Drug metabolism by Escherichia coli expressing human cytochromes P450

Parikh, A, Gillam, EMJ and Guengerich, FP (1997) Drug metabolism by Escherichia coli expressing human cytochromes P450. Nature Biotechnology, 15 8: 784-788. doi:10.1038/nbt0897-784

Author Parikh, A
Gillam, EMJ
Guengerich, FP
Title Drug metabolism by Escherichia coli expressing human cytochromes P450
Journal name Nature Biotechnology   Check publisher's open access policy
ISSN 1087-0156
Publication date 1997-08-01
Year available 1997
Sub-type Article (original research)
DOI 10.1038/nbt0897-784
Open Access Status
Volume 15
Issue 8
Start page 784
End page 788
Total pages 5
Place of publication NEW YORK
Language eng
Subject 1305 Biotechnology
1502 Bioengineering
2402 Applied Microbiology and Biotechnology
1313 Molecular Medicine
2204 Biomedical Engineering
Abstract The broad substrate specificity of the cytochrome P450 (P450) enzyme superfamily of heme-thiolate proteins lends itself to diverse environmental and pharmaceutical applications. Until recently, the primary drawback in using living bacteria to catalyze mammalian P450-mediated reactions has been the paucity of electron transport from NADPH to P450 via endogenous flavoproteins. We report the functional expression in Escherichia coli of bicistronic constructs consisting of a human microsomal P450 enzyme encoded by the first cistron and the auxiliary protein NADPH-P450 reductase by the second. Expression levels of P450s ranged from 35 nmol per liter culture to 350 nmol per liter culture, with expression of NADPH-P450 reductase typically ranging from 50% to 100% of that of P450. Transformed bacteria metabolized a number of typical P450 substrates at levels comparable to isolated bacterial membranes fortified with an NADPH-generating system. These rates compare favorably with those obtained using human liver microsomes as well as those of reconstituted in vitro systems composed of purified proteins, lipids, and cofactors.
Keyword drug metabolism
cytochrome P450
NADPH-cytochrome P450
bicistronic constructs
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID R35 CA44353
P30 ES00267
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads - Archived
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