Cytochrome P450rm from Rhodotorula minuta catalyzes 4-hydroxylation of benzoate.
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.
Fukuda
H
H
Department of Applied Microbial Technology, Kumamoto Institute of Technology.
Nakamura
K
K
Sukita
E
E
Ogawa
T
T
Fujii
T
T
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
D000272:Adipates; D001565:Benzoates; C027316:3-oxoadipic acid; D010649:Phenylalanine; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D050565:Benzoate 4-Monooxygenase