title: Evolving P450pyr hydroxylase for highly enantioselective hydroxylation at non-activated carbon atom.
abstract: Directed evolution of a monooxygenase to achieve very high enantioselectivity for hydroxylation at non-activated carbon atoms is demonstrated for the first time, where a triple mutant of P450pyr hydroxylase is obtained via determination of enzyme structure, iterative saturation mutagenesis, and high-throughput screening with a MS-based ee assay to increase the product ee from 53% to 98% for the hydroxylation of N-benzyl pyrrolidine to (S)-N-benzyl 3-hydroxypyrrolidine.
authors.affiliation: Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117576.
authors.forename: Son Quang
authors.initials: SQ
authors.lastname: Pham
authors.forename: Guillaume
authors.initials: G
authors.lastname: Pompidor
authors.forename: Ji
authors.initials: J
authors.lastname: Liu
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Li
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
mesh_terms: D001665:Binding Sites; D002244:Carbon / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D006900:Hydroxylation; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011759:Pyrrolidines / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism
chemical_list: D011759:Pyrrolidines; D002244:Carbon; D006899:Mixed Function Oxygenases; C032519:pyrrolidine