn khc nhau ca t bo, v d aspartate aminotransferase c dng phn t trong ti th khc vi dng phn t ca enzyme ny bo tng. Nh vy, tnh a dng ca cc phn t enzyme c th th hin nhiu mc , t cc loi khc nhau n cc m hay c quan khc nhau ca cng mt c th v ngay c cc b phn khc nhau ca cng mt t bo. Cc dng phn t khc nhau ca cng mt enzyme, tuy cng c chc nng xc tc mt phn ng ha hc ging nhau nhng v cu trc phn t ca chng u t nhiu c khc nhau do chng c nhng tnh cht khc nhau v ha hc, vt l, min dch, ... thm ch ngay c ng hc v tnh c hiu ca phn ng enzyme.

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Theo kin ngh chnh thc ca y ban thng trc v enzyme ca Hi Ha sinh Quc t, danh t isoenzyme c dng ch nhng dng phn t khc nhau ca mt enzyme tn ti trong mt loi; ngoi danh t ny, danh t isozyme cng c quen dng. Theo mt s tc gi, khi phn loi cc dng phn t khc nhau ca enzyme, phi phn bit gia isoenzyme v heteroenzyme. Danh t isoenzyme ch dnh cho nhng dng phn t ca mt enzyme c ngun gc t cng mt c quan v m cng nh phi c cng hot ng xc tc nh nhau; danh t heteroenzyme dnh cho nhng dng phn t cng c hot ng xc tc ging nhau nhng c th c ngun gc t cc c quan hoc loi khc nhau. Trong cc isoenzyme trnh by, cc isoenzyme lactate dehydrogenase c nghin cu c it k do chng c ngha v mt chn on bnh. Chng c 5 dng phn t tetrame c th phn chia c ring r bng in di thnh cc cu t rt khc nhau.

6.3.5 Phc hp multienzyme

Trong c th sng, ngoi cc enzyme polymer - nhng enzyme c cu trc bc bn, do nhiu n v nh cu to nn; cn tn ti h thng nhiu enzyme hay cn gi l h thng multienzyme. l nhng h thng gm cc enzyme c lin quan vi nhau, xc tc cho dy chuyn phn ng ca mt qu trnh trao i cht xc nh, trong sn phm ca phn ng do enzyme trc xc tc l c cht ca enzyme xc tc cho phn ng tip theo.
E1 E2 E3

C th minh ha h thng gm 3 enzyme E1, E2, E3 xc tc cho dy chuyn phn ng nh sau:

A B C D Trong s ny, A l c cht ca E1, B l sn phm ca phn ng do E1 xc tc nhng li l c cht ca E2, ... Cc h thng nhiu enzyme trong t bo c mc t chc phc tp khc nhau. Cc enzyme trong h thng nhiu enzyme c th tn ti ring r dng ha tan, khng lin kt vi nhau hoc c th kt t vi nhau, lin kt vi nhau kh bn to thnh phc hp nhiu enzyme khi tch ring khi phc hp enzyme, s mt hot tnh xc tc. Ngoi ra, mt s h thng enzyme c th lin kt vi thnh phn cu to ca t bo nh mng ribosome. Cc enzyme ca chui h hp xc tc cho qu trnh chuyn in t t c cht n oxy, c gn cht vo mng trong ca ti th v thc cht l mt phn cu trc ca ti th. Ngi ta gp cc phc hp multienzyme trong nhiu qu trnh chuyn ha: qu trnh kh carboxyl oxy-ha ca pyruvic acid to thnh acetyl coenzyme

89

A, qu trnh tng hp acid bo ngoi ti th nm men, cc loi c v, loi chim, ..., qu trnh tng hp nhng peptide c hot tnh khng sinh nh gramicidin v tyrocidin, qu trnh tng hp tryptophan, ...

6.3.6 Enzyme d lp th
Enzyme d lp th (allosteric enzyme) c tn gi t ting Hilp allos c ngha l khc v stereos c ngha lp th, khng gian. l nhng enzyme c tc dng iu chnh c hiu trn nhng v tr then cht ca mng li chuyn ha, c bit l nhng qu trnh sinh tng hp. Trong phn t ca enzyme d lp th, ngoi trung tm hot ng lm chc nng xc tc, cn c mt s v tr khc c th tng tc vi cc cht khc gi l trung tm allosteric (trung tm iu ha, trung tm d lp th, trung tm d khng gian). Cc cht kt hp vo cc trung tm ny gi l cc cht iu ha allosteric (cht iu ha d lp th). Enzyme d lp th thay i hot xc tc thng qua s thay i cu hnh khng gian ca phn t enzyme, ca trung tm hot ng khi gn vi cc cht iu ha d lp th. Nu lm tng hot gi l cht iu ha dng; nu lm gim hot gi l cc cht iu ha m. Cc cht iu ha ny kt hp vi enzyme nhng khng b chuyn ha di tc dng ca enzyme m n kt hp. Enzyme d lp th thng l nhng enzyme c cu trc bc bn, do cc n v nh cu to nn; trong phn t thng c 2 hay mt s trung tm hot ng, c th kt hp vi 2 hay mt s phn t c cht. C cht c th thc hin chc nng ca cht iu ha - iu ha homotropic (ng hp, ng hng). Cc cht iu ha c cu trc khc c cht - iu ha heterotropic (d hp, d hng). Tuy nhin phn ln cc enzyme d lp th thuc kiu hn hp ng v d hp ngha l hot ng ca chng c th c iu ha nh c cht v cc cht trao i khc.

