重组BbFGF工程菌的发酵条件(英文)

Fermentation Conditions of Recombinant Bovine Fibroblast Growth Factor in Escherichia coli

利用大肠杆菌BL2 1(DE3) pLysS生产重组牛碱性成纤维细胞生长因子 (BbFGF) ,对此工程菌的发酵条件进行了研究 .在前期试验的基础上 ,重点探索培养基、诱导剂及葡萄糖添加方式对T7启动子指导下的bFGF合成的影响 .根据摇瓶试验和分批发酵试验结果 ,建立了一套变速流加葡萄糖的高密度补料分批发酵工艺 .在此工艺下 ,经 11h发酵 ,可得光密度为 30 .7、bFGF产量为 95mg/L的发酵产物 .两步法纯化目的蛋白 ,得到纯度为 95%的重组bFGF ,其生物活性和标准品一致 .

Key factors of the fermentation of a recombinant, E.coli BL21(DE3)pLysS producing basic fibroblast growth factor (bFGF) were studied. Flask_shaking experiments and batch fermentations were performed to investigate the effects of medium, inducer and glucose_feeding mode on bFGF formation directed by a T7 promoter. A fed_batch fermentation condition of high cell density with stepwise increased feeding of glucose was established. After undergoing fermentation for 11 h, the cell optical density was 30.7, and the amount of bFGF produced by E.coli reached 95 mg per liter culture. This target protein was purified to homogeneity (purity of 95%)from the supernatant of bacteria lysate and was found to be biologically identical to bFGF standard.