Heterologous and homologous expression of the arginine biosynthetic argC~H cluster from Corynebacterium crenatum for improvement of (L) -arginine production |
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Authors: | Xu Meijuan Rao Zhiming Yang Juan Xia Haifeng Dou Wenfang Jin Jian Xu Zhenghong |
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Institution: | (1) The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu Province, People’s Republic of China;(2) Laboratory of Pharmaceutical Engineering, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, 214122, People’s Republic of China; |
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Abstract: | The genes involved in l-arginine biosynthesis in Corynebacterium crenatum are organized as the argCJBDFRGH cluster like in Corynebacterium glutamicum. However, the argC~H cluster of the C. crenatum SYPA 5-5, which is an industrialized l-arginine producer, had a lethal mutation occurring in the ArgR repressor encoding gene. The argC~H cluster with an inactive argR was overexpressed in E. coli and C. crenatum. In the recombinant E. coli JM109 enzyme activities were increased, and more l-arginine was found in the supernatants from l-glutamine. When the argC~H cluster was overexpressed in C. crenatum under its native promoter Parg, l-arginine production was increased by 24.9%, but the presence of the recombinant plasmid pJC-9039 had a negative effect on cell growth. Surprisingly, the DO value of the recombinant strain dropped gently and stayed at
a lower level from 24 h to the end of fermentation. The results demonstrated an increasing utilization of oxygen and the distinct
enhancement of unit cell l-arginine yields with the cluster argC~H-bearing in C. crenatum SYPA-9039. This study provides a kind of Corynebacteria with improved l-arginine-producing ability and an efficient elevation for producing amino acid. Moreover, the promoter Parg would be used
as a valid promoter to express objective genes for metabolic engineering in Corynebacteria. |
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