підручники. біохімія / Uchebnik_po_Biokhimii_E_S_Severin

nno cy6c'TpaTOBH npo.uyKTOB H Il03TOMY He MeIDllOT paBHOBeCHe peaKU;HH (PHC.

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•<PepMeHTbl KaTaJIH3HPYIOT 3HepreTttqecKH B03MOlKHbie peaKI.J,tttt;

•3HeprnSI XHMlfqecKOH CHCTeMbl OCTaeTCH noCTOSIHHOH;

•B xo.ue KaTaJitt3a HanpaBJieHHe peaKUttH He H3MeHHeTcH;

•<PepMeHThI He pacXO.llYIOTCSI B npoI.J,ecce peaK-

I.l,HH.

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•<PepMeHTbl 00.lla)J;alOT BhlCOKOH cneI.J,H<PttqHoCThlO;

• <PepMeHTaTHBHaH peaKUHH rrpoXO)lHT B KJieTKe, T.e. npH TeMneparype 37 °C, IlOCTOHHHOM

aTMoc<PepHOM )laBJieHHH " <Ptt3HOJIOfHqecKOM

3HaqeHH.H pH;

• CKOPOCTb <PepMeHTaTHBHOH peaKI.J,HH MOXeT

peryJIHpoBaTbCH.

Ii. 3TAUhl <I>EPMEHTATHBHOfO KATAJIH3A

1. <PopMUJJ06anue fjJepMenm-cyficmpamnow KOM11.ileKca

ToT <PaKT, qTO <PepMeHThI o6na.uaIOT BhICOKOH cneI.J,tt<PHqHoCThlO, IT03BOJIH.II B 1890 r. Bbl)lBHHyTb rnrroTe3y, cornacHO KOTOpOH aKTHBHbIH I.J,eH'Tp<PePMeHTa KOMITJieMeHrnpeH cy6c'Tpary, T.e. COO'FijeTcrnyeT eMy KaK «KJI10q JaMKY». IlocJie BJa.ttMo.ueltcrnHH cy6c'TpaTa ( «KJilOq») c aKTHBHblM I.J,eH'TpOM ( «3aMOK») npottCXO)lSIT XHMW-Iecme npeBpameHHSI cy6c'TparaB rrpo.uyKT. AKT.ttBHhIH I.J,eHTP rrp.tt 3TOM paCCMa'Tp.HBaJICSIKaK CTa6.HJlbHaSI, XeCTKO )leTepMHHttpoBaHHaSI C'TpYKTYPa.

r. 6hm rrpe)l.llo)l(eH .upyrolt Bap.ttaHT rnrroTe3bI «KJI10q-3aMOK», 06bSICHHlOIIl,HH co6hIT.HH B aKTHBHOM I.J,eHTpe <PepMeHTa. Ilo 3TOH rnrroTe3e aKTHBHhIH I.J,eHTP HBJIS1eTcS1 rtt6KoH CTPYKTY-

pOH no OTHOllieHHIO K cy6cTpa'ry.Cy6cTpaT, B3aHMO,UeHCTBYSI c aKTHBHhIM ueHTpOM <PepMeHTa, Bbl3bIBaeT H3MeHeHHe ero KOH$opMaUHH, rrpHBO,USI K <PopMHPOBaHHIO <PepMeHT-cy6cTpaTHOro KOMIIJieKca, 611arorrpmlTHOfO llJlSI XHMH'IeCKHXMO-

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~HJia 3KcrrepHMeHTaJihHOe rro.urnep)l()J.eHHe.

2. llOCAeiJ06ame.;ibnocmb cooblmuii B xoiJe tj>epMenmamuBnow Kamll.llUJa

Ilpouecc <PepMeHTaTHBHOro KaTaJIH3a yc110BHO MO)l(HO pa:meJilITb Ha CJie.uyIOIUHe 3Ta.Ilbl (pHC. 2-12).

IlepBhIH, BTopoif H qernepThIH 3TarrhI KaTaJIH3a Herrpo.uomKHTeJihHhI H 3aBHCfil OT KOHUeHTpaUHH cy6cTpaTa (llJISI rrepBoro 3Tarra) H KOHCTaHT CB.Sl3hIBaHHSI JIHraH,UOB B aKTHBHOM ueHTpe <PepMeHTa

(Jl)I.SI rrepBoro H TpeThero 3TaITOB). lfaMeHeHHSI 3HepreTHKH XHMH'leCKOHpeaK.QHH Ha 3THX CTa,UHSIX He3HaqHTeJibHbl.

TpeTHH 3Tan HaH6onee Me,UJieHHhIH; ,UJIHTeJihHOCTh ero 3aBHCHT OT 3HeprnH aKTHBaUHH XHMHqeCKOH peaKUHH. Ha 3TOH CTa,UHH npoHcxo,U.SIT pa3phrn CBSI3eH B MOJieKYJie cy6cTpaTa, o6pa30BaHHe HOBbIX CBSI3eH H <PopMHpoBaHHe MOJieKYJihl rrpo-

.uyKTa.

3. Po.ab aKmUBHOlO u.enmpa B tj>epMenmamuBHOM Kamll.llUJe

B pe3yJihTaTe Hcc11e.uoBaHHH 6hIJIO rroKaJaHO, ~o

MOJieKYJia <PepMeHTa, KaK rrpaBHJIO, BO MHOfO pa3

6oJihrne MOIIeKYIIhI cy6cTpaTa, rro.uBepra10meroc.S1 XHMHqecKOMY npeBpameHHIO 3THM <PepMeHTOM.

B KOHTaKT c cy6cTpaToM BczyrraeT JIHlllh He6011hrnasi: qacTb MOJieKYJihl <PepMeHTa, o6hNHO OT 5 .uo

10 aMHHOKHCJIOTHbIX OCTaTKOB, <PopMHPYIOIUHX aKTHBHblH ueHTp <PepMeHTa. Po11b OCTaJihHhIX aMHHOKMCJIOTHhIX OCTaTKOB COCTOHT B o6ecrreqemrn npaBHJihHOH KOH<i>OpMauHH MOJieKYJihl <l>epMeHTa llJlSI OITTHMaJibHOro rrpoTeKaHHSI XHMH'leCKOHpeaKUHH.

