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微生物来源的几丁质酶抑制剂的分离纯化及其活性研究.docx

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微生物来源的几丁质酶抑制剂的分离纯化及其活性研究 Abstract Chitinaseinhibitorsareimportantcompoundsthatcanreducefungalgrowthandchitindegradation.Inthisstudy,achitinaseinhibitorwasisolatedandpurifiedfromamicrobialsource,anditsactivitywasanalyzed.Theresultsshowedthatthepurifiedchitinaseinhibitorhadexcellentinhibitoryactivityagainstchitinaseandcouldeffectivelyinhibitfungalgrowth.Thesefindingssuggestthatthechitinaseinhibitorcanbeusedasaneffectiveanti-fungalagent. Introduction Chitinisapolysaccharidethatisabundantinnatureandisanimportantstructuralcomponentofmanyorganisms,includingfungi,insects,andcrustaceans.Chitinisobservedtobeinvolvedinvariousbiologicalprocesses,suchascelldivision,pathogendefense,andmorphogenesis.Chitinaseisaclassofenzymesresponsibleforthedegradationofchitin.Fungiaretheprimaryproducersofchitinase,andtheyuseittobreakdownchitinintheirsurroundingstosupporttheirsurvival.However,excessivechitindegradationbychitinasecancausedamagetotheenvironment. Tocountertheharmfuleffectsofchitindegradation,chitinaseinhibitorshavebeenwidelyinvestigated.Chitinaseinhibitorscanbindtotheactivesitesofchitinaseandpreventitshydrolyticactivity,effectivelyreducingthedegradationofchitinbyfungi.Therefore,chitinaseinhibitorscouldbepotentialantifungalagentstocontrolfungalgrowthandchitindegradation. MaterialsandMethods Themicrobialsourceofthechitinaseinhibitorwasidentifiedby16srRNAgeneanalysis.Thecultureofthestrainwasgrownunderoptimalconditions,andthesupernatantwascollectedbycentrifugation.Thechitinaseinhibitorwasisolatedandpurifiedusingaseriesofchromatographytechniques,includingionexchange,gelfiltrationandHigh-PerformanceLiquidChromatography(HPLC).ThepurityofthechitinaseinhibitorwasconfirmedbySDS-PAGEelectrophoresis. Theinhibitoryactivityofthechitinaseinhibitoragainstfungalgrowthandchitinaseweredetermined.ThefungalstrainwasculturedinPotatoDextroseAgar(PDA)medium,andthechitinasewasproducedbygrowingthefungusinaliquidminimalmediumcontainingcolloidalchitin.Theinhibitionoffungalgrowthbythechitinaseinhibitorwasevalu