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Purification and biochemical characterization of glucose–cellobiose-tolerant cellulases from Scytalidium thermophilum

Folia Microbiologica volume 58, pages 561–568 (2013)Cite this article

Abstract

Two cellulases from Scytalidium thermophilum were purified and characterized, exhibiting tolerance to glucose and cellobiose. Characterization of purified cellulases I and II by mass spectrometry revealed primary structure similarities with an exoglucanase and an endoglucanase, respectively. Molecular masses were 51.2 and 45.6 kDa for cellulases I and II, respectively, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Cellulases I and II exhibited isoelectric points of 6.2 and 6.9 and saccharide contents of 11 and 93 %, respectively. Optima of temperature and pH were 60–65 °C and 4.0 for purified cellulase I and 65 °C and 6.5 for purified cellulase II. Both cellulases maintained total CMCase activity after 60 min at 60 °C. Cysteine, Mn2+, dithiotreitol and ß-mercaptoethanol-stimulated cellulases I and II. The tolerance to cellulose hydrolysis products and the high thermal stabilities of Scytalidium cellulases suggest good potential for industrial applications.

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Acknowledgments

This work was supported by a grant from CNPq. J.C.R.S. received a Ph.D. scholarship from FAPESP, and M.L.T.M.P., R.P.M.F., and J.A.J. are research fellows of CNPq. This work was part of the Doctoral thesis of J.C.R.S (Dept. Bioquímica-FMRP-USP).

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Affiliations

  1. Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil

    Jean Carlos Rodrigues Silva & José Carlos Santos Salgado

  2. Instituto Federal de Educação, Ciência e Tecnologia de São Paulo, Matão, São Paulo, Brazil

    Jean Carlos Rodrigues Silva

  3. Departamento de Biologia e Química, Faculdade de Filosofia, Ciências de Letras de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes, 3900, 14040-901, Ribeirão Preto, São Paulo, Brazil

    Luis Henrique Souza Guimarães, Rosa Prazeres Melo Furriel, Maria Lourdes T. M. Polizeli & João Atilio Jorge

  4. Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    José César Rosa

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  1. Jean Carlos Rodrigues Silva
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  2. Luis Henrique Souza Guimarães
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  3. José Carlos Santos Salgado
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  4. Rosa Prazeres Melo Furriel
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  5. Maria Lourdes T. M. Polizeli
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  7. João Atilio Jorge
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Correspondence to João Atilio Jorge.

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Silva, J.C.R., Guimarães, L.H.S., Salgado, J.C.S. et al. Purification and biochemical characterization of glucose–cellobiose-tolerant cellulases from Scytalidium thermophilum . Folia Microbiol 58, 561–568 (2013). https://doi.org/10.1007/s12223-013-0245-7

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  • DOI: https://doi.org/10.1007/s12223-013-0245-7

Keywords

  • Cellulase
  • Cellobiose
  • Sodium Acetate Buffer
  • CMCase Activity
  • Saccharide Content