Fungal Laccase Efficiently Destains Coomassie Brilliant Blue-R-250 Stained Polyacrylamide Gels

  • Gaurav KumarEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1853)


Fungal laccase-mediated destaining of Coomassie Brilliant Blue-R-250 stained polyacrylamide electrophoretic gel is very sensitive and environment friendly. Fermenting of Cerrena sp. HYB07 yields a broth with laccase activity which when supplemented with distilled water and used as a destaining solution results in a clear gel background in 2 h at 37 °C. Also, separation of the laccase enzyme for efficient functioning is not required. The method utilizes gel destaining and process of decolorizing dye simultaneously with a detection sensitivity of up to 10 ng. This technique avoids the use of hazardous organic compounds, as the destaining solution and the destain solution could be discarded easily without any further treatment.

Key words

Fungal laccase Destaining of gel Sodium dodecyl sulfate–polyacrylamide gel electrophoresis Coomassie Brilliant Blue R-250 


  1. 1.
    Westermeier R (2006) Sensitive, quantitative, and fast modifications for Coomassie Blue staining of polyacrylamide gels. Proteomics 6(Suppl 2):61–64CrossRefPubMedGoogle Scholar
  2. 2.
    Chial HJ, Thompson HB, Splittgerber AG (1993) A spectral study of the charge forms of Coomassie blue G. Anal Biochem 209(2):258–266CrossRefPubMedGoogle Scholar
  3. 3.
    Meyer TS, Lamberts BL (1965) Use of coomassie brilliant blue R250 for the electrophoresis of microgram quantities of parotid saliva proteins on acrylamide-gel strips. Biochim Biophys Acta 107(1):144–145CrossRefPubMedGoogle Scholar
  4. 4.
    Kurien BT, Scofield RH (2012) Accelerated Coomassie Blue staining and destaining of SDS-PAGE gels with application of heat. Methods Mol Biol 869:471–479CrossRefPubMedGoogle Scholar
  5. 5.
    Sreeramulu G, Singh NK (1995) Destaining of Coomassie Brilliant Blue R-250-stained polyacrylamide gels with sodium chloride solutions. Electrophoresis 16(3):362–365CrossRefPubMedGoogle Scholar
  6. 6.
    Majek P et al (2013) Improved coomassie blue dye-based fast staining protocol for proteins separated by SDS-PAGE. PLoS One 8(11):e81696CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Yang J et al (2016) Destaining of Coomassie Brilliant Blue R-250-stained polyacrylamide gels with fungal laccase. Anal Biochem 493:27–29CrossRefPubMedGoogle Scholar
  8. 8.
    Baldrian P (2006) Fungal laccases—occurrence and properties. FEMS Microbiol Rev 30(2):215–242CrossRefGoogle Scholar
  9. 9.
    Pogni R et al (2015) Spectroscopic and computational characterization of laccases and their substrate radical intermediates. Cell Mol Life Sci 72(5):885–896CrossRefPubMedGoogle Scholar
  10. 10.
    Hu MR et al (2009) Laccase-mediator system in the decolorization of different types of recalcitrant dyes. J Ind Microbiol Biotechnol 36(1):45–51CrossRefPubMedGoogle Scholar
  11. 11.
    Singh Arora D, Kumar Sharma R (2010) Ligninolytic fungal laccases and their biotechnological applications. Appl Biochem Biotechnol 160(6):1760–1788CrossRefPubMedGoogle Scholar
  12. 12.
    Chandra R, Chowdhary P (2015) Properties of bacterial laccases and their application in bioremediation of industrial wastes. Environ Sci Process Impacts 17(2):326–342CrossRefPubMedGoogle Scholar
  13. 13.
    Yang J et al (2014) Purification and characterization of a novel laccase from Cerrena sp. HYB07 with dye decolorizing ability. PLoS One 9(10):e110834CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Oklahoma Medical Research Foundation, University of OklahomaOklahoma CityUSA

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