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Regio-and Enantioselective Alkane Hydroxylation with Engineered Cytochromes P450 BM-3
 

Summary: Regio- and Enantioselective Alkane Hydroxylation with
Engineered Cytochromes P450 BM-3
Matthew W. Peters, Peter Meinhold, Anton Glieder, and Frances H. Arnold,*
Contribution from the DiVision of Chemistry and Chemical Engineering, California Institute of
Technology, Pasadena, California 91125
Received June 24, 2003; E-mail: frances@cheme.caltech.edu
Abstract: Cytochrome P450 -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.
Introduction
Cytochromes P450 comprise a superfamily of enzymes with
well over a thousand members that are, as a whole, capable of
oxidizing an immense variety of organic molecules in vivo using
atmospheric dioxygen as an oxidant.1,2 Conversion of even a

  

Source: Arnold, Frances H. - Division of Chemistry and Chemical Engineering, California Institute of Technology

 

Collections: Chemistry; Biology and Medicine