新型海洋源β-半乳糖苷酶在毕赤酵母中的表达与酶学性质  

Expression and characterization of a novelmarine β-galactosidase in Pichia pastoris

作　　者：阎金勇[1] 董敬霞 査根晗 高云[2] 刘小宇[2] 缪明永[2] YAN Jinyong DONG Jingxia ZHA Genhan GAO Yun LIU Xiaoyu MIAO Mingyong(Key Lab of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, China)

机构地区：[1]华中科技大学生命科学与技术学院分子生物物理教育部重点实验室,湖北武汉430074 [2]第二军医大学生物化学与分子生物学教研室,上海200433

出　　处：《生物加工过程》2016年第4期33-36,共4页Chinese Journal of Bioprocess Engineering

基　　金：国家高技术研究发展计划(863计划)(2014AA093513)

摘　　要：从海洋环境β-半乳糖苷酶基因出发,采用毕赤酵母表达体系,构建产β-半乳糖苷酶的基因工程菌,并对重组酶酶学性质进行表征。结果发现:海洋源β-半乳糖苷酶具有优良的乳制品用酶特性,该重组酶的最适pH为7.0~8.0,最适温度为45℃,在50℃以下孵育1 h可保持55%以上的活力,能耐受Fe^(2+)、Na^(2+)、K^(2+)、Ca^(2+)等多种金属离子。宽广的pH、温度、金属离子稳定性,以及低温活性赋予该酶良好的乳制品用酶特性。Using Pichia pastoris expression system, we constructed a recombinant P. pastoris strain producing marine β？galactosidase that was subsequently characterized. The recombinant enzyme had optimum pH between 7. 0 and 8. 0. Its optimum temperature was 45 ℃, and over 50% relative activity below 50 ℃.It was stabile in the presence of Fe^2＋,Na^2＋,K^2＋ and Ca^2＋. Broad stable pH and temperature range and tolerance to metal ions make this recombinant enzyme suitable in various applications.

关 键 词：半乳糖苷酶 毕赤酵母 重组 

分 类 号：Q851[生物学]