玫瑰孢鏈霉菌代謝通量分析及戊糖磷酸途徑改造.pdf

1、天津大學(xué)博士學(xué)位論文玫瑰孢鏈霉菌代謝通量分析及戊糖磷酸途徑改造姓名：王國英申請學(xué)位級別：博士專業(yè)：化學(xué)工程與技術(shù)；生物化工指導(dǎo)教師：聞建平201012ABSTRACTDaptomycinisanoVellipopeptide鋤tibioticwhichw嬲producedby腳幻叼彬Ps加船D印D，淞‘a(chǎn)ndc鋤resistt0mostgrampositiVebacteriaCubicin，thecommercialn鋤eofdapto

2、mycin，hadasalesof538milliondolla體inyear2009TheappHcationofmetabolicflux鋤alysisof勛印幻叼舊鯽m卵D印Dr淞togeneticengineeringhadnotbeenreportedInthisworl【，themetabolicnetworkof泐幻彬)，a絡(luò)加卵D妒D，螂wascon蛐mctedAs仃ategyw嬲deVelopedbasedonmeta

3、bolicflux鋤alysis鋤dageneticengineeringstrainwascons仃uctedwitllmolecularbiologytechnologyThenthee仃ectondaptomycinproductionwaSalsoinVestigatedAndthegeneticengineering舳inofChineseintellectualpropertyri曲ts、)l，asobtainedAnini

4、tialreactionliStwasbuiltbyblastingtlleopenreading仔鋤ecoIle曲甜inthem以bolicnet、)l，orkof勛印幻叼嬲∞P眈D腸，A3(2)againstthegenomesequenceof&，印幻刀紗卯sm船D印D，淞NRRLll379Wimaddingdaptomycinsynthesisrcaction，biomassSynthesisreaction，仃ansportr

5、eaction鋤dgapfilling，themetaboIicne鉚orkof腳幻刀zy卯J加韶。戈印，淞wasconsnIIctedAndthemainnodes鋤dmetabolicpathwayswererecogn泣edbytopological柚alysisBased0ntheme訕olicne觚。止metabolicfluX粕alysisw嬲conductedTheefrectofdilutionra鈀ontheca小on

6、fluxthroughpentosephosphatepathway，Embden—Meyerhof_P啪aspathway觚dtricarboXyIicacidcyclew嬲inVestigatedThe日uxdistributionaroundthenodespyllJVate徹d0【ketoglut撇tew嬲alsostudiedThroughthesimulationofimproVingthespecificd印tomycin

7、productionrate，itw邪foundthatthefluxthroughpentosephosphatepathwaypathway、ⅣasenhancedmostsignificantIyincen們lme詘olismTheassumptionwasmadethattheen巧meiIlpentosephosphatepathwayshouldbeoVerexpressedtogeneratemoreNADPHBysequ

8、enceblast，the6一phosphgluconatcdehydrogenaseencodinggene27，啪、Ⅳasfoundinthegenomesequenceof&，枷朋少cPsrD騮D印口，_淞TheprimerSweredesignedforPcII己ofj邶鋤dthegenewasclonedTheplasmidp1：A2—二彬wasconstTllctedby’I’AcloningandtheexpressVec