生物转化制备D-谷氨酰胺的研究

D-,,,,,(,210093):D-,AS1.505L-D,L-L4--,D-pHL-:37,pH=418,5g/L,-800115g/L,14h,40g/LL-4200U,L-8h4--,D-92%:L-;;D-:TQ93:A:0253-4320(2005)09-0044-03StudyonpreparationofD2glutaminebybiotransformationQIANShao2song,WUXiao2yan,LIUYi,CHENRan,LIJia2you,JIAOQing2cai(StatekeyLaboratoryofPharmaceuticalBiotechnology,SchoolofLifeScience,NanjingUniversity,Nanjing210093,China)Abstract:OpticallypureD2glutamineisinitiallypreparedbybiotransformationsofD,L2glutamineinthepresenceofL2glutaminedecarboxylaseinEscherichiacoliAS1.505.Factorsthataffectedtheenzymaticactivitysuchasconvertingtemperature,pHvalue,etc.,werestudied.Theresultsshowthatthespecificenzymesactivitycancomeupto4200U,andL2glutaminecanbecompletelydegradedbythedecarboxylasefor8hundertheopticalconditionsasfollows:37,418ofpHvalue,5g/Lofcellconcentration,0115g/Loftween280,14hofstrainage,and40g/Lofthesubstrateconcentration.Keywords:L2glutaminedecarboxylase;biotransformation;D2glutamine:2005-06-14:(02CJ-13-01-16):(1968-),,;(1959-),,,,,,,025-83594376,[e-mail]-,L-D-D-,L-D,L-,D-,D-D,L-2:D,L-,L,D-[1-2];D,L-N--D,L-,L-,N--D-D-[3-5];D-N--D,L-D-[6-8]L-,,,L-D,L-,D-L-L-4--,D-D,L-[9],(Escherichiacoli)AS1.505L-,D-1111EscherichiacoliAS1.505,Nexus870FT-IRBruckerDRX-500MarinerAppliedBiosystems(T)WRS-1()UV-3000THZ-CWZZ-2B()CR22E:3g/L,10g/L,5g/L,NaCl3g/L,3g/L,44Sept.2005259ModernChemicalIndustry20059015g/L,18g/LpH712:10g,5g,NaCl3g,3g,015g,L-1g,pH712,1000mL1121121114h,,37170r/min14h,,11212100mL0105g,D,L-014g,012mol/L/(pH418)20mL,-80010015g10mL37170r/min30min2mL,012mol/L(pH910)2mL,,4-37pH4181g1h4-mol,U,1U=110mol/(gh)11213D-D,L-40g1000mLpH418-,510g,378h,,,pH715,D301,,,,4-1816g;011mol/L,,,1814gD-,92%11214D-[]20D=-713(c1,H2O),184185(D2O,500MHz),:2116(m,2H),3146(t,J=713Hz,2H),4102(t,J=712Hz,1H)IR(KBr),:3409,3213,2987,2041,1687,1632,1585,1485cm-1(HRMS):[C5H10O3N2+Na+],16910982,16910991;[C5H10O3N2+H+],14711046,147110542211L-L-,,1L-37,,,37,,1212pHL-,pH,2,pH4155102pH213L-,,,,,,,,,3,,5g/L,,5g/L35420059:D-214L-,-80(Tween-80)(CTAB)(OP)3,4,,-802L-,0115g/L4200U1-80;2OP;3CTAB4215L-,L-(5),14h,,(),5216L-,,,,,40g/L(6)6217L-5g,40g/L,,L-4-2,L-12h,,448h3E1coli11505L-37,pH418,5g/L,-800115g/L,14h,40g/L,4200UL-8h4-,D-92%50%[1]SenumaM,OtsukiO,SakataN,etal.[J].JFermentBioeng,1989,67(4):233-237.[2]MakotoY,AkioO.[J].JMolecularCataB:Enzymatic,1998,4:1-11.[3],,,.[J].(),1998,4:81-84.[4],,,.[J].,2001,3:293-296.[5],,,.[J].,2005,24(5):537-540.[6]SakaiK,ImamuraK,SonodaY,etal.[J].FEBSLetters,1991,289(1):44-46.[7]WakayamaM,KatsunoY,HayashiS,etal.[J].BiosciBiotechBiochem,1995,59(11):2115-2119.[8]WakayamaM,TsutsumiT,YadaH,etal.[J].BiosciBiotechBiochem,1996,60(4):650-653.[9].L-[P].CN20041001473715,2004-04-26.64259