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Expression and Characterization of a GH39 β-Xylosidase II from Caulobacter crescentus

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Abstract

In the present work, the gene xynB2, encoding a β-xylosidase II of the Glycoside Hydrolase 39 (GH39) family, of Caulobacter crescentus was cloned and successfully overexpressed in Escherichia coli DH10B. The recombinant protein (CcXynB2) was purified using nickel-Sepharose affinity chromatography, with a recovery yield of 75.5 %. CcXynB2 appeared as a single band of 60 kDa on a sodium dodecyl sulfate polyacrylamide gel and was recognized by a specific polyclonal antiserum. The predicted CcXynB2 protein showed a high homology with GH39 β-xylosidases of the genus Xanthomonas. CcXynB2 exhibited an optimal activity at 55 °C and a pH of 6. CcXynB2 displayed stability at pH values of 4.5–7.5 for 24 h and thermotolerance up to 50 °C. The K M and V Max values were 9.3 ± 0.45 mM and 402 ± 19 μmol min−1 for ρ-nitrophenyl-β-d-xylopyranoside, respectively. The purified recombinant enzyme efficiently produced reducing sugars from birchwood xylan and sugarcane bagasse fibers pre-treated with a purified xylanase. As few bacterial GH39 family β-xylosidases have been characterized, this work provides a good contribution to this group of enzymes.

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Acknowledgments

This work was supported by grants from the Fundação Araucária (convênio 893/2012), Fundo Paraná SETI, CNPq, and Fundação Parque Tecnológico Itaipu (PTI C&T/FPTI-BR).

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Authors and Affiliations

  1. Centro de Ciências Exatas e Tecnológicas, Universidade Estadual do Oeste do Paraná, Rua Universitária, 2069, Cascavel, Paraná, 85814-110, Brazil

    Juliana Moço Corrêa & Luciana Graciano

  2. Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Rua Universitária, 2069, Cascavel, Paraná, 85814-110, Brazil

    Josielle Abrahão, Rinaldo Ferreira Gandra, Marina Kimiko Kadowaki & Rita de Cássia Garcia Simão

  3. Centro de Ciências Biológicas e da Saúde, Universidade Estadual do Oeste do Paraná, Rua Universitária, 2069, Cascavel, Paraná, 85814-110, Brazil

    Eduardo Alexandre Loth

  4. Central Hidrelétrica de Itaipu, Itaipu Binacional, Foz do Iguaçu, Paraná, Brazil

    Caroline Henn

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  1. Juliana Moço Corrêa
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  2. Luciana Graciano
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  3. Josielle Abrahão
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  6. Marina Kimiko Kadowaki
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  8. Rita de Cássia Garcia Simão
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Correspondence to Rita de Cássia Garcia Simão.

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Corrêa, J.M., Graciano, L., Abrahão, J. et al. Expression and Characterization of a GH39 β-Xylosidase II from Caulobacter crescentus . Appl Biochem Biotechnol 168, 2218–2229 (2012). https://doi.org/10.1007/s12010-012-9931-1

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  • DOI: https://doi.org/10.1007/s12010-012-9931-1

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