应用溶氧反馈控制高密度培养重组大肠杆菌过程中乙酸的产生.pdf

第 20 卷第 2 期 高 校 化 学 工 程 学 报 No.2 Vol.20 2006 年 4 月 Journal of Chemical Engineering of Chinese Universities Apr . 2006 文章编号：1003-9015(2006)02-0233-06 应用溶氧反馈控制高密度培养重组大肠杆菌过程中乙酸的产生 , 郑志永 姚善泾 (浙江大学 化学工程与生物工程学系, 浙江 杭州 310027) 摘 要：在培养重组大肠杆菌的过程中，乙酸的产生会抑制细菌的生长，也不利于重组蛋白的表达。研究了补料培养 重组大肠杆菌过程中，应用脉冲补料方式时溶氧信号的响应特征来辩识是否产生乙酸。当大肠杆菌以0.1 h− 1 的比生长 速率生长时，补料过程中发酵液的乙酸含量很低，溶氧pO2 的响应信号随脉冲补料而振荡。而在诱导表达重组人表皮 生长因子时，大肠杆菌的呼吸能力下降，过量补入的葡萄糖使重组菌产生大量的乙酸，同时pO2 的响应信号失去振荡 的特征。根据pO2 响应信号的特征反馈控制葡萄糖的流加速率，有效地避免了重组蛋白表达过程中乙酸的产生，实现 了重组大肠杆菌的高密度培养，菌体干重达到48 g⋅L− 1，重组人表皮生长因子的表达水平提高了45 % 。 关键词：乙酸；溶氧；过程控制；重组大肠杆菌；高密度培养 中图分类号：Q813.11 文献标识码：A Preventing Acetate Production in High Cell Density Cultivation of Recombinant Escherichia coli by Using Feedback Control of Dissolved Oxygen ZHENG Zhi-yong, YAO Shan-jing (Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China) Abstract: Accumulation of acetate can inhibit cell growth and reduce recombinant protein production during cultivation of Escherichia coli . Relations between the oscillation of the dissolved oxygen response signal and pulses in feed rate in fed-batch cultivation of recombinant E. coli were characterized. The dissolved oxygen response signal was used to detect the acetate production in pulsed fed-batch cultivation of E. coli . In fed-batch cultivation with glucose-limited growth at specific growth rate 0.1 h− 1, the content of acetic acid is kept in low level and dissolved oxygen response signal is oscillated along with the pulses in feed rate in fed-batch phase. The