Tao Yong, Cheng Qiong, Kopatsis Alexander D
Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2012;834:283-303. doi: 10.1007/978-1-61779-483-4_18.
Escherichia coli is the most commonly used microorganism for production of recombinant proteins for different applications. Acetate accumulation during aerobic growth on glucose has significant negative impact on recombinant protein production in Escherichia coli. Various strategies, such as process and genetic approaches have been developed to limit acetate formation to increase the productivity of recombinant proteins. We developed a strategy to combine inactivation of pyruvate oxidase (poxB) and over-expression of acety-CoA synthetase (acs) in E. coli K strain for controlling acetate accumulation. A recombinant peptide was expressed and produced in the engineered strains with a very low acetate -formation in a 10-L fermentation process.
大肠杆菌是用于生产不同用途重组蛋白的最常用微生物。在以葡萄糖为底物进行有氧生长过程中，乙酸盐的积累对大肠杆菌中重组蛋白的生产具有显著负面影响。人们已开发出各种策略，如工艺和基因方法，以限制乙酸盐的形成，从而提高重组蛋白的产量。我们开发了一种策略，通过使丙酮酸氧化酶（poxB）失活并在大肠杆菌K菌株中过表达乙酰辅酶A合成酶（acs）来控制乙酸盐的积累。在10升发酵过程中，一种重组肽在工程菌株中表达并产生，乙酸盐形成量非常低。