AKTHBHhIH ueHTP Ha Bcex 3Tarrax <PepMeHTaTHBHoro KaTaJIH3a HeJih3.SI paccMaTpHBaTb KaK nacCHBHhIH yqaCTOK ,UJI.SI CB.Sl3hIBaHH.SI cy6cTpaTa. 3To KOMIIJieKCHaSI MOJieKYJISIPHaH «MalllHHa»' HCIIOJlb3YIOIUaSI pa3H006pa3Hble XHMHqecKHe MexaHH3MhI, crroco6cTBYJOIUHe rrpeBpameHHIO cy6cTpaTa B rrpo.uyKT.

B aKTHBHOM uempe <PepMema cy6cTpaThI pacrro11ara10Tcsi: TaKHM o6pa30M, ~06hI yqaCTByIOIUHe B peaK.QHH <l>YffKUHOHaJibHbie rpyrrrrhl cy6cTpaTOB HaxO,UHJIHCb B Herrocpe,UCTBeHHOH 6JIH30CTH .upyr K .upyry. 3TO CBOHCTBO aKTHBHOfO ueHTpa Ha3hIBalOT 3$<l>eKTOM c6JIIDKeHHSI H opHeHTaIJ;HH peareHTOB. TaKoe yrropSI,UoqeHHoe pacrronoxeHtte cy6cTpaToB BhI3hIBaeT YMeHhllleHHe 3HTpOIIHH H, KaK CJie,UCTBHe, CHH:lKeHHe 3HeprHH aKTHBaUHH (E.), qTQ onpe.ueJISieT KaTaJIHTHqecKyio 3¢<1JeKTHBHOCTh <IJepMeHTOB.

AKTHBHhIH ueHTp <PepMema TaK:lKe crroco6crnyeT

.uecTa6HJIH3aUHH Me:JKaTOMHbIX CB.Sl3eH B MOJieKYJie cy6cTpaTa, qTo 0611erqaeT rrpoTeKaHHe XHMH11ecKoif peaKUHH H o6pa30Bami:e rrpo.uyKTOB. 3'roCBOHCTBO aKTHBHOro uempa Ha3bIBalOT 3<l><l>eKTOM .ue- <l>opMauHH cy6cTpaTa (pHc.

B. MoJIEKYJHIPHhIE MEXAHH3MhI

<l>EPMEHTATHBHOfO KATAJIH3A

MexaHH3MhI <t>epMeHTaTHBHOro KanUima orrpe-

.uem110TcH poJihIO <l>YHKUHOHaJihHhIX rpyrrrr aKTHBHoro ueHTpa <t>epMeHTa B XHMHqecKOH peaKI..J;HH rrpeBpaiuemrn cy6cTPaTa B rrpo.uyicr. Bhmem110T 2

OCHOBHhIX MeXaHH3Ma <t>epMeHTaTHBHOro KaTa.rrH3a: KHCJIOTHO-OCHOBHOH KaTaJIH3 H KOBaJieHTHbIH KaTaJIH3.

I. KuCAomno-ocnosnoii Ka11UlJlU3

KoHuerruHH KHCJIOTHo-ocHOBHoro KaTaJIH3a 06bHCHHeT <t>epMeHTaTHBJIYIO aKTHBHOCTh yqaCTHeM B XllMHqecKOH peammi: KHCJIOTHbIX rpyrrrr (.uoHOPhI rrpOTOHOB) H/HJIH OCHOBHhIX rpyrrrr (amerrTOpbI rrpoTOHOB). KHcJIOTHo-ocHOBHOH KaTaJIH3 - qacTo BCTPeqa10meecH HBJiem1e. AMHHOKHCJIOTHh!e OCTaTKH, BXOMIUHe B COCTaB aKTHBHOro uempa, HMelOT <l>YHKUHOHa.JlbHbie rpyrrrrhI, rrpoHBJUIIOIUHe CBOHCTBa KaK KHCJIOT'TaK H OCHOBaHHH.

K aMHHOKHCJIOTaM, yqacTBYIOIUHM B KHCJIOTHOocHoBHOM KaTaJIH3e, B rrepB)'IO oqepe)lb OTHOCHT Uttc, Tttp, Cep, J1H3, f.rry, Acrr H fttc. Pa.uttKaJihI 3THX aMHHOKHCJIOT B npOTOHHPOBaHHOH <t>opMe - KHCJIOTbl (.UOHOpb! rrpOTOHa), B .uerrpOTOHHPOBaHHOH -'-OCHOBamrn (amerrTOph! npoTOHa). nJiaro-

.uapH 3TOMY CBOHCTBY <l>YHKUHOHaJihHbIX rpyrrrr aKTHBHOro ueHTPa <t>epMeHTbI CTaHOBHTCH YHHKa.JlhHbIMH 6HOJIOfH.qeCKHMH KaTaJIH3aTopaMH, B OTJIHqHe OT He6HOJIOfH.qeCKHX KaTaJIH3aTOPOB, crroco6HhIX rrpOHBJIHTb JIH60 KHCJIOTHbie, JIH.60 OCHOBHb!e CBOHCTBa.

IlpHMepOM KHCJIOTHO-OCHOBHOfO KaTaJIH3a, B KOTOPOM Ko<t>aKTopaMH HBJUllOTCH HOHbI Zn2+'a B KaqecTBe Ko<t>epMeHTa HCIIOJih3yeTCH MOJieKyJia NAD+, MOlKHO npHBecTH <t>epMeHT aJIKOfOJih):(erH)l- poreHa3y rreqeHH, KaTaJIH3HPYIOIUYIO peaKUHIO OKHCJiemrn CIIHpTa (pHC. 2-13):

C 2HpH + NAD+ 4 CH3-COH + NADH + H+.

