Abstract
The brown-rot basidiomycete Fomitopsis palustris is known to degrade crystalline cellulose (Avicel) and produce three major cellulases, exoglucanases, endoglucanases, and β-glucosidases. A novel β-glucosidase designated as Cel3A was identified from F. palustris grown at the expense of Avicel. The deduced amino acid sequence of Cel3A showed high homology with those of other fungal β-glucosidases that belong to glycosyl hydrolase (GH) family 3. The sequence analysis also indicated that Cel3A contains the N- and C-terminal domains of GH family 3 and Asp-209 was conserved as a catalytic nucleophile. The cloned gene was successfully expressed in the yeast Pichia pastoris and the recombinant protein exhibited β-glucosidase activity with cellobiose and some degree of thermostability. Considering the size and sequence of the protein, the β-glucosidase identified in this study is different from the protein purified directly from F. palustris in the previous study. Our results suggest that the fungus possesses at least two β-glucosidase genes.
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References
Cantarel, B.L., P.M. Coutinho, C. Rancurel, T. Bernard, V. Lombard, and B. Henrissat. 2009. The Carbohydrate-Active EnZymes database (CAZy): An expert resource for glycogenomics. Nucleic Acids Res. 37 (Database issue), D233–D238.
Clamp, M., J. Cuff, S.M. Searle, and G.J. Barton. 2004. The Jalview Java alignment editor. Bioinformatics 20, 426–427.
Cohen, R., M.R. Suzuki, and K.E. Hammel. 2005. Processive endoglucanase active in crystalline cellulose hydrolysis by the brown rot basidiomycete Gloeophyllum trabeum. Appl. Environ. Microbiol. 71, 2412–2417.
Dashtban, M., H. Schraft, and Q. Wensheng. 2009. Fungal bioconversion of lignocellulosic residues; Opportunities & perspectives. Int. J. Biol. Sci. 5, 578–595.
Henrissat, B. and G. Davies. 1997. Structural and sequence-based classification of glycoside hydrolases. Curr. Opin. Struct. Biol. 7, 637–644.
Ito, S. 1997. Alkaline cellulases from alkaliphilic Bacillus: Enzymatic properties, genetics, and application to detergents. Extremophiles 1, 61–66.
Joo, A.R., M. Jeya, K.M. Lee, W.I. Shim, J.S. Kim, I.W. Kim, Y.S. Kim, D.K. Oh, P. Gunasekaran, and J.K. Lee. 2009. Purification and characterization of a β-1,4-glucosidase from a newly isolated strain of Fomitopsis pinicola. Appl. Microbiol. Biotechnol. 83, 285–294.
Kerem, Z., K.A. Jensen, and K.E. Hammel. 1999. Biodegradative mechanism of the brown rot basidiomycete Gleophyllum trabeum: Evidence for an extracellular hydroquinone driven Fenton reaction. FEBS Lett. 446, 49–54.
Lynd, L.R., P.J. Weimer, W.H. van Zyl, and I.S. Pretorius. 2002. Microbial cellulose utilization: Fundamentals and biotechnology. Microbiol. Mol. Biol. Rev. 66, 506–577.
Mach, R.L. and S. Zeilinger. 2003. Regulation of gene expression in industrial fungi: Trichoderma. Appl. Microbiol. Biotechnol. 60, 515–522.
Martinez, D., L.F. Larrondo, N. Putnam, M.D. Gelpke, K. Huang, J. Chapman, K.G. Helfenbein, and et al. 2004. Genome sequence of the lignocellulose degrading fungus Phanerochaete chrysosporium strain RP78. Nature Biotechnol. 22, 695–700.
Nelson, N. 1944. A photometric adaptation of the Somogyi method for the determination of glucose. J. Biol. Chem. 153, 375–380.
Ouyang, J., M. Yan, D. Kong, and L. Xu. 2006. A complete protein pattern of cellulase and hemicellulase genes in the filamentous fungus Trichoderma reesei. Biotechnol. J. 1, 1266–1274.
