具有α葡萄糖苷酶抑制活性海绵共附生微生物及其活性产物的研究

《具有α葡萄糖苷酶抑制活性海绵共附生微生物及其活性产物的研究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、伐一glycosidaseinhibitorderivedfrommarinesponge--associatedmicroorganismsAbstractMarinesponge．associatedmicroorganismsaretheimportantsourceofnatureproductsforpharmaceuticals．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仅一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