2. Kosaiienmnblii KamaitUJ

KonaJieHTHhIH KaTaJIH3 ocHoBaH Ha aTaKe HYK-

Jieo<t>HJihHhIX (OTPHUaTeJibHO 3apmKeHHhIX) HJIM 3JieKTPO<l>HJihHhIX (IlOJIO)l{MTeJibHO 3apH)l{eHHhIX) rpyrrrr aKTHBHOfO ueHTPa <t>epMeHTa MOJieKYJiaMM cy6cTPaTa c <t>opMHPOBaHHeM KOBaJieHTHOH CBH3H Me)l{)ly cy6CTPaTOM M Ko<t>epMeHTOM HJIH <l>YHKUHOHaJihHOH rpyrrnoif aMHHOKHCJIOTHOro OCTaTKa (KaK rrpaBHJIO, O)lHOH) aKTHBHOfO ueHTPa <t>epMeHTa.

,lleifcrntte cepHHOBhIX rrparea3, TaKHX KaK TPHIICHH, XHMOTPHIICHH H TPOM6HH, - npHMep MexaHH3Ma KOBaJieHTHOro KaTaJIH3a, KOf)la KOBaJieHTHaH CBH3b o6pa3yeTCH Me)l{)ly Cy6cTPaTOM H aMHHOKHCJIOTHbIM OCTaTKOM cepHHa aKTHBHOfO ueHTPa <l>epMeHTa. TepMHH «cepHHOBhie rrpOTea3hI» CBH3aH c reM, qTQ aMHHOKHCJIOTHbIB OCTaTOK cepHHa BXO)lHT B coCTaB aKTHBHOro ueHTPa BCex 3THX <t>epMeHTOB H yqaCTBYeT Herrocpe.ucrneHHo B KaTaJIH3e. PacCMOTPHM MeXaHH3M KOBaJieHTHoro KaTaJIH3a Ha rrpHMepe XHMOTPHIICHHa, ocymecTBJIHIOIUero fH.llPOJIH3 rreIITH)l- HhIX CBH3eH IIPH nepenapHBamm 6eJIKOB B )lBeHa)l- uaTHITepCTHOH KHillKe (cM. pa3)leJI 9). ~arnMH

XHMOTPHIICHHa CJIY)KaT rreIITH)lbI, co.uep)l{aII!He aMMHOKHCJIOThI c apoMaTH.qeCKHMH H UHKJIH.qeCKHMH

m,LJ,pocl>o6HhIMH pa,II,HKaJiaMH (<l>eH, Twp, Tpw), qrn yi<a3hIBaeT Ha yqacrne rll,l.l,pocl>o6HhIX c1111 B ct>opMHpoBaHHH ct>epMem-cy6cTpaTHoro KOMIUieKca. MexaHH3M KOBa.TieHTHOfO KaTaJIH3a XHMOTpHTICHHa pacCMOTpeH Ha pwc.

Pa,II,HKaJihI Acn 102 , fwc57 11 Cep195 y1.mcrny10T Henocpe,D,cTBeHHO B aKTe KaTaJIH3a. BcJie,LJ,CTBHe ttyKJieocl>HJihHOM aTaKH nerrTH,ll,HOM CBH3H cy6cTpaTa rrpOHCXO,ll,HT pa3pblB 3TOM CBH3H c o6pa30BaHHeM KOBaJieHTHO-MO,ll,HcPHllHPOBaHHOfO cepHHa - auHJIXHMOTpHilCHHa. llpyroM neITTH,ll,HhIM cl>parMeHT BhlCB060)l(,ll,aeTCH B pe3yJihTaTe pa3pbIBa BO,UOpO,UHOM CBH3M Me)l(,ll,y nenTll,l.l,HhIM ct>parMeHTOM M fttc57 aKTMBHOfO ueHTpa XHMOTpHilCHHa. 3aKJI10qttTeJibHhIM 3TaIT fll,l.l,p0JIM3a rreIITll,l.l,HOM CBH3M 6eJIKOB - ,LJ,ea- UHJIMPOBaHMe XHMOTpMITCMHa B npMCYTCTBMH MOJieKYJihl BO,ll,hl c BhICB060)l(,ll,eHMeM BTOporo cl>parMeHTa rH,ll,pOJIM3yeMOfO 6eJIKa H HCXO,ll,HOM ct>opMbl ct>epMeHrn.

V.OCHOBblK~HET~K~

<DEPMEHTAT~BHblX PEAKl.l~~

KtttteTttKa ct>epMeHTaTHBHhIX peaKUHH - pa3,LJ,eJI

3H3MMOJIOfMM, M3yqa10mttti 3aBMCMMOCTh CKOpo-

CHI XMMmeCKMX peaKUHM, KaTaJIM3MpyeMbIX ct>epMeHTaMH, OT XMMMqeCKOM rrpttpO,ll,hI pearttpy10lll,HX BemecTB, a TaIOKe OT ct>aKTOPOB OKp}')l(aIOIUeti cpe,ahI.

Ll:JIH H3MepeHMH KaTaJIMTMqecKOM aKTMBHOCTM ct>epMeHTOB MCIIOJih3YIOT TaKMe IlOKa3aTeJIH, KaK CKOpOCTh peaKUMM HJIM aKTMBHOCTh ct>epMeHTa. CKopocTh ct>epMeHTaTHBHOM peaKUMM orrpe,D,eJIHeTCH M3MeHeHHeM KOJIMqecTBa MOJieKYJI cy6cTpaTa HJIM rrponyKrn 3a e,LJ,MHMUY BpeMeHM. CKopoCTh ct>epMeHTaTHBHOH peaKUMH - Mepa KaTaJIMTHqecKOM aKTHBHOCTH ct>epMeHTa, ee o603Haqa10T KaK aKTHBHOCTh ct>epMeHTa.

MaTeM.aTttqecKtt cKopocTh ct>epMeHTaTHBHOti peaKI1MH Bblp<DK.aeTCH B H3MeHeHMH KOHUeHTpallMH cy6cTparn (yMeHhrneHtte) HJIH rrpo,Uyi<Ta (YBeJIHqeHtte) 3a e,UMHHllY BpeMeHH:

V= D[S]/t = D[P]/t.

