Cloning, Expressing of the <em>prpC2</em> Gene Encoding Citrate Synthase from <em>Corynebacterium crenatum</em> and Its Effect on L-arginine Synthesis

doi:10.13523/j.cb.20150307

China Biotechnology

2015, Vol. 35

Issue (3): 49-55 DOI: 10.13523/j.cb.20150307

Cloning, Expressing of the prpC2 Gene Encoding Citrate Synthase from Corynebacterium crenatum and Its Effect on L-arginine Synthesis

FANG Zhan¹, XU Mei-juan¹, RAO Zhi-ming¹, MAN Zai-wei¹, XU Zheng-hong², GENG Yan², LU Mao-lin³

1. The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;
2. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China;
3. Jiangsu Institute of Microbiology Corporation Limited, Wuxi 214063, China

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Abstract

Corynebacterium crenatum SYPA5-5 is an L-arginine high-producing industrial strain of mutation breeding. The role of citrate synthase in L-arginine biosynthesis was investigated by overexpressing the citrate synthase (prpC2) gene in C. crenatum SYPA5-5. The resultant 5.37-fold increase in intracellular citrate synthase activity was achieved in the prpC2-overexpressing strain C. crenatum SYPA5-5/pDXW-10-prpC2 . The recombinant strain enhanced the L-arginine yield to 44.7g/L by about 23.1% in 5L fermenter, as compared to the control, with affecting glucose depletion rate slightly. While the L-arginine yield increased in the prpC2-overexpressing strain, the L-lysine yield, the most primary by-product formation during L-arginine fermentation, decreased to 1.21g/L from the original concentration 5.96g/L, correlating with an increase in the tricarboxylic acid cycle (TCA) intermediates (citrate and isocitrate) and an increase in the activity of citrate synthase.

Key words： L-arginine Corynebacterium crenatum Citrate synthase Fermentation

Received: 31 December 2014 Published: 25 March 2015

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FANG Zhan, XU Mei-juan, RAO Zhi-ming, MAN Zai-wei, XU Zheng-hong, GENG Yan, LU Mao-lin. Cloning, Expressing of the prpC2 Gene Encoding Citrate Synthase from Corynebacterium crenatum and Its Effect on L-arginine Synthesis. China Biotechnology, 2015, 35(3): 49-55.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150307 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I3/49

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