Mc lcNG DNG ENZYME TRONG HA SINH21.Tng quan v enzyme21.1.nh ngha enzyme21.1.Lc s nghin cu enzyme21.1.1.Giai on 121.1.2.Giai on 221.1.3.Giai on 331.1.4.Giai on 462.Enzyme v ng dng72.1.ng dng trong ha sinh72.1.1.Tng quan v Ribozyme72.1.2.C ch102.1.3.Tch chit Ribozyme112.1.4.ng dng ca ribozyme122.2.ng dng trong y dc132.3.ng dng trong ha hc132.4.ng dng trong cng nghip142.5.ng dng trong cng nghip thc phm142.6.ng dng trong cng nghip dt162.7.ng dng trong cng nghip thuc da162.8.ng dng trong nng nghip16TI LIU THAM KHO17

NG DNG ENZYME TRONG HA SINHTng quan v enzyme1.1. nh ngha enzymeTrong c th sng (cc t bo) 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 cc phn ng ha hc khc nhau. Cc phn ng ha hc phc tp ny c lin quan cht ch vi nhau v iu chnh ln nhau. Enzyme l cc hp cht protein xc tc cho cc phn ng ha 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 hu ht cc loi t bo ca c th sng. Chnh do nhng tc nhn xc tc c ngun gc sinh hc nn enzyme cn c gi l cc cht xc tc sinh hc (biocatalysators) nhm phn bit vi cc cht xc tc ha hc.Enzyme hc l khoa hc nghin cu nhng cht xc tc sinh hc c bn cht protein. Hay ni cch khc, enzyme hc l khoa hc nghin cu nhng tnh cht chung, iu kin, c ch tc dng v tnh c hiu ca cc enzyme.1.1. Lc s nghin cu enzymeDo enzyme hc c coi nh ct sng ca ha sinh hc nn phn ln cc nghin cu ha sinh t trc n nay u lin quan nhiu n enzyme. V s pht trin ca hc thuyt enzyme, c th chia thnh 4 giai on: Giai on 1: trc th k th XVII Giai on 2: t th k XVII n na u th k XIX Giai on 3: t gia th k XIX n 30 nm u ca th k XX Giai on 4: t nhng nm 30 ca th k XX n nay.1.1.1. Giai on 1Trc th k XVII ngi ta bit s dng cc qu trnh enzyme trong i sng song ch c tnh cht kinh nghim thc t v thng qua hot ng ca vi sinh vt. l cc qu trnh ln men ru, mui da, lm tng v nc chm... thi k ny ngi ta cha hiu v bn cht enzyme v cc qu trnh ln men.1.1.2. Giai on 2 giai on ny cc nh bc hc tin hnh tm hiu bn cht ca cc qu trnh ln men. Thi k ny khi qut hin tng ln men nh l hin tng ph bin trong s sng v enzyme l yu t gy nn s chuyn ha cc cht trong qu trnh ln men.Vo nhng nm 1600 ca th k XVII, Van Helmont l ngi u tin c gng i su tm hiu bn cht ca qu trnh ln men. Van Helmont nhn thy thc cht ca s tiu ha l s chuyn ha ha hc ca thc n v gii thch c ch ca n vi s so snh n vi qu trnh ln men ru. Danh t ferment (t ch Latinh fermentatio - s ln men) c Van Helmont dng ch tc nhn gy ra s chuyn bin cc cht trong qu trnh ln men ru.Vo na cui th k th XVIII, nh t nhin hc ngi Php l Raumur cng nghin cu bn cht ca s tiu ha. Nh t nhin hc ny cho chim qu en nut nhng ming tht t sn trong ng kim loi c thnh c c sn v buc vo dy thp. Sau vi gi khng thy g trong ng. Hin tng ny thc y s nghin cu thnh phn dch tiu ha tm hiu kh nng tiu ha ca dch d dy. Sau th nghim ny mt thi gian, vo nm 1783, nh bc hc ngi l Spalanzani lp li th nghim bng cch ly dch d dy trn vi tht mi v thy c hin tng ha tan xy ra.Vo u th k XIX, cc nh nghin cu tch c cc cht gy ra qu trnh ln men. Nm 1814 Kirchoff, vin s Saint Petercburg pht hin nc chit ca mm i mch c kh nng chuyn ha tinh bt thnh ng nhit thng. y l cng trnh u tin thu c ch phm amylase dng dung dch v lch s enzyme hc thc s c xem nh bt u t y.Mi chn nm sau (nm 1833), hai nh khoa hc ngi Php l Payen v Pessoz chng minh cht c hot ng phn gii tinh bt thnh ng c th tch c dng bt. Th nghim c tin hnh bng cch cho etanol vo dch chit ca la i mch ny mm th thy xut hin kt ta. Kt ta c hnh thnh ny c kh nng chuyn ha tinh bt v nu un kt ta ny s mt tc dng chuyn ha. Danh t diastase (t ch Latinh diastasis - phn ct) l do Payen v Persoz dng gi enzyme amylase lc by gi.Tip ngi ta cng tm ra v tch c nhiu enzyme khc nh enzyme phn gii protein ca dch tiu ha trong d dy nh Pepsin (Emberle v Shwan) - nhng nh khoa hc ngi c, nm 1836)...Sau , l thuyt xc tc ra i. Nm 1835, nh khoa hc Berzelius c quan im cho rng tng tc phn ng l hin tng xc tc. y l mt quan im ng. Song tht ng tic l nh khoa hc ny coi cc cht xc tc ny hot ng c l do "lc sng" khng theo s iu khin ca con ngi. y l quan im duy tm, siu hnh lm tr tr s pht trin ca khoa hc nht l nh hng su sc n s pht trin ca ngnh enzyme hc.1.1.3. Giai on 3Giai on t gia th k XIX n 30 nm u ca th k XX. giai on ny mt s lng rt ln cc enzyme dng ha tan c tch chit.Trong thi k ny, c hai trng phi u tranh vi nhau: l trng