Advertisement

Applied Microbiology and Biotechnology

, Volume 71, Issue 5, pp 646–653 | Cite as

Production and characterization of laccase from Cyathus bulleri and its use in decolourization of recalcitrant textile dyes

  • Salony
  • S. Mishra
  • V. S. Bisaria
Biotechnologically Relevant Enzymes and Proteins

Abstract

Many fungi (particularly the white rot) are well suited for treatment of a broad range of textile dye effluents due to the versatility of the lignin-degrading enzymes produced by them. We have investigated decolourization of a number of recalcitrant reactive azo and acid dyes using the culture filtrate and purified laccase from the fungus Cyathus bulleri. For this, the enzyme was purified from the culture filtrate to a high specific activity of 4,022 IU mg−1 protein, produced under optimized carbon, nitrogen and C/N ratio with induction by 2,6-dimethylaniline. The protein was characterized as a monomer of 58±5.0 kDa with carbohydrate content of 16% and was found to contain all three Cu(II) centres. The three internal peptide sequences showed sequence identity (80–92%) with laccases of a number of white rot fungi. Substrate specificity indicated highest catalytic efficiency (k cat/K M) on guaiacol followed by 2,2′-azino-bis(3-ethylthiazoline-6-sulfonic acid) (ABTS). Decolourization of a number of reactive azo and acid dyes was seen with the culture filtrate of the fungus containing predominantly laccase. In spite of no observable effect of purified laccase on other dyes, the ability to decolourize these was achieved in the presence of the redox mediator ABTS, with 50% decolourization in 0.5–5.4 days.

Keywords

Culture Filtrate Guaiacol Laccase Activity Laccase Production Veratryl Alcohol 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was partially supported by a grant from the Department of Biotechnology, Government of India, to one of the authors (S.M.).

