金枪鱼肉降解用高产蛋白酶菌株的筛选及鉴定.doc

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金枪鱼肉降解用高产蛋白酶菌株的筛选及鉴定

【摘要】：^p：采用透明圈法从舟山普陀白山土壤中筛选出3株能分泌胞外蛋白酶的菌株，并对蛋白酶活力最高的菌株进行分子鉴定及最适宜生长条件分析^p。选择福林酚试剂法测定其蛋白酶活力，对蛋白酶活力最高的菌株进行16SrRNA基因测序，用MEGA软件构建系统发育树，并考察其接种量、温度、时间对该菌株产酶活力的影响。结果表明，zjoub-1菌为高产蛋白酶菌属，并确定为枯草芽孢杆菌。该菌株用于金枪鱼暗色肉发酵时的最适宜条件为接种量2%、pH7.4、温度37℃、时间48h。在此生长条件下，菌株产蛋白酶活力最高，为49.21U/mL，能够有效地降解金枪鱼暗色肉。
【关键词】：^p：金枪鱼肉；透明圈法；福林酚试剂法；筛选；鉴定；生长条件
中图分类号：S965.332文献标识码：A文章编号：0439-8114（20________）13-2506-06
DOI：10.14088/jki.issn0439-8114.20________.13.027
ScreeningandIdentificationofAStrainwithHighProteaseinFermentationofTunaDarkMeat
WULi-na1，ZHAO____iao-hui1，GUOKai-qing1，YANGYu-jie1，FANG____u-bo1，2，CHEN____iao-e1，2，ZHANGYin-zhao3
（1.CollegeofFoodandMedicine，ZhejiangOceanUniversity，Zhoushan316022，Zhejiang，China；2.StateKeyLaboratoryofAquaticProductsProcessingofZhejiangProvince，Zhoushan316022，Zhejiang，China；3.ZhejiangFengyuBiologicalProductsLimitedCompany，Zhoushan316104，Zhejiang，China）
Abstract：UsingThreestrainsofbacteriasecretinge____tracellularproteasewereisolatedfromsoilofBaishaninZhoushanbytransparentcirclemethod，thebacteriumwithhighestproteaseactivitywasidentifiedandthemostsuitablegrowthconditionswasstudied.ThecollectedsleswerescreenedbytransparentcirclemethodtogetthreepurestrainswhoseproteaseactivitywouldbemeasuredbyForintphenolreagentmethod.Thestrainwiththehighestproteaseactivitywasidentifiedbythe16SrRNAsequenceanalysisandthephylogeictreewasconstructedbyMEGAsoftware.Theinoculationamount，temperature，timeandpHhadbeenanalyzedontheinfluenceofitsproteaseactivity.Theresultsshowedthatthestudydemonstratesthatthezjoub-1strainwithhighestproteaseactivitywasdefinedasbacillussubtilis.TheoptimalconditionforfermentationofTunadarkmeatwasasfollows：theinoculationquantitywas2%，theincubationtemperaturewas37℃，theincubationtimewasabout48h，theinitialpHwas7.4.Underthiscircumstance，thestrainhasthehighestproteaseactivity，whichillustratesthatthestrainwithhighproteaseactivitycandegradetheTunadarkmeatefficiently.Keywords：Tunadarkme(