頂頭孢霉優(yōu)良菌株的選育和vgb在頂頭孢霉中的轉(zhuǎn)化與表達.pdf

1、沈陽藥科大學(xué)碩士學(xué)位論文頂頭孢霉優(yōu)良菌株的選育和vgb在頂頭孢霉中的轉(zhuǎn)化與表達姓名：曹棟申請學(xué)位級別：碩士專業(yè)：微生物與生化藥學(xué)指導(dǎo)教師：徐威20090601沈陽藥科大學(xué)碩L學(xué)位論文英文摘要ABSTRACTCephalosporinCisonekindofplactamantibiotics，whichisproducedbytheCephalosporiumacremonium7ACAwhatismadefromCephalospor

2、inCafterchemicalmethodortwo—stepenzymaticCleavageisallimportantrawmaterialofmanysemisyntheticcephalosporinantibioticsWiththedegreeofmarketcompetitionincreases，lowcostandhigh—yieldcephalosporinCwouldbemorecompetitiveinthe

3、marketFirstly，mutantUVcombinedwithresistanceofendproductwereusedforthescreeningofadvantagedCephalosporiumacremoniummutants；Secondly，inordertoincreasetheproductionofCPC，Vitrescillahemoglobin(vgb)genewasintegratedintothege

4、nomeofCephalosporiumacremonium，aimedatincreasingoxygenuptakerateandimprovingcellgrowingconditionCephalosporiumacremoniumNS一1screenedbynaturalseparationWaspureculturewithastableabilityofproducingantibioticsThetiterofCepha

5、losporiumacremoniumNS1Was44436l甥II心1ThemutantUVWasusedtoselecthighCPCproducingstrainThetiterofCephalosporiumacremoniumUV21wasupt0564352J_tgmL～ResistanceofendproductwasusedtostrainCephalosporiumacremoniumUV21forscreeningh

6、ighCPC—producingstrainThetiterofCephalosporiumacremoniumRM8whichWasselectedbythisexperimentis644322I_tgmLl，increasedby45％comparedwithCephalosporiumacremoniumNS一1AplasmidpSY090513一vgbcontainingbothaphleomycinresistancegen

7、eandaVitreoscillahemoglobingeneWastransformedintoCephalosporiumacremoniumTransformantswerescreenedwiththephleomycinresistanceExpressionofVHbintransformantsandthebioactivityoftheexpressedVHbwereverifiedbytheanalysisofaCOd

8、ifferencespectraexperimentsCephalosporiumacremonium(withorwithoutvgbgene)Wasculturedfor72hbysecondaryfermentationmethod，thentransferredtOshakingcultureoffermentationininoculum’SconcentrationoflO％(v／v)ItWasfoundthatthetit