6.3.7 Tin cht ca enzyme (proenzyme, zymogen)

Phn ln cc enzyme c tng hp trong c th thnh nhng phn t enzyme c hot tnh, nhng c nhng enzyme nh cc protease ca tuyn ty, d dy cng nh cc protease xc tc cho qu trnh ng mu thng c tng hp nn qua mt dng trung gian cha c hot tnh xc tc gi l zymogen hoc proenzyme. l nhng tin cht to thnh enzyme, ch khng phi l enzyme thc s. Nhng cht ny khng c hot tnh, phi tri qua mt qu trnh bin i, sp xp li cu trc phn t mi tr thnh enzyme hot ng c. Qu trnh chuyn ha proenzyme thnh enzyme c hot tnh gi l qu trnh hot ha, c thc hin nh s t xc tc hoc do cc enzyme khc xc tc. Qu trnh hot ha zymogen c mt s c im chung nh sau: - L qu trnh thy phn c gii hn protein (limited proteolysis), ct t mt s lin kt peptide gn u N ca phn t zymogen. on peptid c to thnh c th b loi ra hoc vn gn vi phn cn li ca phn t nh cc cu disulfide. - Khi lin kt peptid b ct t thng lm thay i cu hnh khng gian ca phn t theo hng c li cho hot ng xc tc, to thnh phn t enzyme.

90

- Hiu sut hot ha zymogen ph thuc vo iu kin hot ha, nng zymogen, bn cht v nng ca enzyme xc tc cho qu trnh hot ha, nhit , pH v mt s yu t khc. ngi v nhiu loi ng vt c v, cc enzyme thy phn protein (protease) trong ng tiu ha u c tng hp di dng tin cht ca enzyme. V d pepsinogen do nhng t bo chnh ca tuyn d dy tng hp nn v l tin cht ca pepsin, chymotrysinogen v trypsinogen ca tuyn ty theo th t l tin cht ca chymotrysin v trypsin. Cc cht ny u ch c hot ha thnh dng enzyme hot ng sau khi tit vo lng ng tiu ha. Trypsinogen c hot ha thnh trypsin di tc dng ca chnh trypsin hoc enterokinase, cn chymotrypsinogen c hot ha di tc dng ca trypsin v chymotrysin. Hin tng tng hp ra cc zymogen c mt ngha sinh hc quan trng. C th ni rng, cc protease trong ng tiu ha c tng hp qua giai on trung gian nh vy chnh l mt c ch t bo v ca c th. V nu khng nh vy th chnh cc tuyn tng hp nn cc loi enzyme ny s b tiu hy bi chnh nhng enzyme do chng tng hp nn.

6.4 C ch xc tc ca enzyme 6.4.1 C ch ca phn ng c xc tc ni chung

Vn tc phn ng ha hc c xc nh bi gi tr nng lng hot ha tc l mc nng lng ca cc cht tham gia phn ng phi t c ct t lin kt cn thit v hnh thnh cc lin kt mi. Nng lng hot ha cng ln th vn tc phn ng cng chm v ngc li. Do lm gim nng lng hot ha ca phn ng, cc cht xc tc c tc dng thc y vn tc phn ng ha hc. V d, bt platin l mt cht xc tc ha hc c s dng rng ri. V cc cht tham gia phn ng trn b mt platin u c chuyn sang trng thi c kh nng phn ng cao hn, do vy nng lng hot ha s nh hn v tc phn ng s cao hn. Nh vy, trong cc phn ng c xc tc, cht xc tc lm gim nng lng hot ha ca phn ng ha hc, c ngha l n ch tham gia vo cc phn ng trung gian m khng ng vai tr l cht tham gia phn ng. Sau phn ng, cht xc tc li phc hi v trng thi ban u tip tc xc tc.

6.4.2 C ch ca s xc tc do enzyme
Hu nh tt c cc bin i ha sinh trong t bo v c th sng u c xc tc bi enzyme pH trung tnh, nhit v p sut bnh thng, trong khi a s cc cht xc tc ha hc khc li ch xc tc nhit v p sut cao. Chnh nh vic to c mi trng c hiu (bi trung tm hot ng ca enzyme lin kt vi c cht) c li nht v mt nng lng thc hin phn ng m enzyme c c nhng kh nng c bit nu trn.

