Abstract
A cDNA encoding for a laccase was isolated from the white-rot fungus Lenzites gibbosa by RT-PCR and expressed in the Pichia pastoris. The laccase native signal peptide efficiently directed the secretion of the recombinant laccase in an active form. Factors influencing laccase expression, such as pH, cultivation temperature, copper concentration and methanol concentration, were optimized. The recombinant enzyme was purified to electrophoretic homogeneity, and was estimated to have a MW of ~61.5 kDa. The purified enzyme behaved similarly to the native laccase produced by L. gibbosa and efficiently decolorized Alizarin Red, Neutral Red, Congo Red and Crystal Violet, without the addition of redox mediators. The decolorization capacity of this recombinant enzyme suggests that it could be a useful biocatalyst for the treatment of dye-containing effluents. This study is the first report on the synthetic dye decolorization by a recombinant L. gibbosa laccase.
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References
Baldrian P (2006) Fungal laccases-occurrence and properties. FEMS Microbiol Rev 30:215–242
Camarero S, Ibarra D, Martínez MJ, Martínez AT (2005) Lignin-derived compounds as efficient laccase mediators for decolorization of different types of recalcitrant dyes. Appl Environ Microbiol 71:1775–1784
Chen H (2006) Recent advances in azo dye degrading enzyme research. Curr Protein Pept Sci 7:101–111
Claus H, Faber G, König H (2002) Redox-mediated decolorization of synthetic dyes by fungal laccases. Appl Microbiol Biotechnol 59:672–678
Couto SR, Herrera JLT (2006) Industrial and biotechnological applications of laccases: a review. Biotechnol Adv 24:500–513
Dubé E, Shareck F, Hurtubise Y, Daneault C, Beauregard M (2008) Homologous cloning, expression, and characterization of a laccase from Streptomyces coelicolor and enzymatic decolourisation of an indigo dye. Appl Microbiol Biotechnol 79:597–603
Eggert C, Temp U, Eriksson KE (1996) The ligninolytic system of the white-rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl Environ Microbiol 62:1151–1158
Faraco V, Ercole C, Festa G, Giardina P, Piscitelli A, Sannia G (2008) Heterologous expression of heterodimeric laccase from Pleurotus ostreatus in Kluyveromyces lactis. Appl Microbiol Biotechnol 77:1329–1335
Garavaglia S, Cambria MT, Miglio M, Ragusa S, Iacobazzi V, Palmieri F, D’Ambrosio C, Scaloni A, Rizzi M (2004) The structure of Rigidoporus lignosus Laccase containing a full complement of copper ions, reveals an asymmetrical arrangement for the T3 copper pair. J Mol Biol 342:1519–1531
Guo M, Lu F, Liu M, Li T, Pu J, Wang N, Liang P, Zhang C (2008) Purification of recombinant laccase from Trametes versicolor in Pichia methanolica and its use for the decolorization of anthraquinone dye. Biotechnol Lett 30:2091–2096
Hong F, Meinander NQ, Jönsson LJ (2002) Fermentation strategies for improved heterologous expression of laccase in Pichia pastoris. Biotechnol Bioeng 79:438–449
Hoshida H, Fujita T, Murata K, Kubo K, Akada R (2005) Copper-dependent production of a Pycnoporus coccineus extracellular laccase in Aspergillus oryzae and Saccharomyces cerevisiae. Biosci Biotechnol Biochem 69:1090–1097
Khatri NK, Hoffmann F (2006) Impact of methanol concentration on secreted protein production in oxygenlimited cultures of recombinant Pichia pastoris. Biotechnol Bioeng 93:871–879
Lorenzo M, Diego MM, Sanroman A (2006) Effect of heavy metals on the production of several laccase isoenzymes by Trametes versicolor and on their ability to decolourise dyes. Chemosphere 63:912–917
Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM (2005) Heterologous protein production using the Pichia pastoris expression system. Yeast 22:249–270
Madzak C, Mimmi MC, Caminade E, Brault A, Baumberger S, Briozzo P, Mougin C, Jolivalt C (2006) Shifting the optimal pH of activity for a laccase from the fungus Trametes versicolor by structure-based mutagenesis. Protein Eng Des Sel 19:77–84
O’Callaghan J, O’Brien MM, McClean K, Dobson ADW (2002) Optimization of the expression of a Trametes versicolor laccase gene in Pichia pastoris. J Ind Microbiol Biotechnol 29:55–59
Ong E, Pollock WBR, Smith M (1997) Cloning and sequence analysis of two laccase complementary DNAs from the ligninolytic basidiomycete Trametes versicolor. Gene 196:113–119
Otterbein L, Record E, Longhi S, Asther M, Moukha S (2000) Molecular cloning of the cDNA encoding laccase from Pycnoporus cinnabarinus I-937 and expression in Pichia pastoris. Eur J Biochem 267:1619–1625
Paola G (2010) Laccases: a never-ending story. Cell Mol Life Sci 67:369–385
Riva S (2006) Laccases: blue enzymes for green chemistry. Trends Biotechnol 24:219–226
Soares GMB, Costa-Ferreira M, de Amorim MTP (2001) Decolorization of an anthraquinone-type dye using a laccase formulation. Bioresour Technol. 79:171–177
Soden DM, O’Callaghan J, Dobson ADW (2002) Molecular cloning of a laccase isozyme gene from Pleurotus sajor-caju and expression in the heterologous Pichia pastoris host. Microbiology 148:4003–4014
Téllez-Jurado A, Arana-Cuenca A, González Becerra AE, Viniegra-González G, Loera O (2006) Expression of a heterologous laccase by Aspergillus niger cultured by solid-state and submerged fermentations. Enzyme Microb Technol 38:665–669
Wesenberg D, Kyriakides I, Agathos SN (2003) Whiterot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol Adv 22:161–187
Yan HB, Chi YJ (2013) Cloning and characterization of two novel manganese peroxidase genes from the white-rot basidiomycete Lenzites gibbosa. Curr Microbiol 23:224–247
Zumárraga M, Bulter T, Shleev S, Polaina J, Martínez-Arias A, Plou FJ, Ballesteros A, Alcalde M (2007) In vitro evolution of a fungal laccase in high concentrations of organic cosolvents. Chem Biol 14:1052–1064
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This study was financially supported by National Natural Science Foundation of China (No. 30671700, “Study on cloning, characterization and expression of manganese peroxidase genes from white-rot fungi”), which is gratefully acknowledged.
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Zheng, M., Chi, Y., Yi, H. et al. Decolorization of Alizarin Red and other synthetic dyes by a recombinant laccase from Pichia pastoris . Biotechnol Lett 36, 39–45 (2014). https://doi.org/10.1007/s10529-013-1323-2
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DOI: https://doi.org/10.1007/s10529-013-1323-2