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Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products

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Abstract

Two novel GH3 family thermostable β-glucosidases from the filamentous fungus Chaetomium atrobrunneum (CEL3a and CEL3b) were expressed in Trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. Both enzymes were active over a wide range of pH as compared to Neurospora crassa β-glucosidase GH3-3, which was also expressed in T. reesei and purified. The optimum temperature of both C. atrobrunneum enzymes was around 60 °C at pH 5, and both enzymes had better thermal and pH stability and higher resistance to metallic compounds and to glucose inhibition than GH3-3. They also showed higher activity against oligosaccharides composed of glucose units and linked with β-1,4-glycosidic bonds and moreover, had higher affinity for cellotriose over cellobiose. In hydrolysis tests against Avicel cellulose and steam-exploded sugarcane bagasse, performed at 45 °C, particularly the CEL3a enzyme performed similarly to N. crassa GH3-3 β-glucosidase. Taking into account the thermal stability of the C. atrobrunneum β-glucosidases, they both represent promising alternatives as enzyme mixture components for improved cellulose saccharification at elevated temperatures.

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Acknowledgments

This work was supported by jointly by Finland and Brazil in sustainable energy (Academy of Finland-CNPq), decision number 271146 of Academy of Finland, National Council of Scientific and Technological Development (CNPq), Brazil (490249/2012-4 Bilateral agreements/Call number 30/2012 - CNPq/AKA FINLÂNDIA), and the Foundation for Research of São Paulo State (FAPESP), Brazil. The fungal genome sequencing was supported from the project ‘Metagenome’ funded by the Finnish Funding Agency for Innovation, decision number 40148/07. We thank Dr Merja Oja for sequence search, Dr Martina Andberg for assistance with the protein purification and for providing some of the substrates, and Dr. Teun Boekhout, The Fungal Biodiversity Centre (CBS), the Netherlands, for providing the C. atrobrunneum CBS 269.91 strain.

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Authors and Affiliations

  1. VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, 02044, Espoo, Finland

    Ana C. Colabardini, Mari Valkonen, Anne Huuskonen, Matti Siika-aho, Anu Koivula & Markku Saloheimo

  2. Departamento de Ciências Farmacêuticas, Faculdade de Ciências Farmacêuticas de Ribeirão Prêto, Universidade de São Paulo, Ribeirão Prêto, São Paulo, Brazil

    Gustavo H. Goldman

  3. Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Caixa Postal 6192, Campinas, São Paulo, 13083-970, Brazil

    Gustavo H. Goldman

Authors
  1. Ana C. Colabardini
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  2. Mari Valkonen
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Correspondence to Ana C. Colabardini.

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Colabardini, A.C., Valkonen, M., Huuskonen, A. et al. Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products. Mol Biotechnol 58, 821–831 (2016). https://doi.org/10.1007/s12033-016-9981-7

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  • DOI: https://doi.org/10.1007/s12033-016-9981-7

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