胶原蛋白酶产生菌的筛选和应用研究

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1、四川大学博士后学位论文胶原蛋白酶产生菌的筛选和应用研究姓名：谢君申请学位级别：博士后专业：轻工技术与工程指导教师：张铭让;张新申20030601解鲜猪皮的特性进行了对比试验，发现铜绿假单胞菌胶原蛋白酶对鲜猪皮的降解能力远远强于其他两种植物蛋白酶。采用正交分析的方法，研究了铜绿假单胞菌胶原蛋白酶和木瓜蛋白酶协同降解鲜猪皮制备胶原蛋白和肤的工艺条件，其最佳条件为温度45C，液比2，时间24h,pH7,混合酶0.6%。最后评价了所得胶原蛋白的理化性质、卫生指标、氨基酸组成和分子量分布等。关键词:铜绿假单胞菌，火神发光杆菌，胶原蛋白酶，发酵条件，酶解反应。STUDIESONSCRE

2、ENINGANDAPPLICATIONOFSTRAINSPRODUCINGCOLLAGENASEABSTRACT95strainsproducedenzymeswithglutinaseactivitieswerescreenedfromsoilsamplesfromChengdutannery.2strains(CYlandCY2)ofthesestrainscoulddigestcompletelycowhidein24h.UsingcollagenIIassubstrates,theactivitiesofcolagenaseproduced勿the2strainsw

3、eredetermined,andthe-resultsshowedthattheactivitiesofthecollagenasesproducedbythe2strainswereinthesamelevel,whichcouldreach10-16urokinaseunitspermilliliterbroth.CY1wasidentifiedasPseudumonasAeraginosaandCY2wasidentifiedasPhotobacteriumLogei,accordingtothephysiologicalandbiochemicalcharacte

4、rsofthem.Propertiesofcoarseenzymeproduced勿PseudumonasAeraginosawereinvestigated.TheoptimalreactiontemperatureandpHvalueofthecoarseenzymewere--321CandpH7.5.TheactivitiesofthecoarseenzymecouldberestrainedpartiallybyEDTA和EGTA.TheoptimalfermentationconditionsofPseudumonasAeraginosaCY1producing

5、collagenaseweredeterminedbysinglefactoranalysisandorthogonalanalysis.Thebestfermentationmediumwerecomposedof1.5%glutin,4%sucrose,0.20%yeastx102"/oCaC12,0.25oroK2HPOd3H2oand0.05%.21-120,andthebestpHwas6.Thecultivationis37C.Theoptimalshakingspeedwas220rlmin,when100mLflaskmLmediumandinoculums

6、6%.Thecoarse加mPseudumonaspapainandtrypsinwereappliedtohydrolyzefreshpigskinrespectively.TheresultsthattheabilityofhydrolyzingpigskinofthecoarsecollagenasefromPseudumonasAeraginosawasfurtherhigherthantheseoftheothertwokindofplantproteases.Theconditionsofhydrolyzingpigskinusing山ecoarsefromPs

7、eudumonasAeraginosaandthepapainwereinvestigated,andtheoptimalconditionsweretemperature45C,liquorratio2,hydrolysistime24h,pH7andmassof0.69/o.thephysicsandchemicalsaniitionofaminoacidsandmolecularweightoftheproteins加mhydrolysatesofevaluated.尸hotobacteriumLogei,f