91

Trong phn ng c enzyme xc tc, nh s to thnh phc hp trung gian enzyme-c cht m c cht c hot ha. Khi c cht kt hp vo enzyme, do kt qu ca s cc ha, s chuyn dch ca cc electron v s bin dng ca cc lin kt tham gia trc tip vo phn ng s dn ti vic thay i ng nng cng nh th nng, kt qu l lm cho phn t c cht tr nn hot ng hn, nh phn ng c thc hin d dng hn. Nng lng hot ha khi c s xc tc ca enzyme khng nhng nh hn rt nhiu so vi trng hp khng c xc tc m cng nh hn so vi c trng hp c cht xc tc thng thng. V d trong phn ng phn hy H2O2 thnh H2O v O2 nu khng c cht xc tc th nng lng hot ha l 18 Kcal/mol, nu c cht xc tc l platin th nng lng hot ha l 11,7 Kcal/mol, cn nu c enzyme catalase xc tc th nng lng hot ha ch cn 5,5 Kcal/mol. Nhiu dn liu thc nghim cho thy qu trnh to thnh phc hp enzyme-c cht v s bin i phc hp ny thnh sn phm, gii phng enzyme t do thng tri qua ba giai on theo s sau: E + S ES P + E [Trong E l enzyme, S l c cht (substrate), ES l phc hp enzyme-c cht, P l sn phm (product)] - Giai on th nht: enzyme kt hp vi c cht bng lin kt yu to thnh phc hp enzyme-c cht (ES) khng bn, phn ng ny xy ra rt nhanh v i hi nng lng hot ha thp. - Giai on th hai: xy ra s bin i c cht dn ti s ko cng v ph v cc lin kt ng ha tr tham gia phn ng. - Giai on th ba: to thnh sn phm, cn enzyme c gii phng ra di dng t do. Cc loi lin kt ch yu c to thnh gia E v S trong phc hp ES l: tng tc tnh in, lin kt hydrogen, 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. Vi phng php nghin cu bng tia X v phng php ha hc ngi ta lm sng t cch thc gn c cht v c ch hot ng ca mt s enzyme nh lysozyme, chymotrypsin, carboxypeptidase A, ... Sau y s gii thiu chi tit hn c ch phn ng ca carboxypeptidase A. Carboxypeptidase A (EC 3.4.17.1) thuc nhm peptidhydrolase, xc tc cho s thy phn lin kt peptide, phn ng xy ra vi vn tc ln nu amino acid u C l amino acid thm. Enzyme ny cng thy phn lin kt ester. Carboxypeptidase A c khi lng phn t 34,3. KDa cha 1 mol Zn/1 mol E. Zn tham gia trong hot ng xc tc ca enzyme. Khi thay th Zn bng cc kim loi ha tr hai khc s lm thay i hot v c th c tnh

92

c hiu ca enzyme. Trong phn t enzyme, Zn gn b mt phn t, tng tc vi gc His - 69, His - 196 v Glu - 72. Cc gc amino acid c vai tr xc tc trong trung tm hot ng ca enzyme l: Arg-145, Tyr-248 v Glu-270. C ch phn ng xc tc ca Carboxypeptidase A c xc nh trn c s kt qu nghin cu phn ng ca n vi dipeptid glycyltyrosine. Qu trnh phn gii lin kt peptid c th c phn thnh cc bc sau: - To thnh phc ES: Khi tip xc vi c cht, cc nhm trong trung tm hot ng ca enzyme thay i v tr trong khng gian. Nhm guanidin ca Arg-145 cng nh nhm carboxyl ca Glu-270 dch chuyn 2, nhm hydroxyl ca Tyr-248 dch chuyn 12 t ch gn trn b mt phn t chuyn vo trong n vng gn vi lin kt peptid ca c cht. Tng tc gia cc nhm chc ca trung tm hot ng vi glycyltyrosine nh sau (hnh 6.2): - Nhm carboxyl t do ca c cht kt hp vi nhm tch in dng ca Arg-145 ca enzyme qua lin kt ion. - Nhm NH trong lin kt peptide ca c cht to thnh lin kt hydrogen vi nhm -OH ca Tyr-248.

- Oxy trong nhm -CO- ca lin kt peptide tng tc vi Zn, cn carbon trong nhm -CO- ny tng tc vi nhm carboxyl ca Glu-270 qua phn t nc. - Ct t lin kt v gii phng sn phm. Nguyn t Zn phn cc lin kt -CO, tng tnh i in t ca nguyn t carbon, do lm tng tng tc ca n vi nc hoc vi nhm i nhn ca phn t protein enzyme. Gc Glu-270 hot ha phn t nc, nhm -OH c to thnh tn cng trc tip vo nguyn t cacbon ca -CO- (trong lin kt peptide ca c cht), lin kt peptide b ko

93

cng ra v b t. Gc Tyr-248 nhng hydrogen cho nhm -NH trong lin kt peptide cho c cht, gii phng sn phm u tin l tyrosine ca c cht v acyl-enzyme (hnh 6.3). Sau khi lin kt peptide b ct t, trng thi ion ha ca cc nhm acid v base b bin i tng ng vi pH mi trng, Tyr-248 kt hp vi proton, tr v trng thi ban u.

