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Metabolism of mono- and dichloro-dibenzo-p-dioxins by Phanerochaete chrysosporium cytochromes P450

  • Environmental Biotechnology
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Abstract

The white-rot fungus Phanerochaete chrysosporium possesses biodegradative capabilities of polychlorinated dibenzo-p-dioxins (PCDDs). One hundred twenty yeast clones expressing individual P450s of P. chrysosporum (PcCYPs), generated in our previous efforts, were screened for transformation of dioxin, and 40 positive clones were obtained. Of these clones, six clones showed metabolism of 2-chloro-dibenzo-p-dioxin, and a microsomal PcCYP designated as PcCYP11a3 showed much higher activity than any other PcCYPs. The turnover numbers of hydroxylation activities of PcCYP11a3 toward 1-MCDD (58 min−1) and 2-MCDD (13 min−1) are more than 200 times higher than those of previously reported PcCYP65a2. In addition, PcCYP11a3 catalyzes hydroxylation of 2,3-dichloro-dibenzo-p-dioxin. To our best knowledge, PcCYP11a3 has the highest activity toward PCDDs among the known CYPs derived from microorganisms. Although PcCYP11a3 showed no detectable activity toward 2,7-dichloro-dibenzo-p-dioxin and 2,3,7-trichloro-dibenzo-p-dioxin, PcCYP11a3 is promising as a template whose activity would be enhanced by site-directed mutagenesis.

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan

    Noriyuki Kasai, Shin-ichi Ikushiro, Raku Shinkyo, Kaori Yasuda & Toshiyuki Sakaki

  2. Bioresource Laboratories, Mercian Corp., Iwata-shi, Shizuoka, Japan

    Shinji Hirosue & Akira Arisawa

  3. Department of Forest and Forest Products Sciences, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Fukuoka, 812-8581, Japan

    Hirofumi Ichinose & Hiroyuki Wariishi

Authors
  1. Noriyuki Kasai
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  2. Shin-ichi Ikushiro
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  3. Raku Shinkyo
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  4. Kaori Yasuda
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  5. Shinji Hirosue
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  6. Akira Arisawa
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  7. Hirofumi Ichinose
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  8. Hiroyuki Wariishi
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  9. Toshiyuki Sakaki
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Corresponding author

Correspondence to Toshiyuki Sakaki.

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Supplemental Table 1

Primers for cDNAs cloning (A) and construction of expression plasmids (B) of the PcCYPs. (DOC 23 kb)

Table S2

Protein ID of PcCYPs showing DD metabolism (DOC 33 kb)

Fig. S1

Structure of a S. cerevisiae/E. coli shuttle vector pGYR (JPG 36.8 kb)

High resolution image file (EPS 656 kb)

ESM 4

Supplemental information (EPS 656 kb)

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Kasai, N., Ikushiro, Si., Shinkyo, R. et al. Metabolism of mono- and dichloro-dibenzo-p-dioxins by Phanerochaete chrysosporium cytochromes P450. Appl Microbiol Biotechnol 86, 773–780 (2010). https://doi.org/10.1007/s00253-009-2413-x

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  • DOI: https://doi.org/10.1007/s00253-009-2413-x

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