Coexpression of a human P450 (CYP3A4) and P450 reductase generates a highly functional monooxygenase system in Escherichia coli

FEBS Letters

Blake, Jennifer A.R.²  Friedberg, Thomas²  Pritchard, Mike²  Smith, Graeme C.M.²  Burchell, Brian¹  Wolf, C.Roland²  Ding, Shaohong² 

[1] Department of Biochemical Medicine, Ninewells Hospital and Medical School, Dundee, UK;Biomedical Research Centre, Ninewells Hospital and Medical School, Dundee, UK

关键词: Recombinant cytochrome P450;    human;    Drug metabolism;    Bioreactors;    Heterologous expression;    E. coli;   

DOI  :  10.1016/S0014-5793(96)01196-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The catalytic activities of recombinant cytochrome P450s expressed in E. coli have been impeded by the absence of endogenous P450 reductase. To solve this problem, we coexpressed P450 reductase with CYP3A4. Membranes from this strain contained 215 pmol P450/mg protein and a reductase activity of 1315 nmol cytochrome c reduced/min per mg. We detected 6β-hydroxylation of testosterone and oxidation of nifedipine in vivo with turnover numbers of 15.2 and 17.3 min⁻¹, respectively. These values compare favourably with those obtained using an optimally reconstituted system. Our data demonstrate that a catalytically efficient human P450 system can be generated in E. coli.

【 授权许可】

Unknown