phi Pasteur - nh bc hc v i ngi Php v trng phi Liebig - nh bc hc ni ting ngi c. Trng phi Pasteur:Nm 1856 Pasteur cp n bn cht ca qu trnh ln men. ng cho rng khng th tch cc enzyme khi t bo. Tc dng v tnh cht ca enzyme gn lin vi s sng ca t bo v qu trnh ln men ru l kt qu hot ng sng ca t bo nm men ch khng phi l kt qu ca tc dng ca enzyme. ng tin hnh th nghim v nhn thy nu mt dung dch hu c, v d dung dch glucose trong bnh kh trng th khng xy ra qu trnh ln men ru. Chnh v suy ngh y, Pasteur chia cc enzyme thnh 2 loi: "enzyme c t chc" v "enzyme khng c t chc".Theo ng, cc "enzyme c t chc" l nhng enzyme khng th tch khi t bo, khi tch chng s b mt tc dng xc tc nh cc enzyme ca cc t bo nm men thc hin qu trnh ln men ru; cn cc "enzyme khng c t chc" l cc enzyme c th thc hin tnh xc tc ca n ngoi c th nh cc enzyme c trong dch tiu ha (v d Pepsin trong d dy, amylase trong tuyn nc bt, trong mm thc...)Quan im sai lm ny ca Pasteur thng tr ngnh enzyme hc trong mt thi gian di. Nm 1878 Kuhne ngh dng danh t "ferment" (t ting Latinh: fermentatio = ln men) gi cc "enzyme c t chc" v gi cc cht chit c tc dng xc tc cho phn ng ha hc l cc enzyme (t ch Hy Lp: en = bn trong, zyme = men ru, tc l " trong nm men" gi cc enzyme "khng c t chc". Danh t enzyme c xut pht t y. Trng phi Liebig:Chng li quan im trn ca Pasteur, Liebig (trc c c Berzelius) cho rng c th khng c hot ng ca cc t bo vi sinh vt cng c qu trnh ln men. iu c ngha l ng coi enzyme nh l mt cht ha hc gy nn hiu qu tng t nh cc cht xc tc, tc dng c trong v ngoi t bo, khng ph thuc vo hot ng sng ca vi sinh vt.Nhng nm 1871 Liebig tht bi v thc nghim khng chng minh c quan im trn ca mnh. Cc th nghim c tin hnh bng cch ly dch chit t t bo nm men nghin nt u khng c tc dng gy ln men ru. Cng vo nm 1871 Manatxein l mt bc s ngi Nga dng ct thch anh nghin cc t bo nm men v thu c dch chit khng cha t bo c kh nng bin i ng thnh ru. Nhng nhng quan st ny khng c ai ch ti. Chnh v vy, quan im siu hnh ca Pasteur hn ch kh nhiu s pht trin ca ngnh enzyme hc. n nm 1897, H. Bchner - mt nh khoa hc ngi c nhn c dch chit nm men bng cch phn hu t bo hon thin hn. Trong th nghim ny, cc t bo nm men c nghin nt hon ton cng vi bt thy tinh, sau c p bng p sut cao. Dch chit thu c khng cha t bo vn c kh nng gy ra qu trnh ln men (chuyn ha glucose thnh ru). iu chng t qu trnh ln men ru khng phi l kt qu ca hot ng sng ca t bo nm men m l kt qu tc dng ca cc enzyme vn c trong cc t bo. Do , quan im sai lm v enzme "c t chc" v enzyme "khng c t chc" m thc cht l v bn cht ca enzyme n lc ny mi hon ton b nh , m ra mt thi k pht trin mi ca ngnh enzyme hc. Cng t khng c s phn bit v ni dung gia thut ng "ferment" v "enzyme". C th ni rng, cng trnh ca Bchner nh du mt bc ngot quan trng trong lch s pht trin ca enzyme hc. Sau , nhiu loi enzyme trong c th sng c tm ra. V vy vic phn loi v gi tn cc enzyme mt cch thng nht cng cn thit. Nm 1883, Duyclo, nh bc hc Php ra nguyn tc phn loi enzyme theo c cht (substrate) do chng bin i v thm ui tn cng "ase" vo. V d enzyme phn gii tinh bt (amilun) l amylase. Tuy vy, trong thc t cn tn ti nhiu ngoi l v thut ng, v d nhng tn gi enzyme pepsin, trypsin, catalase trc y vn c dng. thi k ny, da vo thnh tu ca ha hc, c bit l ha l v ha keo, cc nh khoa hc hng vo vic nghin cu cc tnh cht ha v l hc ca enzyme cng nh hon thin cc phng php lm thun khit enzyme.Giai on quan trng nht trong thi k ny l cc cng trnh ca nh bc hc v i ngi c E. Fisher. ng t nn mng cho nhng khi nim hin i v tnh c hiu ca enzyme, v s tng tc khng gian gia enzyme v c cht. Gi thuyt ni ting ca ng l gia enzyme v c cht kt hp vi nhau nh " kha vi cha kha". Ri nhng nghin cu ca Bach v Palladin v cc enzyme xy ha kh to nn c s cho vic xy dng hc thuyt xy ha kh sinh hc. Trong thi gian ny ngi ta cng pht hin ra c tnh tc dng thun nghch ca enzyme (anilepski, 1894), cc coenzyme cng c pht hin (Harden v Young, 1906). H l nhng ngi khm ph ra rng, dch chit t bo nm men cha hai loi cht cn thit cho qu trnh ln men l "zymase" v "cozymase". H nhn thy dch chit t bo nm men mt hot tnh xc tc nu b thm tch hoc b un ln n 50. Nhng dch chit b thm tch khng hot ng s hot ng khi c trn vi dch b un nng khng hot ng. Nh vy hot ph thuc vo s c mt ca hai loi cht: thnh phn khng bn vi nhit (heat - labile); khng c th thm tch c (c gi l zymase) v mt