Archives of Pharmacal Research

The Pharmaceutical Society of Korea (대한약학회)
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Temperature Effect on the Functional Expression of Human Cytochromes P450 2A6 and 2E1 in Escherichia coli

  • Yim Sung-Kun (School of Biological Sciences and Technology, Chonnam National University) ;
  • Ahn Taeho (Department of Biochemistry, College of Veterinary Medicine, Chonnam National University) ;
  • Jung Heung-Chae (The National Research Laboratory of Microbial Display, GenoFocus, Inc., and Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology) ;
  • Pan Jae-Gu (The National Research Laboratory of Microbial Display, GenoFocus, Inc., and Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology) ;
  • Yun Chul-Ho (School of Biological Sciences and Technology, Chonnam National University, Hormone Research Institute, Chonnam National University)
  • Published : 2005.04.01
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Abstract

Human cytochromes P450 (GYP) 2A6 and 2E1 are of great interest because of their important roles in the oxidation of numerous drugs and carcinogens. Bacterial expression systems, especially Escherichia coli cells, have been widely used for the production of various GYP enzymes in order to obtain high yield of proteins. The expression methods usually employ longer culture time (30-72 h) at lower temperature (usually under $30^{\circ}C$). Expression levels of GYPs 2A6 and 2E1 at $37^{\circ}C$ were compared to those at $28^{\circ}C$, which is a usual temperature used in most bacterial expression systems for human GYP expression. Within 18 h the expression levels of GYPs 2A6 and 2E1 reached up to 360 and 560 nmol per liter culture at $37^{\circ}C$, respectively, which are compatible with those of 36 h culture at $28^{\circ}C$. The activities of GYPs expressed at $37^{\circ}C$ were also comparable to those expressed at $28^{\circ}C$. The present over-expression system can be useful for rapid production of large amounts of active human GYPs 2A6 and 2E1 in E. coli.

Keywords

References

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