6.5 Tnh c hiu ca enzyme 6.5.1 Khi nim chung

Do cu trc l ha c bit ca phn t enzyme v c bit l ca trung tm hot ng m enzyme c tnh c hiu rt cao so vi nhng cht xc tc thng thng khc. Mi enzyme ch c kh nng xc tc cho s chuyn ha mt hay mt s cht nht nh theo mt kiu phn ng nht nh. c tnh tc dng la chn cao ny gi l tnh c hiu hoc tnh chuyn ha ca enzyme. Tnh c hiu l mt trong nhng c tnh c bn quan trng nht ca enzyme.

6.5.2 Cc hnh thc c hiu

C th phn bit hai kiu c hiu: kiu phn ng v c cht. 6.5.2.1 c hiu kiu phn ng Trong a s trng hp, mi enzyme u c tnh c hiu vi mt loi phn ng nht nh. Nhng cht c kh nng xy ra nhiu loi phn ng ha hc th mi loi phn ng y phi do mt enzyme c hiu xc tc. V d, amino acid c kh nng xy ra phn ng kh carboxyl ha (decarboxylation), phn ng kh amin ha bng cch oxy ha v phn ng vn chuyn nhm amin, v vy mi phn ng ny cn c mt enzyme c hiu tng ng xc tc theo th t l decarboxylase, amino acid oxydase v aminotransferase. 6.5.2.2 c hiu c cht Mi enzyme ch xc tc cho s chuyn ha mt hoc mt s cht nht nh. Mc c hiu c cht ca cc enzyme khc nhau khng ging nhau, ngi ta thng phn bit thnh cc mc nh sau: - c hiu tuyt i Mt s enzyme hu nh ch xc tc cho phn ng chuyn ha mt c cht xc nh v ch xc tc cho phn ng y m thi. V d: urease, arginase, glucoseoxydase, ... i vi cc enzyme ny, ngoi cc c cht c hiu ca chng l urea, arginine, -D-glucose (theo th t tng ng) chng cng c th phn gii mt vi cht khc nhng vi vn tc thp hn nhiu. Chng hn nh urease, ngoi urea n cn c th phn gii hydroxyurea nhng vi tc thp hn 120 ln. Nhng enzyme c tnh c hiu tuyt i thng c dng nh lng chnh xc c cht ca n. 94

- c hiu nhm tuyt i Cc enzyme ny ch tc dng ln nhng cht c cng mt kiu cu trc phn t, mt kiu lin kt v c nhng yu cu xc nh i vi nhm nguyn t phn lin kt chu tc dng. V d: maltase thuc nhm -glucosidase ch xc tc cho phn ng thy phn lin kt glucoside c to thnh t nhm -OH glucoside ca -glucose vi nhm -OH ca mt monose khc. - c hiu nhm tng i Mc c hiu ca cc enzyme thuc nhm ny km hn nhm trn. Enzyme c kh nng tc dng ln mt kiu lin kt ha hc nht nh trong phn t c cht m khng ph thuc vo cu to ca cc phn tham gia to thnh mi lin kt . V d lipase c kh nng thy phn c tt c cc mi lin kt ester. Aminopeptidase c th xc tc cho s thy phn nhiu peptide. - c hiu quang hc (c hiu lp th) Hu nh tt c cc enzyme u c tnh c hiu khng gian rt cht ch, ngha l enzyme ch tc dng vi mt trong hai dng ng phn khng gian ca c cht. Enzyme ch tc dng vi mt trong hai dng ng phn quang hc ca cc cht. V d, phn ng kh nc ca malic acid to thnh fumaric acid di tc dng ca fumarate hydratase ch xy ra i vi L-malic acid m khng tc dng ln D-malic acid. Enzyme cng th hin tnh c hiu ln mt dng ng phn hnh hc cis hoc trans. V d: enzyme fumarate hydratase ch tc dng ln dng trans ca fumaric acid m khng tc dng ln dng cis to thnh L-malic acid. Trong t nhin cng c cc enzyme xc tc cho phn ng chuyn ha tng h gia cc cp ng phn khng gian tng ng. V d, lactate racemase ca vi khun xc tc cho phn ng chuyn ha ln nhau gia D- v Llactic acid, aldo-1-epimerase xc tc cho phn ng ng phn ha -D-glucose thnh -Dglucose, maleinate cis-trans isomerase ca vi khun xc tc cho phn ng ng phn ha gia maleic acid (dng cis) v fumaric acid (dng trans), ... Cc enzyme ny c vai tr quan trng khi sn xut cc cht dinh dng bng phng php ha hc, v chng c th chuyn cc cht t dng c th khng th s dng c thnh dng c th hp thu. Enzyme cn c kh nng phn bit c 2 gc i xng trong phn t ging nhau hon ton v mt ha hc. V d, hai nhm -CH2OH trong phn t glycerol, glycerophosphate kinase xc tc cho phn ng chuyn v gc phosphate t ATP n C3 ca glycerol (ch khng phi C1).

