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Effect of glycosylation on the biochemical properties of β-xylosidases from Aspergillus versicolor

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Abstract

Aspergillus versicolor grown on xylan or xylose produces two β-xylosidases with differences in biochemical properties and degree of glycosylation. We investigated the alterations in the biochemical properties of these β-xylosidases after deglycosylation with Endo-H or PNGase F. After deglycosylation, both enzymes migrated faster in PAGE or SDS-PAGE exhibiting the same Rf. Temperature optimum of xylan-induced and xylose-induced β-xylosidases was 45°C and 40°C, respectively, and 35°C after deglycosylation. The xylan-induced enzyme was more active at acidic pH. After deglycosylation, both enzymes had the same pH optimum of 6.0. Thermal resistance at 55°C showed half-life of 15 min and 9 min for xylose- and xylan-induced enzymes, respectively. After deglycosylation, both enzymes exhibited half-lives of 7.5 min. Native enzymes exhibited different responses to ions, while deglycosylated enzymes exhibited identical responses. Limited proteolysis yielded similar polypeptide profiles for the deglycosylated enzymes, suggesting a common polypeptide core with differential glycosylation apparently responsible for their biochemical and biophysical differences.

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References

  • Andrade, S.V., M.L.T.M. Polizeli, H.F. Terenzi, and J.A. Jorge. 2004. Effect of carbon source on the biochemical properties of β-xylosidases produced by Aspergillus versicolor. Proc. Biochem. 39, 1931–1938.

    Article  CAS  Google Scholar 

  • Bütner, R. and R. Bode. 1992. Purification and characterization of β-xylosidase activities from the yeast Arxula adeninivorans. J. Basic Microbiol. 32, 159–166.

    Article  Google Scholar 

  • Carmona, E.C., M.R. Brocheto-Braga, A.A. Pizzirani-Kleiner, and J.A. Jorge. 1998. Purification and biochemical characterization of an endoxylanase from Aspergillus versicolor. FEMS Microbiol. Lett. 166, 311–315.

    Article  CAS  Google Scholar 

  • Carmona, E.C., A.A. Pizzirani-Kleiner, R.T.R. Monteiro, and J.A. Jorge. 1997. Xilanase production by Aspergillus versicolor. J. Basic Microbiol. 37, 387–394.

    Article  Google Scholar 

  • Davis, B.J. 1964. Disc electrophoresis. II. Method and application to human serum proteins. Ann. N. Y. Acad. Sci. 121, 404–427.

    Article  PubMed  CAS  Google Scholar 

  • Dey, N.B., P. Bounelis, T.A. Fritz, D.M. Bedewell, and R.B. Marchase. 1994. The glycosylation of phosphoglumutase is modulated by carbon source and heat shock in Saccharomyces cerevisiae. J. Biol. Chem. 269, 27143–27148.

    PubMed  CAS  Google Scholar 

  • Dobberstein, J. and C.C. Emeis. 1991. Purification and characterization of β-xylosidase from Aureobasidium pullulans. Appl. Microbiol. Biotechnol. 35, 210–215.

    Article  CAS  Google Scholar 

  • Harrison, M.J., A.S. Nouwens, D.R. Jardine, N.E. Zachara, A.A. Gooley, and H. Nevalainen. 1998. Modified glycosylation of cellobiohydrolase I from a high cellulase producing mutant strain of Trichoderma reesei. Eur. J. Biochem. 256, 119–127.

    Article  PubMed  CAS  Google Scholar 

  • Hui, J.P.M., P. Lanthier, T.C. White, S.G. McHugh, M. Yaguchi, R. Roy, and P. Tribault. 2001. Characterization of cellobiohydrolase I (Cel7A) glycoforms from extracts of Trichoderma reesei using capillary isoelectric focusing and electrospray mass spectrometry. J. Chrom. B 752, 349–368.

    Article  CAS  Google Scholar 

  • Kern, G., N. Schülke, F.X. Schmid, and R. Jaenicke. 1992. Stability, quaternary structure, and folding of internal, external, and core-glycosylated invertase from yeast. Protein Sci. 1, 120–131.

    Article  PubMed  CAS  Google Scholar 

  • Klarskov, K., K.K. Piens, J. Stahlberg, P.B. Hoj, J.M. Van Beeumen, and M. Claeyssens. 1997. Cellobiohydrolase I from Trichoederma reesei: Identification of an active-site nucelophile and additional information on sequence including the glycosylation pattern for the core protein. Carbohydr. Res. 304, 143–154.

    Article  PubMed  CAS  Google Scholar 

  • Kristufek, D., S. Zellinger, and C.P. Kubicek. 1995. Regulation of β-ylosidase formation by xylose in Trichoderma reesei. Appl. Microbiol. Biotechnol. 42, 713–717.

    Article  CAS  Google Scholar 

  • Kulkarni, N., A. Shendye, and M. Rao. 1999. Molecular and biotechnolological aspects of xylanase. FEMS Microbiol. Rev. 23, 411–456.

    Article  PubMed  CAS  Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227, 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lige, B., S. Ma, and R.B. Van Huystee. 2001. The effects of the site-directed removal of N-glycosylation from cationic peanut peroxidase on its function. Arch. Biochem. Biophys. 386, 17–24.

