乳酸片球菌细菌素的发酵与纯化分析-fermentation and purification of bacteriocin from pediococcus lactis.docx

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2018-07-29 发布于上海
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乳酸片球菌细菌素的发酵与纯化分析-fermentation and purification of bacteriocin from pediococcus lactis.docx

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乳酸片球菌细菌素的发酵与纯化分析-fermentation and purification of bacteriocin from pediococcus lactis

ABSTRACTBacteriocins,whichsharemanyvarietiesanddifferentproperties,arerecognizedaspotentialbiopreservatives.However,onlyNisiniscurrentlylicencedasafoodadditiveinover60countries.Therefore,todevelopnewbacteriocinfromlacticacidbacteriawitheffortsandtostudybacteriocinwhichhavebeenfoundindepthisthecurrentimportanttaskforthemajorityofscientists.Intheearlywork,ourlaboratoryhasisolatedastrainfromtheChinesetraditionalfermentedcabbage,whichcouldproducebacteriocin.Afterwards,ithasbeenidentifiedasPediococcusacidilactici.ToourKnowledge,veryfewpapershavebeenpublishedonPediocininChina,thisworkaimedtostudyfermentationandpurificationprocessofPediocinfromP.acidilacticiPA003.Specifically,itisdividedintothefollowingfourparts1、Aefficientlaboratory-scalepurificationmethodwasdevelopedtopurifyapediocinproducedbyP.acidilacticiPA003,andthenithasbeenidentificated.PediocinproducedbyP.acidilacticiPA003waspurifiedbyammoniumsulfateprecipitation,gelfiltrationchromatography,ion-exchangechromatography,andreverse-phasehigh-performanceliquidchromatography.Thisprocedureallowedtherecoveryof3.8%ofthebacteriocinpresentintheculturesupernatantwithpurityhigherthan99%andanapproximately461foldincreaseinspecificactivity.Themolecularmassofthepurifiedpediocinwasabout4624Da,whichwasidenticaltopediocinPA-1?AcH,asdeterminedbyMatrix-assistedlaserdesorptionionization-timeofflightmassspectroscopy.Bytheway,wealsofoundP.acidilacticiPA003canproducemorethanonekindofbacteriocins.2、TheeffectivemembranesystemtoseparatebacteriocinwasdevelopedThisstudydevelopedaneffectivecombinedultrafiltration(UF)–nanofiltration(NF)membranesystemforultrafilteringbacteriocinfermentationbrothandconcentratingthefiltratetoimprovethedownstreamefficiency.WithoptimalPS-10membrane,UFprocesseswasproposeduntilvolumeconcentrationfactor(VCF)reached2.5,thendiafiltrationprocessesofultrafiltrationwasproposed.ThepermeateobtainedfromUFwasthenconcentratedbyNF.WhentheVCFofNFreached4.5,1000mLofUFpenetrantcanbeconcentratedto220mL.Thistwo-stagemembraneprocessemploy