6.6 Cc yu t nh hng n vn tc phn ng enzyme

Phn ng do enzyme xc tc ph thuc vo nhiu yu t nh: nng enzyme, bn cht v nng cc cht phn ng (c cht), nhit , pH ca mi trng, cc ion kim loi, cc cht v c v hu c khc, ... iu ng lu l cc yu t ha l khng ch nh hng n phn ng enzyme theo kiu ging nh cc phn ng ha hc thng thng m 95

cn nh hng n vn tc phn ng thng qua tc dng ca chng i vi cu trc phn t enzyme.

6.6.1 Nng enzyme

Ni chung trong iu kin tha c cht, vn tc phn ng ph thuc tuyn tnh vo nng enzyme, v = k[E] Trong : v l vn tc phn ng; [E] l nng enzyme. Cng c trng hp khi nng enzyme qu ln, vn tc phn ng tng chm.

6.6.2 Nng c cht, m hnh Michaelis - Menten

Nm 1913, L. Michaelis v M. Menten a ra m hnh gii thch tnh cht ng hc ca phn ng enzyme v lp c phng trnh biu din mi quan h vn tc phn ng (v) vi nng c cht ca enzyme. c im quan trng nht ca m hnh ny l: m u phn ng cn thit phi to thnh phc trung gian enzyme - c cht (ES). Sau phc ES chuyn ha tip to thnh sn phm cui cng ca phn ng v enzyme t do, enzyme li kt hp vi phn t c cht khc bt u vng xc tc mi. Trng hp n gin nht, phn ng ch c mt c cht S, enzyme (E) xc tc cho s chuyn ha n ch to thnh mt sn phm P, phn ng xy ra nh sau:
K3

Trong : k1, k2, k3 l cc hng s vn tc cc phn ng tng ng v1, v2, v3. Vn tc phn ng chuyn ha phc ES to thnh sn phm v enzyme quyt nh vn tc xc tc phn ng chuyn S P. Vn tc phn ng t l vi nng ca phc ES, nng ES cng cao, vn tc phn ng cng ln. K hiu [E0] l nng enzyme ban u, [ES] nng phc trung gian enzyme-c cht v [E] l nng enzyme t do khi phn ng t n cn bng. Do [E] = [ ][ES] . [S] l nng c cht ban u v cng c xem l nng c cht lc phn ng t n cn bng v nng c cht trong phn ng lun ln gp nhiu ln nng enzyme do c th b qua lng [S] trong phc ES. Hn na, khi nghin cu ng hc 96

phn ng enzyme thng xc nh vn tc ban u, lng c cht b chuyn ha cha ng k so vi nng ban u ca n. T phng trnh phn ng (1) c th thit lp cc phng trnh vn tc phn ng nh sau: - Vn tc phn ng to thnh ES l: k1 ([E0] - [ES]) [S] (2) - Vn tc phn li phc ES l tng vn tc ca 2 phn ng: + Phn ng phn li to thnh E v S (ngc vi phn ng kt hp) + Phn ng bin i thnh P v E Do vn tc phn li phc ES l bng: (k2 + k3)[ES] (3). Khi h thng phn ng t n cn bng, vn tc to thnh ES bng vn tc phn li ES: k1 ([E0] - [ES]). [S] = (k2 + k3) [ES] (4) n gin hn, dng hng s

v sp xp li cc s hng trong phng trnh (4) s c:

Nh trn ni, ES cng ln vn tc phn ng cng ln. Khi nng c cht ln, tt c cc phn t enzyme c trong phn ng u tham gia trong phc ES, vn tc phn ng s t n cc i (V). Do c th thit lp t l sau:

(6)

Tnh

t phng trnh (5) v thay vo (6) s c:

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(7) y l phng trnh Michaelis-Menten c Holden v Briggx hon thin. Km c gi l hng s Michaelis. th biu din nh hng nng c cht n vn tc phn ng (theo phng trnh (7)), trong v l hm s ca [S] c dng ca nhnh hyperbol vung gc. xc nh Km cng nh V c th da vo th biu din mi quan h gia vn tc phn ng vi nng c cht: thay i nng c cht, vi mi nng c cht xc nh vn tc ban u ca phn ng. Theo cch bin din phng trnh (7), cn tin hnh kh nhiu th nghim mi v c chnh xc. V vy ngi ta ci bin phng trnh ny thnh dng c ng biu din thng. Cch n gin nht l ly s nghch o ca c 2 v trong phng trnh (7) (Lineaweaver v Burk, 1934), do s c phng trnh sau:

(8) Phng trnh (8) c dng Y - ax + b, ng biu din ca n l thng, ct trc tung im v ct trc honh im . Theo cch ny, ch cn tin hnh th nghim vi ba, bn nng c cht c th v c th kh chnh xc v xc nh c cc tr s Km v V. iu quan trng cn nhn mnh l khi nghin cu ng hc phn ng enzyme cn xc nh vn tc ban u ca phn ng, s dng ch phm enzyme tinh khit. Ngoi ra, nng c cht cng khng nn qu ln v n c th lm gim vn tc phn ng enzyme.

