硕士论文-酶解乳铁蛋白制取乳铁素的工艺方法研究.docVIP

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硕士论文-酶解乳铁蛋白制取乳铁素的工艺方法研究.doc

浙江大学生物系统工程与食品科学学院 硕士学位论文 酶解乳铁蛋白制取乳铁素的工艺方法研究 姓名:金月亭 申请学位级别:硕士 专业:食品科学 指导教师:应铁进 浙江大学硕上学位论文Abstract Abstract ThispaperwasmainlyaboutanovelmethodofextractinglactoferricinB (LFcinB)fromlactoferrinB(LFB)enzymolysisdigestandfurtherpurification.A refinedtechnologyofenzymolysisWasstudied,andarelatedmathematicalmodelof theenzymolysisprocesswasestablished.Themainresultswerelistedasfollows: 1.Themainparametersofionexchangechromatography(IEC)forfurther purificationoflactoferricinB(LFcinB)were-(/)chromatographiccolumn:HiTrapQ HP(1mL),startbuffer(portA):20mmolGlycin-NaOH,pill0.8;elutionbuffer (portB):20mmolGlycin-NaOH,1.0molNaCI,pill0.8.@gradientofelution:theratio ofstartbufferfrom25%atthebeginningto75%in20min.@columntemperature: 25℃@flowrate:lmL/min. 2.Theefficienciesoffivedifferentproteolyticenzymeswerecompared,i.e.pepsin, alkalineproteinase,serrapeptase,flavourzyme,andpapain,inextracting lactoferricinB(LFcinB)fromlactoferrinB(LFB)enzymolysisdigest.Theresuhs showedthatpepsinwasfarmoreefficientintheperformancethananyotherrivals, withhigherefficiencyofultrafiltrationandloweroutputofhybridproteinas by—product. 3.Throughsinglefactorexperiment,abettercombinationofconditionsforpepsin hydrolysissystemWaSfound,withtheparametersasfollows:37"C,substrate concentration5edl00mL,enzymedosage【E]/[S】=1500u/g,pH2,120min. 4.Throughorthogonalexperiment,onerefinedcombinationofhydrolysisconditions Wasfound,theparameterswere:29/100mL,1250u/g,75min,andpH2.3.Withthe yieldoflactoferricinB,1.72%. 5.Throughorthogonalexperiment,onerefinedcombinationofhydrolysisconditions wasfound,theparameterswere:49/100mL,750u/g,75min,pH2.5,withpurityof lactoferricinB,43.50%. 6.Atrialemployingcombinationdesignofquadraticregressionrotationwas 浙江丈学硕l:学位论文Abstract conducted.Theresultleadtotheestablishmentofamodelequation: Y=I.63-0.04X]+0.09X3+0.14X4—0.044Xj2.0.04XiX3—0.08Xlx4—0.I3X22 —0.29X2X3+0.1OX3z—O.15X3X4—0.05X42. Fromthemathematicalmodel,atheoreticallyoptimizedcombinationofhydrolysis conditionswaspredicted:s

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