Yu Bing-Qi, Shen Wei, Wang Zheng-Xiang, Zhuge Jian
Key Laboratory of Industrial Biotechnology of Ministry of Education, Southern Yangtze University, Wuxi 214036, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Mar;21(2):270-4.
The glyoxylate cycle was hypothesed to be indispensable for glutamate overproduction in coryneform bacteria, for it was thought to fulfill anaplerotic functions and to supply energy during the growth phase. During glutamate overproduction phase, however, it has been noted that the high level of the cycle is detrimental to the glutamate production. In order to clarify the relationship between the glutamate production and the glyoxylate cycle, a chromosomal aceA-disrupted mutant of wild-type C. glutamicum ATCC 13032 was constructed. The isocitrate lyase (ICL) activity of the parental strain was 0.011 u/mg of protein and reached 1.980 u/mg of protein after acetate induction; the mutant strain WTdeltaA, however, had no detectable ICL activity and was no longer able to grow on minimal medium with acetate as the sole carbon source. Compared with the wild-type C. glutamicum WT, the mutant strain WTdeltaA, exhibited the same growth rate with glucose as the sole carbon source, indicating glyoxylate cycle is not required for its growth on glucose. On the contrary, the glutamate production in WTdeltaA was severely impaired and more residual glucose was found in the fermentation broth at the end of fermentation with the mutant strain than with the wild-type strain. Further investigations into the relationship between the glutamate production and the glyoxylate cycle are under the way, which may help to elucidate the mechanism of glutamate overproduction.
乙醛酸循环被认为对于棒状杆菌过量生产谷氨酸是不可或缺的,因为它被认为在生长阶段发挥回补功能并提供能量。然而,在谷氨酸过量生产阶段,已经注意到该循环的高水平对谷氨酸生产是有害的。为了阐明谷氨酸生产与乙醛酸循环之间的关系,构建了野生型谷氨酸棒杆菌ATCC 13032的染色体aceA缺失突变体。亲本菌株的异柠檬酸裂解酶(ICL)活性为0.011 u/mg蛋白质,经乙酸盐诱导后达到1.980 u/mg蛋白质;然而,突变菌株WTdeltaA没有可检测到的ICL活性,并且不再能够在以乙酸盐作为唯一碳源的基本培养基上生长。与野生型谷氨酸棒杆菌WT相比,突变菌株WTdeltaA以葡萄糖作为唯一碳源时表现出相同的生长速率,这表明乙醛酸循环对于其在葡萄糖上的生长不是必需的。相反,WTdeltaA中的谷氨酸生产严重受损,并且在突变菌株发酵结束时,发酵液中发现的残留葡萄糖比野生型菌株更多。关于谷氨酸生产与乙醛酸循环之间关系的进一步研究正在进行中,这可能有助于阐明谷氨酸过量生产的机制。