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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (01): 29-34    DOI:
    
Activity Analysis of L-serine Dehydratase and Effect of the Gene Knockout on L-serine Accumulation in Corynebacterium glutamicum SYPS-062
JIN Xue-xia1,2, ZHANG Xiao-mei1, DOU Wen-fang1, XU Hong-yu1, XU Zheng-hong1,2
1. Lab of Pharmaceutical Engineering, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122, China;
2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China
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Abstract  

Corynebacterium glutamicum SYPS-062, a strain isolated from soil sample can directly product L-serine from sugar substances. The sdaA gene encoding L-serine dehydratase (EC 4.3.1.17, L-SerDH) from C.glutamicum SYPS-062 was amplified and sequenced. The plasmid pET-28a- sdaA was constructed and transfromed into E.coli BL21 (DE3). SDS-PAGE showed that the gene sdaA was expressed successfully in recombinant E.coli BL21.Then L-SerDH was purified by Ni-NTA affinity chmmatography. The results showed a single band about 46 kDa on SDS-PAGE gel, and the specified activity was about 0.57 U/mg. However, compared the sdaA genes between C.glutamicum SYPS-062 and ATCC13032, it was appeared that the two sdaA cannot lead to the differences of activity of L-SerDH. To further investigate the mechanism of L-serine accumulation by SYPS-062. A sdaA deleted mutant of C.glutamicum SYPS-062 was successfully constructed. The resulting plasmid,pK18mobsacB-△sdaA was constructed and introduced into C.glutamicum SYPS-062 by electroporation .The Recombinant bacteria C.glutamicum SYPS-062△sdaA was conformed by PCR. The deletion of gene sdaA in C.glutamicum SYPS-062 yielded 15.13% increase of L-serine production with slow growth.



Key wordsL-serine      Corynebacterium glutamicum      sdaA      L-serine dehydratase     
Received: 08 September 2010      Published: 25 January 2011
ZTFLH:  Q786  
Cite this article:

JIN Xue-xia, ZHANG Xiao-mei, DOU Wen-fang, XU Hong-yu, XU Zheng-hong. Activity Analysis of L-serine Dehydratase and Effect of the Gene Knockout on L-serine Accumulation in Corynebacterium glutamicum SYPS-062. China Biotechnology, 2011, 31(01): 29-34.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I01/29


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