    Article  PubMed  CAS  Google Scholar 

  • Lowry, O.H., N.J. Rosebrough, A.L. Farr, and R.J. Randall. 1951. Protein measurement with folin phenol reagent. J. Biol. Chem. 193, 265–275.

    PubMed  CAS  Google Scholar 

  • Mandels, G.R. 1953. Localization of carbohydrases at surface of fungus spores by acid treatment. Exp. Cell Res. 5, 48–55.

    Article  PubMed  CAS  Google Scholar 

  • Maras, M., A. De Bruyn, J. Schraml, P. Herdewijn, M. Claeyssens, W. Fiers, and R. Contreras. 1997. Structural characterization of N-linked oligosaccharides from cellobiohydrolase secreted by filamentous fungi Trichoderma reesei Rut-C-30. Eur. J. Biochem. 245, 617–625.

    Article  PubMed  CAS  Google Scholar 

  • Meldgaard, M. and I. Svendsen. 1994. Different effects of N-glycosylation on the thermostability of highly homologous bacterial (1,3−1,4)-β-glucanases secreted from yeast. Microbiology 140, 159–166.

    Article  PubMed  CAS  Google Scholar 

  • Polizeli, M.L.T.M., A.C. Rizzati, R. Monti, H.F. Terenzi, J.A. Jorge, and D.S. Amorin. 2005. Xylanases from fungi: properties and industrial applications. Appl. Microbiol. Biotechnol. 67, 577–591.

    Article  PubMed  CAS  Google Scholar 

  • Poutanen, K., P. Ratto, and L. Viikari. 1987. Evaluation of different microbial xylanolytic systems. J. Biotechnol. 6, 49–60.

    Article  CAS  Google Scholar 

  • Stals, I., K. Sandra, B. Devreese, J. Van Beeumen, and M. Claeyssens. 2004a. Factors influencing glycosylation of Trichoderma reesei cellulases. II: N-glycosylation of Cel7A core protein isolated from different strains. Glycobiology 14, 725–737.

    Article  PubMed  CAS  Google Scholar 

  • Stals, I., K. Sandra, S. Geysens, R. Contreras, J. Van Beeumen, and M. Claeyssens. 2004b. Factors influencing glycosylation of Trichoderma reesei cellulases. I: Postsecretorial changes of the O- and N-glycosylation pattern of Cel7A. Glycobiology 14, 713–724.

    Article  PubMed  CAS  Google Scholar 

  • Tsuji T., K. Yamamoto, and T. Tochikura. 1990. Formation of deglycosylated α-L-fucosidase by endo-β-N-acetylglucosaminidase in Fusarium oxysporum. Appl. Environ. Microbiol. 56, 928–933.

    PubMed  CAS  Google Scholar 

  • Varki, A. 1993. Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3, 97–130.

    Article  PubMed  CAS  Google Scholar 

  • Venturi, L.L., M.L.T.M. Polizeli, H.F. Terenzi, R.P.M. Furriel, and J.A. Jorge. 2002. Extracellular β-D-glucosidase from Chaetomium thermophilum var. coprophilum: production, purification and some biochemical properties. J. Basic Microbiol. 42, 55–66.

    Article  PubMed  CAS  Google Scholar 

  • Vogel, H.J.A. 1956. A convenient growth medium for Neurospora (medium N). Microbial Genet. Bull. 37, 387–394.

    Google Scholar 

  • Wong, K.K.Y., L.U.L. Tan, and J.N. Saddler. 1998. Multiplicity of β-1,4-xylanases in microorganisms: functions and applications. Microbiol. Rev. 52, 305–317.

    Google Scholar 

  • Wray, W., T. Boulikas, V.P. Wray, and R. Hancock. 1981. Silver staining of proteins in polyacrylamide gels. Anal. Biochem. 118, 197–203.

    Article  PubMed  CAS  Google Scholar 

  • Zanoelo, F.F., M.L.T.M. Polizeli, H.F. Terenzi, and J.A. Jorge. 2004. Purification and biochemical properties of a thermostable xylose-tolerant β-D-xylosidase from Scytalidium thermophilum. J. Ind. Microbiol. Biotechnol. 31, 170–176.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Departamentos de Biologia e Química, Universidade de São Paulo, Avenida Bandeirantes, 3900, CEP 14040-901, Ribeirão Preto, SP, Brasil

    Alexandre Favarin Somera, Marita Gimenez Pereira, Luis Henrique Souza Guimarães, Maria de Lourdes Teixeira de Moraes Polizeli, Héctor Francisco Terenzi & João Atílio Jorge

  2. Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes, 3900, CEP 14040-901, Ribeirão Preto, SP, Brasil

    Rosa Prazeres Melo Furriel

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  1. Alexandre Favarin Somera
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  2. Marita Gimenez Pereira
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  3. Luis Henrique Souza Guimarães
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  4. Maria de Lourdes Teixeira de Moraes Polizeli
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  5. Héctor Francisco Terenzi
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  7. João Atílio Jorge
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Correspondence to João Atílio Jorge.

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Somera, A.F., Pereira, M.G., Souza Guimarães, L.H. et al. Effect of glycosylation on the biochemical properties of β-xylosidases from Aspergillus versicolor . J Microbiol. 47, 270–276 (2009). https://doi.org/10.1007/s12275-008-0286-9

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