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

Jennifer A. R. Blake, Mike Pritchard, Shaohong Ding, Graeme C. M. Smith, Brian Burchell, C.Roland Wolf, Thomas Friedberg

    Research output: Contribution to journalArticle

    87 Citations (Scopus)

    Abstract

    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 6ß-hydroxylation of testosterone and oxidation of nifedipine in vivo with turnover numbers of 15.2 and 17.3 min , 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.
    Original languageEnglish
    Pages (from-to)210-214
    Number of pages5
    JournalFEBS Letters
    Volume397
    Issue number2-3
    DOIs
    Publication statusPublished - 18 Nov 1996

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