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Expression of laccase gene lcc1 in Coprinopsis cinerea under control of various basidiomycetous promoters

  • Applied Genetics and Molecular Biotechnology
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Abstract

Coprinopsis cinerea laccase gene lcc1 was expressed in this basidiomycete under naturally non-inductive conditions using various homologous and heterologous promoters. Laccase expression was achieved in solid and liquid media with promoter sequences from the C. cinerea tub1 gene, the Agaricus bisporus gpdII gene, the Lentinus edodes priA gene and the Schizophyllum commune Sc3 gene. As measured by enzyme activity in liquid cultures, a 277-bp gpdII promoter fragment, followed by a 423-bp priA fragment, was most efficient. A shorter priA sequence of 372 bp was inactive. tub1 promoter fragments were reasonably active, whereas the S. commune Sc3 promoter sequence was less active, in comparison. Irrespective of the promoter used, addition of copper to the medium increased enzymatic activities for highly active transformants by 10- to 50-fold and for less active transformants for 2- to 7-fold. The highest enzymatic activities (3 U/ml) were reached with the gpdII promoter in the presence of 0.1 mM CuSO4.

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Acknowledgements

We are very grateful to Han Wösten for the idea of using the S. commune Sc3 promoter and for its sequence. We thank Mojtaba Zomorrodi for technical assistance in peptide sequencing. The laboratory in Göttingen is funded by the Deutsche Bundesstiftung Umwelt.

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  1. Claire Burns

    Present address: Department of Biology, Indiana University, Bloomington, IN, 47405, USA

Authors and Affiliations

  1. Molecular Wood Biotechnology, Institute of Forest Botany, Georg-August-University Göttingen, Büsgenweg 2, 37077, Göttingen, Germany

    Sreedhar Kilaru, Patrik J. Hoegger, Andrzej Majcherczyk & Ursula Kües

  2. School of Biological Sciences, University of Bristol, Woodland Road, Bristol, BS8 1UG, UK

    Claire Burns, Andy Bailey & Gary D. Foster

  3. Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan

    Kazuo Shishido

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  1. Sreedhar Kilaru
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Kilaru, S., Hoegger, P.J., Majcherczyk, A. et al. Expression of laccase gene lcc1 in Coprinopsis cinerea under control of various basidiomycetous promoters. Appl Microbiol Biotechnol 71, 200–210 (2006). https://doi.org/10.1007/s00253-005-0128-1

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