SOD-肝素结合结构域融合蛋白的毕赤酵母重组菌株构建及表达  

Construction and expression of recombinant Pichia pastoris for the fusion protein of SOD and heparin binding domain

作　　者：叶芬 周建森 郭静科 刘树滔[1] YE Fen;ZHOU Jiansen;GUO Jingke;LIU Shutao(Institute of Biotechnology,Fuzhou University,Fuzhou,Fujian 350002,China;Department of Food and Bioengineering,Zhicheng College,Fuzhou University,Fuzhou,Fujian 350002,China)

机构地区：[1]福州大学生物工程研究所,福建福州350002 [2]福州大学至诚学院食品与生物工程系,福建福州350002

出　　处：《福州大学学报（自然科学版）》2022年第1期132-138,共7页Journal of Fuzhou University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(31500685);浙江省自然科学基金资助项目(LY16C050002)。

摘　　要：利用基因工程技术将人源Cu,Zn-SOD与EC-SOD的C末端肝素结合结构域(HBD)的编码序列进行密码子优化并人工合成,然后电转化至毕赤酵母进行表达.单因素实验表明,与现有研究的一些重组EC-SOD相比,其比活力提高了1.30~3.78倍.正交分析得最佳摇瓶条件为诱导时间为5 d,初始pH值为5.2,诱导剂量的体积分数为1.0%,酶活可达1120.2 U·mL^(-1),是初始酶活的2.94倍;这些结果表明,SOD-HBD融合蛋白在一定程度上解决了重组EC-SOD表达量低的问题,为重组EC-SOD蛋白的推广与应用提供重要的物质基础条件.Using genetic engineering technology,the coding sequence of human Cu,Zn-SOD and the heparin binding domain(HBD)of EC-SOD were optimized and synthesized,and then electrically transformed into Pichia pastoris for expression.The single factor experiment showed that compared with the recombinant EC-SOD in previous studies,its specific activity in this study increased by 1.30~3.78 folds.By orthogonal analysis,the optimal shaking conditions were as follows:induction time 5 d,initial pH=5.2,induction dose 1.0%(volume fraction),enzyme activity up to 1120.2 U·mL^(-1),which was 2.94 folds of the initial enzyme activity.These results of constructed SOD-HBD fusion protein solved the problem of low expression of recombinant EC-SOD to a certain extent,and provided important material basis for the promotion and application of recombinant EC-SOD.

关 键 词：超氧化物歧化酶 肝素结合结构域 巴斯德毕赤酵母 构建 表达 

分 类 号：Q812[生物学—生物工程]