Ha HalfaJihHOM 3Tane [O - t0 ] CKopocTh peaKUMM rrpHMO rrporropuwoHaJibHa BpeMeHH M MMeeT JIHHeMHYIO 3aBMCMMOCTh. f pacl>ttlfeCKH M3MeHeHMe CKOpOCTM ct>epMeHTaTHBHOH peaKUMM orrpe,D,eJIHeTCH TaHreHCOM yrJia HaKJIOHa KacaTeJibHOH K KPHBOti rrpocl>ttJIH peaKUMM. qeM 6oJihrne yroJI

HaKJIOHa, TeM 60Jibllie M3Mettemie CKOpOCTM peaKUMM (pttc.

c Te'!eHtteMBpeMeHM M3Mettetttte CKOpOCTM cpepMeHTaTMBHOH peaKUHM B 3KCnepHMeHTaJibHblX ycJIOBHHX YMeHbIIIaeTcH, 06 3TOM CBM,D:eTeJibCTByeT YMeHbIIIetttte YTJia ttaKJiotta KacaTeJihtto:H B MOMeHT BpeMetttt t. CttIDKetttte cKopocTM cpepMettTaTMBtto:H peaKUMM MO)KeT npOHCXO)J,HTb 3a C'ifapH)J,a cpaKTOPOB: yMeHbIIIeHMH KOHUeHTpaUMH cy6CTpaTa, yBeJIH'ieHMH KOHUeHTpaum-1 npo)J,yKTa, KOTOpbIH MO)KeT OKa3bIBaTb HHrn6ttpyiomee )J,eHCTBMe, MO- ryr npOMCXO)J,MTb M3MeHeHHH pH paCTBOpa, HHaK-

THBaUIDI cpepMeHTa M T.)J,.

Ha 3Tane [t 1 - tJ cKopocTb peaKUHM M3MeHHeTCH HeJIMHeHHO B 3aBMCMMOCTM OT BpeMeHtt. Tio3TOMY )J,J1H onpe)J,eJieHHH CKOpOCTH cpepMeHTaTMBHOH peaKUMM ':lameBcero ttccne)J,YJOT H3MeHetttte

CKOPOCTM Ha Ha'iaJibHOM3TaTie ti], f)J,e Ha-

6JIIO)J,aI<n JIMHeHHOe H3MeHeHHe KOHUeHTpautttt npo)J,yi<.Ta (HJIM cy6cTparn).

CKopocTb cpepMettTaTttBHoH: peaKUMM 3aBMCMT OT pH)J,a cpaKTOPOB, TaKMX KaK KOJIM'ieCTBO11 aKTMBHOCTb cpepMeHTOB, KOHUeHTPaUMH cy6CTparn, TeMnepaTypa cpe)J,bI, pH pacrnopa, npttcyrcrn11e

A

[P]

peryJrnTOpHbIX MOJieKYJI (aKTMBaTOpOB M 11Hrtt6M-

TOPOB). PaccMOTPHM BJnrntttte 3TMX cpaKTOpOB Ha

CKOPOCTb cpepMeHTaTMBHOH peaKUHM.

A 3ABHCHMOCTb CKOPOCrn <DEPMEHTATHBHOH

PFAKl.UIH OT KOJIWIECTBA <DEPMEHTOB

Tiptt npoBe)J,eHHM cpepMeHTaTHBHOH peaKUMM B ycnoBttHx H36bITKa cy6cTparn cKopocTb peaKUMM 6y)J,eT 3aBHCeTb OT KOHUeHTpaUMM cpepMeHTa. fpa-

cpM'ieCKaSI3aBHCMMOCTb TaKOH peaKUMM ttMeeT BH)J,

npHMOH JIMHMM (pttc. 0)1,HaKO KOJilf'-IeCTBO

cpepMeHTa '!aCTOtteB03MO)l(HO onpe)J,eJIMTh B a6coJIIOTHhIX BeJIM'ittHax,Il03TOMY Ha npaKTttKe IlOJib3ylOTCH ycnoBHhIMtt Bentt'ittttaMtt,xapaKTeptt3YJOIIIttMtt aKTHBHOCTb cpepMeHTa: 0)1,Ha Me)K)J,yHapO)J,HaH e)l,MHMI!a aKTHBHOCTM (ME) COOTBeTCTByeT TaKOMY KOJIM'ieCTBYcpepMeHTa, KOTOpoe KaTaJIH3MpyeT npeBpameHtte 1 MKMOJih cy6cTPaTa 3a 1 MMH nptt OITTHMaJibHbIX ycJIOBMHX npoBe)l,eHHH cpepMeHTaTHBHOH peaKUMM. OnTMMaJibHbie ycnoBHH ttH)J,11BH)l,yaJihHhI MH Ka)K)J,OfO cpepMeHTa 11 3aBttCSIT OT TeMnepazypbr cpe)J,hl, pH pacrnopa, np11 OTCyTCTBl1M aKTttBaTopoB Ji[ 11Hrtt611TOpOB.

5

[S]

1 MKMOilb npeapa~eHHOro cy6cTpaTa

1 ME=

1 MHH

Kom1qecrno e;::urnm.1, aKTHBHOCTH nME onpenemnoT no <PopMYJie:

Ko11114ecTao npeapa~eHHoro cy6cTpaTa (MKMOilb)

n ME=

BpeMsi (MHH)

B 1973 r. 6bma npttmITa HOBIDI enttHHUa aKTHBHOCTll <PepMeHTOB: 1 KaTaJI (KaT), COOTBeTCTBYJOIUllH TaKOMY KOJIWiecrny KaTaJill3aTopa, KOTOpoe npeBparuaeT 1 MOJih cy6cTPaTa 3a I c. KonWJecrno KaTaJIOB onpenemnoT no <PopMyne:

Ko11114ecTao npeapa~eHHoro cy6cTparn {MOilb) n Kenan= -----------------

BpeMsi (c)

Me)l()zyHapoMIDI enttHttua <PepMeHTaTHBHOH aKTHBHOCTH ME CBH3aHa c KaTaJIOM cnenyioruttMH paBeHCTBaMtt:

1 KaT = 1 MOJib S/c = 60 MOJib S/MHH =

60xl06 MKMOJlb/MHH = 6x107 ME,

1 ME = 1 MKMOJlb/MHH = 1/60 MKMOJlb/C = 1/60

MKKaT = 16,67 HKaT.