phn on bn vi nhit (heat - stable), c th thm tch c (c gi l cozymase). Ngy nay chng ta bit rng "zymase" bao gm tt c enzyme, cn "cozymase" bao gm cc ion kim loi, ATP, ADP v cc coenzyme nh NAD+. Thi gian ny ngi ta cng hiu bit c tc dng km hm v hot ha ca mt s enzyme (Sorensen 1909). Vo u th k XX, pht sinh ra c s ng hc trong tc ng ca enzyme da vo nhng nghin cu ca nh bc hc Anh l Brown v nh bc hc Php l Henri. n nm 1913, Michaelis v Menten pht trin cc cng trnh trn v nu ln thuyt ng hc ca s xc tc enzyme. Sau i chin th gii ln th nht nh bc hc ni ting ngi c l Willstatter c rt nhiu cng hin trong vic tm hiu bn cht ha hc ca enzyme. l cng trnh khoa hc 5 nm ca ng v cc cng s (1922) nhm lm thun khit enzyme bng phng php hp th chn lc. Qua t nhn xt thy l nhng giai on cui ca qu trnh lm thun khit enzyme, thng b mt i nhng cht cha c bit no do, enzyme b mt tnh xc tc, cho php Willstatter nu ln ln u tin gi thuyt v enzyme hai cu t (enzyme hai thnh phn). Nhm hot ng (coenzyme, coferment, agon) ch c kh nng xc tc khi kt hp vi phn protein c hiu (apoferment, apoenzyme, feron = protein) n xc nh cc c tnh ca enzyme v ng vai tr ch o trong vic th hin tc dng xc tc ca enzyme. Willstatter coi feron (protein) l cht tr ch c tc dng g. Agon l cht c hp ph trn cht ny. V vo nm 1926, trong mt dp thuyt trnh, ng cho rng enzyme khng thuc mt trong cc hp cht bit, tc l enzyme khng phi l protein, khng phi l glucid, m chng l nhng "cht c bit". chnh l quan nim sai lm ca Willstatter. ng l ngi tm ra c nhiu phng php lm sch enzyme cng nh lm sng t nhiu tnh cht c hiu enzyme. Nhng mc ch chnh l lm sng t bn cht ha hc ca enzyme th ng li khng t c. Ngy nay ngi ta quan nim nu l enzyme hai thnh phn th phn coenzyme quy nh kiu phn ng v chu trch nhim lm bn. Cn apoenzyme quy nh tnh c hiu ca enzyme cng nh tng hiu sut xc tc.Coenzyme + apoenzyme = (holo) enzyme = (enzyme hon chnh)Coferment + apoferment = (holo) fermentCoenzyme ch dng ch phn khng phi protein ca enzyme trong trng hp khi n 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 "prostetic" khi n lin kt cht ch vi phn protein ca enzyme.1.1.4. Giai on 4Bn cht ha hc ca enzyme ch c xc nh ng n t sau khi kt tinh c enzyme. Nm 1926 nh ha sinh M tr tui Sumner (39 tui) thnh cng trong vic chng minh protein c kt tinh t ht u tng l cht ging enzyme xc tc cho phn ng thy phn ur. y cng chnh l enzyme u tin c kt tinh. Bn nm sau (1930) M Northrop tch c pepsin dng tinh th, v vo nm 1931 Northrop v Kunitz cng tch c trypsin dng tinh th.Trong thi k ny J.B.S Hardane vit quyn "Enzymes". Mc d lc bn cht phn t ca Enzyme hu nh vn cn l b mt, nhng tc gi a ra d on tuyt vi v vai tr ca cc tng tc v lin kt yu gia enzyme v c cht trong c ch hot ng ca enzyme. iu ny vn gi nguyn tnh thi s trong thi i ca chng ta.Cc cng trnh ca Sumner v Northrop m ra mt chng mi trong lch s pht trin ca enzyme hc hin i. Nhng kt qu t c cho php xc nh c mt cch dt khot bn cht ha hc ca enzyme l protein. Phi ni rng bn cht ha hc ca phn ln enzyme l protein v nh ngha c tnh cht kinh in v Enzyme phi xem li t sau pht hin ca T. R. Cech nm 1981. Cech pht hin mt RNA c hot tnh xc tc nh enzyme v gi l ribozyme (xut pht t cc tn ribose v enzyme). Ribozyme xc tc cho qu trnh chuyn ha tin cht. RNA thng tin (pre - m RNA) thnh m-RNA. Do enzyme khng nht thit phi l protein! y l mt pht minh c ngha rt ln. Tc gi ca pht minh ny c gii Nobel nm 1989. Cho n nay khong 100 ribozyme c bit. C th ni rng, nhng cng trnh ni trn m mn cho giai on th t ca lch s pht trin enzyme hc ko di cho n hin nay.T gia th k th XX, nht l thi gian gn y enzyme hc pht trin rt mnh. Nh ng dng cc phng php mi, hin i nh: in di, sc k, quang ph, ng v phng x... cho php nghin cu cu trc cng nh c ch tc dng ca nhiu enzyme, c ch ca qu trnh sinh tng hp enzyme v s iu ha hot ng ca enzyme trong t bo.Ngi ta xc nh c cu to ca coenzyme. xc nh c mi lin h ca enzyme v cc vitamin (nhiu vitamin l thnh phn cu to ca coenzyme v phn ln cc vitamin tan trong nc l thnh phn cu to ca cc coenzyme).Ngi ta cng xc nh c cc enzyme xc tc cho cc qu trnh trao i cht nh: h thng enzyme ng phn, Embden - Meyerhof - Parnas nm 1933, h thng enzyme ca chu trnh Kreps - Szent Gyorgy nm 1937 (chu trnh citric acid), chu trnh ornithrin trong trao i cht ca protein nm 1932 (Krebs - Henseleit). Nh nhng phng php mi