Original Research Paper

Biotechnology Letters

, Volume 31, Issue 10, pp 1589-1593

Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression

  • Zhong-Liu WuAffiliated withChengdu Institute of Biology, Chinese Academy of Sciences Email author 
  • , Jing QiaoAffiliated withChengdu Institute of Biology, Chinese Academy of SciencesGraduate School of the Chinese Academy of Sciences
  • , Zhi-Gang ZhangAffiliated withChengdu Institute of Biology, Chinese Academy of SciencesGraduate School of the Chinese Academy of Sciences
  • , F. Peter GuengerichAffiliated withDepartment of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine
  • , Yan LiuAffiliated withChengdu Institute of Biology, Chinese Academy of Sciences
  • , Xiao-Qiong PeiAffiliated withChengdu Institute of Biology, Chinese Academy of Sciences

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Abstract

To elucidate the effects of codon optimization and chaperone coexpression on the heterologous expression of mammalian cytochrome P450s (P450) in Escherichia coli, the expression of P450s 2B1, 2S1, 2U1, 2W1, and 27C1 were investigated. With codon optimization for N-terminus or the entire gene, the expression levels of P450 27C1, 2U1 and 2W1 increased 22-fold, 3.6-fold and 2.1-fold, respectively, while those for P450s 2B1 and 2S1 remained unchanged. With coexpression of E. coli molecular chaperones GroEL/ES, the expression level increased up to 14-fold for P450 27C1, and 3- to 5-fold for P450s 2B1, 2S1, and 2W1. Simultaneous application of these two techniques resulted in synergetic effects.

Keywords

Chaperon Codon Cytochrome P450 Escherichia coli Expression