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i vi cc enzyme allosteric, ng biu din s ph thuc ca v vo [S] khng c dng hyperbol. Trong nhiu trng hp, ng biu din quan h gia vn tc ban u ca phn ng vi nng c cht c dng gn nh ch S (dng sigmoid). iu c ngha l khi phn t c cht u tin kt hp vo mt trung tm hot ng ca enzyme lm tng nhanh vic kt hp cc phn t c cht tip theo vo cc trung tm hot ng khc trong phn t enzyme. iu ny cng ging nh khi mt phn t O2 kt hp vo Hb lm tng nhanh s tng tc ca cc phn t O2 tip theo. ng biu din c dng ch S cng cho thy ch cn nng c cht tng ln rt nh cng c th lm tng vn tc xc tc nhiu ln ln hn so vi cc enzyme tun theo phng trnh Michaelis - Menten.

6.6.3 nh hng ca cc cht km hm

Hot ca enzyme c th b thay i di tc dng ca mt s cht c bn cht ha hc khc nhau. Cc cht lm gim hot enzyme gi l cc cht km hm hoc cc cht c ch (inhibitor), thng k hiu l I. Cc cht ny c th l nhng ion, cc phn t v v, hu c k c cc protein. Cc cht c ch tham gia trong s iu ha, kim tra cc qu trnh trao i cht trong h thng sng.

(9) Cc cht gy bin tnh protein u l nhng cht km hm khng c hiu enzyme. Nhiu cht khc khng lm bin tnh protein nhng vn lm gim hot xc tc ca n theo c ch khc. Cc cht ny c th km hm thun nghch hoc khng thun nghch enzyme. Nu l km hm thun nghch, phn ng kt hp gia enzyme v cht km hm (I) 99

nhanh chng t n cn bng. Trong trng hp km hm khng thun nghch, K-i rt b, c th xem nh bng 0. I kt hp vi E bng lin kt ng ha tr hoc kt hp rt cht n mc kh lng tch khi E, s phn li phc EI rt chm. - Cc cht km hm cnh tranh Cc cht km hm cnh tranh l nhng cht km hm thun nghch enzyme, c cu trc tng t vi cu trc ca c cht, do c kh nng kt hp vo trung tm hot ng ca E chim ch kt hp ca c cht. S kt hp ca I v S vo trung tm hot ng ca enzyme c tnh cht loi tr ln nhau. Nh vy, I cnh tranh lm gim vn tc phn ng xc tc l do lm gim s lng phn t enzyme c kh nng kt hp vi c cht. V d, cht km hm cnh tranh ca succinate dehydrogenase l malonic acid (HOOC-CH2COOH), c cu to kh ging vi c cht ca enzyme l succinic acid (HOOC-CH2-CH2COOH). Do ch I v S c khuynh hng y nhau ra khi phc vi enzyme cho nn nu nng c cht rt ln so vi nng cht km hm, c th loi tr hon ton tc dng km hm ca n. Trong trng hp n gin nht (phng trnh phn ng (1)), khi c cht km hm cnh tranh, c cc phn ng xy ra nh sau: E + S ES E + P E+I EI

Nh trn ni, cht km hm cnh tranh cng kt hp vo trung tm hot ng ca enzyme, nhng khc vi c cht ch bn thn cht km hm khng b chuyn ha di tc dng ca enzyme.

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Cht km hm ny kt hp vi enzyme ch khc vi trung tm hot ng (hnh 6.5) lm thay i cu trc khng gian ca phn t enzyme, do theo hng khng c li cho hot xc tc ca n lm gim vn tc phn ng xc tc. Sau khi kt hp vi cht km hm khng cnh tranh, enzyme vn c th kt hp vi c cht to thnh phc EIS. C th vit phng trnh phn ng (1) khi c cht km hm khng cnh tranh nh sau: E + S ES E + P E + I EI EI + S EIS Khi c cht km hm khng cnh tranh vn tc cc i b gim dn trong khi Km khng thay i, vn tc phn ng b gim tng ng. Di tc dng ca cht km hm khng cnh tranh, mc km hm khng ph thuc vo tng quan nng gia S v I, nng c cht ln cng khng loi tr c tc dng km hm. Trong mt s trng hp sn phm phn ng c th tc dng nh cht km hm khng cnh tranh ca enzyme. Ngoi ra cng c trng hp cht km hm ch kt hp vi ES m khng kt hp vi E t do, trong trng hp ny c V v Km u b gim dn, do dc ca ng biu din khng i. Cc cht km hm, nht l cc cht km hm c tnh c hiu cao c ngha ln trong nghin cu khoa hc cng nh trong thc t. V d: disopropilphosphofluoridate (DIPF hoc DFP), iodoacetamid, p-cloromerurbenzoate, ... thng c dng pht hin cc nhm chc nng trong trung tm hot ng ca enzyme. DIPF ch phn ng vi gc Ser c vai tr xc tc trong trung tm hot ng ca enzyme acetylcholinesterase, trypsin, chymotrysin, to thnh phc khng hot ng. Cc cht km hm protein thng c tnh c hiu kh cao, km hm thun nghch enzyme, chng c vai tr quan trng trong vic iu ha, kim tra qu trnh trao i cht trong h thng sng. Thuc loi ny, cc protein iu ha hot proteinase c nghin cu nhiu hn c v c s dng trong y hc, trong nghin cu khoa hc.

