桑树内生菌micrococcusluteus-y2发酵产dnj的研究-study on dnj production by mulberry endophytic bacteria micrococcus - y2 fermentation.docxVIP

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2018-07-30 发布于上海
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桑树内生菌micrococcusluteus-y2发酵产dnj的研究-study on dnj production by mulberry endophytic bacteria micrococcus - y2 fermentation.docx

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桑树内生菌micrococcusluteus-y2发酵产dnj的研究-study on dnj production by mulberry endophytic bacteria micrococcus - y2 fermentation

one-factor-at-a-timeexperiment.TheoptimizedfermentationconditionsforDNJproductionwere:inoculumvolume2%(v/v),solutionamount50mL/250mL,fermentationtemperature30℃,initialpH7.0,fermentationtime30h.Basedontheresultsabove,theeffectsofnutritiononproductionofDNJwerestudiedinvolvingtheoptimalcarbon,theoptimalnitrogen,inorganicsaltcombinationbyone-factor-at-a-timeexperiment.Intheoptimizationofmedium,theinfluencesofpeptone,maltose,MgSO4,KH2PO4andNaClonDNJproductionwerefirstevaluatedusingPlackettBurmandesign.Amongofthetestcomponents,theeffectofpeptonereachextremelysignificantlevelandtheKH2PO4showssignificanteffectontheDNJproduction.However,maltose,MgSO4andNaClhavenosignificanteffectsonDNJproduction.Thepathofsteepestascentwasusedtoapproachtheoptimalregionofthemediumcomposition.Inthelaststep,theoptimalconcentrationsoftestcomponentsweredeterminedbycentralcompositedesignandresponsesurfaceanalysis.ThecomponentsofoptimummediumforproductionofDNJwere:maltose20.0g/L,peptone36.53g/L,KH2PO42.22g,MgSO40.5g/L,NaCl4g/L,demineralizedwater1000mL.Basedontheoptimalfermentationconditionandmedium,theDNJconcentrationoffermentationbrothafterRP-HPLC-UVreached213.18μg/ml,184.63%higherthanthatofbasalmedium.Theinitialextraction,purificationofDNJproducedbystrainY2wasstudiedbyRep-HPLC-UVanalysis.StrainY2fermentationbrothwasconcentrated,precipitatedbyethanol,separatedbyNKA-9andduolite,concentratedagain,andfreezedtodehydration.DeterminedbyRP-HPLC-UVanalysisafterderivatization,theDNJconcentrationwasupto0.337%,enhancedby5.3times,thesampleproductwaspurifiedeffective.Determinationoftheα-glucosidaseinhibitionactivityoffermentationcrudeextractbyα-glucosidaseinhibitionmodelinvitro.Theresultsareasfollows:Thesampleproductsbothbetterthanacarbosehaveeffectα-glucosidaseinhibitionactivity.TheIC50ofstrainY2is111.58mg/L,andY1is2307.56mg/L.Itsactivityispositivelyrelativewithconcentrationincertainrange,andexhibitsdose-dependentinhibition.DeterminationofthegermfreemulberryDNJconcentration.Thegermfreemulberryweredetectedwithb