title: Hydroxylation activity of P450 BM-3 mutant F87V towards aromatic compounds and its application to the synthesis of hydroquinone derivatives from phenolic compounds.
abstract: Cytochrome P450 BM-3 from Bacillus megaterium is a fatty acid hydroxylase exhibiting selectivity for long-chain substrates (12-20 carbons). Replacement of Phe87 in P450 BM-3 by Val (F87V) greatly increased its activity towards a variety of aromatic and phenolic compounds. The apparent initial reaction rates of F87V as to benzothiophene, indan, 2,6-dichlorophenol, and 2-(benzyloxy)phenol were 227, 204, 129, and 385 nmol min(-1) nmol(-1) P450, which are 220-, 66-, 99-, and 963-fold those of the wild type, respectively. These results indicate that Phe87 plays a critical role in the control of the substrate specificity of P450 BM-3. Furthermore, F87V catalyzed regioselective hydroxylation at the para position of various phenolic compounds. In particular, F87V showed high activity as to the hydroxylation of 2-(benzyloxy)phenol to 2-(benzyloxy)hydroquinone. With F87V as the catalyst, 0.71 mg ml(-1) 2-(benzyloxy)hydroquinone was produced from 1.0 mg ml(-1) 2-(benzyloxy)phenol in 4 h, with a molar yield of 66%.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
authors.forename: Woro Triarsi
authors.initials: WT
authors.lastname: Sulistyaningdyah
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
authors.forename: Qing-Shan
authors.initials: QS
authors.lastname: Li
authors.forename: Chiharu
authors.initials: C
authors.lastname: Maeda
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yano
authors.forename: Rolf D
authors.initials: RD
authors.lastname: Schmid
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
mesh_terms: D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D006873:Hydroquinones / Q000378:metabolism*; D006900:Hydroxylation; D009154:Mutation; D009249:NADP / Q000378:metabolism; D010636:Phenols / Q000737:chemistry / Q000378:metabolism*; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism*
chemical_list: D006873:Hydroquinones; D010636:Phenols; D011084:Polycyclic Aromatic Hydrocarbons; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; C031927:hydroquinone