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Applied and Environmental Microbiology, September 2009, p. 5570-5580, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.02942-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Role of P450 Monooxygenases in the Degradation of the Endocrine-Disrupting Chemical Nonylphenol by the White Rot Fungus Phanerochaete chrysosporium{triangledown}

Venkataramanan Subramanian and Jagjit S. Yadav*

Environmental Genetics and Molecular Toxicology Division, Department of Environmental Health, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0056

Received 24 December 2008/ Accepted 14 June 2009

The white rot fungus Phanerochaete chrysosporium extensively degraded the endocrine disruptor chemical nonylphenol (NP; 100% of 100 ppm) in both nutrient-limited cultures and nutrient-sufficient cultures. The P450 enzyme inhibitor piperonyl butoxide caused significant inhibition (~75%) of the degradation activity in nutrient-rich malt extract (ME) cultures but no inhibition in defined low-nitrogen (LN) cultures, indicating an essential role of P450 monooxygenase(s) in NP degradation under nutrient-rich conditions. A genome-wide analysis using our custom-designed P450 microarray revealed significant induction of multiple P450 monooxygenase genes by NP: 18 genes were induced (2- to 195-fold) under nutrient-rich conditions, 17 genes were induced (2- to 6-fold) in LN cultures, and 3 were induced under both nutrient-rich and LN conditions. The P450 genes Pff 311b (corresponding to protein identification number [ID] 5852) and Pff 4a (protein ID 5001) showed extraordinarily high levels of induction (195- and 167-fold, respectively) in ME cultures. The P450 oxidoreductase (POR), glutathione S-transferase (gst), and cellulose metabolism genes were also induced in ME cultures. In contrast, certain metabolic genes, such as five of the peroxidase genes, showed partial downregulation by NP. This study provides the first evidence for the involvement of P450 enzymes in NP degradation by a white rot fungus and the first genome-wide identification of specific P450 genes responsive to an environmentally significant toxicant.

* Corresponding author. Mailing address: Environmental Genetics and Molecular Toxicology Division, Department of Environmental Health, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0056. Phone: (513) 558-4806. Fax: (513) 558-4397. E-mail: Jagjit.Yadav{at}uc.edu

{triangledown} Published ahead of print on 19 June 2009.

Applied and Environmental Microbiology, September 2009, p. 5570-5580, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.02942-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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