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Deletion of cg1360 Affects ATP Synthase Function and Enhances Production of L-Valine in Corynebacterium glutamicum

Journal of Microbiology and Biotechnology 2019년 29권 8호 p.1288 ~ 1298
 ( Wang Xiaochen ) - University of Science and Technology of China

 ( Yang Hongyu ) - Chinese Academy of Sciences Tianjin Institute of Industrial Biotechnology
 ( Zhou Wei ) - Chinese Academy of Sciences Tianjin Institute of Industrial Biotechnology
 ( Liu Jun ) - Chinese Academy of Sciences Tianjin Institute of Industrial Biotechnology
 ( Xu Ning ) - Chinese Academy of Sciences Tianjin Institute of Industrial Biotechnology

Abstract


Bacterial ATP synthases drive ATP synthesis by a rotary mechanism, and play a vital role in physiology and cell metabolism. Corynebacterium glutamicum is well known as an industrial workhorse for amino acid production, and its ATP synthase operon contains eight structural genes and two adjacent genes, cg1360 and cg1361. So far, the physiological functions of Cg1360 (GenBank CAF19908) and Cg1361 (GenBank CAF19909) remain unclear. Here, we showed that Cg1360 was a hydrophobic protein with four transmembrane helices (TMHs), while no TMH was found in Cg1361. Deletion of cg1360, but not cg1361, led to significantly reduced cell growth using glucose and acetic acid as carbon sources, reduced F1 portions in the membrane, reduced ATP-driven proton-pumping activity and ATPase activity, suggesting that Cg1360 plays an important role in ATP synthase function. The intracellular ATP concentration in the Δcg1360 mutant was decreased to 72% of the wild type, while the NADH and NADPH levels in the Δcg1360 mutant were increased by 29% and 26%, respectively. However, the Δcg1361 mutant exhibited comparable intracellular ATP, NADH and NADPH levels with the wild-type strain. Moreover, the effect of cg1360 deletion on L-valine production was examined in the L-valine-producing V-10 strain. The final production of L-valine in the V-10-Δcg1360 mutant reached 9.2 ± 0.3 g/l in shake flasks, which was 14% higher than that of the V-10 strain. Thus, Cg1360 can be used as an effective engineering target by altering energy metabolism for the enhancement of amino acid production in C. glutamicum.

키워드

ATP synthase; cg1360; C. glutamicum; L-valine production
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