均匀设计法优化重组大肠杆菌产酮基还原酶培养基

Optimization of Fermentation Medium for Keto Reductase Production from Recombinant Escherichia coli Using Uniform Design

系统研究了典型碳源、氮源、金属离子、磷酸盐以及初始p H值等因素对重组大肠杆菌产酮基还原酶的影响。单因素试验结果表明,甘油、酵母浸粉FM888、酵母蛋白胨FP101、Mg SO4·7H2O以及离子浓度为0.1 mol·L-1 p H值为T7的磷酸缓冲液对菌体生长以及酶活的表达有一定的促进作用。通过均匀设计试验及分析得到培养基的最优配方:甘油6.86 g·L-1,FM888 19.53g·L-1,FP101 8.37 g·L-1,Mg SO4·7H2O 2.50 g·L-1,K2HPO414.96 g·L-1,KH2PO47.34 g·L-1。优化条件下酶活为431.21 U·m L-1,比优化前(70.25 U·m L-1)提高了5.13倍。此研究可为工业化大生产提供理论指导。

The effects of a number of factors on the production of keto reductase were systematically studied, typically including carbon source, nitrogen source, phosphate and initial p H. The research results of single factor experiment showed that the growth and enzyme activity were promoted by glycerin, yeast extract FM888, yeast peptone FP101, Mg SO4·7H2O, and phosphate buffer with p H 7and 0.1 mol·L^-1 ion concentration. Through uniform design experiments, the optimal conditions were obtained as follows： Glycerol6.86 g·L^-1, FM888 19.53 g·L^-1, FP101 8.37 g·L^-1, Mg SO47H2O 2.50 g·L^-1, K2HPO414.96 g·L^-1, KH2PO47.34 g·L^-1. Under the optimal conditions, the activity of keto reductase reached 431.21 U·m L^-1, which was 5.13 times greater than that of the basic fermentation conditions previously（70.25 U·m L^-1）. This research could provide theoretical guidance for large-scale production of keto reductase.