title: Regioselective hydroxylation of omeprazole enantiomers by bacterial CYP102A1 mutants.
abstract: A large set of Bacillus megaterium CYP102A1 mutants are known to metabolize various drugs to form human metabolites. Omeprazole (OMP), a proton pump inhibitor, has been widely used as an acid inhibitory agent for the treatment of gastric acid hypersecretion disorders. It is primarily metabolized by human CYP2C19 and CYP3A4 to 5'-OH OMP and a sulfone product, respectively. It was recently reported that several CYP102A1 mutants can oxidize racemic and S-OMP to 5'-OH OMP and that these mutants can further oxidize 5'-OH racemic OMP to 5'-COOH OMP. Here, we report that the S- and R-enantiomers of OMP are hydroxylated by 26 mutants of CYP102A1 to produce 1 major metabolite (5'-OH OMP) regardless of the chirality of the parent substrates. Although the binding of R-OMP to the CYP102A1 active site caused a more apparent change of heme environment compared with binding of S-OMP, there was no correlation between the spectral change upon substrate binding and catalytic activity of either enantiomer. The 5'-OH OMP produced from racemic, S-, and R-OMP could be obtained with a high conversion rate and high selectivity when the triple R47L/F87V/L188Q mutant was used. These results suggest that bacterial CYP102A1 mutants can be used to produce the human metabolite 5'-OH OMP from both the S- and R-enantiomers of OMP.
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Ryu
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Bo-Yeon
authors.initials: BY
authors.lastname: Park
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: So-Young
authors.initials: SY
authors.lastname: Kim
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Sun-Ha
authors.initials: SH
authors.lastname: Park
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Jung
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Min
authors.initials: M
authors.lastname: Park
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Ki Deok
authors.initials: KD
authors.lastname: Park
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Taeho
authors.initials: T
authors.lastname: Ahn
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.).
authors.forename: Hyung-Sik
authors.initials: HS
authors.lastname: Kang
authors.affiliation: School of Biological Sciences and Technology (S.H.R., B.-Y.P., S.-Y.K, S.-H.P., H.-J.J., M.P., H.-S.K., C.-H.Y.), and Department of Biochemistry, College of Veterinary Medicine (T.A.), Chonnam National University, Gwangju, Republic of Korea; and Gwangju Center, Korea Basic Science Institute, Gwangju, Republic of Korea (K.D.P.) chyun@jnu.ac.kr.
authors.forename: Chul-Ho
authors.initials: CH
authors.lastname: Yun
mesh_terms: D001410:Bacillus megaterium / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D020134:Catalytic Domain / Q000502:physiology; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D006418:Heme / Q000378:metabolism; D006900:Hydroxylation / Q000502:physiology*; D009154:Mutation / Q000235:genetics; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism*; D009853:Omeprazole / Q000378:metabolism*; D010084:Oxidation-Reduction; D013237:Stereoisomerism
chemical_list: D001426:Bacterial Proteins; D006418:Heme; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases; D009853:Omeprazole