Liu Zhijie, Zhou Li, Hua Qiang
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Sheng Wu Gong Cheng Xue Bao. 2012 May;28(5):565-76.
Gene expression is regulated by different transcriptional regulators. The transcriptional regulator isocitrate lyase regulator (IclR) of Escherichia coli represses the expression of the aceBAK operon that codes for the glyoxylate pathway enzymes. In this study, physiological and metabolic responses of the deletion of the ic1R gene in E. coli BW25113 were investigated based on the quantification and analysis of intracellular metabolic fluxes. The knockout of the iclR gene resulted in a decrease in the growth rate, glucose uptake rate and the acetate secretion rate, but a slight increase in biomass yield. The latter could be attributed to the lowered metabolic fluxes through several CO2 generating pathways, including the redirection of 33% of isocitrate directly to succinate and malate without CO2 production as well as the reduced flux through the pentose phosphate pathway. Furthermore, although the glyoxylate shunt was activated in the iclR mutant, the flux through phosphoenolpyruvate (PEP) carboxykinase kept almost unchanged, implying an inactive PEP-glyoxylate cycle and no extra loss of carbon atoms in the mutant strain. Both the reduced glucose uptake rate and the active glyoxylate shunt were responsible for the minor decrease in acetate secretion in the ic1R knockout strain compared to that in the wild-type E. coli strain.
基因表达受不同转录调节因子的调控。大肠杆菌的转录调节因子异柠檬酸裂解酶调节因子(IclR)抑制编码乙醛酸途径酶的aceBAK操纵子的表达。在本研究中,基于细胞内代谢通量的定量和分析,研究了大肠杆菌BW25113中ic1R基因缺失的生理和代谢反应。iclR基因的敲除导致生长速率、葡萄糖摄取率和乙酸分泌率降低,但生物量产量略有增加。后者可归因于通过几种产生CO2的途径的代谢通量降低,包括33%的异柠檬酸直接重定向到琥珀酸和苹果酸而不产生CO2,以及通过磷酸戊糖途径的通量降低。此外,尽管乙醛酸旁路在iclR突变体中被激活,但通过磷酸烯醇式丙酮酸(PEP)羧激酶的通量几乎保持不变,这意味着PEP-乙醛酸循环不活跃,且突变菌株中没有额外的碳原子损失。与野生型大肠杆菌菌株相比,葡萄糖摄取率降低和乙醛酸旁路激活均导致ic1R敲除菌株中乙酸分泌略有减少。