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Characterisation of cellobiose dehydrogenases from the white-rot fungi Trametes pubescens and Trametes villosa

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Abstract

Cellobiose dehydrogenase (CDH) is an extracellular haemoflavoenzyme that is produced by a number of wood-degrading and phytopathogenic fungi and it has a proposed role in the early events of lignocellulose degradation and wood colonisation. In the presence of a suitable electron acceptor, e.g. 2,6-dichloro-indophenol, cytochrome c, or metal ions, CDH oxidises cellobiose to cellobionolactone. When screening 11 different Trametes spp. for the formation of CDH activity, all the strains investigated were found to secrete significant amounts of CDH when cultivated on a cellulose-containing medium. Amongst others, Trametes pubescens and Trametes villosa were identified as excellent, not-yet-described, producer strains of this enzyme activity that has various potential applications in biotechnology. CDH from both strains was purified to apparent homogeneity and subsequently characterised. Both monomeric enzymes have a molecular mass of approximately 90 kDa (gel filtration) and a pI value of 4.2–4.4. The best substrates are cellobiose and cellooligosaccharides; additionally, lactose, thiocellobiose, and xylobiose are efficiently oxidised. Glucose and maltose are poor substrates. The preferred substrate is cellobiose with a K m value of 0.21 mM and a k cat value of 22 s−1 for CDH from T. pubescens; the corresponding values for the T. villosa enzyme are 0.21 mM and 24 s−1, respectively. Both enzymes showed very high activity with one-electron acceptors such as ferricenium, ferricyanide, or the azino-bis-(3-ethyl-benzthiazolin-6-sulfonic acid) cation radical.

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Acknowledgements

We sincerely thank Hansjörg Prillinger for some of the fungal strains. This work was supported by the Austrian Science Fund (Fonds zur Förderung der wissenschaftlichen Forschung), project FWF P14537-B11.

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Authors and Affiliations

  1. Division of Biochemical Engineering, Institute of Food Technology, BOKU–University of Natural Resources and Applied Life Sciences Vienna, Muthgasse 18, 1190, Vienna, Austria

    R. Ludwig, A. Salamon, J. Varga, M. Zámocky, C. K. Peterbauer, K. D. Kulbe & D. Haltrich

  2. Institute of Molecular Biology, Slovak Academy of Sciences, 845 51, Bratislava, Slovak Republic

    M. Zámocky

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  1. R. Ludwig
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  2. A. Salamon
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  3. J. Varga
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  4. M. Zámocky
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  5. C. K. Peterbauer
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  6. K. D. Kulbe
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  7. D. Haltrich
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Correspondence to D. Haltrich.

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Ludwig, R., Salamon, A., Varga, J. et al. Characterisation of cellobiose dehydrogenases from the white-rot fungi Trametes pubescens and Trametes villosa . Appl Microbiol Biotechnol 64, 213–222 (2004). https://doi.org/10.1007/s00253-003-1501-6

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  • DOI: https://doi.org/10.1007/s00253-003-1501-6

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