Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Westermeier R (2006) Sensitive, quantitative, and fast modifications for Coomassie Blue staining of polyacrylamide gels. Proteomics 6(Suppl 2):61–64
Chial HJ, Thompson HB, Splittgerber AG (1993) A spectral study of the charge forms of Coomassie blue G. Anal Biochem 209(2):258–266
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):144–145
Kurien BT, Scofield RH (2012) Accelerated Coomassie Blue staining and destaining of SDS-PAGE gels with application of heat. Methods Mol Biol 869:471–479
Sreeramulu G, Singh NK (1995) Destaining of Coomassie Brilliant Blue R-250-stained polyacrylamide gels with sodium chloride solutions. Electrophoresis 16(3):362–365
Majek P et al (2013) Improved coomassie blue dye-based fast staining protocol for proteins separated by SDS-PAGE. PLoS One 8(11):e81696
Yang J et al (2016) Destaining of Coomassie Brilliant Blue R-250-stained polyacrylamide gels with fungal laccase. Anal Biochem 493:27–29
Baldrian P (2006) Fungal laccases—occurrence and properties. FEMS Microbiol Rev 30(2):215–242
Pogni R et al (2015) Spectroscopic and computational characterization of laccases and their substrate radical intermediates. Cell Mol Life Sci 72(5):885–896
Hu MR et al (2009) Laccase-mediator system in the decolorization of different types of recalcitrant dyes. J Ind Microbiol Biotechnol 36(1):45–51
Singh Arora D, Kumar Sharma R (2010) Ligninolytic fungal laccases and their biotechnological applications. Appl Biochem Biotechnol 160(6):1760–1788
Chandra R, Chowdhary P (2015) Properties of bacterial laccases and their application in bioremediation of industrial wastes. Environ Sci Process Impacts 17(2):326–342
Yang J et al (2014) Purification and characterization of a novel laccase from Cerrena sp. HYB07 with dye decolorizing ability. PLoS One 9(10):e110834
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kumar, G. (2018). Fungal Laccase Efficiently Destains Coomassie Brilliant Blue-R-250 Stained Polyacrylamide Gels. In: Kurien, B., Scofield, R. (eds) Protein Gel Detection and Imaging. Methods in Molecular Biology, vol 1853. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8745-0_28
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8745-0_28
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8744-3
Online ISBN: 978-1-4939-8745-0
eBook Packages: Springer Protocols