响应面法优化胶原蛋白发酵培养基和培养条件

目的利用统计学方法对重组大肠杆菌发酵培养基进行优化,提高胶原蛋白产量。方法应用Plackett-Burman试验设计法和响应面法,对发酵培养基6种组分配比和2个初始发酵条件进行优化;用Design-Expert软件对实验数据进行多元回归分析,并建立3种主要因素(葡萄糖、混合氮源和K2HPO4)与胶原蛋白产量之间的函数关系。用最终优化的配方进行5次验证试验。结果培养基3个最佳浓度为:葡萄糖为14.69 g/L、混合氮源为15.04 g/L、K2HPO4为11.91 g/L,胶原蛋白表达率可达33.95%。5次验证试验所测得的胶原蛋白平均表达率为34.02%,与预测结果基本一致。结论优化的重组大肠杆菌发酵培养基可显著提高胶原蛋白产量。

Optimization of medium and condition for fermentation of collagen by response surface method

Objective To optimize the medium for fermentation of recombinant E.coli by statistical method so as to increase the yield of collagen.Methods The proportions of six components of medium and two initial conditions for fermentation were optimized by Plackett-Burman experimental design and response surface method.The experimental data were subjected to multiple regression analysis by using Design-Expert software,based on which the functional relationship between three primary components(glucose,mixed nitrogen sources and dipotassium hydrogen phosphate)and the yield of collagen was established.The final optimal medium was subjected to five validation tests.Results The optimal concentrations of glucose,mixed nitrogen sources and dipotassium hydrogen phosphate in medium were 14.69 g / L,15.04 g / L and 11.91 g / L respectively,by which the expression rate of collagen was 33.95%.The mean expression rate of collagen in five validation tests was 34.02%,which was basically consistent with that expected.Conclusion The yield of collagen increased significantly by using the optimized medium for fermentation of recombinant E.coli.