trong vic tch v lm sch enzyme, ngi ta xc nh c vai tr rt quan trng ca kim loi trong s xc tc ca enzyme v tc dng hot ha ca chng. xc nh c s phn b ca cc enzyme trong t bo. nghin cu c ch tc dng cng nh cu to cc protein enzyme. Bng phng php Rhengen, ngi ta nghin cu cu trc ca ca phn t enzyme, nh cu trc ca ribonuclease (1960, Stein).Enzyme v ng dngEnzyme c xem nh l mt k thut quan trng ca cng ngh sinh hc do c cc chc nng sau: Enzyme l cht xc tc cho mi bin i vt cht trong cng ngh sinh hc. Enzyme v nhiu hot cht sinh hc khc l sn phm ca cng ngh sinh hc. Chng c th dng lm cng c mi ca cng ngh sinh hc, hay s dng trong cc lnh vc khc. Enzyme c xem l thuc th c tnh chuyn ha cao m khng c enzyme th cc qu trnh cng ngh sinh hc khng th ti u ha c.Hin nay, vic sn xut ch phm enzyme cc loi v ang pht trin mnh m trn qui m cng nghip. Thc t c hng nghn ch phm enzyme bn trn th trng th gii, cc ch phm ny c khai thc v tinh ch c mc tinh khit theo tiu chun cng nghip v ng dng. Cc ch phm enzyme ph bin nh amylase, protease, catalase, cellulase, lipase, glucoseoxydaseCh phm enzyme khng ch c ng dng trong y hc m cn c ng dng trong nhiu lnh vc cng nghip khc nhau, trong nng nghip, trong ha hc" ngha ca vic s dng enzyme trong cc lnh vc thc t khng km so vi ngha ca vic s dng nng lng nguyn t".1.2. ng dng trong ha sinh1.2.1. Tng quan v RibozymeMt s ARN c kch thc nh c hot tnh xc tc ging nh enzyme c gi l ribozyme.V vai tr tham gia cc qu trnh phin m v dch m thng tin di truyn, RNA c chia lm ba loi chnh:

ARN thng tin (k hiu: mARN, vit tt t messengerARN): gm mt si Polynucleotit dng thng, c chc nng sao chp thng tin di truyn t gen cu trc em n Riboxom l ni tng hp protein.ARN vn chuyn (k hiu: tARN, vit tt t transportARN): l mt mch polynucleotit c t 80 n 100 ribonucleotit t xon nh hnh chc ba nn mt s on c nguyn tc b sung A-U; G-X. C nhng on khng c nguyn tc b sung, nhng on ny to thnh cc thy trn. Mt trong cc thy trn mang b ba i m. Mt si mt ca si ARN 3' gn vi mt acid amin v u mt t do 5'. Mi tARN ch vn chuyn mt loi acid amin ng vi b ba i m m mang. Cc acid amin s c vn chuyn n riboxom tng hp protein.ARN riboxom (k hiu: rARN): gm mt mch dng xon tng t tARN.Ngoi ra, mt s ARN c vai tr iu khin hot ng gien hoc c chc nng tham gia cc qu trnh pht trin, bit ho t bo nh RNAi (interfering RNA) hay microARN.Khng ch c ngha trong nghin cu c bn pht minh ny cng cung cp mt cng c mi trong k thut di truyn vi tim nng ha hn trong ngn cn s ly nhim virus.Trong mt cng trnh nghin cu hai nh khoa hc Sidney Altman v Thomas Cech pht hin ra rng RNA c th xc tc cho phn ng t nhn i v tng hp nn nhng phn t RNA khc (ribozyme), m u cho lung kin cho rng RNA l nguyn liu di truyn u tin trn tri t. Mt "th gii" RNA c tin l tn ti trc khi DNA n nhn vai tr l nguyn liu di truyn then cht, v RNA chuyn n gi vai tr lin lc gia DNA v protein. S khm ph cht xc tc RNA khng nhng lin h mt thit vi tin ha m cn xut c RNA gi vai tr hot ng nhiu hn trong biu hin gene nhn ra t sm. RNA ribosome xc tc hnh thnh lin kt peptide trong dch m.

Trc khi cng trnh nghin cu ca Altman and Cech, tt c cc enzyme u c coi l cc protein. Cc tnh cht v chc nng c th ca protein c xc nh bng cc nhn t di truyn hay cn gi l gene. Thng tin di truyn trn ADN quy nh cu trc v chc nng ca protein, tuy nhin trc khi nhng thng tin c th c s dng trc tip tng hp nn protein, cc m di truyn cn phi c sao m thnh mt dng phn t acid nuleic khc l RNA (ribonucleic acid). Altman v Cech tin hnh nhng nghin cu c lp v pht hin ra rng RNA khng ch l mt phn t c chc nng di truyn m cn c vai tr nh l mt cht xc tc sinh hc. Khng ch c ngha trong nghin cu c bn pht minh ny cng cung cp mt cng c mi trong k thut di truyn vi tim nng ha hn trong ngn cn s ly nhim virus.Nhng khm ph ca Altman and Cech rng ARN khng ch c chc nng di truyn m cn c chc nng xc tc sinh hc nh l cc enzyme gy ra mt bt ng rt ln v t lu cc nh sinh hc tin rng axit nucleic l cc phn t sinh hc gi vai tr trong di truyn cn protein l cc phn t chc nng cu trc v chc nng xc tc. Cc enzyme l cc cht xc tc sinh hc m n c vai tr quyt nh trong gn nh l tt c cc phn ng xy ra trong c th sng, cc enzyme c kh nng tng tc phn ng ca phn ng ln rt nhiu ln. Cc phn ng trong c th sng thc s rt kh xy ra nu nh vng mt ca cc enzyme, hay ni mt cch n gin, nu khng c enzyme th s khng c s sng.

