title: Microbial Flavoprotein Monooxygenases as Mimics of Mammalian Flavin-Containing Monooxygenases for the Enantioselective Preparation of Drug Metabolites.
abstract: Mammalian flavin-containing monooxygenases, which are difficult to obtain and study, play a major role in detoxifying various xenobiotics. To provide alternative biocatalytic tools to generate flavin-containing monooxygenases (FMO)-derived drug metabolites, a collection of microbial flavoprotein monooxygenases, sequence-related to human FMOs, was tested for their ability to oxidize a set of xenobiotic compounds. For all tested xenobiotics [nicotine, lidocaine, 3-(methylthio)aniline, albendazole, and fenbendazole], one or more monooxygenases were identified capable of converting the target compound. Chiral liquid chromatography with tandem mass spectrometry analyses of the conversions of 3-(methylthio)aniline, albendazole, and fenbendazole revealed that the respective sulfoxides are formed in good to excellent enantiomeric excess (e.e.) by several of the tested monooxygenases. Intriguingly, depending on the chosen microbial monooxygenase, either the (R)- or (S)-sulfoxide was formed. For example, when using a monooxygenase from Rhodococcus jostii the (S)-sulfoxide of albendazole (ricobendazole) was obtained with a 95% e.e. whereas a fungal monooxygenase yielded the respective (R)-sulfoxide in 57% e.e. For nicotine and lidocaine, monooxygenases could be identified that convert the amines into their respective N-oxides. This study shows that recombinantly expressed microbial monooxygenases represent a valuable toolbox of mammalian FMO mimics that can be exploited for the production of FMO-associated xenobiotic metabolites.
authors.affiliation: Analytical Biochemistry, Groningen Research Institute of Pharmacy (T.G., H.P.P., R.B.), Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute (M.K., M.W.F.), and Interfaculty Mass Spectrometry Center (H.P.P.), University of Groningen, Groningen, the Netherlands.
authors.forename: Turan
authors.initials: T
authors.lastname: Gul
authors.affiliation: Analytical Biochemistry, Groningen Research Institute of Pharmacy (T.G., H.P.P., R.B.), Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute (M.K., M.W.F.), and Interfaculty Mass Spectrometry Center (H.P.P.), University of Groningen, Groningen, the Netherlands.
authors.forename: Marzena
authors.initials: M
authors.lastname: Krzek
authors.affiliation: Analytical Biochemistry, Groningen Research Institute of Pharmacy (T.G., H.P.P., R.B.), Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute (M.K., M.W.F.), and Interfaculty Mass Spectrometry Center (H.P.P.), University of Groningen, Groningen, the Netherlands.
authors.forename: Hjalmar P
authors.initials: HP
authors.lastname: Permentier
authors.affiliation: Analytical Biochemistry, Groningen Research Institute of Pharmacy (T.G., H.P.P., R.B.), Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute (M.K., M.W.F.), and Interfaculty Mass Spectrometry Center (H.P.P.), University of Groningen, Groningen, the Netherlands m.w.fraaije@rug.nl.
authors.forename: Marco W
authors.initials: MW
authors.lastname: Fraaije
authors.affiliation: Analytical Biochemistry, Groningen Research Institute of Pharmacy (T.G., H.P.P., R.B.), Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute (M.K., M.W.F.), and Interfaculty Mass Spectrometry Center (H.P.P.), University of Groningen, Groningen, the Netherlands.
authors.forename: Rainer
authors.initials: R
authors.lastname: Bischoff
mesh_terms: D015766:Albendazole / Q000737:chemistry / Q000378:metabolism; D000814:Aniline Compounds / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D005273:Fenbendazole / Q000737:chemistry / Q000378:metabolism; D008012:Lidocaine / Q000737:chemistry / Q000378:metabolism; D009538:Nicotine / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000378:metabolism*; D012240:Rhodococcus / Q000201:enzymology*; D013379:Substrate Specificity; D013454:Sulfoxides / Q000737:chemistry / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D015262:Xenobiotics / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000814:Aniline Compounds; D001426:Bacterial Proteins; D013454:Sulfoxides; D015262:Xenobiotics; D005273:Fenbendazole; D009538:Nicotine; D008012:Lidocaine; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); D015766:Albendazole