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Molecular breeding of white rot fungus Pleurotus ostreatus by homologous expression of its versatile peroxidase MnP2

  • Environmental Biotechnology
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Abstract

Using a DNA-mediated transformation technique, a molecular breeding approach to isolate Pleurotus ostreatus strains with enhanced productivity of its versatile peroxidase MnP2 was conducted. A recombinant mnp2 construct under the control of P. ostreatus sdi1 expression signals was introduced into the wild-type P. ostreatus strain by cotransformation with a carboxin-resistant marker plasmid. A total of 32 transformants containing the recombinant mnp2 sequence were isolated in a screening with specific amplification by PCR. Productivity of MnP2 in the recombinants was evaluated by the decolorization ability of Poly R-478 on agar plates in the absence of Mn2+. Recombinant P. ostreatus strains with elevated manganese peroxidase (MnP) productivity were successfully isolated. One of the recombinants, TM2-10, was demonstrated to secrete recombinant MnP2 predominantly on a synthetic medium containing 15 mM ammonium oxalate, which was confirmed by reverse transcription PCR (RT-PCR) and isozyme profile analysis using anion-exchange chromatography. The benzo[a]pyrene-removing activity by fungal treatment was also analyzed using the isolated recombinant strains.

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Acknowledgements

This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, Japan.

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Authors and Affiliations

  1. Research Institute for Sustainable Humanosphere, Kyoto University, Gokasho Uji, Kyoto, 611-0011, Japan

    Takahisa Tsukihara, Yoichi Honda, Takahito Watanabe & Takashi Watanabe

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  1. Takahisa Tsukihara
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  2. Yoichi Honda
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  3. Takahito Watanabe
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  4. Takashi Watanabe
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Correspondence to Yoichi Honda.

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Tsukihara, T., Honda, Y., Watanabe, T. et al. Molecular breeding of white rot fungus Pleurotus ostreatus by homologous expression of its versatile peroxidase MnP2. Appl Microbiol Biotechnol 71, 114–120 (2006). https://doi.org/10.1007/s00253-005-0136-1

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  • DOI: https://doi.org/10.1007/s00253-005-0136-1

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