Trong ribosome chnh thnh phn c vai tr xc tc hnh thnh lin kt peptit (enzyme peptidyl transferase) l ARN. im khc bit gia peptidyl transferase vi cc loi ribozyme khc (nh l ARN antisen, P RNAase ) l cc ribozyme khc u gy tc ng xc tc ln trung tm phn ng l photpho cn peptidyl transferase gy hiu ng xc tc ln trung tm phn ng l Cacbon qua hnh thnh lin kt peptit. C th ribozyme ca ribosome l mt du vt st li ca dng sng nguyn thu vn ch c cc enzyme ARN.Chc nng xc tc ca RNA cng cung cp mt cng c hon ton mi trong cng ngh gen. c bit l cc cng ngh c s dng ribozyme s c th c s dng trong vic phn ct ARN c th ph hu nhng phn t ARN c nhng c im gy hi v khng mong mun trong cc c th sinh vt nht nh. Ni mt cch c th hn, cc nh khoa hc hy vng rng kh nng ct xn gene bng phng thc ct v ph hu ARN ca virus s l c ch bo v c th chng li s ly nhim virus, cng ngh lin quan n cc ribozyme cng gip to ra cc thc vt khng virus v gp phn iu tr c hiu qu cc bnh do nhim virus nh l bnh cm lnh, HIV, v cc bnh truyn nhim do virus khc, do cn c cc nghin cu tip theo sau Altman v Cech v chc nng xc tc ca ARN1.2.2. C chS dng ribozyme ngn chn s sao chp ca virus hoc phin m gen virus. Ribozyme l cc phn t RNA c mt bn phn xc tc c kh nng phn ct cc phn t RNA ch xung quanh cc cnh tay RNA c kh nng gn vi cc trnh t ch bi cc cp base b cu tng t antisense oligonucleptide. V l thuyt th mt ribozyme c th phn ct c nhiu phn t RNA ch. Khi thm nhim mt vect c cha trnh t ca mt ribozyme th c th to ra nhiu bn phin m phn t ribozyme tr liu trong cc t bo ch. Mt chin lc khng virus khc li s dng cc polypeptide khng tri c thit k tng tc vi bn phin m t nhin, v th m ngn chn c s lp rp ca virus hoc lm mt chc nng enzyme ca chng.1.2.3. Tch chit RibozymeRibozyme l cc phn t acid ribonucleic (RNA) c kh nng tc ng nh cc enzyme ngay c khi vng mt hon ton cc protein. Chng c hot tnh xc tc b gy v hoc hnh thnh cc lin kt cng ha tr vi tnh c hiu v gia tng tc cc phn ng mt cch phi thng.Qua cc ngun nguyn liu ng, thc vt chnh c th t chit xut cc ch phm enzyme, chng ta thy rng hai ngun nguyn liu ny khng th dng sn xut cc ch phm enzyme vi quy m ln bi cc nhc im sau y: Chu k sinh trng ca chng di Ngun nguyn liu ny khng ci to c. Nhiu nguyn liu dng lm thc phm (dng n) khng th dng lm nguyn liu sn xut vi quy m ln cc ch phm enzyme nhm tho mn cc nhu cu ca nn kinh t quc dn. Dng vi sinh vt lm ngun nguyn liu sn xut cc ch phm enzyme c nhiu u im ni bt v c tnh cht c o vt xa so vi ngun nguyn liu t ng vt, thc vt, cng nh s khc phc c mi kh khn v hn ch trn.Trc ht vi sinh vt l ngun nguyn liu v tn sn xut enzyme vi s lng ln. y cng l ngun nguyn liu m con ngi ch ng to ra c. Chu k sinh trng ca vi sinh vt ngn (t 16 - 100 gi) v vy c th nui cy hng trm ln trong nm.Enzyme vi sinh vt c hot tnh rt mnh, vt xa cc sinh vt khc. V vy ch cn mt lng nh enzyme c th chuyn ha mt lng ln c cht. S liu tnh ton cho bit, trong vng 24 gi, vi sinh vt c kh nng chuyn ha mt lng thc n gp 30 - 40 ln so vi trng lng c th chng. Trong khi , h enzyme ca con ln trn 50 kg ch c th chuyn ha c vi kg thc n trong ngy. H enzyme vi sinh vt v cng phong ph. Vi sinh vt c kh nng tng hp nhiu loi enzyme khc nhau, trong c nhng enzyme ng, thc vt khng tng hp c. V d cellulase, raxemase...Phn ln cc thc n nui vi sinh vt li d kim v gi r. Nhiu vi sinh vt cho enzyme thng c kh nng pht trin trn cc mi trng n gin, gi r, d kim nh cc ph liu ca cc ngnh sn xut. Hn na, c th dng nhng nguyn liu khng phi thc phm, nhng dung dch mui v c nui vi sinh vt. V vy dng vi sinh vt lm ngun thu enzyme s mang li gi thnh r, thi gian nhanh v hiu qu kinh t cao.Vi sinh vt sinh sn pht trin vi tc cc k nhanh chng, khi lng li nh, kch thc b, nhng t l enzyme trong t bo tng i ln nn quy trnh sn xut ch phm enzyme kh d dng, hiu sut thu hi cao. Lng enzyme