B MenttUHHCKOH tt <PapMaueBTWiecKoii npaKTHKe WU! oueHKll aKTllBHOCTll <PepMeHTOB qacTO HCnOJih3YIOT Me)l(,llyHaponHhie enttHllUbI aKTllBHOCTll - ME. .IJ:mi oueHKH KOJittqecrna MOJieKyn <PepMeHTa cpentt npymx 6eJIKOB naHHOH TKaHH onpeneAAIOT yneJibHYJO aKTHBHOCTh (yn. aK.) <PepMeHTa, qllcJieHHO paBHYIO KOJillqecTBY enttHllU aKTllBHOCTll $epMeHTa (nME) B o6pa3ue TKaHH, neneHHOMY Ha Macey (Mr) 6enKa B 3TOH TKaHH:

Ko11114ecTao npeepa~eHHoro cy6cTparn (MKMOilb)

Yp,.aK. =

BpeMsi (MHH) x 1<011114eCTBO 6e111<a (Mr)

Ilo ynenhHOH aKTHBHOCTH cynHT 06 oqttcTKe <PepMeHTa: qeM MeHbllle nOCTOpOHHllX 6eJIKOB, TeM Bhlllle yneJihHIDI aKTHBHOCTh.

B. 3ABHCHMOCTb CKOPOCTH (J>EPMEHfATHBHOit

PEAKUHH Of .:rEMIIEPATYPhl CPJUU>I

IloBhillleHtte TeMnepaTyphI no onpeneneHHhIX npenenoB oKa3hrnaeT BJIIDIHtte Ha cKopocTh <Pep-

KOH4eHTpa4111si cpepMeHTa

PMC. 2-16. 3aBMCMMOCTb CKOpocTM 41epMeHTaTMBHOM peaK- l.IMM (V) OT KOHl.leHTPBl.IMM <iiepMeHTa.

MeHTaTHBHOH peaKUllll, nono6HO BJIHHHHIO TeMnepazypbl Ha JII06yio XHMWieCK)'IOpeaKUttIO. C noBhillleHtteM TeMneparyphl ycKOpHeTCH nemtceHtte MOJieK)'JI,qro npttBOnITT K nOBhillleHHIO BepoHTHOCTll B3allMoneiicTBHH pearnpyioIUHX eeruecTB. KpoMe roro, TeMnepazypa MoxeT noBhilllaTh 3HepmIO peampy10ruttx MOJieK)'JI,qro TaKXe npllBO):(llT K YCKOpeHllIO peamtttt. OnttaKo cKopocTh XHMWieCKOH peaKUtttt, KaTaJIH3ttpyeMIDI <PepMeHTaMH, ttMeeT ceoM: TeMnepazypHblH OnTHMYM, npeBbillleHtte KOTOporo conpoBO)l()laeTC.sI nOHll)ICeHtteM <PepMeHTaTllBHOH aKTllBHOCTll, B03HllKaIOIUllM ll3-3a TepMttqecKOH neHaTYPaUllll 6eJIKOBOH MOJieK)'JibI(pttc.

.IJ:mi 60JiblllllHCTBa <t>epMeHTOB qenoBeKa OTITllMaJibHa TeMneparypa 37-38 °C. OnHaKo B npttpone cyruecrnyioT tt TepMocTa6HJihHhie <PepMeHThI. HanpttMep, Taq-noJillMepaJa, BhmeneHHIDI tt3 MHKpoopraHH3MOB, )ICllBYIUllX B ropj{qllX llCTOqHHKax, He llHaKTllBllpyeTC.sI npll nOBhillleHllll TeMneparypbl no 95 °C. 3roT <PepMeHT llCnOJib3YJOT B HayqHOnpaKTWJeCKOH Me):(HUHHe WU! MOJieKyJI.sipHOH arnocTHKll Ja6oneeaHttH c ttcnonh30BaHtteM Merona nOJIHMepa3HOH uenHOH peaKUHll

PMC. 2-17. 38BMCMMOCTb CKOPOCTM 41epMeHTaTMBHOM peaK- l.IMM (V) OT TeMnep81ypbl.

B. 3ABHCHMOCTh CKOPOCTH

<I>EPMEHTATHBHOH PEAKil,HH OT PH CPE,Uhl

AKTIIBHOCTh ¢epMeHTOB JaBMCHT OT pH paCTBOpa,

B KOTOpoM rrpcrreKaer ¢epMeHTaTMBHaH peaI<IJJIJI.

Ka)K)loro ¢epMettTa cy:w;ecTByer 3HaqeHMe pH, rrpM KOTOpoM Ha6Jno.n,aercH ero MaKCMMaJibHaH aICTMBHOCTb. On<noHeHMe OT OITTMMaJibHOI'O3HaqeHMH pH rrpMBO- )J,l1T K IIOHIDKeHMIO ¢epMeHTaTMBHOM aICTMBHOCTM.

Bmurnwe pH Ha aKTIIBHOCTh ¢epMettTOB CBH3aHo c MOHM3aUMei1 ¢YHKUl10HaJibHbIX rpyrrrr aMllHOKMCJIOTHbIX OCTaTKOB )J,aHHOI'O6eJIKa, o6ecrreqHBaIOIUMX OIITMMaJibttylO KOH¢opMaumo aKTMBHOfO ueHTpa ¢epMeHTa. TipM M3MeHeHMM pH OT OIITMMaJibHbIX 3HaqeHMi1 rrpoMCXO)J,MT M3MeHeHMe MOHM3aUl1M ¢YHKUMOHaJibHbIX rpyrrrr MOJieKyJihl 6eJIKa. HarrpMMep, rrpM 3aKMCJieHl1M cpe)J,hl IIPOMCXO)J,JIT IlpoTOHMpoBaHMe coo6o)J,HhIX aMJmorpyrrrr (NHn, a rrpw 3aruenaqwBaHMM rrpOMCXO)J,MT OTIUeITJieHMe rrpOTOHa OT Kap6oKCHJibHbIX rpyrrrr 3-ro rrpMBO)J,MT K M3MeHeHl1lO KOH¢opMaUMM MOJieKyJibI ¢epMeHTa M KOH¢opMauMM aKTIIBHOfO uempa; CJie)J,OBaTeJibHO, HapyrnaeTcH rrpwcoe)J,MHeHMe cy6crpaTa, Ko¢aKTOpoB M KO¢epMeHTOB K aKTIIBHOMY uempy. KpoMe Toro, pH cpe)J,bI MO)K.eT BJIMHTb Ha CTerreHb MOHM3aUMM MJIM rrpocrpattcTBeHHYIO opraHmaumo cy6crpaTa, qro TaK)K.e BJiliHeT Ha cpo.n:cTBo cy6crpara K aKTMBHOMY uempy. Tipw 3HaqwreJibHOM CITKJIOHeHMM

v

OT OIITMMaJibHOI'O3HaqeHMH pH MO)K.eT rrpottCXO)J,l1Tb )J,eHazypaUMH 6eJIKOBOM MOJieKyJibl c TIOJIHOM IIOTepei1 ¢epMeHTaTl1BHOH aKTl1BHOCTtt.

