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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (09): 49-55    DOI: Q78
    
Cloning, Expression and Analysis of the argH Gene Encoding Argininosuccinate  Lyase from Corynebacterium crenatum
RAO Zhi-ming1,XU Mei-juan1,LU Yuan-xiu1,ZHOU Chen1,LAN Chun-yan1,DOU Wen-fang2,ZHANG Xiao-mei2,XU Hong-yu2,XU Zheng-hong1,2
1.Key Laboratory of Industrial Biotechnology, Ministry of Education,Wuxi 214122,China
2.Lab of Pharmaceutical Engineering, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122,China
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Abstract  

 Argininosuccinate lyase (EC 4.3.2.1; AL) genes from L-arginine producing mutant Corynebacterium crenatum SYPA were cloned and sequenced. Analysis of argH sequences revealed that only one ORF existed , which used ATG as the initiation codon and coded a peptide of 476 amino acids with acalculated molecular weight of 50.8 kDa. Only 10 nucleotide difference was found in the structure gene and the difference caused 3 change of amino acid by comparision of the gene sequences between C. crenatum SYPA and the Corynebacterium glutamicum ATCC 13032. The ORF sequence of argHs showed homologies of 99.4%. The argH gene from C. crenatum was expressed both in E.coli and C. crenatum SYPA. Then AL was purified by Ni-NTA affinity chromatography and the enzymatic characterization of it was determines. The expression vector pJC1-tac-argH was transducted to C. crenatum SYPA. The AL was expressed well and the activity was improved by 66.8%. The fermentive character of CCH1(pJC1-tac-argH) was also primary analysed. The result shows that the acid producing ability of recombinant strain is improved by 14.2%.

Key words： L-arginine      Corynebacterium crenatum      Arginieosuccinate Lyase      Characterization of enzyme      Fermentation     
Received: 09 April 2010      Published: 25 August 2010
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RAO Zhi-Meng
XU Mei-Juan
LIU Yuan-Xiu
ZHOU Chen
LA Chun-Yan
DOU Wen-Fang
ZHANG Xiao-Mei-Hu
HONG Yu
HU Zheng-Hong
Cite this article:

RAO Zhi-Meng, XU Mei-Juan, LIU Yuan-Xiu, ZHOU Chen, LA Chun-Yan, DOU Wen-Fang, ZHANG Xiao-Mei-Hu, HONG Yu, HU Zheng-Hong. Cloning, Expression and Analysis of the argH Gene Encoding Argininosuccinate  Lyase from Corynebacterium crenatum. China Biotechnology, 2010, 30(09): 49-55.

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https://manu60.magtech.com.cn/biotech/Q78     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I09/49

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