c th c sn xut ra trong mt thi gian ngn. i vi mt s trng hp c th dng 100% sinh khi vi sinh vt lm ngun enzyme.Vi sinh vt rt nhy cm i vi tc ng ca mi trng, thnh phn dinh dng nui chng cng nh mt s tc nhn l ha, c hc khc. Do c th thay i nhng iu kin nui cy chn ging to nhng chng t bin cho ta hm lng enzyme ng k vi hot tnh xc tc cao. C th ni rng, nh ngun enzyme vi sinh vt, ngi ta c th iu khin s tng hp enzyme d dng hn cc ngun nguyn liu khc tng lng enzyme c tng hp hoc tng hp nh hng enzyme. Tuy vy trong qu trnh chn ngun nguyn liu t vi sinh vt, cn lu mt s vi sinh vt c kh nng sinh c t c bin php x l thch hp. Ni chung cc vi sinh vt mun c s dng lm ngun nguyn liu tch enzyme cn phi tho mn cc iu kin sau: Kh nng tng hp enzyme mnh trong mt thi gian ngn. D tch enzyme v khng sinh c t.1.2.4. ng dng ca ribozymeCc ribozyme c ng dng nh cc tc nhn khng virus, dng iu tr ung th v cc ri lon di truyn v cng l cng c lm sng t cc con ng v ph chun ch. Khi u s dng ribozyme l nhm vo khng virus, ch yu l iu tr virus gy thiu ht min dch trn ngi (HIV). Cc virus m c th tc ng thng qua sn phm trung gian RNA h gen trong chu k ti bn ca chng nh HIV, virus gy vim gan B, C l nhng ch hp dn bi v cc mu n ribozyme c th nhm ti RNA h gen virus v c cc mRNA. Ribozyme cng c s dng rng ri i vi cc gen t bo ch bao hm c nhng gen biu hin mt cch lm lc trong ung th. ch ribozyme u tin l bn sao tng hp bcr-abl c kin to t nhim sc th Philadelphia lin quan ti bnh bch cu dng tu mn tnh. c trng ca nhim sc th ny l bng s chuyn v (translation) biu hin c protein tng hp bcr-abl ti np. Trong trng hp ny, ribozyme c thit k nhm vo mRNA tng hp mt cch c hiu ch khng phi l nhm vo bcr hay abl bnh thng ngn chn chc nng ca gen gy ung th bcr-abl. Khi t bin codon 12 trong c-H-ras t GGU thnh GUU th to nn mt v tr cho s phn ct qua trung gian ribozyme u ba. Mt ribozyme biu hin ni sinh nhm vo ch ny c hiu ng ngn nga s hnh thnh tiu im (focus formation) vi khong 50% t bo NIH3T3 c thm nhim vi gen ras hot ha. Tri li, cc t bo biu hin ribozyme tng t ny nhng li thm nhim vi ras hot ha trong c s thay i codon v tr 61 thay v v tr 12 th khng ngn nga c s hnh thnh tiu im. Cc ribozyme nhm vo HER-2/neu c biu hin qu mc trong ung th v lm gim ng k s to u ca cc t bo ny trn chut. Hn na, nhm trc tip vo cc gen gy ung th, cc ribozyme cng c p dng mt cch gin tip hn vi t cch l cc tr liu khng ung th. Chng hn nh cc riobozyme nhm vo cc gen khng a thuc 1 (multiple drug resistance gene 1) hoc fos mRNA trong cc dng t bo ung th to c cc t bo nhy cm hn vi cc tc nhn ha tr liu. Mt khc, ribozyme nhm vo bcl-2 lm bng pht apoptosis trong cc t bo ung th vng ming. Cc yu t i hi cho s di cn cng l ch hp dn cho ribozyme. Cc ribozyme nhm mc tiu chng li CAPL/mts, metalloprotein gian bo (matrix), pleiotrophin v VLA-6 integrin u lm gim tim nng di cn ca cc t bo khi u cn lu tm. S to mch cng l ch quan trng cho gen tr liu ung th v qu trnh ny c ngn chn chut bi ribozyme ch vo protein gn yu t tng trng nguyn bo si v pleiotrophin. Cc tr liu vi c s ribozyme c test ng vt nhm c ch cc ri lon tng sinh khc nh ti hp ng mch vnh. Antitelomerase RNA ribozyme cng c test p dng trong gen tr liu ung th. D nhin vic s dng ribozyme nh cc tc nhn tr liu cn c s thay i ph hp vi tng loi ung th.1.3. ng dng trong y dcEnzyme c mt v tr quan trng trong y hc. c bit l cc phng php nh lng v nh tnh enzyme trong ha hc lm sng v phng th nghim chn on. Do , hin nay trong y hc xut hin lnh vc mi gi l chn on enzyme, c nhim v:- Phn tch xc nh nng c cht nh glucose, ure, cholesterol vi s h tr ca enzyme. - Xc nh hot tnh xc tc ca enzyme trong mu sinh vt.- Xc nh nng c cht vi s h tr ca thuc th enzyme nh du. Dng enzyme nh lng cc cht, phc v cng vic xt nghim chn on bnh, v d dng kim tra glucose nc tiu rt nhy.

