The catalytic activity of cytochrome P450cam towards styrene oxidation is increased by site-specific mutagenesis.
The styrene oxidation activity of cytochrome P450cam, has been greatly improved by rational protein engineering. Compared to the wild-type enzyme, the active-site mutants Y96A and Y96F bound styrene more tightly, consumed NADH more rapidly, and were more efficient at utilising reducing equivalents for product formation. Styrene oxide formation rates were enhanced 9-fold in the Y96A mutant relative to wild-type, and 25-fold in the Y96F mutant, thus demonstrating the effectiveness of active-site redesign in improving the activity of a haem monooxygenase towards an unnatural substrate.
Nickerson
D P
DP
Inorganic Chemistry Laboratory, Oxford, UK.
Harford-Cross
C F
CF
Fulcher
S R
SR
Wong
L L
LL
D001665:Binding Sites / Q000235:genetics; D019475:Camphor 5-Monooxygenase / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010084:Oxidation-Reduction; D015202:Protein Engineering; D013053:Spectrophotometry; D020058:Styrene; D013343:Styrenes / Q000378:metabolism*; D013379:Substrate Specificity
D013343:Styrenes; D020058:Styrene; D019475:Camphor 5-Monooxygenase