title: Hydroxylation of Compactin (ML-236B) by CYP105D7 (SAV_7469) fromStreptomyces avermitilis.
abstract: Compactin and pravastatin are competitive cholesterol biosynthesis inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase and belong to the statin drugs; however, the latter shows superior pharmacokinetic characteristics. Previously, we reported that the bacterial P450, CYP105D7, fromStreptomyces avermitiliscan catalyze the hydroxylation of 1-deoxypentalenic acid, diclofenac, and naringenin. Here, we demonstrate that CYP105D7 could also catalyze compactin hydroxylation in vitro. In the presence of both bacterial and cyanobacterial redox partner systems with an NADPH regeneration system, the reaction produced two hydroxylated products, including pravastatin (hydroxylated at the C6 position). The steady-state kinetic parameters were measured using the redox partners of putidaredoxin and its reductase. TheKmandkcatvalues for compactin were 39.1 ± 8.8 µM and 1.12 ± 0.09 min-1, respectively. Thekcat/Kmvalue for compactin (0.029 min-1·µM-1) was lower than that for diclofenac (0.114 min-1·µM-1). Spectroscopic analysis showed that CYP105D7 binds to compactin with aKdvalue of 17.5 ± 3.6 µM. Molecular docking analysis was performed to build a possible binding model of compactin. Comparisons of different substrates with CYP105D7 were conclusively illustrated for the first time.
authors.affiliation: Ocean College, Zhejiang University, Dinghai, Zhoushan, Zhejiang 316021, P.R. China.
authors.forename: Qiuping
authors.initials: Q
authors.lastname: Yao
authors.affiliation: Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, P.R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.affiliation: Ocean College, Zhejiang University, Dinghai, Zhoushan, Zhejiang 316021, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Liu
authors.affiliation: Kitasato Institute for Life Sciences, Kitasato University, Kanagawa 252-0373, Japan.
authors.forename: Haruo
authors.initials: H
authors.lastname: Ikeda
authors.affiliation: Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Fushinobu
authors.affiliation: Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, P.R. China.
authors.forename: Shengying
authors.initials: S
authors.lastname: Li
authors.affiliation: Ocean College, Zhejiang University, Dinghai, Zhoushan, Zhejiang 316021, P.R. China.
authors.forename: Lian-Hua
authors.initials: LH
authors.lastname: Xu
mesh_terms: D001709:Biotechnology / Q000379:methods; D020134:Catalytic Domain; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D005288:Ferredoxins / Q000378:metabolism; D006900:Hydroxylation; D007700:Kinetics; D008148:Lovastatin / Q000031:analogs & derivatives* / Q000378:metabolism; D062105:Molecular Docking Simulation; D010084:Oxidation-Reduction; D017035:Pravastatin / Q000737:chemistry / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013302:Streptomyces / Q000201:enzymology / Q000378:metabolism*
chemical_list: D005288:Ferredoxins; C012258:mevastatin; C100137:putidaredoxin; D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin; D017035:Pravastatin
keywords: CYP105D7; Compactin; Streptomyces avermitilis; hydroxylation; pravastatin