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Oligomeric state and structural stability of two hyperthermophilic β-glucosidases from Thermotoga petrophila

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Abstract

The β-glucosidases are enzymes essential for several industrial applications, especially in the field of plant structural polysaccharides conversion into bioenergy and bioproducts. In a recent study, we have provided a biochemical characterization of two hyperthermostable β-glucosidases from Thermotoga petrophila belonging to the families GH1 (TpBGL1) and GH3 (TpBGL3). Here, as part of a continuing investigation, the oligomeric state, the net charge, and the structural stability, at acidic pH, of the TpBGL1 and TpBGL3 were characterized and compared. Enzymatic activity is directly related to the balance between protonation and conformational changes. Interestingly, our results indicated that there were no significant changes in the secondary, tertiary and quaternary structures of the β-glucosidases at temperatures below 80 °C. Furthermore, the results indicated that both the enzymes are stable homodimers in solution. Therefore, the observed changes in the enzymatic activities are due to variations in pH that modify protonation of the enzymes residues and the net charge, directly affecting the interactions with ligands. Finally, the results showed that the two β-glucosidases displayed different pH dependence of thermostability at temperatures above 80 °C. TpBGL1 showed higher stability at pH 6 than at pH 4, while TpBGL3 showed similar stability at both pH values. This study provides a useful comparison of the structural stability, at acidic pH, of two different hyperthermostable β-glucosidases and how it correlates with the activity of the enzymes. The information described here can be useful for biotechnological applications in the biofuel and food industries.

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Acknowledgments

We would like to thank the National Synchrotron Light Laboratory (LNLS, Brazil) and CENAPAD-SP (Centro Nacional de Processamento de Alto Desempenho em São Paulo). The authors would like to thank FAPESP and CNPq for the financial support for this work via grants # 2012/21054-9, # 478900/2012-0, # 2008/58037-9 and # 2011/13242-7; and fellowships # 2012/03503-0 (VMS) and # 501037/2012-8 (FC).

Conflict of interest

The authors declare that they do not have competing interests.

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    Authors and Affiliations

    1. Centro de Ciências Naturais e Humanas (CCNH), Universidade Federal do ABC (UFABC), Santo André, SP, Brazil

      Francieli Colussi, Viviam M. da Silva, Ian Miller & Wanius Garcia

    2. Departamento de Física e Biofísica, Instituto de Biociências, UNESP Univ Estadual Paulista, Botucatu, SP, Brazil

      Mário de Oliveira Neto

    3. Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil

      Junio Cota & Fábio M. Squina

    4. Departamento de Física, Instituto de Biociências, Letras e Ciências Exatas, UNESP Univ Estadual Paulista, São José do Rio Preto, SP, Brazil

      Leandro C. de Oliveira

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    Correspondence to Wanius Garcia.

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    Handling Editor: C. Schiene-Fischer.

    F. Colussi and V. M. da Silva have contributed equally to this work.

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    Colussi, F., da Silva, V.M., Miller, I. et al. Oligomeric state and structural stability of two hyperthermophilic β-glucosidases from Thermotoga petrophila . Amino Acids 47, 937–948 (2015). https://doi.org/10.1007/s00726-015-1923-3

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