重组人胸腺肽α1工程菌的高密度发酵  

High Cell Density Fermentation of Recombinant Human Thymosin α1

作　　者：颜真[1] 张英起[1] 李民[2] 陈常庆[2] 

机构地区：[1]第四军医大学生物技术中心,西安710032 [2]中国科学院生物工程研究中心,上海200233

出　　处：《药物生物技术》2000年第4期209-212,共4页Pharmaceutical Biotechnology

摘　　要：为实现重组人胸腺肽α1工程菌E .coliTop10 /pThioHis Tα的高密度高表达发酵 ,首先进行了培养条件的摸索 ,确定了该工程菌的最佳培养条件、诱导剂IPTG的浓度、诱导起始和结束时间 ,然后用 5L自控发酵罐进行分批补料培养 ,发酵中采用分阶段限制性流加氮、碳源 ,保持溶解氧在 35 %左右 ,结果经 3mmol/LIPTG诱导 5h ,3批重复发酵 ,最终菌体密度均达到 6 0A60 0 以上 (相当于干菌 2 5 .6 g/L) ,保持并超过了该重组蛋白在试管和摇瓶中的表达量 ,融合蛋白ThioHis Tα的表达占菌体总蛋白的 42 .4%左右 ,含量达到 5 .7g/L ,相当于胸腺肽α12 .4g/L 。In order to reach the goal of high cell density fermentation and high level expression of recombinant human thymosin α 1 ( E.coli Top10/pThio His Tα). Pilot test of tube culture and flask shaking culture were done to get optimized culture conditions, including induction time, induction duration and inducer concentration of IPTG. Then fed batch culture was carried out on 5 L automatic fermentor. During that time, nitrogen and carbohydrate were controlled at specific speed. Dissolved oxygen was maintained around 35%. Bacterium was harvested after 5 h induction of 3 mmol/L IPTG. In three times repeated fermentation, final cell density were all more than 60 A 600 , (relative to 25.6 g dry cell weight /L). The average expression was 42.4% of total bacterial proteins. The content of fused protein was about 5.7 g/L, equal to 2.4 g/L thymosin α . These results were no doubt the useful bases for further purification and large scale production of thymosin α.

关 键 词：高密度发酵 工程菌 重组人胸腺肽 生产工艺 

分 类 号：TQ464.7[化学工程—制药化工]