Urease nh lng ureDng enzyme lm thuc v d protease lm thuc tc nghn tim mch, tiu m vt thng, lm thng ng h hp, chng vim, lm thuc tng tiu ha protein, thnh phn ca cc loi thuc dng trong da liu v m phmTrong y hc cc protease cng c dng sn xut mi trng dinh dng nui cy vi sinh vt sn xut ra khng sinh, cht khng c Ngoi ra ngi ta cn dng enzyme protease c c v tinh ch cc huyt thanh khng c cha bnh.Amylase c s dng phi hp vi coenzyme A, cytocrom C, ATP, carboxylase ch thuc iu tr bnh tim mch, bnh thn kinh, phi hp vi enzyme thy phn cha bnh thiu enzyme tiu ha.1.4. ng dng trong ha hcCho n nay, vic ng dng enzyme trong ha hc l do enzyme c cm ng cao i vi nhit , pH v nhng thay i khc ca mi trng.Mt trong nhng ng dng ch phm enzyme ng c ch nht trong thi gian gn y l dng cht mang gn phc enzyme xc tc cho phn ng nhiu bc. V d tng hp glutathion, acid bo, alcaloid, sn xut hormoneCng bng cch to phc, ngi ta gn vi sinh vt s dng trong cng ngh x l nc thi, sn xut alcohol, amino acidTrong nghin cu cu trc ha hc, ngi ta cng s dng enzyme, v d dng protease nghin cu cu trc protein, dng endonuclease nghin cu cu trc nucleic acidDng lm thuc th trong ha phn tch.1.5. ng dng trong cng nghipVic s dng enzyme trong cng nghip l a dng, phong ph v t c nhiu kt qu to ln. Th nhn thng k s b sau y v cc lnh vc dng protease ta c th thy c s a dng: cng nghip tht, cng nghip ch bin c,cng nghip ch bin sa, cng nghip bnh m, bnh ko, cng nghip bia, cng nghip sn xut sa kh v bt trng, cng nghip hng phm v m phm, cng nghip dt, cng nghip da, cng nghip phim nh, cng nghip y hcVi amylase, c dng trong sn xut bnh m, cng nghip bnh ko, cng nghip ru, sn xut bia, sn xut mt,glucose, sn xut cc sn phm rau, ch bin thc n cho tr con, sn xut cc mt hng t qu, sn xut nc ngt, cng nghip dt, cng nghip giyTrong phm vi gio trnh ny chng ta ch cp n vic ng dng ch phm enzyme trong mt s lnh vc.1.6. ng dng trong cng nghip thc phmProtease vi cng nghip thc phm: Vic s dng trong ch bin lm mm tht l ng dng c tnh truyn thng. Nhn dn ta t rt lu dng thm nu canh tht b; dng rau sng v chui cht, v kt hp thc n nhiu tht; u trong chng to bnm thc cht l s dng papain, bromelain, fixin. Ngi Nga cn dng protease t ht u tng ny mm lm mm tht.Ngoi kh nng phn gii lm mm tht, to thc n d tiu ha, cng ngh sn xut cc loi dch thy phn giu protein c p dng mt cch c hiu qu tnh nng ca protease.Enzyme l mt cng c ch bin cc ph liu ca cng nghip thc phm thnh thc n cho ngi v vt nui.Ngi ta cn khai thc tnh ng t nh ca renin, pepsin vo cng nghip thc phm nh trong sn xut phomat.Pectinase vi cng nghip thc phm: Pectinase c dng trong mt s ngnh cng nghip thc phm sau: Sn xut ru vang. Sn xut nc qu v nc ung khng c ru. Sn xut cc mt hng t qu: qu c c, mt. Sn xut nc gii kht. Sn xut c ph.Ch phm pectinase c s dng trong sn xut nc qu t cc nguyn liu qu nghin hay lm trong nc qu p. Bi v khi c pectin th khi qu nghin s c trng thi keo, do khi p dch qu khng that ra c. Nh pectinase m nc qu trong sut, d lc, hiu sut tng.Pectinase cn gp phn chit rt cc cht mu, tanin v cc cht ha tan khc, do lm tng cht lng ca thnh phm.Nhng nghin cu khi p nho c x l bng pectinase khng nhng lm tng hiu sut m cn lm tng mu sc.Trong sn xut mt nh, mt ng nh pectinase m dch qu c nng m c hn.Cellulase vi cng nghip thc phm: Cellulose l thnh phn c bn ca t bo thc vt, v vy n c mt trong mi loi rau qu cng nh trong cc nguyn liu, ph liu ca cc ngnh trng trt v lm nghip. Nhng ngi v ng vt khng c kh nng phn gii cellulose. N ch c gi tr lm tng tiu ha, nhng vi lng ln n tr nn v ch hay cn tr tiu ha.Ch phm cellulase thng dng : Tng cht lng thc phm v thc n gia sc. Tng hiu sut trch ly cc cht t nguyn liu thc vt.ng dng trc tin ca cellulase i vi ch bin thc phm l dng n tng hp thu, nng cao phm cht v v v lm mm nhiu loi thc phm thc vt. c bit l i vi thc n cho tr con v ni chung cht lng thc phm c tng ln.Mt s nc dng cellulase x l cc loi rau qu nh bp ci, hnh, c rt, khoai ty, to v lng thc nh go. Ngi ta cn x l c ch, cc loi to binTrong sn xut bia, di tc dng ca cellulase hay phc h citase trong c cellulase, thnh t bo ca ht i mch b ph hy to iu kin tt cho tc ng ca protease v ng ha.Trong sn xut agar-agar, tc dng ca ch phm cellulase