Aromatic C-H bond hydroxylation by P450 peroxygenases: a facile colorimetric assay for monooxygenation activities of enzymes based on Russig's blue formation.
Aromatic C-H bond hydroxylation of 1-methoxynaphthalene was efficiently catalyzed by the substrate misrecognition system of the hydrogen peroxide dependent cytochrome P450BSbeta (CYP152A1), which usually catalyzes hydroxylation of long-alkyl-chain fatty acids. Very importantly, the hydroxylation of 1-methoxynaphthalene can be monitored by a color change since the formation of 4-methoxy-1-naphthol was immediately followed by its further oxidation to yield Russig's blue. Russig's blue formation allows us to estimate the peroxygenation activity of enzymes without the use of high performance liquid chromatography, gas chromatography, and nuclear magnetic resonance measurements.
Shoji
Osami
O
Department of Chemistry, Graduate School of Science,Nagoya University, Furo-cho, Chikusa-ku,Nagoya 464-8602, Japan.
Wiese
Christian
C
Fujishiro
Takashi
T
Shirataki
Chikako
C
Wünsch
Bernhard
B
Watanabe
Yoshihito
Y
D020134:Catalytic Domain; D003124:Colorimetry / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D006841:Hydrocarbons, Aromatic / Q000737:chemistry* / Q000378:metabolism; D006900:Hydroxylation; D008958:Models, Molecular; D015394:Molecular Structure; D009281:Naphthalenes / Q000737:chemistry / Q000378:metabolism; D010544:Peroxidases / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D006841:Hydrocarbons, Aromatic; D009281:Naphthalenes; D003577:Cytochrome P-450 Enzyme System; C448759:1-methoxynaphthalene; D010544:Peroxidases; C437763:cytochrome P-450 CYP152A1 (Bacillus subtilis)