Applied Biochemistry and Biotechnology

, Volume 169, Issue 1, pp 100–109 | Cite as

Extracellular Enzymes of the White-Rot Fungus Fomes fomentarius and Purification of 1,4-β-Glucosidase

  • Tomáš Větrovský
  • Petr Baldrian
  • Jiří GabrielEmail author


Production of the lignocellulose-degrading enzymes endo-1,4-β-glucanase, 1,4-β-glucosidase, cellobiohydrolase, endo-1,4-β-xylanase, 1,4-β-xylosidase, Mn peroxidase, and laccase was characterized in a common wood-rotting fungus Fomes fomentarius, a species able to efficiently decompose dead wood, and compared to the production in eight other fungal species. The main aim of this study was to characterize the 1,4-β-glucosidase produced by F. fomentarius that was produced in high quantities in liquid stationary culture (25.9 U ml−1), at least threefold compared to other saprotrophic basidiomycetes, such as Rhodocollybia butyracea, Hypholoma fasciculare, Irpex lacteus, Fomitopsis pinicola, Pleurotus ostreatus, Piptoporus betulinus, and Gymnopus sp. (between 0.7 and 7.9 U ml−1). The 1,4-β-glucosidase enzyme was purified to electrophoretic homogeneity by both anion-exchange and size-exclusion chromatography. A single 1,4-β-glucosidase was found to have an apparent molecular mass of 58 kDa and a pI of 6.7. The enzyme exhibited high thermotolerance with an optimum temperature of 60 °C. Maximal activity was found in the pH range of 4.5–5.0, and K M and V max values were 62 μM and 15.8 μmol min−1 l−1, respectively, when p-nitrophenylglucoside was used as a substrate. The enzyme was competitively inhibited by glucose with a K i of 3.37 mM. The enzyme also acted on p-nitrophenylxyloside, p-nitrophenylcellobioside, p-nitrophenylgalactoside, and p-nitrophenylmannoside with optimal pH values of 6.0, 3.5, 5.0, and 4.0–6.0, respectively. The combination of relatively low molecular mass and low K M value make the 1,4-β-glucosidase a promising enzyme for biotechnological applications.


Cellulose 1,4-β-glucosidase Glycosyl hydrolase Saprotrophic basidiomycetes Wood-rotting fungi Fomes fomentarius 



This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic (OC08050, LA10001).


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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Tomáš Větrovský
    • 1
  • Petr Baldrian
    • 1
  • Jiří Gabriel
    • 1
    Email author
  1. 1.Laboratory of Environmental MicrobiologyInstitute of Microbiology of the ASCRPraha 4Czech Republic

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