重组人白细胞介素-6融合蛋白的大肠杆菌发酵工艺

Studies on Fermentation of Recombinant E. coli Harboring Interleukin-6 Fusion protein

研究了重组大肠杆菌表达人白细胞介素-6(rhIL-6)融合蛋白的发酵工艺。摇瓶实验对比了不同培养基对重组菌密度和rhIL-6表达量的影响,确定了以含0.1%葡萄糖的2×YT培养基为最适培养基。同时,还对诱导剂异丙基疏基半乳糖苷(IPTG)加入量进行优化,按每升发酵液加入0.02mmol,就能有效诱导rhIL-6的表达。在10 L发酵罐间歇实验中,以含0.1%葡萄糖的2×YT为培养基,比较了溶氧、pH、诱导前培养时间、诱导后培养时间对rhIL-6的表达量和菌密度的影响,得到最优培养条件为pH为6.8～7.0,接种量4%,温度37℃,诱导前溶氧量30%,在O.D._(600)为1.2～1.8时加入IPTG诱导,控制诱导后溶氧为5%～10%,再培养5 h,最终rhIL-6表达量为38%,菌体密度为5.8 g/L。

In this paper, fermentation conditions for recombinant E. coli JM109(pHZTM181) were investigated. In the shake flask experiments,2×YT containing 0.1% glucose was determined as the most suitable medium The optimum inducer concentration was only 0.02 mmol/L,which was far below the recommending value in documents. The pilo-scale experiments were carried out in a 10 L fermentor. Conditions such as dissolved oxygen,pH,preinduction time and postinduction time were investigated. The expression of foreign protein was 40% and the cell density was up to 5.8 under the optimum condition