Nhng yu t nh hng n tc phn ng ca enzyme

1.Trc ht ta cn bit Enzyme l g? Enzyme l protein c kh nng xc tc c hiu cho cc phn ng ha hc. Chng thc y mt phn ng xy ra m khng c mt trong sn phm cui cng. Enzyme c trong nhiu i tngsinh hc nh thc vt, ng vt v mi trng nui cy vi sinh vt.Hin nay ngi ta thuc nhiu loi ch phm enzyme khc nhau v s dng rng ri trong nhiu lnh vc nh y hc, nng nghip, cng nghip Enzyme lcht xc tcsinh hcc thnh phn c bn lprotein. Enzyme, cng nh nhng protein khc, c trng lng phn t khong 12.000 n hn1000.000.Mt s enzyme cu to gm ton nhng phn t L amino acid lin kt vi nhau tothnh, gi l enzyme mt thnh phn. a s enzyme l nhng protein phc tp gi l enzyme haithnh phn. Phn khng phi protein gi l nhm ngoi hay coenzyme . Mt coenzyme khi kthp vi cc apoenzyme khc nhau (phn protein) th xc tc cho qu trnh chuyn ha cc chtkhc nhau nhng chng ging nhau v kiu phn ngTrong cuc sngsinh vtxy ra rt nhiu phn ng ha hc, vi mt hiu sut rt cao, mc d iu kin bnh thng v nhit , p sut, pH. S d nh vy v n c s hin din cacht xctcsinh hc c gi chung lenzyme. Nh vy, enzyme l ccproteinxc tc cc phn ngha hc.Trong ccphn ngny, cc phn tlc bt u ca qu trnh c gi l c cht (substrate), enzyme s bin i chng thnh cc phn tkhc nhau. Tt c cc qu trnh trong t bo u cn enzym. Enzym c tnh chn lc rtcao i vi c cht ca n.Hu ht phn ng cxc tcbi enzym u c tc cao hn nhiu so vi khi khng cxc tc. C trn 4 000 phn ng sinh hac xc tc bi enzymHat tnh ca enzym chu tc ng bi nhiu yu t. Cht c ch l cc phn t lm gim hot tnh caenzym, trong khi yu t hot ha l nhng phn t lm tng hot tnh ca enzym Vi du:Cu trc proteinca enzyme TIM. TIM l mt enzyme cc k hiu qu trong qu trnhchuyn ingthnh nng lng cho c the 2.Cc yu t nh hng n tc ca phn ng enzyme C nhiu yu t nh hng n tc phn ng enzyme : 1.nh hng ca nng enzyme 2 . n h h n g c a n n g c c h t [ S ] 3.nh hng ca cht km hm (inhibitor)4.nh hng ca cht hot ha (activato)5 . n h h n g c u n h i t 6 . n h h n g c a p H 7.nh hng ca cc yu t khc 1.nh hng ca nng enzyme

Trong iu kin d tha c cht, ngha l [S] >>>[E] th tc phn ng ph thuc vo [S], v=K[E] c dng y = ax. Nh ngi ta o [E] bng cch o vn tc phn ng do enzyme xc tc.C nhiu trng hp trong mi trng c cha cht km hm hay hot ho th vn tc phnng do enzyme xc tc khng ph thuc tuyn tnh vi [E] . Hnh: S ph thuc ca vn tc phn ng vo [E] 2.nh hng ca nng c cht [S] Ta kho st trng hp n gin nht : ch mt c cht.Gi v 1 l vn tc ca phn ng to thnh phc cht ES.Gi v -1 l vn tc ca phn ng phn ly phc cht ES to thnh E v SGi v 2 l vn tc ca phn ng to thnh E v P (sn phm). V i:v 1 =k 1 [E][S]v -1 =k -1 [ES]v 2 =k 2 [ES]Khi h thng t trng thi cn bng ta c:k -1 [ES]+k 2 [ES] = k 1 [E][S](k -1 +k 2 )[ES] = k +1 [E][S] (2)Gi E 0 l nng ban u:[E 0

