摘要
目的:优化嘧啶核苷磷酸化酶(PyNPase)基因工程菌的发酵条件。方法:通过工程菌摇瓶培养、测定吸光度D值、凝胶灰度扫描分析表达量。结果:当起始pH值为7.0～7.2,以60/500mL的装液量、5%的接种量,于32℃培养3h,加入终浓度为0.2g/L的L-阿拉伯糖诱导6h后收获菌体,可得到较高的生物量和重组蛋白表达量。L-阿拉伯糖诱导开始时间和菌体收获时间对蛋白表达有显著影响。结论:为PyNPase结构研究和工业化生产奠定了基础。
Objective： To obtained the optimized fermentation conditions of reconbinant pyrimidine nucleoside phosphorylase （PyNPase）, Methods： Through the engineering bacterium culture in shake-flask, D values were determined and the amount of protein expression was analysized with gray gel scan. Results： It was found that higher expression of reconbinant PyNPase could be achieved under the following conditions： 5% inoculum amount was added to 60 mL/500 mL media at initial pH7.0-7.2. After 3 h of culture, the final concentrationin of 0.2 g/L L-arabinose was added to culture recombinant PyNPase expression. The recombinant E.coli was harvested after 6 h of induction. The results showed that the times of induction and harvest were important factors for the expression of recombinant PyNPase. Conclusion： These studies will build a solid foundation for the structural research and industrial production of PyNPase.
出处
《生物技术通讯》
CAS
2007年第2期252-254,共3页
Letters in Biotechnology
关键词
嘧啶核苷磷酸化酶
基因工程菌
发酵
优化
pyrimidine nucleoside phosphorylase
recombinant bacteria
fermentation
optimization
