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Improvement of l-Arginine Production by Overexpression of a Bifunctional Ornithine Acetyltransferase in Corynebacterium crenatum

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Abstract

Ornithine acetyltransferase (EC 2.3.1.35; OATase) gene (argJ) from the l-arginine-producing mutant Corynebacterium crenatum SYPA5-5 was cloned, sequenced, and expressed in Escherichia coli BL21 (DE3). Analysis of the argJ sequence revealed that the argJ coded a polypeptide of 388 amino acids with a calculated molecular weight of 39.7 kDa. In this study, the function of the OATase (argJ) of C. crenatum SYPA5-5 has been identified as a conserved ATML sequence for the autolysis of the protein to α- and β-subunits. When the argJ regions corresponding to the α- and β-subunits were cloned and expressed separately in E. coli BL21, OATase activities were abolished. At the same time, a functional study revealed that OATase from C. crenatum SYPA5-5 was a bifunctional enzyme with the functions of acetylglutamate synthase (EC 2.3.1.1, NAGS) and acetylornithine deacetylase (EC 3.5.1.16, AOase) activities. In order to investigate the effects of the overexpression of the argJ gene on l-arginine production, the argJ gene was inserted into pJCtac to yield the recombinant shuttle plasmid pJCtac-argJ and then transformed into C. crenatum SYPA5-5. The results showed that the engineered strains could not only express more OATase (90.9%) but also increase the production of l-arginine significantly (16.8%).

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Acknowledgments

This work was supported by Programs for New Century Excellent Talents in University (no. NCET-07-0380, NCET-10-0459), National Basic Research Program (973 Program) (no. 2007CB707804), and the National High-Tech Programs of China (no. 2006AA020104, 2006AA020301, 2007AA02Z207).

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Authors and Affiliations

  1. Laboratory of Pharmaceutical Engineering, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Wenfang Dou, Dongmei Cai, Xiaomei Zhang & Zhenghong Xu

  2. The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu Province, 214122, People’s Republic of China

    Meijuan Xu, Zhiming Rao & Zhenghong Xu

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  1. Wenfang Dou
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  2. Meijuan Xu
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  3. Dongmei Cai
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  4. Xiaomei Zhang
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Correspondence to Zhenghong Xu.

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Dou, W., Xu, M., Cai, D. et al. Improvement of l-Arginine Production by Overexpression of a Bifunctional Ornithine Acetyltransferase in Corynebacterium crenatum . Appl Biochem Biotechnol 165, 845–855 (2011). https://doi.org/10.1007/s12010-011-9302-3

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