1、DB61 jf f 1ft DB61/T 410-2007 2007-09-03 2007-09-03 I DB6ur 410-2007*t;jfttJ Il1f:i.RA1:6.t 4t1:Il1HK 0*ft$AR WA.WfrD.*.:$AWA.*ft.A;m*,*,*ffili,a.*tiftlnj(:ffl.!t I I I t t I I I1 i l DB6tff 410-2007 1 II*m.ffM*oo.mmfff*.o,2 1115Im3t1tj:-fjitl)(fi:00*-.-:ilI1:i1*5mtr.J 51 flHffi J)!l*5moo t.kc Lff 8
2、 W:I iJ 51 m)(IF,ltmIJfiM1.:riJ ft:&(:ftliWJ00 P;J tP!IX1iTAOC:ffjm T*5m,rtffi,)jjJ*JRft.i6Jt;1)ixEfJ nEff1:1iiJf1J:l3:i!)(1!I:ooliUTAOC*o L:fff8 Jt)j iJ 51 m)(1*,JtliiTAOC*Jfiffi T*;fm0 IFU 76(2000:iF1jffD-iFfi11iJg ATP 5-=M1r ADP 5-=J11J 6-PGDH.6-MtJlmttll NADP+1:Ii;mUJ1pnt-tr NADPH JJjjm:mJM;IJg(
3、Wolnt=f.1l.U ji1tt:EI Fi/:,1 25C iJf;Jmt.kf!l:-F1fJ:minIU iJ.i5#J$tt1tJ 1 Jl mol iJwfPi A rJ&7t&ffi C wfPitrJrRlt 4 JI tE.4t1.iJfHR-f-,D-.4t1.flTH&5-:.J1tt(ATP).1t;JD-6-.1ii1U:;f115-.:.JlIHr(ADP)JJlA;J:0 illititj,Hl31MD-lii+ATP.D-6-M.Hm.+ADP t 6-riiij)Uii(6-PGDH)iJftF8 I,D-6-.ijmHBz:mJf:liUHn1l7-;f1
4、J.(NADP)1t)tt)!l 5-.t.WIiij:fllJJw)HtlM;H1i:JlInIl7-11.(NADPH)o 1JtA7g:6-PGDH D-6-+NADP+.5-.+NADPH+CO,+H t.tEZ.fY-NADPH Jf5J1J,8,3 T D-ij,8,1$t.:ilI1:i1itEZ.Jlljjf5(1 340nm tlJP&7t7tltm itNADPH Jf5J00,tr!,a;j-JiijtiIJl),fi!tiJE D-ijiJ,8,iio,5.1 NADP:-15C-20C1f l 5.2 ATP:-15C-2oCHf1:1fI 5.3 MgSO,7H10
5、 DB61ff 410-2007 5.4 6-PGDH 5.5 1iiUfU;5.6.=.ZM:1ni:5.7 Hel 5.8 NaOH 5.9 KOH 5.10:i:*5.11 tB 1.rem tlj,fb t*Uif 6 iii!6.1 l$1flm(5 mL,1 mL 50-200 ilL,10-50 II L)6.2 ltJIll;1;:&L Ocm 6.3 9r7t1tYtllir.*,340nm U!E 6.4!1T!ttJ,fL1 10 II m 6.5,c.m 6.6.c.,1IEtf,:lJm,:ffi:!G!k,ii4,J$J*,7 jmtJ!I7 7.1 iitfflJ
6、IlltilJ 7.1.1=ZD/$Hi:NADP(g60mg),ATP(150mg),Imol/L MgSO.7H 20 4ml,-r:27mL:7k,pH7.6o 7.1.2 6-PGDH I i1=i!f1li.J;j22 U I 100J.1LIfF.7.1.3 1i1i1J!(.Ri i!1li.J;j26U 11.OmL!H.7.1.4 He:2 moIlL 7.1.5 NaOH:2mollL.7.1.6 KOH:mollL 7.2 lit*J*7.2.1 jff:it:,7C,!,q:it1fii*X1m,x13,tpIi1*,fb,1:itt1TtftWlIJ,1tffll.O
7、OOl.800mUfH&:itt1Ttft.7.2.2 i!I!i3l:jEUf,fb,fflNaOHnt,KOHmtfJpH8ftti,1tffl L OOOl.800mLm1fl:itt1TtftWltl 0 7.2.3*1*jt llt;fftptfJCOl;:1*,fflNaOHmKOHmpH8ftti,tfPnfjd15min,1tffll.OOO1.800mL m:ittlJ0 7.2.4 fi,&,.tafii*fflNaOHffi:WI;KOHmiaJpH8ftti,tfrfjFfjd15min,1tffll.OOOl.800mL m:itt1TtftW!1o 7.3 J 01