]=[E]+[ES]=>[E]=[E 0 ]-[ES] (3)Thay tr s [E] t (3) vo (2) ta c:(k -1 +k 2 )[ES] = k 1 ([E 0 ]-[ES]) [S]k 1 [E 0 ] [S] [ES] = --------------k -1 +k 2 +k 1 [S] Nu t Km= k -1 +k 2 /k 1 (Km: gi l hng s Michaelis Menten).Ta c : [ES] = [E0][S]/ Km+ [S]Mt khc vn tc phn ng to thnh sn phm P l:V = k 2 [ES]Thay [ES] bng gi tr trn ta thu c:k 2 [E 0 ] [S]v = ----------------- (4)K m + [S]Qua y ta thy nng enzyme cng cao th vn tc phn ng enzyme cng ln. Vn tc tcc i khi ton b enzyme lin kt vi c cht, ngha l:V max =k 2

[E 0 ]Thay vo phng trnh (4) ta c: Nhng yu t nh hng n tc phn ng ca enzyme [S]v = V max ---------- (5)K m + [S]Phng trnh (5) gi l phng trnh Michaelis Menten.Km gi l hng s Michaelis Menten c trng cho mi enzymeKm c trng cho i lc ca enzyme vi c cht, Km c tr s cng nh th i lc ca enzymevi c cht cng ln, ngha l vn tc ca phn ng do enzyme xc tc cng ln. Hnh: Bin thin vn tc phn ng theo nng c cht. Khi tng [S] th v phn ng tng, tng [S] n mt gi tr no th v t n gi tr vmax v skhng tng na nu ta vn tip tc tng [S].Khi Km=[S] th v =1/2 Vmax Nm 1934. Lineweaver v Burk, trn c s ca phng trnh (5) nghch o bin thnhdng ng thng y=ax+b, n c ngha ln i vi vic nghin cu km hm enzyme. Hnh : S ph thuc ca tc phn ng vo nng c cht theo LineweaverBurk 3.nh hng ca cht km hm ( inhibitor ) L cht c tc dng lm gim hot hay lm enzyme khng cn kh nng xc tc bin c cht thnh sn phm. Km hm enzyme c th thc hin bng nhiu cch khc nhau (thunnghch hay khng thun nghch). Thun nghch c 3 cch : Trang 4

Cch 1: Km hm cnh tranh (competitive inhibition) Trong trng hp km hm cnh tranh l c cht v cht km hm u tc dung ln trung tmhot ng ca enzyme, Cht km hm chon ch o ca c cht enzyme. Hnh : Kiu km hm cnh tranh Khi c cht d tha, nng cht km hm thp th c th loi b tc dng ca cht km hm,cn nng c cht thp v nng cht km hm cao th li c tc dng km hm hon ton. 1/v= ( K m /V max ) 1/S +1/V max = 1+[I]/K I Trang

Nhng yu t nh hng n tc phn ng ca enzyme Hnh : S ph thuc ca tc phn ng vo nng c cht theo LineweaverBurk khic km hm cnh tranh Ngi ta thy km hm nh vy phn ln xy ra gia cht km hm v c cht c s tngng v mt ho hc. v d: malic acid c cu trc gn ging vi succinic acid nn km hmcnh tranh enzyme succinatedehydrogenase , l enzyme xc tc cho s bin i succinic acidthnh acid fumaric acid.Trng hp c bit ca km hm cnh tranh l km hm bng sn phm. Trng hp ny xyra khi mt sn phm phn ng tc dng tr li

enzyme v chon v tr hot ng phn tenzyme.ng thng c cht km hm th c xin ln hn v ct trc tung mt im l 1/Vmax Cch 2: Km hm phi cnh tranh ( uncompetitive inhibition )c trng ca kiu km hm ny l cht km hm ch lin kt vi phc hp ES, m khng linkt vi enzyme t do. Trang 6

