title: A single active site mutation inverts stereoselectivity of 16-hydroxylation of testosterone catalyzed by engineered cytochrome P450 BM3.
abstract: Inversion of stereoselectivity: screening of a minimal mutant library revealed a cytochrome P450 BM3 variant M01 A82W S72I capable of producing 16 α-OH-testosterone. Remarkably, a single active site mutation S72I in M01 A82W inverted the stereoselectivity of hydroxylation from 16 β to 16 α. Introduction of S72I mutation in another 16 β-OH-selective variant M11 V87I, also resulted in similar inversion of stereoselectivity.
authors.affiliation: Department of Chemistry and Pharmaceutical Sciences, Division of Molecular and Computational Toxicology, Vrije Universiteit, De Boelelaan 1083, 1081 HV Amsterdam, Netherlands.
authors.forename: Harini
authors.initials: H
authors.lastname: Venkataraman
authors.forename: Stephanie B A de
authors.initials: SB
authors.lastname: Beer
authors.forename: Laura A H van
authors.initials: LA
authors.lastname: Bergen
authors.forename: Nick van
authors.initials: Nv
authors.lastname: Essen
authors.forename: Daan P
authors.initials: DP
authors.lastname: Geerke
authors.forename: Nico P E
authors.initials: NP
authors.lastname: Vermeulen
authors.forename: Jan N M
authors.initials: JN
authors.lastname: Commandeur
mesh_terms: D055162:Biocatalysis; D020134:Catalytic Domain; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006900:Hydroxylation; D009154:Mutation; D015202:Protein Engineering; D013237:Stereoisomerism; D013739:Testosterone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System