产耐高温Fe-SOD(H171A)重组大肠杆菌发酵条件的优化  

Optimization of fermentation process for expression of a hyperthermostable Fe-SOD(H171A) in Recombinant E.coli BL21(DE3)

作　　者：陈伟 李伟[1] 朱虎 曲剑波 刘建国 

机构地区：[1]中国石油大学化学工程学院,青岛266580

出　　处：《食品科技》2014年第12期46-51,共6页Food Science and Technology

基　　金：国家自然科学基金项目(21276280);中国石油大学(华东)自主创新科研计划项目(13CX02062A)

摘　　要：超氧化物歧化酶因其在药物和化妆品等行业体现出重要应用价值,近年来为世人所关注。为了提高该酶的表达量,我们针对E.coli BL21(DE3)/p ET28a(+)-SOD重组表达系统,运用单因素实验及正交实验相结合的手段,确定了产耐高温Fe-SOD(H171A)重组大肠杆菌的最优培养基成分及最优诱导条件。实验结果表明,重组细菌在当向培养基中加入亮氨酸至终浓度为1.20g/L、氮源为30 g/L酵母粉并且调整碳氮比为1时,达到了最优诱导效果。另外当诱导条件为诱导剂浓度0.15 mmol/L,诱导温度37℃,诱导时间10 h时,重组菌达到了最优诱导效果。在优化条件下,菌体密度达到最高并且目的蛋白表达量为全菌体蛋白的52.90%,比基础培养基发酵条件高出0.68倍。The fermentation process of Fe-superoxide dismutase（H171A） expressed by E. coli BL21（DE3）/p ET28a（＋）-SOD, which induced with IPTG was studied. The investigation of optimization contains variety and content of amino acid and organic nitrogen source, as well as ratio of C：N by using of one factor method and orthogonal design. Besides, concentration of IPTG, induction temperature and time was also investigated. The results list as follows： leucine 1.20 g/L, yeast extract 30 g/L, C：N 1：1, IPTG 0.15 mmol/L,induce temperature 37 ℃, induction time 10 h. The expression level has reached at 52.90% under these optimum conditions, which was 0.68 times more than that in LB medium.

关 键 词：超氧化物歧化酶 重组大肠杆菌 发酵 优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]