title: Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression.
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.
authors: Chengdu Institute of Biology, Chinese Academy of Sciences, P.O. Box 416, 610041, Chengdu, China. wuzhl@cib.ac.cn
authors: Zhong-Liu
authors: ZL
authors: Wu
authors: Jing
authors: J
authors: Qiao
authors: Zhi-Gang
authors: ZG
authors: Zhang
authors: F Peter
authors: FP
authors: Guengerich
authors: Yan
authors: Y
authors: Liu
authors: Xiao-Qiong
authors: XQ
authors: Pei
mesh_terms: D000818:Animals; D003062:Codon; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D008322:Mammals; D018832:Molecular Chaperones / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D003062:Codon; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System