Nhng yu t nh hng n tc phn ng ca enzyme Hnh : Kiu km hm phi cnh tranhHnh: S ph thuc ca tc phn ng vo nng c cht theo Lineweaver-Burk khic km hm phi cnh tranh Cch 3: Km hm hn tp ( mixed inhibition ) Trang 7

Nhng yu t nh hng n tc phn ng ca enzyme Hnh: Kiu km hm hn tp Trong , cht km hm khng nhng lin kt vi enzmye t do m cn lin kt vi c phchp ES to thnh phc hp EIS khng to c sn phm P.Tng t nh trn ta c phng trnh : Hnh: S ph thuc ca tc phn ng vo nng c cht theo LineweaverBurk khic km hm hn tp Cc gi tr , c nh ngha nh trn. Trng hp

= gi l km hm khng cnhtranh (noncompetitive inhibition).Mt trng hp km hm cn gp na l k m hm enzyme bng nng cao ca c cht gi lkm hm c cht nh km hm urease khi nng ure cao, ngoi ra cn c cc enzymekhc nh lactatdehydrogenase, carboxypeptidase, lipase, pyrophotphatase, photphofructokinase(i vi ATP). Nguyn nhn ca nhng hin tng ny cn cha c bit r. c th l:+ Tn ti nhiu trung tm lin kt vi c cht bng cc i lc khc nhau. Khi nng c chtthp th enzyme c th ch lin kt vi mt phn t c cht, cn khi nng c cht cao nlin kt vi nhiu c cht dn n hnh thnh phc hp ES khng hot ng.+ C cht cng c th c lin kt nh nhng v tr c bit ca enzyme. l mt nhmenzyme quan trng (enzyme d lp th) bn cnh trung tm xc tc cn c trung tm iuchnh.+ C cht c th lin kt vi mt cht hot ho v bng cch ny n tch khi E.+ C cht c th chon ch (ngn cn) mt cofactor ( ng yu t ) hay mt coenzyme. Nhng yu t nh hng n tc phn ng ca enzyme + C cht ho ca enzyme.c th nh hng n ion lc ca mi trng v qua lm mt itnh chuyn 4.nh hng ca cht hot ha (activator ) L cht lm tng kh nng xc tc nhm chuyn ha c cht thnh sn phm. Thng thng lnhng cation kim loi hay nhng hp cht hu c nh cc vitamin tan trong n c.V d: Mg ++ hot ha cc enzyme m c cht c phosphoryl ha nh pyrophosphatase(c cht l pyrophosphate), adenosinetriphosphatase (c cht l ATP). Cc cation kim loi cth c tnh c hiu, tnh i khng v tc dng cn tu thuc vo nng . 5.nh hng cu nhit Ta c th tng vn tc ca mt phn ng ha hc bng cch tng nhit mi trng, hintng ny tun theo quy lut Vant-Hoff. iu ny c ngha khi tng nhit ln 10 0 C th tc phn ng tng ln khang 2 ln.i vi phn ng do enzyme xc tc cng c th p dng c quy lut ny nhng ch trongmt phm

