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

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An Erratum to this article was published on 09 June 2010

Abstract

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.

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Fig. 1
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Abbreviations

P450:

Cytochrome P450

HPLC:

High performance liquid chromatography

HRP:

Horseradish peroxidase

MALDI:

Matrix-assisted laser deposition ionization

TOF:

Time-of-flight

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Acknowledgements

This work was supported by a Grant-in-Aid for Scientific Research (S) to Y.W. from the Ministry of Education, Culture, Sports, Science, and Technology (Japan) and the International Research Training Group (DFG and JSPS) “Complex Functional Systems in Chemistry” (Münster and Nagoya). We thank Prof. Isamu Matsunaga for his kind gift of the expression system of P450BSβ and its mutants. T.F. is supported by JSPS Research Fellowships for Young Scientists.

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Authors and Affiliations

  1. Department of Pharmaceutical and Medicinal Chemistry, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan

    Osami Shoji, Takashi Fujishiro & Chikako Shirataki

  2. Research Center for Materials Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan

    Yoshihito Watanabe

  3. Department of Pharmaceutical and Medicinal Chemistry, University of Münster, Hittorfstr. 58-62, 48149, Münster, Germany

    Christian Wiese & Bernhard Wünsch

Authors
  1. Osami Shoji
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  2. Christian Wiese
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  3. Takashi Fujishiro
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  4. Chikako Shirataki
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  6. Yoshihito Watanabe
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Corresponding author

Correspondence to Yoshihito Watanabe.

Additional information

An erratum to this article can be found at http://dx.doi.org/10.1007/s00775-010-0679-1

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Shoji, O., Wiese, C., Fujishiro, T. et al. Aromatic C–H bond hydroxylation by P450 peroxygenases: a facile colorimetric assay for monooxygenation activities of enzymes based on Russig’s blue formation. J Biol Inorg Chem 15, 1109–1115 (2010). https://doi.org/10.1007/s00775-010-0671-9

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  • DOI: https://doi.org/10.1007/s00775-010-0671-9

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