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.
作者
叶芬
周建森
郭静科
刘树滔
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)
出处
《福州大学学报(自然科学版)》
CAS
北大核心
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)。