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| The Activity Study of Aminodeoxychorismate Synthase of |Differernt Corynebacterium glutamicum |
| REN Jian-Hong1, ZHANG Xiao-Mei2, DOU Wen-Fang2, XU Hong-Yu2, HU Zheng-Hong2 |
1. Laboratory of Pharmaceutical Engineering,School of Medicine and Pharmaceutics,Jiangnan University,Wuxi214122,China
2. Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi214122,China |
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Abstract The two pabAB genes encoding aminodeoxychorismate synthase(ADC synthase) from a Lserine producing strain Corynebacterium glutamicum SYPS062 and model strain Corynebacterium glutamicum ATCC 13032 were ampilified by PCR. The result of nucleotide sequence analysis showed that both pabAB fragments were 1863bp, encoding 620 amino acids. 16 bases differences that resulted in the changes of 7 amino acids were found in the pabAB of SYPS062.The two pabAB were inserted into pET28a to yield the recombinant expression vector pET28apabAB and then transfromed into BL21(DE3).Upon IPTG induction,soluble ADC synthase was overproduced by E.coli BL21(DE3) harboring the expression construct.Recombinant ADC synthase purified by NiNTA affinity chromatography showed a single band about 67kDa on SDSPAGE gel, and activity of aminodeoxychorismate synthase analysis show that the enzyme specific activity of SYPS062 is 46.6% lower than ATCC 13032.
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Received: 19 March 2009
Published: 28 July 2009
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