s lm tng cht lng agar-agar hn so vi phng php dng acid ph v thnh t bo. c bit l vic s dng ch phm cellulase tn thu cc ph liu thc vt em thy phn, dng lm thc n gia sc v cng ngh ln men.Nhng ng dng ca cellulase trong cng nghip thc phm c kt qu rt tt. Tuy nhin hn ch ln nht l rt kh thu c ch phm c cellulase hot cao.Amylase vi cng nghip thc phm: Ch phm amylase c dng ph bin trong mt s lnh vc ca cng nghip thc phm nh sn xut bnh m, glucose, ru, bia...Trong sn xut bnh m, ch phm amylase lm thay i hon tan cht lng ca bnh m c hng v, mu sc, xp...Ch phm amylase sch cho cht lng bnh m tt hn dng phc hp vi protease.Trong sn xut bnh ko ngi ta thng dng maltose l sn phm thy phn tinh bt bng amylase v glucose bng glucoamylase. Chnh glucoamylase, l yu t lm tng hiu sut trong sn xut ru.Trong sn xut bia, vic s dng amylase c trong cc ht ny mm thay th la mch (malt) gp phn ng k trong vic gim gi thnh.1.7. ng dng trong cng nghip dtTrong cng nghip dt, ch phm amylase c dng r h vi trc khi ty trng v nhum. Amylase c tc dng lm vi mm, c kh nng nhng t, ty trng v bt mu tt. R h bng enzyme khng nhng nhanh, khng hi vi, mao dn tt m cn m bo v sinh, do tng c nng sut lao ng.Trong sn xut t tm, ngi ta dng protease lm sch si t. Vi cng on x l bng enzyme sau khi x l bng dung dch x phng s gip la c tnh n hi tt, bt mu ng u v d trang tr trn la.1.8. ng dng trong cng nghip thuc daTrong cng nghip da, enzyme protease c dng lm mm da, lm sch da, rt ngn thi gian, trnh nhim mi trng. Vic x l c tin hnh bng cch ngm da trong dung dch enzyme, hay pht dch enzyme ln b mt da. Enzyme s tch cc cht nhn v lm t mt s lin kt trong phn t collagen lm cho da mm hn.Thc t cho thy khi x l da bng ch phm protease t vi sinh vt c th rt ngn thi gian lm mm v tch lng xung nhiu ln. iu quan trng l cht lng lng tt hn khi ct. So vi phng php ha hc th vic x l bng enzyme c s lng lng tng 20-30%. Lng khng cn x l thm sau khi ngm trong dch enzyme.1.9. ng dng trong nng nghipC th s dng cc loi ch phm enzyme khc nhau chuyn ha cc ph liu, c bit l cc ph liu nng nghip ci to t phc v nng nghip. Nht hng nm sn xut hng vn tn ch phm cellulase cc loi dng trong nng nghip. C ch phm cha c cellulase, hemicellulase, protease v amylase.Cng ngh ny kh ph bin nhiu quc gia. nc ta vic dng enzyme vi sinh vt gp phn trong sn xut phn hu c ang c khai thc thay th cho phn ha hc.Tm li, c th ni rng, vic nghin cu ng dng cc ch phm enzyme ngy cng c ch trng cc lnh vc khc nhau. Trong 20 nm cui th k XX v cc nm u ca th k XXI cc enzyme khc nhau c ng dng. Vit Nam bc u c nhiu nghin cu ng dng cc enzyme trong ch bin nng sn, thc phm, nht l trong lnh vc sn xut bia, ru, ch bin tinh bt (Vin cng nghip thc phm, Vin cng ngh sinh hc cng ngh thc phm, i hc Bch khoa H Ni). Vic nghin cu cc enzyme phc v nng nghip, cng nghip cng c quan tm v c cc kt qu ng khch l. V d, ch phm enzyme mi ra i phc v nng nghip E2001 c tc dng tng ph nhiu t, tng nng sut cy trng. c cc nghin cu ng dng protease trong sn xut ru bia, rt ngn thi k ln men cng nh sn xut nc mm ngn ngy bng cng ngh enzyme protease. Enzyme amylase cng c nghin cu ng dng rng ri trong sn xut ng bt, maltodextrin, nha glucose, siro, glucose fructose quy m cng nghip.TI LIU THAM KHOTi liu ting Vit1. Nguyn Hu Chn, 1983. Enzyme v xc tc Sinh hc. Nxb Y hc, H Ni.2. Phm Th Trn Chu, Trn Th ng, 2000. Ha sinh hc. Nxb Gio dc, H Ni.3. Ngc Lin, Phm Th Trn Chu, 1972. Enzyme I, II. i hc Tng hp, H Ni.4. Nguyn Tin Thng, Nguyn nh Huyn, 1998. Gio trnh sinh ha hin i. Nxb Gio dc, H Ni.5. Nguyn Xun Thng, o Kim Chi, Phm Quang Tng, Nguyn Vn ng, 2004. Ha sinh hc. Nxb Y hc, H Ni.6. L Ngc T, La Vn Ch, Phm Trn Chu, Nguyn Ln Dng, 1982. Enzyme vi sinh vt. Nxb KH&KT, H Ni.7. L Ngc T (ch bin), L Vn Ch, ng Th Thu, Phm Quc Thng, Nguyn Th Thnh, Bi c Hi, Lu Dun, L Don Din, 2000. Ha sinh Cng nghip, Nxb KH&KT, H Ni.8. Phan Tng Lc, Nguyn Hu H, Nguyn Th Thanh, 2012. Biu hin gen HIV-1 p24 cy thuc l chuyn gen lc lp. Vin sinh hc nhit i Vin khoa hc cng ngh Vit Nam.Ti liu ting Anh9. Eur. J. Biochem. 271, 10151024 (2004) FEBS 2004. Hydrolytic cleavage by a group I intron ribozyme is dependent on RNA structures not important for splicing.3