血管生成抑制素基因工程大肠杆菌的高密度发酵研究

High Cell Density Culture of the Engineered Escherichia coli Strain Haboring hAGN Gene

用 5L发酵罐研究了E .coliTG1 pBVA2和E .coliTG1 pBVK1 3的高密度培养工艺 ,确定了诱导及补料策略 ,在不降低外源基因表达量的前提下 ,工程菌TG1 pBVA2高密度发酵菌体干重为 1 6 8g L ,hAGN(K1 - 4)的表达量为菌体总蛋白的 2 4 1 % ,相当于 1 39g L ;同样的方法 ,工程菌TG1 pBVK1 3菌体干重可达 1 6g L ,hAGN(K1 - 3)占菌体总蛋白 2 5 8% ,相当于1 45g L。

The high cell density cultivation techniques of E.coli strains TG1/pBVA2 and TG1/pBVK13 was studied and the induction and fed batch conditions were determined. Without decreasing the expression level of foreign genes, the cell density of TG1/pBVA2 reaches OD 600 =0 42×100, the expressed hAGN(K1 4) constitutes 24 1% of the total bacterial protein corresponding to 1 39g/L. With the same method, the biomass of TG1/pBVK13 is OD 600 =0 40×100, and hAGN(K1 3) was estimated at 25 1% of the total bacterial protein that corresponds to 1 45g/L.