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Engineering Cytochrome P450 BM3 for Terminal Alkane Hydroxylation
Author(s) -
Meinhold Peter,
Peters Matthew W.,
Hartwick Adam,
Hernandez Alisha R.,
Arnold Frances H.
Publication year - 2006
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.200505465
Subject(s) - chemistry , hydroxylation , regioselectivity , alkane , saturated mutagenesis , octane , bacillus megaterium , cytochrome p450 , stereochemistry , biocatalysis , organic chemistry , enzyme , biochemistry , mutant , catalysis , bacteria , reaction mechanism , gene , genetics , biology
Enzymes that catalyze the terminal hydroxylation of alkanes could be used to produce more valuable chemicals from hydrocarbons. Cytochrome P450 BM3 from Bacillus megaterium hydroxylates medium‐chain fatty acids at subterminal positions at high rates. To engineer BM3 for terminal alkane hydroxylation, we performed saturation mutagenesis at selected active‐site residues of a BM3 variant that hydroxylates alkanes. Recombination of beneficial mutations generated a library of BM3 mutants that hydroxylate linear alkanes with a wide range of regioselectivities. Mutant 77‐9H exhibits 52% selectivity for the terminal position of octane. This regioselectivity is octane‐specific and does not transfer to other substrates, including shorter and longer hydrocarbons or fatty acids. These results show that BM3 can be readily molded for regioselective oxidation.