人溶菌酶在毕赤酵母中的表达及体外抑菌活性研究  

Expression of Human Lysozyme in Pichia Pastoris and Its Antibarterial Activity in vitro

作　　者：李学优 曹丁 肖宏艳 黄秀敏 甘祥武 LI Xue-you;CAO Ding;XIAO Hong-yan(Guangzhou lnstitute of Microbiology,Guangzhou,Guangdong 510663)

机构地区：[1]广州市微生物研究所

出　　处：《安徽农业科学》2020年第4期100-102,105,共4页Journal of Anhui Agricultural Sciences

基　　金：广州市科技计划项目(201806010022)

摘　　要：对重组人溶菌酶真核表达载体的构建筛选、表达与活性鉴定,经过双酶切和测序鉴定表明,重组载体正确。电转化毕赤酵母GWDL菌株,通过G418抗性平板筛选得到高表达高效价菌株,经诱导表达,检测蛋白含量,体外抑菌检测,表达产物15 ku和预期结果一致,筛选得到效价为1907 U/mL,编号为SMD1168-pPIC9k-HLZ的高效价菌株;采用重组人溶菌酶制剂,检测样品GWDL对5种菌的最小抑菌浓度(MIC),藤黄微球菌的最小抑菌浓度较好,表明重组人溶菌酶在毕赤酵母中成功表达,活性较为稳定。The screening,expression and activity identification of constructed recombinant human lysozyme eukartic expression vectors were studied.The recombinant vectors were shown to be correct through double enzyme digestion and sequencing identification.To electro-transform the pichia pastoris GWDL strain,through G418 resistance flat screening to obtain high expression of high-efficiency strains,the expression product(15 ku)was consistent with the expected results after induced expression,detection of protein content and in vitro antiseptic detection,we obtained No.SMD1168-pPIC9k-HLZ high titer strain with a titer of 1907 U/mL;To detect the sample GWDL minimum bacterial concentration(MIC)for five bacteria by using Recombinant human lysozyme preparation,the minimum concentration of micrococcus luteus was better,indicating that recombinant human lysozyme in pichia pastoris successfully expressed,the activity was more stable.

关 键 词：毕赤酵母 菌株筛选 高效表达 抑菌活性 

分 类 号：S188[农业科学—农业基础科学]