Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression.
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.
Wu
Zhong-Liu
ZL
Chengdu Institute of Biology, Chinese Academy of Sciences, P.O. Box 416, 610041, Chengdu, China. wuzhl@cib.ac.cn
Qiao
Jing
J
Zhang
Zhi-Gang
ZG
Guengerich
F Peter
FP
Liu
Yan
Y
Pei
Xiao-Qiong
XQ
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
D003062:Codon; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System