References

  1. Abadulla E, Tzanov T, Costa S, Robra K-H, Cavaco-Paulo A, Gubitz GM (2000) Decolorization and detoxification of textile dyes with a laccase from Trametes hirsuta. Appl Environ Microbiol 66:3357–3362CrossRefPubMedGoogle Scholar
  2. Abbott TA, Wicklow DT (1984) Degradation of lignin by Cyathus species. Appl Environ Microbiol 47:585–587Google Scholar
  3. Arora DS, Sandhu DK (1985) Laccase production and wood degradation by white rot fungus Daedalea flavida. Enzyme Microb Technol 7:405–408CrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  5. Chivukula M, Renganathan V (1995) Phenolic azo dye oxidation by laccase from Pyricularia oryzae. Appl Environ Microbiol 61:4374–4377Google Scholar
  6. Dhawan S, Lal R, Kuhad RC (2003) Ethidium bromide stimulated hyper laccase production from bird's nest fungus Cyathus bulleri. Lett Appl Microbiol 36:64–67CrossRefPubMedGoogle Scholar
  7. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  8. Eggert C, Temp U, Eriksson KEL (1996) The ligninolytic system of white rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl Environ Microbiol 62:1151–1158PubMedGoogle Scholar
  9. Gallagher S, Winston SE, Fuller SA, Hurrell JGR (1989) Analysis of proteins. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology. Greene/Wiley, New York, unit 10.8Google Scholar
  10. Glenn JK, Gold MH (1985) Purification and characterization of an extracellular Mn(II)-dependant peroxidase from the lignin-degrading basidiomycete Phanerochaete chrysosporium. Arch Biochem Biophys 242:329–341CrossRefPubMedGoogle Scholar
  11. Heinfling A, Martinex MJ, Martinex AT, Bergbaner M, Szewyk U (1998) Transformation of industrial dyes by manganese peroxidase from Bjerkandera adusta and Pleurotus eryngii in a manganese-independent reaction. Appl Environ Microbiol 64:2788–2793PubMedGoogle Scholar
  12. Hünig S, Balli H, Conrad H, Schott A (1964) Ber zweistufige Redoxsyteme, III Polarographie von 2,2′ Azinenaromatischer Heterocyclen. Liebigs Ann Chem 676:52–65CrossRefGoogle Scholar
  13. Kundu SS, Kundu S, Prasad DN, Jakhmola RC (1988) Lignin biodegradation to improve digestibilities of straws. Int J Anim Sci 3:1–16Google Scholar
  14. Paszczynski A, Pasti MB, Goszczynski S, Crawford DL, Crawford RL (1991) New approach to improve degradation of recalcitrant azo dyes by Streptomyces sp. and Phanerochaete chrysosporium. Enzyme Microb Technol 13:378–384CrossRefGoogle Scholar
  15. Piontek K, Antorini M, Choinowski T (2002) Crystal structure of a laccase from the fungus Trametes versicolor at 1.90-Å resolution containing a full complement of coppers. J Biol Chem 277:37663–37669CrossRefPubMedGoogle Scholar
  16. Robinson T, Chandran B, Nigam P (2001) Studies on the production of enzymes by white-rot fungi for the decolorization of textile dyes. Enzyme Microb Technol 29:575–579CrossRefGoogle Scholar
  17. Ronne H (1995) Glucose repression in fungi. Trends Genet 11:12–17CrossRefPubMedGoogle Scholar
  18. Saxena A, Kuhad RC, Saxena RK, Gupta R (1994) Production and characterisation of a xylanase from Cyathus stercoreus. World J Microbiol Biotechnol 10:293–295CrossRefGoogle Scholar
  19. Schliephake K, Lonergan GT, Jones CL, Mainwaring DE (1993) Decolourisation of a pigment plant effluent by Pycnoporus cinnabarinus in a packed‐bed bioreactor. Biotechnol Lett 15:1185–1188CrossRefGoogle Scholar
  20. 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 27:100–107CrossRefPubMedGoogle Scholar
  21. Sethuraman A, Akin DE, Eisele JG, Eriksson KEL (1998b) Effect of aromatic compounds on growth and ligninolytic enzyme production of two white rot fungi: Ceriporiopsis subvermispora and Cyathus stercoreus. Can J Microbiol 44:872–885CrossRefGoogle Scholar
  22. Swamy J, Ramsay JA (1999) The evaluation of white rot fungi in the decoloration of textile dyes. Enzyme Microb Technol 24:130–137CrossRefGoogle Scholar
  23. Thakker G, Evans CS, Rao KK (1992) Purification and characterization of laccase from Monocillium indicum Saxena. Appl Microbiol Biotechnol 37:321–327CrossRefGoogle Scholar
  24. Tien M, Kirk TK (1983) Lignin degrading from Phanerochaete chrysosporium: purification, characterization and catalytic properties of unique H2O2-requiring oxygenase. Proc Natl Acad Sci U S A 81:2280–2284CrossRefGoogle Scholar
  25. Vasdev K, Kuhad RC (1994a) Decolorization of polyR-478 (polyvinylamine sulfonate anthrapyridone) by Cyathus bulleri. Folia Microbiol (Praha) 39:61–64CrossRefGoogle Scholar
  26. Vasdev K, Kuhad RC (1994b) Induction of laccase production in Cyathus bulleri under shaking and static culture conditions. Folia Microbiol (Praha) 39:326–330CrossRefGoogle Scholar
  27. Vasdev K, Dhawan S, Kapoor RK, Kuhad RC (2005) Biochemical characterization and molecular evidence of a laccase from the bird's nest fungus Cyathus bulleri. Fungal Genet Biol 42:684–693CrossRefPubMedGoogle Scholar
  28. Wood DA (1980) Production, purification and properties of extracellular laccase of Agaricus bisporus. J Gen Microbiol 117:327–338Google Scholar
  29. Xu F (1999) Laccase. In: Flickinger MC, Drew SW (eds) Encyclopedia of bioprocess technology: fermentation, biocatalysis and bioseparation, vol 3. Wiley, New York, pp 1545–1554Google Scholar
  30. Zollinger H (1987) Colour chemistry-synthesis, properties of organic dyes and pigments VCH, New York, pp 92–100Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiNew DelhiIndia

Personalised recommendations