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《四個(gè)腸桿菌植酸酶基因的克隆、表達(dá)及性質(zhì)研究》由會員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、中國農(nóng)業(yè)科學(xué)院博士學(xué)位論文四個(gè)腸桿菌植酸酶基因的克隆、表達(dá)及性質(zhì)研究姓名：谷維娜申請學(xué)位級別：博士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：姚斌20070601AbstractPhytases(myo-inositolhexakisphosphatephosphohydrotases)hydrolysephytatetolowermyo-inositolphospharesandinorganicphosphate．Asatypeofanimalfeedadditives，additionofphytaseinanimalf
2、eedallowsmonogastricanimaltoutilizethephytinphOSphomsinfeedandreducestheenvironmentalphosphomspoHufiomPhyta∞has麗dccoralapplicationsinindustryproccs$essuchas．foodstuffandpharmaceuticalindustry．Microorganismisanimportantsourceofphytase．Screeningofnewandgoodnaturep
3、hytasesfromthemicroorganismisanimportantresearchdirection．Apairofdegeneratepfinl∞(FIandRI)wasdesignedthroughidentityanalysisofpublishedanthlOacidsequencesofphytase．UsingpriI礎(chǔ)fs，fragmentsofthreephytasegeneswmobtainedbyPCRmethodfromgenomeofstrainsHafniaalveiB125，D
4、ickeyadadamiiB187andDickeyaparadisiacaB188．耵”full—lengthofthreegeneswereobtainedbymethodofTAILrPCRandnameriasB125appA,B187appAandB188appA．AnovelphytasegenerevealedbygenomeanalysisofYersiniapestisWaSobtainedbyPCR．namod硒YlappA．ThehighestidentityofnucleicacidWas75％
5、amongthesefourgenes∞187appAandB188appA)，thelowestidentityofnucleicacidWas48％(B188appAandB125appA，B188appAandYlappA)．IncomparisonwiththesequencesofpreviouslyisolatedphytasesbyBLASTsearch，thededucedaminoacidsequenceofBl25appAshowed95％identitytophytasefromObesumbac
6、teriumproteus．thededucedaminoacidsequenceofBl87appAandBl88appAshowed53％and52％identitiestoaphytasefromKlebsiellapneumoniae，reSpectively．TheresultprovedthatBl87appAandBl88appAaretwonewphytasegenes．11lededucedaminoacidsequenceofYlappAshowed81％identitytophytasefromY
7、ersiniainrermedia．ThegeneB125appA，8187appAandB188appAwereexpressedinEscherichiacoliBL21(DE3)．ThegeneylappAwasexpressedinPichiapastoris．Recombinantproteinsw啪purifiedandtheirenzymaticpropertieswefedetermined．TheoptimumpHfortheB125r-APPAWas4．5andtheoptimumtemperatu
8、reWas60℃．TherelativephytaseactivityWasabove80％aftertreatedinhlffersofpH2．0-pH10．0．硼1especificactivity0fBl25r-APPAWas356．7l-O／rag．11峙‰andK峨valuesforBl25r-APPAw粥0．49lnl
