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《具有α葡萄糖苷酶抑制活性海綿共附生微生物及其活性產(chǎn)物的研究》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、伐一glycosidaseinhibitorderivedfrommarinesponge--associatedmicroorganismsAbstractMarinesponge．a(chǎn)ssociatedmicroorganismsaretheimportantsourceofnatureproductsforpharmaceuticals．Itisapotentialwayfordevelopingnewdiabetesdrug(0【。glycosidaseinhibitor)·Inthisstudy,thea。glucosidaseinhibitoryactivityscreeningm
2、odelisbuilt—uptosearchfortheactivecrudeextractfromthemicroorgainismmetabolitesamplespreparedfrommarinespongesinFridayharborofUS．Thereactionsystemofthe0【-glucosidaseinhibitoryactivityscreeningisperformedona96wellsplates．Thereactioncondition，includingtheconcentrationofenzymes，issetat0．008U／mL，pH6．8，3
3、7。C，andwavelength405nm．Moreover,thescreeningmodelwasoptimizedwithreactantPNPGconcentration10mmol／L，reactiontimeof25minandthesolventDMSOcontentof2％(∥功UsedAcarboseasapositivecontrolledsample，212crudeextractssamplesfrommarinespongeassiciatedmicroorganismswerescreenedwiththeaboveoptimizedmodel．Theresul
4、tsindicatedthatmetabolitesof19strainshadhighinhibitioryactivity(>50％)．Amongthem，5strainsshowedinhibitoryeffecthigherthan70％against僅一glucosidaseatconcentration0．4，0．6，0．8me／mL．TheinhibitoryeffectswashigherthanthatofAcarbose(58％)atconcentration1．6me／mE．Asresult，thestrainHY936，withhighinhibitoryactivi
5、ty(79．3％)atconcentration0．4me／mL，wasidentifiedasBacillus．firmusbythemeansof16SrDNAsequenceThecultivatingconditionofstrainswasoptimizedforahigherproductionofactivecomponents．Singlefactorexperimentaldesignwasperformedtoanalysistheinfluenceofcultivatingtime，rotatingspeed，inoculationvolume．Theorthogona
6、ltestL9(34)tookpeptone，yeastextract，andNaClconcentrationsintheculturemediaaswellasthepHvalueintoaccount．Aftertheoptimizationofcultureconditions，thefermentationprocessusingstrainsofBacillus。firmuswasconductedtoaccumulate2．8gcrudeextract，whichwasseperatedandpurifiedthroughsilicacollumnofTLCorHPLC．Fr．
7、1andFr．4wereobtainedbyusingsilicagelcolumnandtheyhavehighinhibitoryactivityagainst0【一glucosidase．theinhibitaryratesof35．5％and19．6％．respectively．Thensubfraction11wasisolatedfromFt．1showedinhibitaryrateof82．3