Pozzo, T., J.L. Pasten, E.N. Karlsson, and D.T. Logan. 2010. Structural and functional analyses of β-glucosidase 3B from Thermotoga neapolitana: a thermostable three-domain represent-tative of glycoside hydrolase 3. J. Mol. Biol. 397, 724–739.
Rabinovich, M.L., M.S. Melnick, and A.V. Bolobova. 2002. The structure and mechanism of action of cellulolytic enzymes. Biochemistry (Moscow). 67, 1026–1050.
Riou, C., J.M. Salmon, M.J. Vallier, Z. Gunata, and P. Barre. 1998. Purification, characterization, and substrate specificity of a novel highly glucose-tolerant beta-glucosidase from Aspergillus oryzae. Appl. Environ. Microbiol. 64, 3607–3614.
Somogyi, M. 1952. Note on sugar determination. J. Biol. Chem. 195, 19–23.
Song, B.C., K.Y. Kim, J.J. Yoon, S.H. Shim, K. Lee, Y.S. Kim, Y.K. Kim, and C.J. Cha. 2008. Functional analysis of a gene encoding endoglucanase that belongs to glycosyl hydrolase family 12 from the brown-rot basidiomycete Fomitopsis palustris. J. Microbiol. Biotechnol. 18, 404–409.
Steenbakkers, P.J.M., H.R. Harhangi, M.W. Bosscher, M.M.C. van der Hooft, J.T. Keltjens, C. van der Drift, G.D. Vogels, and H.J.M. Op Den Camp. 2003. β-Glucosidase in cellulosome of the anaerobic fungus Piromyces sp. strain E2 is a family 3 glycoside hydrolase. Biochem. J. 370, 963–970.
Varghese, J.N., M. Hrmova, and G.B. Fincher. 1999. Threedimensional structure of a barley β-d-glucanexohydrolase, a family 3 glycosyl hydrolase. Structure 7, 179–190.
Vuong, T.V. and D.B. Wilson. 2010. Glycoside hydrolases: catalytic base/nucleophile diversity. Biotechnol. Bioeng. 107, 195–205.
Wymelenberg, A.V., G. Sabat, D. Martinez, A.S. Rajangam, T.T. Teeri, J. Gaskell, P.J. Kersten, and D. Cullen. 2005. The Phanerochaete chrysosporium secretome: database predictions and initial mass spectrometry peptide identifications in cellulose-grown medium. J. Biotechnol. 118, 17–34.
Yoon, J.J., C.J. Cha, Y.S. Kim, and W. Kim. 2008a. Degradation of cellulose by the major endoglucanase produced from the brownrot fungus Fomitopsis pinicola. Biotechnol. Lett. 30, 1373–1378.
Yoon, J.J., C.J. Cha, Y.S. Kim, D.W. Son, and Y.K. Kim. 2007. The brown-rot basidiomycete Fomitopsis palustris has the endoglucanases capable of degrading microcrystalline cellulose. J. Microbiol. Biotechnol. 17, 800–805.
Yoon, J.J., K. Igarashi, T. Kajisa, and M. Samejima. 2006. Purification, identification and molecular cloning of glycoside hydrolase family 15 glucoamylase from the brown-rot basidiomycete Fomitopsis palustris. FEMS. Microbiol. Lett. 259, 288–294.
Yoon, J.J. and Y.K. Kim. 2005. Degradation of crystalline cellulose by the brown-rot basidiomycete Fomitopsis palustris. J. Microbiol. 43, 487–492.
Yoon, J.J., K.Y. Kim, and C.J. Cha. 2008b. Purification and characterization of thermostable-β-glucosidase from the brownrot basidiomycete Fomitopsis palustris grown on microcrystalline cellulose. J. Microbiol. 46, 51–55.
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Ji, HW., Cha, CJ. Identification and functional analysis of a gene encoding β-glucosidase from the brown-rot basidiomycete Fomitopsis palustris . J Microbiol. 48, 808–813 (2010). https://doi.org/10.1007/s12275-010-0482-2
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DOI: https://doi.org/10.1007/s12275-010-0482-2