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Cytochrome P450 monooxygenases involved in anthracene metabolism by the white-rot basidiomycete Phanerochaete chrysosporium

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Abstract

Cytochrome P450 monooxygenases (P450s) involved in anthracene metabolism by the white-rot basidiomycete Phanerochaete chrysosporium were identified by comprehensive screening of both catalytic potentials and transcriptomic profiling. Functional screening of P. chrysosporium P450s (PcCYPs) revealed that 14 PcCYP species catalyze stepwise conversion of anthracene to anthraquinone via intermediate formation of anthrone. Moreover, transcriptomic profiling explored using a complementary DNA microarray system demonstrated that 12 PcCYPs are up-regulated in response to exogenous addition of anthracene. Among the up-regulated PcCYPs, five species showed catalytic activity against anthracene. Based upon both catalytic and transcriptional properties, these five species are most likely to play major roles in anthracene metabolic processes in vivo. Thus, the combination of functional screening and a microarray system may provide a novel strategy for obtaining a thorough understanding of the catalytic functions and biological impacts of PcCYPs.

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Acknowledgment

This research was supported in part by a Grant-in-Aid (#21688013) for Young Scientists (A) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) (to H. I.).

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Authors and Affiliations

  1. Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Nomathemba Loice Chigu, Sinji Hirosue, Chie Nakamura, Hiroshi Teramoto, Hirofumi Ichinose & Hiroyuki Wariishi

  2. Bioresource Laboratories, Mercian Corporations, Iwata, 438-0078, Japan

    Sinji Hirosue

  3. Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Hiroyuki Wariishi

  4. Innovation Center for Medical Redox Navigation, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Hiroyuki Wariishi

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  1. Nomathemba Loice Chigu
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  2. Sinji Hirosue
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  3. Chie Nakamura
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  5. Hirofumi Ichinose
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Correspondence to Hiroyuki Wariishi.

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Table S1

Microarray expression profiling of PcCYP genes induced by anthrone (DOC 42 kb)

Figure S1

cDNA and deduced amino acid sequences of PcCYPs responsive for anthracene metabolism (DOC 86 kb)

Figure S2

Conversion of anthracene by purified LiP (JPG 4.36 KB)

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Chigu, N.L., Hirosue, S., Nakamura, C. et al. Cytochrome P450 monooxygenases involved in anthracene metabolism by the white-rot basidiomycete Phanerochaete chrysosporium . Appl Microbiol Biotechnol 87, 1907–1916 (2010). https://doi.org/10.1007/s00253-010-2616-1

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