Applied Microbiology and Biotechnology

, Volume 63, Issue 5, pp 560–563

Purification and characterization of laccase from the white-rot fungus Daedalea quercina and decolorization of synthetic dyes by the enzyme

Original Paper


The white-rot fungus Daedalea quercina produced the ligninolytic enzymes laccase and Mn-dependent peroxidase. Laccase was purified using anionexchange and size-exclusion chromatographies. SDS-PAGE showed the purified laccase to be a monomeric protein of 69 kDa (71 kDa using gel filtration) with an isoelectric point near 3.0. The optimum pH for activity was bellow 2.0 for 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (Km=38 μM), 4.0 for 2,6-dimethoxyphenol (Km=48 μM), 4.5 for guaiacol (Km=93 μM) and 7.0 for syringaldazine (Km=131 μM). The temperature optimum was between 60 and 70 °C depending on the pH and buffer used. The enzyme was stable up to 45 °C, and stability was higher at alkaline pH. Enzyme activity was increased by the addition of Cu2+ and inhibited by Mn2+, sodium azide, dithiothreitol, and cysteine. Laccase from Daedalea quercina was able to decolorize the synthetic dyes Chicago sky blue, poly B-411, remazol brilliant blue R, trypan blue and reactive blue 2.


  1. Baldrian P (2003) Interactions of heavy metals with white-rot fungi. Enzyme Microb Technol 32:78–91CrossRefGoogle Scholar
  2. Baldrian P, Gabriel J (2003) Lignocellulose degradation by Pleurotus ostreatus in the presence of cadmium. FEMS Microbiol Lett 220:235–240CrossRefPubMedGoogle Scholar
  3. Chet I, Trojanowski J, Hüttermann A (1985) Decolourisation of the dye poly B-411 and its correlation with lignin degradation by fungi. Microbios Lett 29:37–43Google Scholar
  4. Eggert C, Temp U, Eriksson K-EL (1996) The lignolytic system of the white rot fungus Pycnoporus cinnabarinus: Purification and characterization of the laccase. Appl Environ Microbiol 62:1151–1158PubMedGoogle Scholar
  5. Hatakka A (1994) Lignin-modifing enzymes from selected white-rot fungi: production and role in lignin degradation. FEMS Microbiol Rev 13:125–135Google Scholar
  6. Katagiri N, Tsutsumi Y, Nishida T (1995) Correlation of brightening with cumulative enzyme activity related to lignin biodegradation during biobleaching of kraft pulp by white rot fungi in the solid-state fermentation system. Appl Environ Microbiol 61:617–622PubMedGoogle Scholar
  7. Leonowicz A, Matuszewska A, Luterek J, Ziegenhagen D, Wojtas-Wasilewska M, Cho N-S, Hofrichter M, Rogalski J (1999) Biodegradation of lignin by white rot fungi. Fungal Genet Biol 27:175–185CrossRefPubMedGoogle Scholar
  8. Lo SC, Ho YS, Buswell JA (2001) Effect of phenolic monomers on the production of laccases by the edible mushroom Pleurotus sajor-caju, and partial characterization of a major laccase component. Mycologia 93:413–421Google Scholar
  9. Munoz C, Guillen F, Martinez AT, Martinez MJ. Induction and characterization of laccase in the ligninolytic fungus Pleurotus eryngii. Curr Microbiol 34:1–5PubMedGoogle Scholar
  10. Nagai M, Sato T, Watanabe H, Saito K, Kawata M, Enei H (2002) Purification and characterization of an extracellular laccase from the edible mushroom Lentinula edodes, and decolorization of chemically different dyes. Appl Microbiol Biotechnol 60:327–335CrossRefPubMedGoogle Scholar
  11. Ngo TT, Lenhoff HM (1980) A sensitive and versatile chromogenic assay for peroxidase and peroxidase-coupled reactions. Anal Biochem 105:389–397PubMedGoogle Scholar
  12. Niku-Paavola ML, Raaska L Itävaara M (1990) Detection of white rot fungi by a non-toxic stain. Mycol Res 94:27–31Google Scholar
  13. Novotný Č, Vyas BRM, Erbanová P, Kubátová A, and Šašek V (1997) Removal of PCBs by various white rot fungi in liquid cultures. Folia Microbiol 42:136–140Google Scholar
  14. Paszczynski A, Crawford RL (2000) Recent advances in the use of fungi in environmental remediation and biotechnology. Soil Biochem 10:379–422Google Scholar
  15. Schliephake K, Mainwaring DE, Lonergan GT, Jones IK, Baker WL (2000) Transformation and degradation of the disazo dye Chicago Sky Blue by a purified laccase from Pycnoporus cinnabarinus. Enzyme Microb Technol 27:100–107PubMedGoogle Scholar
  16. Thurston CF (1994) The structure and function of fungal laccases. Microbiology 140:19–26Google Scholar
  17. Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Methods Enzymol 161B:238–248Google Scholar
  18. Xu F (1997) Effects of redox potential and hydroxide inhibition on the pH activity profile of fungal laccases. J Biol Chem 272:924–928PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Laboratory of Biochemistry of Wood-rotting FungiInstitute of Microbiology AS CRPrague 4Czech Republic

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