摘要
对重组肠激酶在大肠杆菌中的高密度发酵表达条件、纯化方法及其生物活性测定进行了研究。结果表明:在E.coli表达系统中,控制温度为37℃,培养基pH 7.4,当A600为8.0时,加入0.3mmol/LIPTG进行诱导,温度降低至30℃,连续诱导3 h,蛋白表达量达25%。经SDS-PAGE测定,纯化的重组肠激酶纯度达80%以上,并能高效切开融合蛋白。
The optional high cell-density fermentation conditions, purification methods and biological activity of the recombinant enterokinase in E. coli were studied. In E.coli expressing system, the optimized conditions were temperature 37℃ and pH 7.4. When OD600 is 8.0, with 0.3 mmol/L IPTG induced at temperature 30℃, and continued 3h, the recom- binant enterokinase was up to 25% of the total amount of the proteins in E.coli. The purified enterokinase reached a purity of above 80% by SDS-PAGE analysis and showed high activity at fusion protein.
出处
《科技通报》
2006年第6期767-770,共4页
Bulletin of Science and Technology
基金
苏州大学第七批大学生课外学术科研基金
关键词
重组肠激酶
大肠杆菌
发酵
纯化
生物活性
recombinant enterokinase
E.coli
fermentation
purification
biological activity
