【摘要】 通过响应曲面法(RSM)达到优化重组人神经生长因子发酵培养基组分含量的目的.对发酵基础培养基采用四因素三水平实验设计,优化了葡萄糖、酵母粉、蛋白胨和KH2PO4的最佳浓度,得到了最大预测值并进行了验证.最终优化培养基组分为:葡萄糖7.95 g/L、蛋白胨19.06 g/L、酵母粉13.51 g/L和KH2PO43.87g/L,此时预测最大菌体量Ymax=2.398 g/L.验证试验结果表明:使用优化后的培养基表达重组人神经生长因子,其菌体量的平均值为(2.357±0.083)g/L,实验值与预测值基本相符,均高于优化前菌体量,且其表达量也得到相应提高.更多还原

【Abstract】 A four-factor,three-level Box-Behnken factorial design combined with response surface methodology(RSM) was used to optimize the medium composition for the rhNGF production in Escherichia coli cells,which carry a plasmid encoding NGF DNA as a part of the fermentation strategy for recombinant human nerve growth factor.The concentrations of glucose,yeast extract,tryptone and KH2PO4 in medium were changed according to the design consisting of 27 experiments,and the biomass yield was calculated.Estimated optimum conditions of the factors for the growth of Escherichia coli cells were as follows: 7.95 g/L glucose,19.06 g/L tryptone,13.51 g/L yeast extract and 3.87 g/L KH2PO4.The model prediction of 2.398 g/L biomass at optimum conditions was verified experimentally as(2.357±0.083) g/L,much higher than any value obtained in initial experiments as well as in studies carried out previously.And the expression yield of the rhNGF product was improved.更多还原