Aromatic C-H bond hydroxylation by P450 peroxygenases: a facile colorimetric assay for monooxygenation activities of enzymes based on Russig's blue formation

• Published in: Journal of biological inorganic chemistry Vol. 15; no. 7; pp. 1109 - 1115
• Main Authors: Shoji, Osami, Wiese, Christian, Fujishiro, Takashi, Shirataki, Chikako, Wünsch, Bernhard, Watanabe, Yoshihito
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-09-2010; Springer-Verlag
• Subjects: Biochemistry; Biomedical and Life Sciences; Catalytic Domain; Colorimetric assay; Colorimetry - methods; cytochrome P-450; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - metabolism; Enzyme catalysis; Hydrocarbons, Aromatic - chemistry; Hydrocarbons, Aromatic - metabolism; hydrogen peroxide; Hydroxylation; Life Sciences; Microbiology; Models, Molecular; Molecular Structure; Naphthalenes - chemistry; Naphthalenes - metabolism; Original Paper; Peroxidases - chemistry; Peroxidases - metabolism; Protein Structure, Tertiary; Colorimetric assay; Hydrogen peroxide; Hydroxylation; Enzyme catalysis; Cytochrome P450
• ISSN: 0949-8257; 1432-1327
• DOI: 10.1007/s00775-010-0671-9

Aromatic C-H bond hydroxylation of 1-methoxynaphthalene was efficiently catalyzed by the substrate misrecognition system of the hydrogen peroxide dependent cytochrome P450BSβ (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.