Fibrolase表达菌自诱导摇瓶发酵研究

Research on auto induction Culture of Recombinant E.coli Expressed Fibrolase

[目的]解决Fibrolase表达时的渗漏问题,实现高密度发酵。[方法]先在培养基中培养Fibrolase大肠杆菌胞内可溶表达工程菌,再按1%的接种量接种到ZYM-5052培养基中培养,定时取样,研究可溶表达工程菌表达量、菌体量与培养时间的关系,进行纯化和活性分析。[结果]自诱导表达能明显提高菌体收获量和目的蛋白表达量,在培养14 h时表达量和菌体量达到最大值,纯化的重组Fibrolase具有明显的纤溶活性。[结论]该方法能有效抑制表达渗漏问题,实现高密度发酵。

[ Objective] The aims of the research were to increase the cell density and to resolved the problem of leaked expression of Fibrolase. [ Method] Incubate E. coli Origami B（DE3） expressing of Fibrolase in ZYM-505 media with stroking at 37 ℃, addition of 1% culture to 100 ml medium of ZYM- 5052 and shake the culture at 37 ~C. Then , the relation between of the incubate time and recombinant protein proportion or biomass were studied. Finally, recombinant Fibrolase was purified and their fthrinolytic activity was evaluated. [ Result ] The cell density and expression levels of Fibrolase were markedly increased by expression through auto induction, they were achieved their maximal level after auto induction at 14 h, and purified Fibrolase has the obviously activity of fthrinolytic. [ Conclusion ] The method of auto induction can resolved the problem of leaked expression of Fibrolase and increase the cell density.