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Cloning and Optimizing Expression of Enoate Reductase Gene from Clostridium acetobutylicum in Escherichia coli(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2011 01
Page:
87-90
Research Field:
Articles
Publishing date:

Info

Title:
Cloning and Optimizing Expression of Enoate Reductase Gene from Clostridium acetobutylicum in Escherichia coli
Author(s):
LÜ Tong;  LIU Yang;  ZHAO Qing;  REN Jie;  WANG Min;  WU Qiaqing & ZHU Dunming
(1College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China)
(2Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China)
Keywords:
enoate reductase;  optimizing expression;  asymmetric reduction;  C=C bond;  Clostridium acetobutylicum
CLC:
Q936 : Q78
PACS:
DOI:
10.3724/SP.J.1145.2011.00087
DocumentCode:

Abstract:
Enone reductase is a class of biocatalysts which catalyze the reduction of C=C bond of enones and/or enoates. The gene which encodes enone reductase from Clostridium acetobutylicum ATCC824 was cloned. The cloned sequence consisted of 1 995 bp and encoded a protein of 664 amino acids, with a molecular weight of 73 ×103. The gene was cloned into the expression vector pET-32a(+), and the resulting plasmid DNA was transformed into the Escherichia coli Rosetta (DE3) pLysS strain. The heterologous expression was optimized and the best results were obtained for induction at an D600 nm of 0.6~0.8 with 0.1 mmol/L IPTG 12 h at 18 ℃. The SDS-PAGE analysis indicated that the ERs was expressed in the supernatant. Fig 5, Ref 16

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Last Update: 2011-02-28