6.6.4 Cc cht hot ha (activator)

Cht ny lm tng hot xc tc ca enzyme. Cc cht ny c bn cht ha hc khc nhau, c th l cc anion, cc ion kim loi hoc cc cht hu c c cu to phc tp hn. V d, tc dng ca anion chloride, brom, iode n hot ca -amylase ng vt; tc dng ca mt s ion kim loi nh Mn++, Zn++, ... i vi hot ca cc protease ... Tuy nhin tc dng kch hot ch gii hn nhng nng xc nh, vt qu gii hn ny c th lm gim hot enzyme. Gluthation dng kh (Glu-Cys-Gly) c tc dng kh lin kt disulfide thnh nhm sulhydryl t do (-SH) cho nn cng c tc dng hot ha nhiu enzyme. Cc cht nu thng kt hp trc tip vi phn t enzyme, lm thay i cu to khng gian ca n theo hng c li cho hot ng xc tc ca enzyme. Mt s cht khc c th tc dng theo cch gin tip, v d loi tr cc yu t km hm khi mi trng phn ng.

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6.6.5 Nhit
Vn tc phn ng do enzyme xc tc ch tng theo nhit trong mt gii hn xc nh m phn t enzyme vn cn bn cha b bin tnh. i lng c trng cho nh hng nhit n vn tc phn ng ha hc cng nh phn ng enzyme l h s nhit Q10. l t l gia hng s vn tc phn ng nhit no so vi hng s vn tc phn ng nhit thp hn 10oC. H s ny cng ln, phn ng cng kh xy ra nhit bnh thng. H s Q10 ca phn ng enzyme l t 1 n 2 (ca phn ng ha hc bng 2 n 3). T h s Q10 c th tnh c nng lng hot ha ca phn ng enzyme, nh gi qu trnh xc tc.

Hnh 6.6: nh hng ca nhit n hot xc tc ca enzyme

ng biu din nh hng ca nhit n vn tc phn ng ca nhiu enzyme c dng nh hnh 6.6. Nhit ng vi hot enzyme cao nht gi l nhit ti thch ca enzyme (topt). a s enzyme c topt vo khong 40-50oC. Tuy nhin Topt ca 1 enzyme khng c nh m c th thay i ty c cht, pH mi trng, thi gian phn ng, ... Nhit m enzyme b mt hon ton hot tnh xc tc gi l nhit ti hn, thng vo khong trn 70oC. nhit ti hn, enzyme b bin tnh, t khi c kh nng c hi

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phc li hot . Ngc li, nhit di 0oC, hot enzyme tuy b gim nhng li c th tng ln khi a v nhit bnh thng. bn ca phn t enzyme tng khi c c cht, coenzym, Ca2+, ...

6.6.6 pH mi trng
pH mi trng nh hng r rt n phn ng enzyme v n nh hng n mc ion ha c cht, enzyme v nh hng n bn ca protein enzyme. a s enzyme bn trong gii hn pH gia 5 v 9, bn ca enzyme c th tng ln khi c c cht, coenzyme, Ca2+ , ... ng biu din nh hng pH n vn tc phn ng ca nhiu enzyme c dng nh hnh 6.7. pH ti thch (pHopt) cho hot ng ca nhiu enzyme vo khong 7. Tuy nhin, cng c mt s enzyme c pHopt rt thp (pepsin, protease acid ca vi sinh vt, ...) hoc kh cao (subtilysine, pHopt > 10). pHopt ca enzyme cng khng c nh m ph thuc nhiu vo cc yu t khc nh c cht, tnh cht dung dch m, nhit , ...
pH