0IITMMYM 3HaqeHMH pH y pa3HblX ¢epMeHTOB pa.3-

JllfqHbIH (pttc. <I>epMeHThl, pa60TalOIUHe B

KMCJibIX ycJIOBHHX cpe)J,bI (HanpMMep, rrerrcMH B )K.e- JIY)J,Ke 11JIH Jil130COMaJibHbie ¢epMeHTbI), 3BOJIIOUHOHHO npwo6peTaIOT KOH¢opMaUHIO, o6ecneqMBalO-

IUYIO pa6ory ¢epMema np11 KMCJibIX 3Ha'-leHHHXpH. 0)1,HaKo 6oJlbIIIaH qacTu ¢epMeHTOB oprattM3Ma '-le- JIOBeKa MMeer OIITlfMYM pH, 6Jil13KMM K Heti.rpaJibHOMY, c0Brra.n:a10ru11i1 c ¢w3HOJior11qecK11M 3HaqeHMeM pH (Ta6JI.

Ta6nM4a 2-1. OnTMManbHb1e aHa'!eHMRpH AllR HeKoTopblX

cpepMeHTOB

f. 3ABHCIIMOCTh CKOPOCTH <l>EPMEHTATIIBHOM PEAKIUIB OT KOJIWIECTBA CYBCTPATA

EcJIM KOHUeHTpaumo cpepMeHTOB ocrnBMTh rroCTOHHHOM, M3MeHAA TOJibKO KOJIH'IeCTBOcy6cTpaTa, TO rpacpMK CKOpOCTM cpepMeHTaTMBHOM peaK-

UMl1 OIIMCbIBaIOT mrrep60JIOM (p11c.

Ilptt yBeJiwieHMM KOJIMqecrna cy6cTparn HaqaJihHM cKopocTh Bo3pacrneT. Korn:a cpepMeHT crnHoBMTCH IIOJIHOCThlO HaCblllleHHhlM cy6cTpaTOM, T.e. rrpo11cxon:11T MaKCMMaJihHO B03MO)l(HOe rrptt n:aH-

HOM KOHUeHTpauHM cpepMeHTa cpopMMPOBaHMe

¢epMeHT-cy6cTpaTHOfO KOMTIJieKca, Ha6JIIOJl:aIOT

Ha1160JihlllYIO CKOPOCTb o6pa30BaHM5l rrpon:yKTa.

,lJ,aJibHetilUee TIOBhIWeHMe KOHll,eHTpaUMM cy6cTpaTa He rrpMBOJ:J:MT K yBeJittqeHmo o6pa3oBaHl15l rrpo- n:YKTa, T.e. CKOPOCTb peaKUl1M He B03pacTaeT. ,lJ,aHHOe COCTOHHMe COOTBeTCTByeT MaKCl1MaJihHOM CKOpOCTM peaKU11M Vmax·

TaKMM o6pa3oM, KOHUeHTpaUM cpepMeHTa - JIMMMTl1PYIOmttll: cpaKTop B o6pa3oBaHMM rrpoJ:J:YKTa. 3TO Ha6JIIOJ:J:eHMe Jierno B OCHOBY cpepMeHTaTMBHOM KMHeTttKM, pa3pa6ornHHoti yqeHhIMM JI. Mttxa3JittcoM M M. MeHTeH B 1913 r.

Cl>epMeHTaTl1BHhIM rrpouecc MO)((HO Bbipa311Th CJieJ:J:YIOill,MM ypaBHeHMeM:

E+S ES ___... E + P

f)le kl - KOHCTaHTa CKOPOCTM o6pa30BaHM5l ¢ep- MeHT-cy6cTpaTHOf0 KOMIIJieKca; k_I - KOHCTaHTa CKOpOCTM o6paTHOM peaK11,11M, pacrrarra ¢epMeHTcy6cTpaTHOfO KOMIIJieKca; k2 - KOHCTaHTa CKOpoCTM o6pa30BaHM5l rrpon:yKTa peaK11,11M.

CJien:y10mee COOTHOIIIeH11e KOHCTaHT CKOpOCTeM

(k_I + k2)/k1 Ha3bIBaIOT KOHCTaHTOM M11Xa:mnca M 060JHaqa10T K m.

CKopocTh peaKUJrn nponoputtoHaJihHa KOHUeH- Tpaum-r ¢epMeHT-cy6cTpaTttoro KOMJUieKca ES, a cKopocTh o6pa3oBaHMH ES JaB11c11T OT KOHUeHTpau1111 cy6cTpaTa 11 KOHUeHTpaUM11 CB06on:Horo cpepMeHrn. Ha KOH11,eHTpaun10 ES BmrneT CKOpoCTh ¢opM11poBaH11H 11 pacna.u.a ES.

Ha1160JihWM cKopoCTh peaKu1111 Ha6JI10n:aeTCH B TOM CJiyqae, KOf)::{a Bee MOJieKyJihl cpepMeHTa HaxoMTCH B KOMIIJieKce c cy6cTpaTOM, T.e. B ¢epMeHTcy6cTpaTHOM KOMIIJieKce ES, T.e. [E] = [ES].

3aBMCl1MOCTh CKOPOCTl1 ¢epMeHTaTl1BHOM peaKU1111 OT KOHUeHTpa1111M cy6cTparn Bblpa)((aeTCH CJieJl:YlOill,11M ypaBHem1eM (MaTeMaT11qecKoe BhrneneHtte 3TOM cpOpAfYJibl MO)((HO HaMTM B noco6HHX no cpepMeHTaTMBHOM Kl1HeT11Ke):

V = Vmax(S]

Km+ [SJ

3To ypaBHeH11e no11yq11JI0 Ha3BaHMe ypaBHeHHH Mttxa3JI11ca-MeHTeH.

