title: Enzymatic hydroxylation of an unactivated methylene C-H bond guided by molecular dynamics simulations.
abstract: The hallmark of enzymes from secondary metabolic pathways is the pairing of powerful reactivity with exquisite site selectivity. The application of these biocatalytic tools in organic synthesis, however, remains under-utilized due to limitations in substrate scope and scalability. Here, we report how the reactivity of a monooxygenase (PikC) from the pikromycin pathway is modified through computationally guided protein and substrate engineering, and applied to the oxidation of unactivated methylene C-H bonds. Molecular dynamics and quantum mechanical calculations were used to develop a predictive model for substrate scope, site selectivity and stereoselectivity of PikC-mediated C-H oxidation. A suite of menthol derivatives was screened computationally and evaluated through in vitro reactions, where each substrate adhered to the predicted models for selectivity and conversion to product. This platform was also expanded beyond menthol-based substrates to the selective hydroxylation of a variety of substrate cores ranging from cyclic to fused bicyclic and bridged bicyclic compounds.
authors.affiliation: Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Alison R H
authors.initials: AR
authors.lastname: Narayan
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Gonzalo
authors.identifier: 0000000301054337
authors.initials: G
authors.lastname: Jiménez-Osés
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Peng
authors.identifier: 000000028188632X
authors.initials: P
authors.lastname: Liu
authors.affiliation: Department of Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Solymar
authors.initials: S
authors.lastname: Negretti
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Wanxiang
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Gilbert
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Raghunath O
authors.initials: RO
authors.lastname: Ramabhadran
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Yun-Fang
authors.initials: YF
authors.lastname: Yang
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Lawrence R
authors.initials: LR
authors.lastname: Furan
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Li
authors.affiliation: Skaggs School of Pharmacy &Pharmaceutical Sciences, University of California, San Diego, California 92093, USA.
authors.forename: Larissa M
authors.initials: LM
authors.lastname: Podust
authors.affiliation: 1] Department of Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA [2] Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: John
authors.initials: J
authors.lastname: Montgomery
authors.affiliation: Department of Chemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: K N
authors.initials: KN
authors.lastname: Houk
authors.affiliation: 1] Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, USA [2] Department of Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA [3] Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA [4] Department of Microbiology &Immunology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: Sherman
mesh_terms: D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism; D006900:Hydroxylation; D008610:Menthol / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D010084:Oxidation-Reduction; D011789:Quantum Theory; D013237:Stereoisomerism; D013302:Streptomyces / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D008610:Menthol; D003577:Cytochrome P-450 Enzyme System