摘要
为便于下游中试规模生产,采用温度敏感型启动子PRPL构建rhTNFα工程菌,并对影响工程菌发酵培养和表达的因素进行了初步研究。结果显示,采用RTB培养基,50℃热水快速升温,诱导培养4~4.5h,湿菌体收获率和表达量均达到较高水平;50L发酵罐连续发酵3批,14000r/min连续离心,菌体收获率湿重达16.gg/L培养物,rhTNFα表达量占菌体总蛋白的10.5%,活性达1.35×107U/mg蛋白。.
After pRL-rhTNF/JM013 engineering bacterial strain was constructed usingtemperature-sensitive promotor PRPL, the factors influencing the fermentation of it and theexpression of rhTNF α were preliminarily studied. The result showed that, by using RTBmedium, the recovery and expression rates of the strain reached high levels after being heatedwith water (50℃ ) and inducing at 42℃ for 4~4. 5h. Three lots of the engineering productswere separated by continuous centrifugation (14000r/min). The average recovery rate of rhTNF α reached 16. 9g/L cultrue, and the activity of it reached 1. 35 × 107U/mg protein, and10. 5% of total somatic protein was expressed.
出处
《中国生物制品学杂志》
CAS
CSCD
1998年第1期14-17,共4页
Chinese Journal of Biologicals
基金
国家"863"高技术计划资助