title: Cytochrome P450rm from Rhodotorula minuta catalyzes 4-hydroxylation of benzoate.
abstract: Rhodotorula minuta, a red yeast, produces a cytochrome P450, tentatively named P450rm, catalyzing the formation of isobutene from isovalerate. We found that P450rm interacted with benzoate and the dissociation constant of P450rm for benzoate was 36 microM. A reconstituted system that consisted of purified P450rm and cytochrome P450 reductase catalyzed the 4-hydroxylation of benzoate in addition to the formation of isobutene; the turnover rate was approximately 40 nmol/min/nmol P450rm. The P450rm-monooxygenase system was specific for benzoate and did not catalyze hydroxylation of other aromatic carboxylates. Since only a benzoate 4-hydroxylase that requires tetrahydropteridine has been isolated to date, P450rm appears to be the first isolated cytochrome P450 that acts as a benzoate 4-hydroxylase. The P450rm-monooxygenase system in microsomes of R. minuta might function in the degradation of L-phenylalanine on the pathway to beta-ketoadipate.
authors.affiliation: Department of Applied Microbial Technology, Kumamoto Institute of Technology.
authors.forename: H
authors.initials: H
authors.lastname: Fukuda
authors.forename: K
authors.initials: K
authors.lastname: Nakamura
authors.forename: E
authors.initials: E
authors.lastname: Sukita
authors.forename: T
authors.initials: T
authors.lastname: Ogawa
authors.forename: T
authors.initials: T
authors.lastname: Fujii
mesh_terms: D000272:Adipates / Q000378:metabolism; D050565:Benzoate 4-Monooxygenase; D001565:Benzoates / Q000378:metabolism*; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006900:Hydroxylation; D008861:Microsomes / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism; D012248:Rhodotorula / Q000201:enzymology*; D013057:Spectrum Analysis; D013379:Substrate Specificity
chemical_list: D000272:Adipates; D001565:Benzoates; C027316:3-oxoadipic acid; D010649:Phenylalanine; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D050565:Benzoate 4-Monooxygenase