响应面法优化一种新型类人胶原蛋白发酵培养基  

Optimization of medium for a new type human-like collagen fermentation by response surface methodology

作　　者：张驰[1,2] 范代娣[1] 米钰[1] 马晓轩[1] 骆艳娥[1] 薛文娇[1] 

机构地区：[1]西北大学陕西省可降解生物医用材料重点实验室/陕西省生物材料与发酵工程技术研究中心 [2]西北大学生命科学学院,陕西西安710069

出　　处：《西北大学学报（自然科学版）》2009年第2期230-236,共7页Journal of Northwest University（Natural Science Edition）

基　　金：国家自然科学基金资助项目(20776119);国家高技术研究发展计划"863"计划基金资助项目(2007AA03Z456A);陕西省教育厅科研计划基金资助项目(07JK417)

摘　　要：目的借助于MINITAB统计学软件,采用Plackett-Burman设计和响应面优化法对基因工程菌Escherichia coliBL21发酵生产类人胶原蛋白的原始培养基进行优化研究。方法首先利用Plackett-Burman设计筛选出对类人胶原蛋白产量影响显著的3个因素,即葡萄糖甘油1∶4混合碳源、酵母粉和硫酸铵。在此基础上用最陡爬坡实验逼近最大响应区域,找到后续实验的中心点。再利用Box-Behnken实验设计及响应面分析法进行回归分析,确定主要影响因素的最佳水平。结果优化后的3种主要影响成分为葡萄糖甘油1∶4,混合碳源12.24 g/L,硫酸铵7.12 g/L,酵母粉5.53g/L。结论方程拟合良好,优化后的培养基发酵产量经摇瓶和发酵罐验证,产量分别为5.91g/L和6.12g/L,较优化前分别提高了306%和296%。Aim To improve human-like collagen production by optimizing its culture medium. Methods The Plackett-Burman design （PBD） was used to evaluate the effects of eight composites on the production of collagen. The steepest ascent experiment was adopted to determine the optimal region of the medium composition. The optimum composition of the fermentation medium for maximum collagen yield, as determined on the basis of a three- level three-factor Box-Behnken design, was obtained by response surface methodology （RSM）. Results The optimal concentrations of the variables are glucose and glycerol mixed carbon resource 12. 24 g/L, ammonium sulfate 7. 12 g/L and yeast powder 5.53 g/Lo The predicted collagen yield was 6. 05 g/L. Conclusion The high correlation between the predicted and observed values showed the validity of the model. Collagen yield was increased to 5.91 g/L and 6. 12 g/L respectively in Erlenmeyer flasks and 12. 8 L fermenter.

关 键 词：类人胶原蛋白 重组大肠杆菌 发酵 响应面优化法 

分 类 号：TQ929[轻工技术与工程—发酵工程] TU311.2[建筑科学—结构工程]