8、i 1 ilIff)7.3.1 li:S;i;jlffi?DB61IT 410-2007 ItHtilll,1t1JnA 1.OOOmL-=z.IJ$C!1k,2.000 mL m:*,O.020mL 6-flfj.Jlltfj,mii:l5min FnIJ&YtJ.l1i(AI)W1JnA 0.020 mL fjlUfmM1&n,#fmii&mz%pj(;(*tJ.2Omin),IJ&Ytll1i(A2),:!m*&mz1 20min FnfIlt,4ft1i3Ji 2min tHUIJ&Ytll1i,1I 2min IJ&YtJ.l1iW:it1Jno sIJ&Ytll1i=(fu-AJ
9、8 0 7.3.2*$8l:t*ll ltilll,1f(1JnA 1.OOOmL:=z.IJ$C!1k,O.100mL tHMtf,1.900 mL m:*,O.020mLI 6-PGDH IftHi,mii:l5min FnIJ&Ytllfi(AI),1J11A O.020 mL fjlUfmiU;1.r.twnii&mz%pj(;(*g 20min),IJ&Ytll1i(A2)n*&mztE 20min FnfIit,4ij;fiijlfRj 2min mttJIJ&Yt r lt1i;2min I*J I1IJ&Yt1l1i:it1Jn tFJi,IJ&Ytllfi=(A2-AI)n&
10、.0 i 1*i-IJfJ:r 7.tlllj!fJi,s tf.JIJ&Yt1l1i,ffltFJi,IJ&Yt!.t1iSIJ&Ytlt1iWI2LjIJ0A.(1):6.A=(ArA)nz,-(ArAI)e.(l)1IJ IJ&Ytll1imz*T 0.100.1JfjjIJlIflYliJD*.8 it.I i 1JnAbtilllrf1m sxtmt tF,Ji,1&tl :z.lJ$Cgl9 1.00OmL 1.00OmL tFJi,m-O.100mL m:*2.000mL 1.900mL 6-PGDH I 11:0.020mL 0.020mL 1:,5minFnmI1IJ&Ytl
11、tUl(AI)1Jnl2Lrpj(;7.tJtttr&mz fjlUttMn 0.020mL 0.020mL Im0:J,m&mz%Fn(*g 20min)IJ&YtJ.lfi(A2).*Jimz1 20min FnfIit,4ftla:ii 2min mtlIJ&Ytll1i,1I 2min I*J I1IJ&Ytll1i:tt1Jn.ttt11i!i0i(2)it_*J.l:V/XMW.c-XLlA X/.(2)eXdXV2 X 1000 f D-fjlUtttf.J*lt;7g:i,.3.040X 196.1 c=-XAAX f eX1.OOXO.100X1000 5.961I I c=
12、XL1AX f A.rf1:C 1&W)!J1:*ll:D-fjfm giL fJi,mHi MW:fft4m)!J1:I1JJt:l:D-fjfm)!J1:ib 196.1g1mo,D-fj.1iIL 3 DB61(f 410-2007 jJJjIHI:*J 195.1g1mol Vj tElIll.fti!JWi.ifHIJ,1*fQ,ljift mL V EE i.i;f4:f!5 IrffJIM.tf IJ I1*fQ,ljill:mL f*fffldiflJff*d bElIll.IJYtf,lji1ll:em e NADPH IJmJt:tE 340 nm:6.31 x mmor1
13、 x em-9 D-IiIiJIJRtl.!Milit!1fi tEJtHfD-tin3t1M,itHfJ9H5!i%.9.1 19tmRlI.!/1t*fijifjf*67mg D-tiiJWHifl,72mg D-tiIt,:1.1O;.$IJ 100mL IJ.:F.JlljJ*(ffi 0.6gD-til1!/L).9.2 19tJ2im 9.2.1:tJomjflljJiJ2ii*,9.2.2 _iitfM:ii1:1Jr:iE:l:JiJ2i;f4:m&BJ4L AdtB9;Q,:no O.050mL IJm:nnjUtlYlm.i?ft,&fL!t!J(30min),m ILUf
14、lfiJJ:t(1:1-fu)it*lt,JZt&J1*fQQ 9.2.3 pgtF m5.ifjIJ%ttf*H.,;f4:mftAmttQ 9.2.4 19tlitiV(m*2):nu;.tE#ftIJm B;f4:fiF1tE;f4:+f*m z.f1P7t;f4:m mm:I:f7.K 6-til1!IDiltftfj 1.OOOmL-2.000mL O.200mL 1.OOOmL O.100mL-1.900mlmL O.020mL 1.OOOmL-O.100mL 1.900mL 0,020mL 1.OOOmL O.050mL O.050mL 1.900mL 0.020mL mi?,*J6min*Il1Ytltm(AI),Jffi 7.3tJ;!JI fn tf 1U:jto 2 X.1 AIIJHiifr.1 A#/1,X 100%.,.,(3).1 A.tiKi 4 DB611T 410-2007,r 1 I I I t 1I I I 1 1 I I I 5 1 I I
