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Expression and Characterization of CYP51, the Ancient Sterol 14-demethylase Activity for Cytochromes P450 (CYP), in the White-Rot Fungus Phanerochaete chrysosporium

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Lipids

Abstract

Phanerochaete chrysosporium was the first fungal genome to exhibit more than one hundred cytochrome P450 (CYP) genes for a fungus within its genome (~150). It can metabolize lignocellulose and a wide range of environmental xenobiotics including many carcinogens and pollutants where cytochromes P450 may be involved. In the present paper we describe the heterologous expression and characterization of an ancestral CYP form, sterol 14α-demethylase (CYP51—EC1.14.13.70), from this organism. CYP51 was cloned from a cDNA library and expressed in both Escherichia coli, where it exhibited high affinity for azole antifungals, and Saccharomyces cerevisiae. Proof of function was observed by complementation of a conditional knock-down mutant of yeast CYP51. The CYP51 gene was found to be 1,956 bases long and contained 7 exons and 6 introns coding for a polypeptide 550 amino acids long (62 kDa). The CYP51 protein exhibited high affinity (k d 0.25–0.45 μM) for azole antifungal compounds.

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Acknowledgments

We are grateful to the Biotechnology and Biological Science Research Council of the UK for support.

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Authors and Affiliations

  1. Institute of Life Science, School of Medicine, Swansea University, Singleton Park, Swansea, SA2 8PP, UK

    Andrew Warrilow, Cynthia Ugochukwu, David Lamb, Diane Kelly & Steven Kelly

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  1. Andrew Warrilow
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  2. Cynthia Ugochukwu
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  3. David Lamb
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  4. Diane Kelly
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  5. Steven Kelly
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Corresponding author

Correspondence to Steven Kelly.

Additional information

P. chrysosporium CYP51 Accession Number: GenBank FJ174578.

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Warrilow, A., Ugochukwu, C., Lamb, D. et al. Expression and Characterization of CYP51, the Ancient Sterol 14-demethylase Activity for Cytochromes P450 (CYP), in the White-Rot Fungus Phanerochaete chrysosporium . Lipids 43, 1143–1153 (2008). https://doi.org/10.1007/s11745-008-3239-5

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  • DOI: https://doi.org/10.1007/s11745-008-3239-5

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