THE CATALYTIC ACTIVITY OF CYTOCHROME P450(CAM) TOWARDS STYRENE OXIDATION IS INCREASED BY SITE-SPECIFIC MUTAGENESIS

The styrene oxidation activity of cytochrome P450(cam) has been greatl y improved by rational protein engineering. Compared to the wild-type enzyme, the active-site mutants Y96A and Y96F bound styrene more tight ly, consumed NADH more rapidly, and were more efficient at utilising r educing equivalents for product formation. Styrene oxide formation rat es were enhanced 9-fold in the Y96A mutant relative to wild-type, and 25-fold in the Y96F mutant, thus demonstrating the effectiveness of ac tive-site redesign in improving the activity of a haem monooxygenase t owards an unnatural substrate. (C) 1997 Federation of European Biochem ical Societies.