6.6.7 Tc dng ca cht chng chuyn ha

T lu ngi ta nhn thy c hin tng cnh tranh gia nhng cht chuyn ha v nhng cht c lin quan v mt cu trc i vi chng. Nhng cht ny c gi l cc cht chng chuyn ha. Trn thc t, c nhiu cht c hot tnh sinh l b cnh tranh bi nhng cht c lin quan v mt cu trc, v nhiu cht c hot tnh dc l c kh nng lm bin i qu trnh chuyn ha nh tc dng ca chng i vi cc enzyme v cc h enzyme c lin quan. Tt c ng vt bc cao v nhiu vi sinh vt khng c kh nng t tng hp mt s hp cht hu c vn rt cn thit cho s sng cn v pht trin ca c th, cc cht ny thng phi c a t bn ngoi vo. Ty theo ging, loi, nhu cu i hi cc cht ny c khc nhau. Cc cht ny thng bao gm mt s loi amino acid, mt s acid bo, cc vitamin, mt s purine v pyrimidine, ... v thng c c th s dng tng hp protein, nucleic acid, ... hoc c bin i thnh cc cht cn thit khc ca t bo. Vic s dng cc cht ny phi nh vo tc dng ca enzyme v nhng enzyme ny c th b c ch bi nhng cht c cu trc lin quan vi cc cht y. Nhng cht c tc dng c ch nh vy c gi l cht chng chuyn ha. Cc cht chng chuyn ha cng c th lm tr ngi cho vic s dng cc cht chuyn ha do chnh c th tng hp nn. Trong khi nghin cu cc con ng chuyn ha, ngi ta c th dng cc cht chng chuyn ha c ch qu trnh chuyn ha v ngc li, ngi ta c th khc phc hin tng c ch s pht trin ca c th bng cch cho thm nhng cht chuyn ha cn thit. Nh vy, s nghin cu nhng cht chng chuyn ha s gip ta pht hin c nhng cht chuyn ha quan trng v nhng yu t pht trin cn thit cho cc chng loi khc nhau v ng thi gip ta tm kim nhng cht c ch hiu nghim i vi s pht trin ca cc vi sinh vt gy bnh v cc u c. Cc 103

cht chng chuyn ha ly c t cc c th sng nh vi khun, nm actinomyces, v.v... thng c gi l cht khng sinh. Mt hin tng c ch cnh tranh quen thuc gia cht chuyn ha v cht chng chuyn ha l s cnh tranh gia paraaminobenzoic acid (vit tt l PABA) v cc cht sulfonamid c cu trc tng qut l NH2-C6-H4-SO2NHR nh sulfanilamid, sulfaguanidin, sulfiazol, sulfadiazin. - Paraaminobenzoic acid l mt trong nhng tin cht quan trng tng hp folic acid. Acid ny l mt coenzyme quan trng trong cc phn ng ha sinh vn chuyn v s dng cc mnh c mt carbon. Nhng phn ng ny c vai tr rt quan trng i vi s pht trin ca c th. Vai tr ca cc cht sulfonamid l ch c ch (cnh tranh vi paraaminobenzoic acid) enzyme c lin quan trong qu trnh tng hp folic acid t tin cht paraaminobenzoic acid v nhng cht c lin quan khc, lm cho qu trnh tng hp ny khng thc hin c.

- Nhng vi khun i hi paraaminobenzoic acid lm yu t pht trin l nhng vi khun c kh nng dng paraaminobenzoic acid lm tin cht t tng hp folic acid, s pht trin ca nhng vi khun ny thng b c ch bi sulfonamid v ngc li, c th khc phc hin tng c ch ny bng cch cho thm paraaminobenzoic acid. Nhng c th khng c kh nng t tng hp folic acid, thng i hi phi a cht ny t ngoi vo lm yu t pht trin v khng i hi paraaminobenzoic acid tng hp folic acid cho nn s pht trin ca nhng loi c th ny khng chu tc dng c ch ca sulfonamid. C th ngi khng i hi paraaminobenzoic acid lm yu t pht trin (m dng trc tip folic acid), do khi dng sulfonamid, cht ny s c ch cnh tranh vi paraaminobenzoic acid ca vi khun trong c th, lm cho vi khun khng pht trin c, nhng khng nh hng n s pht trin ca ngi mang vi khun.

Ti liu tham kho

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Phm Thi Trn Chu, Trn Th ng, 1999. Ha sinh hc. NXB Gio dc, trang 62-76. Nguyn Xun Thng (ch bin), o Kim Chi, Phm Quang Tng, Nguyn Vn ng, 2004. Ha sinh hc. NXB Y hc, H Ni. nh H (ch bin), ng Th Hoi An, Nguyn Th Ho, nh H, Phm Th Mai, Trn Thanh Lan Phng, Th Thanh Thy, L Xun Trng, 2005. Ha sinh y hc. NXB Y hc, tp. H Ch Minh. Hong Quang (ch bin), Nguyn nh , Trng Th Minh c, Bch Vng Hi, Phan Hi Nam, on Trng Ph, 2000. Ha sinh y hc. NXB Qun i nhn dn, H Ni. Nelson D. L., Cox M. M., 2005. Lehninger Principles of Biochemistry, Fourth Edition. Freeman and Company, New York, USA.

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