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带鱼蛋白酶解工艺条件的响应面优化.docx

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带鱼蛋白酶解工艺条件的响应面优化 Title:OptimizationofProteaseHydrolysisProcessforBelt-FishusingResponseSurfaceMethodology Abstract: Theobjectiveofthisstudywastooptimizethehydrolysisprocessofbelt-fishproteinusingproteaseenzymethroughresponsesurfacemethodology.Theeffectofthreeindependentvariables,namelyenzymeconcentration,hydrolysistime,andtemperaturewereinvestigatedontheresponsevariable,degreeofhydrolysis(DH),tomaximizetheproteinhydrolysisefficiency.Acentralcompositedesign(CCD)wasemployedtodesigntheexperimentsandgenerateamathematicalmodeltopredicttheoptimalconditionsformaximumDH.TheresultsshowedthattheenzymeconcentrationandhydrolysistimehadsignificanteffectsontheDH,whilethetemperaturehadaminoreffect.Theoptimizedconditionsobtainedwereanenzymeconcentrationof2.5%,hydrolysistimeof4.5hours,andtemperatureof55°C,whichyieldedamaximumDHof24.56%. 1.Introduction: Proteinhydrolysisisgainingattentionduetoitsabilitytoenhancetheflavor,texture,andnutritionalqualityoffoodproducts.Proteaseenzymeshavebeenwidelyusedinthehydrolysisofvariousproteinsourcestoimprovetheirfunctionalproperties.Belt-fish(Trichiurusspp.)isavaluableseafoodresourcewithhighproteincontent,andhydrolysisofbelt-fishproteincanleadtotheproductionofpeptideswithbioactiveproperties.Inthisstudy,weaimedtooptimizetheproteasehydrolysisprocessforbelt-fishproteinusingresponsesurfacemethodology. 2.Materialsandmethods: 2.1.Materials: -Belt-fishsampleswereobtainedfromalocalseafoodmarketandstoredat-20°Cuntilfurtheruse. -Alcalase2.4L(Novozymes,Denmark)wasusedastheproteaseenzyme. 2.2.Experimentaldesign: Acentralcompositedesign(CCD)with3factorsand20runs,including6axialpoints,wasemployedtoinvestigatetheeffectsofenzymeconcentration(X1:1.5-3.5%),hydrolysistime(X2:2-5hours),andtemperature(X3:40-60°C)onthedegreeofhydrolysis(DH).TheexperimentaldesignmatrixandcorrespondingexperimentalresponsesareshowninTable1. 2.3.Proteinhydrolysis: Briefly,belt-fishsampleswerethawed,washed,andgroundtoafinepowder.Theenzymeconcentration,hydrolysistime,andtemperatureweresetacco