B cJiyqae, Korn:a cKopocTh peaK111111 paBHa noJIOB11He MaKCl1MaJihHOM, Km= [SJ (p11c. 2-19). TaKMM o6pa30M, KOHCTaHTa MttXa3JIMCa 411CJieHHO paBHa KOHll,eHTpa111111 cy6cTparn, rrp11 KOTOPOM Jl:OCTHraeTC5l IIOJIOBl1Ha MaKCl1MaJibHOM CKOPOCTl1.

YpaBHeHne MHXa3JI11ca-MeHTeH - ocHoBHoe ypaBHeH11e ¢epMeHTaTl1BHOM Kl1HeTMKM, OIIl1CblBaJOill,ee 3aBHCl1MOCTh CKOPOCTl1 ¢epMeHTaT11BHOM peaKUM11 OT KOHUeHTpau1111 cy6cTparn.

EcJIM KOHUeHTpauttH cy6cTpaTa JHaq11TeJihHO 6oJibllle Km (S>>Km), TO yBeJI11qeH11e KOHUeHTpa11Ml1 cy6cTparn Ha BeJinq11Hy Km npaKT11qecKM He BJll15leT Ha CYMMY (Km + S) 11 ee MO)l(HO C4MTaTb paBHOM KoH11eHTpau1111 cy6cTparn. CJien:oBaTeJihHo, CKOpOCTb peaKUM11 CTaHOBMTC5l paBHOM MaKCl1MaJibHOM cKopocni:: V = Vmax. B 3Tl1X yc110B11Hx peaKUM MMeeT HyJieBOM TIOPMOK, T.e. He 3aBl1CMT OT KoHueHTpau1111 cy6cTparn. Mo)((HO cn:eJiaTh BblBOJl:, qTo Vmax - BeJil14MHa IIOCT05lHHa5l Jl]15l )laHHOM KOHUeHTpau11M cpepMeHTa, He 3aBMCHill,a5l OT KOHUeHTpauMM cy6cTparn.

EcJIH KOHUeHTpauttH cy6cTpaTa JHaq11TeJihHO MeHbllle Km (S<<Km), TO CYMMa (Km+ S) rrpttMepHO paBHa Km, CJien:oBaTeJibHO, v = vm)S]/Km, T.e. B n:aHHOM CJiyqae CKOpOCTb peaKUHM npHMO rrpo-

~

:c

..0

~

:r

!ti

::c

[S]

P1tc. 2-19. 3aBltCltMOCTb CKOPOCTlt peaKl.lltlt (V) OT KOHL1eHTpa1.11t1t cy6CTpaTa S. Vmax - MaKCMManbHafl CKOpOCTb peaKL.11<1"1 npi.i AaHHO~ KOHL1eHTpa1.1MM cpepMeHTa B onrnManbHblX ycnOBMflX npoeeAeHMfl peaK1.1i.i111. Km - KOHCTaHTa Mi.ixasni.ica.

ITOPUlfOH<UlbHa KOHUeHTpau1rn: cy6CTPaTa (peaKUIDI 11MeeT rrepBbIH rropSl)J,OK).

Vmax 11 K m - KHHeT11qecKHe xapaKTep11cT11KH 3cpcpeKTl1BHOCTl1 cpepMeHTa.

)laeT xapaKTep11CTHKY KaTaJIJITwqecKOH aKTl1BHOCTl1 cpepMeHTa 11 11MeeT pa3MepHOCTb CKopocTl1 cpepMeHTaTl1BHOH peaKU1111 MOJib/JI, T.e. orrpe)leIDieT MaKCl1MaJihtty10 B03M0)KH0CTh o6pa30BaHIDI rrpo.rzyKTa rrp11 )laHHOH KOHUeHTpau1111 cj:>epMeHTa 11 B ycnoB11HX m6bITKa cy6cTpaTa.

K m xapaKTepmyeT cpo)lCTBO )laHHoro cpepMeHTa K )laHHOMY cy6cTpazy 11 HBJI.HeTCH BeJittql1HOH rroCTOHHHOH, He 3aBl1CHW,eH OT KOHUeHTpau1111 cpepMeHTa. qeM MeHbIIIe Km, TeM 6oJibllle cpo)l- crno cpepMeHTa K )laHHOMY cy6cTpazy'TeM BbIIIIe HaqaJihHM CKOpOCTh peaKU1111 11 Hao6opoT'qeM 6onbIIIe Km, TeM MeHbIIIe Haqa.rrbHaH CKOpOCTb peaKU1111, TeM MeHbllle CpO)lCTBO cpepMeHTa K cy6cTpaTy.

Ha p11c. rrpe)lCTaBJieHa 3aB11c11MOCTb CKOpocT11 )lByx cpepMeHTaTHBHhlX peaKU11H (1 11 2) OT KOHUeHTpaum1 cy6crparn. KoHCTaHTa M11xa3mrca rrepBoro cpepMeHTa MeHbIIIe KOHCTaHThI M11xa3n11ca BTOporo cpepMeHTa (Km 1<KmJ Cne)lOBaTeJibHO, cpO)lCTBO rrepBoro cpepMeHTa K cy6cTpaTY Bblllle, qeM y BTOporo cpepMeHTa, 11 HaqaJibHM CKOpOCTh peaKil,1111, KaTaJil13MpyeMOH rrepBbIM cpepMeHTOM, BbIIIIe B cpaBHeHl111 co BTOpbIM cpepMeHTOM.

VI. ~Hf~S~POBAH~E

Cl>EPMEHTAT~BHO~

AKT~BHOCT~

Ilo)l TepMMHOM «MHrM611pOBaHMe cpepMeHTaTHBHOH aKTl1BHOCTH» ITOHHMaIOT CHH)l(eH11e Ka-

v

<l>epMeHT 1

Vmax

1/2 Vmax

[S]

PMC. 2-20. BnMRHMe pa311M'4HblXKOHLieHTpa~M cy6crpara Ha CKOpocTb pe8KL.IMM, KaT8nM3MpyeMOM cj>epMeHT8MM 1 M 2.

