1、DB61 if fJji 1ft DB61/T 414-2007 I I I f I l t I , I J 2007-09-03fj 2007-09-03:Mt! f I DB61fT 44-2007 *1fEIJ !lftAj; j!i*1-11 lft0 *1fE$AOOMfiWA.Wo *1fEm: $AOOMfiWAo *1fE.A: .*ffi,.m*,*Vili,. *f1fE*1tIJ(1t;fi 0 1 I I I DB61ff 414-2007 , ! 1 m: :$: * m .m 1 Jt7t;W;Etx*4 if L-1ft3IHl1f.6trlJ7t; a :$:t
2、3FfiE m ttNifL-1ft*.W1;Eo 2 ;tAl,m:I1.1 fflJt1tJ: . F 3itl)(itl: 91J 3l!1i1:$: f,f,fiE IJ CJ IJ:l:lffij px;:fiJ :$:,;j; miJ 0 ;tJl!:;Ii BW:l 851 F8 Xi:, JtI%R fri JilT 1:f IJ .a.(ftmmIJW).rrm:$:m,ffij,.*:$:nmIJ Ji: WJ Ie mis: Xi: IJ ltiNf Yli BJtJj Er-J CJ ImX1 , :!tlfUT:$: J?i mT :$: f5F 1to EN 113
3、8-1995 7JJtHt*UlirutifL-:ijL*:lVJl;E L-MDH NAD NADH GOT U A C 4 JmJl L-1jt !1H (L-1jt L1jt*!bJli.tl mlpt=tfM f3 _- z.!b $tt1UW IW 1ffi ID ,f, Jfl-1.fl:, 1 25 C IJ fj; m 1 11 mol iJ!jtJJJ9f Pltft;:.=. oxaloacetate + NADH + H+ (1) L-Malate + NAD+ t z.mi LU1JZ#lJttll)5t, r$NtJ.! L:*q:.1ZUtH2) aNADH ,ff
4、dltWJ!J-;tjJJ L i !P:m,e.j;jt, :fW:Fni 340nm i;:x:=. L-aspartate + 2-oxoglutarate 5 i.itjf 5, 1 m.tfiil.Jltti/lP 5.2 L i;titM 5,3 NAD DB61 rr 414-2007 5. 4 It.:e:.Z.*lii. 5.5 L-MDH 5.6 HCl 5.7 NaOH 5.8 KOH 5.9 tii* 5. 1OteiiIitIf)iii8f.lit1lJj* 6 it. 6. 1 Uji (5 mL 1 mL 50-200L 10-50L) 6. 2 t1!.no.:
5、)tt LOcm 6. 3 9:)t:)tJllt, iiIsjtiE:, f 340nm IJ:iE 6. 4 . mAi.jm, :fLf 1 0 6.5 il,l.tIL 6.6 ilui, mF, -3.Mi, iilr,.fiIj!k, .Ja. 7 ljiIT. 7. 1 iitif1jlr,*IJ 7.1. 1 tfllfU 0.88g L-.Il, ffl NaOH iJj pHlO.O, fflm:)tcIEW 60mLo 7.1.2 NAD If1:Hl 1fffla1, fIt ImL m:7JCtfdlf 35mg TfiP NAD Itf.o 7.1.3 1l.-:e
6、:.Z.!HHl itffl a1, ffl 0.4mL .m1A7JCfdlf 160U (fJ.Il-1imIlT*5IlP .Il-1itm;Iifa:IjI tf.o 7.1.4 L-MDH ;ii. 600 U/100L 7.1.5 HCl: 2 mollL 7.1.6 NaOH: 2molIL 7.1.7 KOH: 2molIL 7. 2 lit*IJ. 7.2. 1 7;1!i1!jt ffl2mol/L KOH iaJ:lHOmL*rrpH11I78, 1JQ:l:li7JC, 1fffll. 0001. 800 mLjHtilo 7. 2. 2 j.rtl8f. u:ff,
7、2500r/min /L.15min, IOC.tiPt10mL. ffl2mollL KOH iJj10mUIHtpH1Fi78, 1JQ :lJ7JC, 1fffll. 0001. 800mL:i1tHtili0 7.2. 3 *91*8f. :ff9=J(fJCC11*, ffl2mollL KOH iaJ10mL*tpH1H78. *JIlf:kif.J15min. f.tffl1. OOO 1. 800mL:i1tlftltwW 7. 3 )Y!lj(j.i 1 iHT) 7.3. 1 itS;jmt IOCtlllI. itak1JQA 1. OOOmL tt;(HtIf, O.
8、200mL J(Il-1il1tIll!:.;i; O. OlOmL 1. 000 mL It!7j(. mil-JI 3min JI5IOCI! (j(if.J 20min). ij;Jt)t!l!JOA 1. OOOmL x.ztfH*tp, 0.200mL NAD IfF, tt-MZ.Itltlfi;f! - O. OlOmL 0.100 mL Uf, m:*O. 900 mL. mil. 3min J8i1t;It!JoA L-MDH m0.050 mL. fflmilIt!JOAt.IIIlJm S fj! x.z1tltfttp NADlfF tt-MZ.flfj;f!r fm
9、:I:* 1.000mL 0.200mL 0.010 mL 1.000 mL 1.000mL 0.200mL O.OlOmL 0.100 mL 0.900 mL ?J, Agf.j 3min J8Itl1r4ijt-fRn IJItfIJS Itjili* (2) ii1tj: I ilnIJ!lmJ9tIJJf$J1it:lt. L :SY:.!itHt32: $J1it:l: 134.09 glmol E!X; L :sy:*lttxll$ 1 , i MW J1it:l: 132.07 glmol) , I VI t.IIIl9=Ij!lJ.mIJJ,1*f, mL V 2 lJ.f.mIJJ9Hllt J 9. 1 L .J9HifiHf-1-tF;l L .*ifiji I 9.2 Ilem I 9.2. 1 ii1JnMtifiHfIJ!JIJ 0.050mL 1JJ1itlJOiUmi19=I. ji1iOOIJ.(AT)*.*lf.mLdJ81*ItJ1JF, xtjfMJ8JJ1NjtHflKiij1ij, m4J;l!Nff*,mTtt. , :tmAt4J8lM. 5. l!:l:lW!&:ftJJlllA1H (l. I I j 5
