响应面法优化重组工程菌的发酵工艺条件

Optimization of Fermentation Conditions for Genetically Engineered Bacterial Using Response Surface Methodology

目的:对基因改造运动发酵单胞菌的发酵工艺条件进行优化,提高重组菌发酵乙醇产量。方法:使用分子克隆实验操作技术构建重组运动发酵单胞菌,以单因素实验为基础,利用Box-Behnken中心组合实验和响应面分析法,确定了影响重组菌高产乙醇的三个重要因素。结果:成功构建含有YfdZ、MetB基因和Hsp基因的重组菌Zymomonas mobilis HYM,发酵主要影响因素的最佳条件分别为温度28℃,葡萄糖浓度24%(W/V),pH7.4。在此优化条件下,Zymomonas mobilis HYM的乙醇产量可高达105.0735g/l,比原始菌株乙醇产量提高16.4%。结论:用中心组合设计和响应面分析法优化重组运动发酵单胞菌的发酵工艺条件,显著提高乙醇产量。

Objective：Optimizing the fermentation conditions for the ethanol production of the genetically engineered bacterial to increase the ethanol production.Method：Zymomonas mobilis HYM was constructed by genetic engineering.Based on the single-factor experiments,the three factors which had significant impact on the ethanol production were obtained through Box-Behnken center united experiment design and Response Surface Methodology.Result：The Z.mobilis HYM constructed in our laboratory is genetically engineered by introducing three genes （ YfdZ,MetB and Hsp） from other sources.The optimum conditions for maximum ethanol concentrations were obtained at a temperature of 28.1℃,initial pH of 7.4 and initial sugar concentration of 240 g/l.A maximum ethanol concentration of 105.07 g/l for Zymomonas mobilis HYM was obtained at these optimum parameters,and the production was increased by 16.4% compared with its original Z.mobilis counterpart.Conclusion：The RSM with BBD can be successfully used to determine the accurate values of the fermentation conditions for the maximum ethanol production using the Zymomonas mobilis HYM.