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The functional properties of a xyloglucanase (GH12) of Aspergillus terreus expressed in Aspergillus nidulans may increase performance of biomass degradation

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Abstract

Filamentous fungi are attractive hosts for heterologous protein expression due to their capacity to secrete large amounts of enzymes into the extracellular medium. Xyloglucanases, which specifically hydrolyze xyloglucan, have been recently applied in lignocellulosic biomass degradation and conversion in many other industrial processes. In this context, this work aimed to clone, express, and determine the functional properties of a recombinant xyloglucanase (AtXEG12) from Aspergillus terreus, and also its solid-state (SSF) and submerged (SmF) fermentation in bioreactors. The purified AtXEG12 showed optimum pH and temperature of 5.5 and 65 °C, respectively, demonstrating to be 90 % stable after 24 h of incubation at 50 °C. AtXEG12 activity increased in the presence of 2-mercaptoethanol (65 %) and Zn+2 (45 %), while Cu+2 and Ag+ ions drastically decreased its activity. A substrate assay showed, for the first time for this enzyme’s family, xylanase activity. The enzyme exhibited high specificity for tamarind xyloglucan (K M 1.2 mg mL−1) and V max of 17.4 μmol min−1 mg−1 of protein. The capillary zone electrophoresis analysis revealed that AtXEG12 is an endo-xyloglucanase. The heterologous xyloglucanase secretion was greater than the production by wild-type A. terreus cultivated in SmF. On the other hand, AtXEG12 activity reached by SSF was sevenfold higher than values achieved by SmF, showing that the expression of recombinant enzymes can be significantly improved by cultivation under SSF.

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Acknowledgments

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho de Desenvolvimento Científico e Tecnológico (CNPq), National System for Research on Biodiversity (Sisbiota-Brazil, CNPq 563260/2010-6/FAPESP n° 2010/52322-3), and LNBio (Mass Spectrometry staff) and CTBE facilities (Capillary Electrophoresis). J.A.J. and M.L.T.M.P. are Research Fellows of CNPq. G.L.V. and L.F.C.R. were recipient FAPESP Fellowship (2013/16563-4; 2010/10184-3, respectively). A.R.L. Damasio received grants from FAPESP 2012/20549-4, and F. Segato received grants from FAPESP 2014/18714-2. We gratefully acknowledge the CNPEM facility MAS at Brazilian Biosciences National Laboratory, Ricardo Alarcon and Mauricio de Oliveira for the technical assistance, and Mariana Cereia for the language review.

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Authors and Affiliations

  1. Departamento de Bioquímica e Imunologia, FMRP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    Gabriela Leal Vitcosque, Liliane Fraga Costa Ribeiro, Rosymar Coutinho de Lucas & Lucas Ferreira Ribeiro

  2. Chemical Biochemical Environmental Engineering Department, University of Maryland, Baltimore County, MD, USA

    Liliane Fraga Costa Ribeiro

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

    Rosymar Coutinho de Lucas, Tony Marcio da Silva, Aline Zorzetto Lopes Gonçalves, João Atilio Jorge & Maria de Lourdes T. M. Polizeli

  4. Instituto de Ciências da Saúde, Agrárias e Humanas do Centro Universitário do Planalto de Araxà (UNIARAXÀ), Araxà, MG, Brazil

    Tony Marcio da Silva

  5. Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA

    Lucas Ferreira Ribeiro

  6. Departamento de Bioquímica e Biologia Tecidual, Instituto de Biologia, Universidade de Campinas, Campinas, SP, Brazil

    André Ricardo de Lima Damasio

  7. Empresa Brasileira de Pesquisa Agropecuária, EMBRAPA Instrumentação, São Carlos, SP, Brazil

    Cristiane Sanchez Farinas

  8. Departamento de Biotecnologia, Escola de Engenharia de Lorena, Universidade de São Paulo, São Paulo, SP, Brazil

    Fernando Segato

  9. Departamento de Botanica, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil

    Marcos Silveira Buckeridge

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  1. Gabriela Leal Vitcosque
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  2. Liliane Fraga Costa Ribeiro
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  3. Rosymar Coutinho de Lucas
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  5. Lucas Ferreira Ribeiro
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  6. André Ricardo de Lima Damasio
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  7. Cristiane Sanchez Farinas
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  8. Aline Zorzetto Lopes Gonçalves
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  9. Fernando Segato
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  10. Marcos Silveira Buckeridge
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  11. João Atilio Jorge
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  12. Maria de Lourdes T. M. Polizeli
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Corresponding author

Correspondence to Maria de Lourdes T. M. Polizeli.

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Gabriela Leal Vitcosque, Liliane Fraga Costa Ribeiro, and Rosymar Coutinho de Lucas equally contributed to this work.

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Vitcosque, G.L., Ribeiro, L.F.C., de Lucas, R.C. et al. The functional properties of a xyloglucanase (GH12) of Aspergillus terreus expressed in Aspergillus nidulans may increase performance of biomass degradation. Appl Microbiol Biotechnol 100, 9133–9144 (2016). https://doi.org/10.1007/s00253-016-7589-2

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