TY - JOUR T1 - Efficient screening of P450 BM3 mutants for their metabolic activity and diversity towards a wide set of drug-like molecules in chemical space JF - Drug Metabolism and Disposition JO - Drug Metab Dispos DO - 10.1124/dmd.111.039461 SP - dmd.111.039461 AU - Jelle Reinen AU - Jolanda S. van Leeuwen AU - Yongmin Li AU - Lifang Sun AU - Peter D.J. Grootenhuis AU - Caroline J. Decker AU - John Saunders AU - Nico P.E. Vermeulen AU - Jan N.M. Commandeur Y1 - 2011/01/01 UR - http://dmd.aspetjournals.org/content/early/2011/06/14/dmd.111.039461.abstract N2 - In the present study the diversity of a library of drug metabolizing bacterial cytochrome P450 BM3 mutants was evaluated by an LC-MS based screening method. A strategy was designed to identify a minimal set of BM3 mutants which displays differences in regio- and stereoselectivities and is suitable to metabolize a large fraction of drug chemistry space. We first screened the activities of six structurally diverse BM3 mutants towards a library of 43 marketed drugs (encompassing a wide range of human P450 phenotypes, cLogP values, charges, and molecular weights) using a rapid LC-MS method with an automated method development and data processing system. Significant differences in metabolic activity were found for the mutants tested and based on this drug library screen, nine structurally diverse probe drugs were selected which were subsequently used to study the metabolism of a library of fourteen BM3 mutants in more detail. Using this alternative screening strategy we were able to select a minimal set of BM3 mutants with high metabolic activities and diversity with respect to substrate specificity and regiospecificity that could produce both human relevant and BM3 unique drug metabolites. This panel of four mutants (M02, MT35, MT38, and MT43) was capable of producing P450-mediated metabolites for 41 of the 43 drugs tested while metabolizing 77% of the drugs by more than 20%. This we see as the first step in our approach to use bacterial P450 enzymes as general reagents for lead diversification in the drug development process and the biosynthesis of drug(-like) metabolites. ER -