vi nht nh,v bn cht enzyme l protein.Khi ta tng nhit ln trn 40-50 0 C xyra qu trnh ph hu cht xc tc. Sau nhit ti u tc phn ng do enzyme xc tc sgim. Nh tn ti nhit ti u ngi ta phn bit phn ng ho sinh vi cc phn ng v c thng thng.Mi enzyme c mt nhit ti u khc nhau, phn ln ph thuc ngun cung cpenzyme, thng thng trong khong t 40-60 0 C , cng c enzyme c nhit ti u rt caonh cc enzyme ca nhng chng a nhit. Cc chng vi sinh vt a nhit, c bit cc vikhun chu nhit c cha enzyme chu nhit cao. nh hng ca nhit ln hat enzyme Nhng yu t nh hng n tc phn ng ca enzyme 6.nh hng ca pH S phn li khc nhau ca mt phn t protein cc gi tr pH khc nhau lm thay i tnh chtca trung tm lin kt vi c cht v tnh cht hot ng ca phn t enzyme.iu ny dn ngi tr xc tc khc nhau ph thuc vo gi tr pH. Nh bit mi enzyme c mt pH tiu,mi enzyme c ng biu din nh hng pH ln vn tc phn ng do chng xc tc.ng biu din c dng nh hnh sau: Hnh : nh hng ca pH ln hat enzyme nhhngcagitrpHntcdngenzymecthdocccssau:a/ Enzyme c s thay i khng thun nghch phm vi pH cc hp. b/ hai sn ca pH ti u c th xy ra s phn ly nhm prosthetic hay coenzyme.c/ Lm thay i mc ion ho hay phn ly c cht.d/ Lm thay i mc ion ho nhm chc nht nh trn phn t enzyme dn n lm thay ii lc lin kt ca enzyme vi c cht v thay i hot tnh cc i. Nh xc nh Vmax v Km ph thuc gi tr pH cho php nhn nh li bn cht ca cc nhmtham gia vo lin kt c cht v qu trnh t xc tc. Trang 10 Nhng yu t nh hng n tc phn ng ca enzyme 7. Cc yu t khc + nh sng: C nh hng khc nhau n tng loi enzyme, cc bc sng khc nhau c nhhng khc nhau, thng nh sng trng c tc ng mnh nht, nh sng c tc ng yunht.nh sng vng t ngoi cng c th gy nn nhng bt li, enzyme trng thi dung dch bnhn khi c kt tinh dng tinh th, nng enzyme trong dung dch cng thp th cng km bn, tc ng ca tia t ngoi s tng ln khi nhit cao. V d di tc ng ca tia t ngoi nhit cao enzyme

amylase nhanh chng mt hot tnh.+ S chiu in: in chiu vi cng cng cao th tc ng ph hu cng mnh. Tc ngs mnh hn i vi dch enzyme c nng thp. C th do to thnh nhng gc t do, t tn cng vo phn ng enzyme+ Sng siu m: Tc ng rt khc nhau i vi tng loai enzyme, c enzyme b mt hot tnh,c enzyme li khng chu nh hng. Nhn xt chung: bn ph thuc vo trang thi tn ti ca enzyme, cng tinh khit thenzyme cng km bn, dch cng long th bn cng km, tc ng ca mt s ion kim loitrong dch vi nng khong 10 -3 M nh Ca ++ lm tng tnh bn. Enzyme allosteric ( Enzyme d lp th, d khng gian) Cho n nay, ngi ta m t enzyme m hat tnh enzyme ph thuc nng c cht khng cdng hyperbol m c dng sigmoid l enzyme allosteric Hnh . Bin thin vn tcphn ng theo nng c cht i vi enzyme ny, khi nng c cht thp th tc phn ng tng chm, sau tip tctng nng th tc nhanh chng t gi tr cc i. Trang 11

Nhng yu t nh hng n tc phn ng ca enzyme Nh ta bit, enzyme tun theo ng hc Michaelis-Menten th 1 hay nhiu c cht cng chlin kt vo 1 v tr trn phn t enzyme, iu ny s dn n enzyme bo ha c cht.Cn enzyme c ng cong tc sigmoid ch xut hin khi enzyme l mt oligomer, nn cth lin kt vi nhiu phn t c cht. iu ny c ngha trn enzyme allosteric c nhiu trungtm lin kt, mi monomer c 1 trung tm lin kt. Ngi ta cho rng, trong trng hp ny c tnh hp tc gia cc v tr lin kt c cht trong phn t enzyme oligomer.Cc enzyme oligomer ny c Monod gi l allosteric enzyme d lp th (allosteric enzyme).ng cong tc sigmoid c th b cht iu ha (modulator) y v pha tri hay phi.

Chtiu ha dng tc lm tng i lc ca enzyme allosteric vi c cht, ngc li l cht iuha m. Cc cht iu ha c th lm nh hng khc nhau n cc thng s ng hc, lmthay i gi tr ring l mt trong hai gi tr Km hay Vmax. Hnh : Minh ha khi c modulator Trang 12