TaJIHTMqecKOH aKTHBHOCTH B rrpttcyrcTBHH orrpe- )leJieHHhlX Bew,ecTB - MHrtt611TopoB. K 11Hrtt611TOpaM CJie)lyeT OTHOCHTh Bew,eCTBa, Bhl3bIBaIOW,Me cHmKeH11e aKTl1BHOCTM cpepMeHTa. Cne)lyeT oTMeTHTb, qTo BCe )leHazyp11pYJOW,l1e areHThl Tal()l(e Bhl3hIBalOT yMeHbIIIeH11e CKOPOCTl1 JII06oif cpepMeHTaTHBHOH peaKUHH, BCJie)lCTBMe Hecrreu11cp11qecKOH )leHaTypail,MH 6eJIKOBOH MOJieKYJibl, rro3TOMY )leHaTyp11py10w,11e areHThl K HHTH6HTOpaM He OTHOCHT.

11HrM6MTOpbl Bhl3bIBalOT 60JiblllOH MHTepec )lJlH

BhlHCHeHIDI MexaHH3MOB cpepMeHTaTHBHOfO KaTaJlM3a, ITOMOraIOT yCTaHOBMTh POJih OT)leJibHhIX cpepMeHTOB B MeTa6onttqecKMX ITYTHX opraHH3Ma.

B OCHOBe )leHCTBIDI MHOfHX JieKapcTBeHHhIX rrperrapaTOB H Sl)J,OB Jie)l(HT MHrn6MpOBaHMe aKTHBHOCTH cpepMeHTOB, I103TOMY 3HaHHe MexaHH3MOB 3TOro rrpouecca KpaifHe Baxtto )lJIH MoneKynHpHoif cpapMaKOJIOfl111 H TOKCHKOJIOfHH.

11HrH611TOphl crroco6Hhl B3al1MO)leikTBOBaTb c cpepMeHTaMH c paJttoif cTerreHbIO rrpoqHoCTH. Ha OCHOBaHHH 3TOro pa3JIMqa10T o6paTHMOe H Heo6paTHMOe MHrn611poBaH11e. Ilo MeXaHH3MY )leHCTBIDI HHrH6HTOpbl I10)lpa3)leJIHIOT Ha KOHKYpeHTHhie H HeKOHKypeHTHbie.

A. OJiPATHMOE HHfHliHPOBAHHE

06paTHMbie HHrM6HTOPbl CBH3hIBalOTCH c cpepMeHTOM cna6bIMl1 HeKOBaJieHTHhlMH CBH3HMH J1 rrp11 orrpe.n:eneHHbIX YCJIOBHHX nerKO OT)leJIHIOTCH OT cpepMeHTa. 06paTl1Mbie HHrH6MTOpbl 6bIBalOT KOHKYPeHTHhlMH H HeKOHKYPeHTHhIMM.

1. KonKypenmnoe unzu6upooanue

K KOHKypeHTHOMY MHr11611poBaHMIO OTHOCHT

o6paTHMOe CHH)l(eHMe CKOPOCTH cpepMeHTaTHBHOH peaKU11H, Bhl3BaHHOe 11Hr11611TOPOM, CBH3hIBaIOllJ,HMCH c aKTHBHhIM ueHTpOM cpepMeHTa J1 rrpeITHTCTBYIOW,11M o6pa30BaH1110 cpepMeHT-cy6cTpaT- HOro KOMrrJieKCa. TaKOH THil HHr11611poBaHMH Ha6n10.n:a10T, KOr)la 11Hr11611Top - cTpyKTYPHbIH aHanor cy6cTparn, B pe3ynhTaTe B03HMKaeT KOHKYPeHUMH MOJieKyn cy6cTpaTa 11 11ttr116ttTopa 3a MecTo B aKTHBHOM ueHTpe cpepMeHTa. B 3TOM cnyqae c cpepMeHTOM B3aHMO)leifcrnyeT JIM6o cy6CTpaT' JIM60 HHfi16HTOP' o6pa3yH KOMITJieKCbl cpepMeHT-cy6cTpaT (ES) 11Jll1 cpepMeHT-11HrH6H-

TOP IlpM cpopMMpoBaHMH KOMITJleKca cpepMeHTa 11 HHm6MTOpa (El) rrpO)lYKT peaKU11H He 06pa3yeTCH (p11c.

,il.JISI KOHK)lpeHTHOI'Orn:rra HHrH6HpOBaHIDI crrpaBe)l)IHBbl cne.uyiomne ypaBHeHttSI:

E + S ¢:::} ES --t E + P,

E + I ¢:::} EI.

K.riaccw-i:ecKllli rrpHMep KOH~HTHoro HHm6HpoBattHSI - HHrn6ttpoBaHHe cyKUHHar.ue:rn,n.poremnHOM peaKI.J,llH MaJIOHOBOM KHCJIOTOH (pttc. 2-22). MaJIOHOBaSI KHCJIOTa - CTpYJCTYPHbIB attaJIOf CyKUHHa'ra (HaJIWIHe .usyx Kap6oKCHJihHbIX rpyrrrr) H MO)[(et TIU<-

)!(e B3illfMO)leHCTBOBaTb c aJCTHBHbIM I.1,empoM CYKI.l,H- HaT .uern.uporeHaJhI. 0)lHaKO OTII.1,eIIJieHHe .usyx aroMOB oo.uopo.ua OT MaJIOHOBOH KHCJIOTbl HeB03M0)[(HO; CJie)lOBaTeJlbHO, CKOpocrb peaKI.J,llH CIDl)[(aete.SI.

KlmeTH'lecKHe3aBHCHMOCTH

KottK)lpettTHhie Httrn6tttopb1 yMeHblllaIOt CKOpocTh XMMttqecKoil: peaKI.1,HM. KoHK)lpeHTHhIH HHm-

6HTOp IIOBhilllaet Km )lJlSI .UaHHOro cy6crpaTa (YMeHhlllaer cpo.ucTBo cy6crpara K cpepMettry). 3to 03Haqaer, qro B rrpHCytCTBHH KOHK)lpeHTHOro MH-