Skip to main content
Log in

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

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • 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 Trameteshirsuta. Appl Environ Microbiol 66:3357–3362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Abbott TA, Wicklow DT (1984) Degradation of lignin by Cyathus species. Appl Environ Microbiol 47:585–587

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arora DS, Sandhu DK (1985) Laccase production and wood degradation by white rot fungus Daedalea flavida. Enzyme Microb Technol 7:405–408

    Article  CAS  Google Scholar 

  • 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 72:248–254

    CAS  PubMed  Google Scholar 

  • Chivukula M, Renganathan V (1995) Phenolic azo dye oxidation by laccase from Pyricularia oryzae. Appl Environ Microbiol 61:4374–4377

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dhawan S, Lal R, Kuhad RC (2003) Ethidium bromide stimulated hyper laccase production from bird's nest fungus Cyathus bulleri. Lett Appl Microbiol 36:64–67

    Article  CAS  PubMed  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  • 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 62:1151–1158

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 10.8

    Google Scholar 

  • 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 242:329–341

    Article  CAS  PubMed  Google Scholar 

  • 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 64:2788–2793

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hünig S, Balli H, Conrad H, Schott A (1964) Ber zweistufige Redoxsyteme, III Polarographie von 2,2′ Azinenaromatischer Heterocyclen. Liebigs Ann Chem 676:52–65

    Article  Google Scholar 

  • Kundu SS, Kundu S, Prasad DN, Jakhmola RC (1988) Lignin biodegradation to improve digestibilities of straws. Int J Anim Sci 3:1–16

    CAS  Google Scholar 

  • 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 13:378–384

    Article  CAS  Google Scholar 

  • 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 277:37663–37669

    Article  CAS  PubMed  Google Scholar 

  • 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 29:575–579

    Article  CAS  Google Scholar 

  • Ronne H (1995) Glucose repression in fungi. Trends Genet 11:12–17

    Article  CAS  PubMed  Google Scholar 

  • Saxena A, Kuhad RC, Saxena RK, Gupta R (1994) Production and characterisation of a xylanase from Cyathus stercoreus. World J Microbiol Biotechnol 10:293–295

    Article  CAS  PubMed  Google Scholar 

  • 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 15:1185–1188

    Article  CAS  Google Scholar 

  • 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 27:100–107

    Article  CAS  PubMed  Google Scholar 

  • 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 44:872–885

    Article  CAS  Google Scholar 

  • Swamy J, Ramsay JA (1999) The evaluation of white rot fungi in the decoloration of textile dyes. Enzyme Microb Technol 24:130–137

    Article  CAS  Google Scholar 

  • Thakker G, Evans CS, Rao KK (1992) Purification and characterization of laccase from Monocillium indicum Saxena. Appl Microbiol Biotechnol 37:321–327

    Article  CAS  Google Scholar 

  • 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 81:2280–2284

    Article  Google Scholar 

  • Vasdev K, Kuhad RC (1994a) Decolorization of polyR-478 (polyvinylamine sulfonate anthrapyridone) by Cyathus bulleri. Folia Microbiol (Praha) 39:61–64

    Article  CAS  Google Scholar 

  • Vasdev K, Kuhad RC (1994b) Induction of laccase production in Cyathusbulleri under shaking and static culture conditions. Folia Microbiol (Praha) 39:326–330

    Article  CAS  Google Scholar 

  • 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 42:684–693

    Article  CAS  PubMed  Google Scholar 

  • Wood DA (1980) Production, purification and properties of extracellular laccase of Agaricus bisporus. J Gen Microbiol 117:327–338

    CAS  Google Scholar 

  • Xu F (1999) Laccase. In: Flickinger MC, Drew SW (eds) Encyclopedia of bioprocess technology: fermentation, biocatalysis and bioseparation, vol 3. Wiley, New York, pp 1545–1554

    Google Scholar 

  • Zollinger H (1987) Colour chemistry-synthesis, properties of organic dyes and pigments VCH, New York, pp 92–100

Download references

Acknowledgements

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

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to S. Mishra.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Salony, Mishra, S. & Bisaria, V.S. Production and characterization of laccase from Cyathus bulleri and its use in decolourization of recalcitrant textile dyes. Appl Microbiol Biotechnol 71, 646–653 (2006). https://doi.org/10.1007/s00253-005-0206-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-005-0206-4

Keywords

Navigation