《A marked enhancement in production of amylase by Bacillus amyloliquefaciens in flask fermentation using statistical methods》.pdfVIP

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《A marked enhancement in production of amylase by Bacillus amyloliquefaciens in flask fermentation using statistical methods》.pdf

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10.1007/s11771-011-0803-6 Amarkedenhancementinproductionofamylaseby Bacillusamyloliquefaciensinflaskfermentationusingstatisticalmethods ZHAOWei1,2 ZHENGJia2 WANGYu-guang2 ZHOUHong-bo1,2 1.KeyLaboratoryofBiometallurgyofMinistryofEducation,CentralSouthUniversity, Changsha410083,China; 2.SchoolofMineralsProcessingandBioengineering,CentralSouthUniversity,Changsha410083,China Abstract:Atotalof126bacterialstrainswereisolatedfromsoilsamples.Amongthem,11isolateswerefoundpositiveforamylase production.StrainYLproducedthelargestzoneofclearanceonplateassay.TheisolateYLwasidentifiedasBacillussp.basedon morphologicalandphysiocheimcalcharacterization.Accordingto16SrRNAgenesequencingdata,theclosestphylogenetic neighborofstrainYLwasBacillusamyloliquefaciens(99.54％).Afterthat,anoptimizationofcultureconditionswascarriedoutfor theimprovementofa-amylaseproduction.Responsesurfacemethodology(RSM)wasappliedtoevaluatetheeffectofmedium componentsincludingwheatbran,cottonseedextract,yeastextract,starch,NaClandCaCl2.Threevariables(wheatbran,cottonseed extract,andstarch),whichwereidentifiedtosignificantlyaffectamylaseproductionbyPlackett-Burmandesignwerefurther optimizedusingresponsesurfacemethodologyofBox-Behnkendesign(BBD).Theoptimalconcentrationsestimatedforeach variablerelatedtothemaximumofamylaseactivity(86kU/mL)were10.80g/Lwheatbran,9.90g/Lcottonseedextract,0.5g/L starch,2.0g/Lyeastextract,5.00g/LNaCland2.00g/LCaCl2.Thefermentationusingoptimizedculturemediumalloweda significantincrcaseinamylaseproduction(by3-fold).Theimprovementinthea-amylaseproductionafteroptimizationprocesscan beconsideredadequateforlarge-scaleapplications. responsesurfacemethodology;amylase;mediumoptimization;Bacillusamyloliquefaciens ProjectsupportedbytheNationalNaturalScienceFoundationofChina;Project(2010CB630902)supp