Regio- and Enantioselective Alkane Hydroxylation with Engineered Cytochromes P450 BM-3
Abstract
Cytochrome P450 ΒΜ-3 from Bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant. BM-3 variant 9-10A-A328V hydroxylates octane at the 2-position to form S-2-octanol (40% ee). Another variant, 1-12G, also hydroxylates alkanes larger than hexane primarily at the 2-position but forms R-2-alcohols (40−55% ee). These biocatalysts are highly active (rates up to 400 min-1) and support thousands of product turnovers. The regio- and enantioselectivities are retained in whole-cell biotransformations with Escherichia coli, where the engineered P450s can be expressed at high levels and the cofactor is supplied endogenously.
Additional Information
© 2003 American Chemical Society. Received 24 June 2003. Published online 11 October 2003. Published in print 1 November 2003. The authors thank Dr. Nathan Dalleska for his assistance with the gas chromatography. This work is supported by the National Science Foundation (BES-9981770).Attached Files
Published - ja0303790.pdf
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Additional details
- Eprint ID
- 77460
- Resolver ID
- CaltechAUTHORS:20170515-134122486
- NSF
- BES-9981770
- Created
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2017-05-16Created from EPrint's datestamp field
- Updated
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2023-06-01Created from EPrint's last_modified field