葉酸途徑對谷氨酸棒桿菌SYPS-062積累L-絲氨酸的影響分析.pdf_第1頁
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1、江南大學(xué)碩士學(xué)位論文葉酸途徑對谷氨酸棒桿菌SYPS062積累L絲氨酸的影響分析姓名:任建洪申請學(xué)位級別:碩士專業(yè):發(fā)酵工程指導(dǎo)教師:許正宏20090801AbstractAbstractAsoneofthemostimportantcoenzymeinthedegradationpathwayofLserine,folatealsoplaysallimportantroleinonecarbonunitcycleandcellproli

2、ferationInthispaperapreliminarystudyhasbeencarriedontheeffectoffolatemembolismonthegrowthandL—serineaccumulationofCorynebacteriumglutamicumSYPS一062,andapproachedtheimpactofexpressionoffolatemetabolismrelatedgeneonCglutam

3、icumSYPS062FolatemetabolismpathwayinCglutamicumSYPS一062wasregulatedthroughbothendogenousandexogenousregulationSulfamethoxazole,methotrexate,folateandPABAwereaddedtoinvestigatethefunctionoffolatemembolismtoCglutamicumSYPS

4、062Asendogenousregulation,antisenseRNAandoverexpressiontechniqueswereappliedtegulatetheexpressionofthekeyenzymeinfolatesynthesisDistarbanceofsulfamethoxazoleandmethotrexaterevealedthattheycoulddepressthegrowthofCglutamic

5、umSYPS062andincreasetheyieldofL—serineIncomrastPABAacceleratedthegrowthofCglutamicumSYPS062whilegreatlydecreasedtheLserineproductioncapacityHoweverthegrowthofCglutamicumSYPS一062wash’tacceleratedbyfolateduringtheadditionc

6、oncentrationrange,insteadofoverdosagefolatedepressedthegrowthofCglutamicumSYPS一062ItindicatedthatCglutamicumSYPS一062withthecapacityofdenovofolatesynthesiscail’tuptakeexogenousfolateMoreovertheanalysisoffluxdistributionce

7、rtificatedthatfolatemetabolismascontrollablepathwaywasveryimportantinCglutamicumSYPS一062growthandLserineproductionForfurtherinvestigation,wecomparedthedifferencesofpabABbetweenCglutamicumATCC13032andCglutamicumSYPS一062Th

8、esequencehomologyofpabABinthesetwostrainsis9914%,16basepairsdifferencesthatresultedinthechangesof7aminoacidswerefoundTheexpressionofpabABwasdown—regulatedbyantisenseRNAtechnique,theresultsshowthatslowergrowthrateandhighe

9、rLserineoutputhaveoccurredinthegeneticengineeringbacterianlepabABgenesfromdifferentsourceswereexpressedinEscherichiacoliandthespecificactivityofaminodeoxychorismatesynthasethatpabABencodeweremeasuredSpecificactivityofami

10、nodeoxychorismatesynthaseinCglutamicumSYPS062Was466%lowerthanthatinCglutamicumATCC13032,whichindicatedthattheconcentrationofcoenzymeofSHMTinCglutamicumSYPS062mightbelowerthanmodelstrainThedecreaseofdegradationmightbethem

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