Abstract
A recombinant laccase from Trametes versicolor in Pichia methanolica was produced constitutively in a defined medium. The recombinant laccase was purified using ultrafiltration, anion-exchange chromatography, and gel filtration. The molecular weight of the purified laccase was estimated as 64 kDa by SDS-PAGE. The purified recombinant laccase decolorized more than 90% of Remazol Brilliant Blue R (RBBR) initially at 80 mg l−1 after 16 h at 45°C and pH 5 when 25 U laccase ml−1 was used. The purified recombinant laccase could efficiently decolorize RBBR without additional redox mediators.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bulter T, Alcalde M, Sieber V, Meinhold P, Schlachtbauer C, Arnold FH (2003) Functional expression of a fungal laccase in Saccharomyces cerevisiae by directed evolution. Appl Environ Microbiol 69:987–995
Couto SR, Herrera JLT (2006) Industrial and biotechnological applications of laccases: a review. Biotechnol Adv 24:500–513
Guo YJ (2005) Experimental techniques in protein electrophoresis, 2nd edn. Science Press, Beijing, pp 53–56
Guo M, Lu FP, Pu J, Bai DQ, Du LX (2005) Molecular cloning of the cDNA encoding laccase from Trametes versicolor and heterologous expression in Pichia methanolica. Appl Microbiol Biotechnol 69:178–183
Guo M, Lu FP, Du LX, Pu J, Bai DQ (2006) Optimization of the expression of a laccase gene from Trametes versicolor in Pichia methanolica. Appl Microbiol Biotechnol 71:848–852
Hong F, Meinander NQ, Jönsson LJ (2002) Fermentation strategies for improved heterologous expression of laccase in Pichia pastoris. Biotechnol Bioeng 79:438–449
Hong YZ, Xiao YZ, Zhou HM, Fang W, Zhang M, Wang J, Wu LJ, Yu ZL (2006) Expression of a laccase cDNA from Trametes sp. AH28-2 in Pichia pastoris and mutagenesis of transformants by nitrogen ion implantation. FEMS Microbiol Lett 258:96–101
Jönsson L, Saloheimo M, Penttila M (1997) Laccase from the white-rot fungus Trametes versicolor: cDNA cloning of lcc1 and expression in Pichia pastoris. Curr Genet 32:425–430
Koschorreck K, Richer SM, Ene AB, Roduner E, Schmid RD, Urlacher VB (2008) Cloning and characterization of a new laccase from Bacillus licheniformis catalyzing dimerization of phenolic acids. Appl Microbiol Biotechnol 79:217–224
Liu W, Chao Y, Liu S, Bao H, Qian S (2003) Molecular cloning and characterization of a laccase gene from the basidiomycete Fome lignisus and expression in Pichia pastoris. Appl Microbiol Biotechnol 63:174–181
Morozova OV, Shumakovich GP, Shleev SV, Yaropolov YI (2007) Laccase-mediator systems and their applications: a review. Appl Biochem Microbiol 43:523–535
Schliephake K, Lonergan GT (1996) Laccase variation during dye depolarization in a 200 L packed-bed bioreactor. Biotechnol Lett 18:881–886
Soares GMB, Costa-Ferreira M, de Pessoa Amorim MT (2001) Decolorization of an anthraquinone-type dye using a laccase formulation. Bioresour Technol 79:171–177
Wesenberg D, Kyriakides I, Agathos SN (2003) White-rot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol Adv 22:161–187
Wong YX, Yu J (1999) Laccase-catalyzed decolorization of synthetic dyes. Water Res 33:3512–3520
Acknowledgements
The authors would like to acknowledge the financial support from Tianjin Natural Sciences Foundation of China (07JCZDJC03100), Tianjin Municipal Science and Technology Commission.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Guo, M., Lu, F., Liu, M. et al. Purification of recombinant laccase from Trametes versicolor in Pichia methanolica and its use for the decolorization of anthraquinone dye. Biotechnol Lett 30, 2091–2096 (2008). https://doi.org/10.1007/s10529-008-9817-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-008-9817-z