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Biochemical characterization of a thermostable β-1,3-xylanase from the hyperthermophilic eubacterium, Thermotoga neapolitana strain DSM 4359

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Abstract

The biochemical properties of a putative β-1,3-xylanase from the hyperthermophilic eubacterium Thermotoga neapolitana DSM 4359 were determined from a recombinant protein (TnXyn26A) expressed in Escherichia coli. This enzyme showed specific hydrolytic activity against β-1,3-xylan and released β-1,3-xylobiose and β-1,3-xylotriose as main products. It displayed maximum activity at 85 °C during a 10-min incubation, and its activity half-life was 23.9 h at 85 °C. Enzyme activity was stable in the pH range 3–10, with pH 6.5 being optimal. Enzyme activity was significantly inhibited by the presence of N-bromosuccinimide (NBS). The insoluble β-1,3-xylan K m value was 10.35 mg/ml and the k cat value was 588.24 s−1. The observed high thermostability and catalytic efficiency of TnXyn26A is both industrially desirable and also aids an understanding of the chemistry of its hydrolytic reaction.

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Acknowledgments

This work was supported by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative BioProduction, Kobe), Ministry of Education, Culture, Sports, Science and Technology, Japan. FO would like to thank Mika Okazaki for useful discussions.

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Authors and Affiliations

  1. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Fumiyoshi Okazaki & Nanami Nakashima

  2. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Chiaki Ogino & Akihiko Kondo

  3. Department of Life Sciences, Graduate School of Bioresources, Mie University, 1577 Kurimamachiya, Tsu, Mie, 514-8507, Japan

    Yutaka Tamaru

  4. RIKEN Biomass Engineering Program, 1-7-22 Suehiro, Tsurumi, Yokohama, 230-0045, Japan

    Akihiko Kondo

Authors
  1. Fumiyoshi Okazaki
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  2. Nanami Nakashima
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  3. Chiaki Ogino
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  4. Yutaka Tamaru
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  5. Akihiko Kondo
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Correspondence to Akihiko Kondo.

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Okazaki, F., Nakashima, N., Ogino, C. et al. Biochemical characterization of a thermostable β-1,3-xylanase from the hyperthermophilic eubacterium, Thermotoga neapolitana strain DSM 4359. Appl Microbiol Biotechnol 97, 6749–6757 (2013). https://doi.org/10.1007/s00253-012-4555-5

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  • DOI: https://doi.org/10.1007/s00253-012-4555-5

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