摘要
In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables(induction time,inoculum age and pH),which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows:induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.
In order to improve the production of human-like collagen III（HLC III）by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables（induction time,inoculum age and pH）,which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows：induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.
基金
Supported by the National Natural Science Foundation of China（20776119）
the National High Technology Research and Development Program of China（2007AA03Z456A）
the Special Research Program of the Education Department of Shaanxi Province（07JK417）
