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Hyaluronate Lyase of a Deep-Sea Bacillus niacini

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Abstract

A hyaluronate lyase (BniHL) was purified to homogeneity from a culture of a deep-sea Bacillus niacin strain JAM F8. The molecular mass of purified BniHL was approximately 120 kDa. The purified enzyme degraded hyaluronan as well as chondroitin sulfates A and C by a β-elimination mechanism. The optimal pH and temperature were around pH 6 and 45 °C for hyaluronan degradation. The enzyme required optimally 2, 50, and 100 mM calcium ions for degradation of hyaluronan, chondroitin sulfate C, and chondroitin sulfate A, respectively. Calcium ions slightly increased the thermal stability of the enzyme. In a genome analysis of strain JAM F8, a BniHL coding gene was identified on the bases of the molecular mass and N-terminal and internal amino acid sequences. The gene consisted of 3411 nucleotides and coded 1136 amino acids. The deduced amino acid sequence showed the highest similarity to the hyaluronate lyase of a Bacillus sp. A50 with 89 % identity.

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Author notes

    Authors and Affiliations

    1. Department of Applied Biological Chemistry, Graduate School of Agriculture, Kinki University, 3327-204 Nakamachi, Nara City, Nara, 631-8505, Japan

      Atsushi Kurata, Mioko Matsumoto & Noriaki Kishimoto

    2. Research and Development Center for Marine Biosciences, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan

      Tohru Kobayashi & Shigeru Deguchi

    Authors
    1. Atsushi Kurata
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    2. Mioko Matsumoto
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    3. Tohru Kobayashi
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    4. Shigeru Deguchi
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    5. Noriaki Kishimoto
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    Corresponding authors

    Correspondence to Atsushi Kurata or Tohru Kobayashi.

    Additional information

    Atsushi Kurata and Tohru Kobayashi contributed equally to this work.

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    Fig. S1

    ESI-MS/MS spectrograms of degradation products from HA and ChS-C. A, spectrogram of degradation products from HA; B, that of authentic ∆Di-HA; C, that of degradation products from ChS-C; D that of authentic ∆Di-6S. (PPTX 134 kb)

    Fig. S2

    Diagram of domain structure of BniHL. Amino acid numbers are shown above diagram. Each domain is shown as follows:, CBM_4_9 superfamily domain; tandem Big_2 domains (bacterial Ig-like domain); glycosaminoglycan (GAG)-lyase superfamily domain; PL family 8N-terminal alpha helical domain; PL family 8 super- sandwich domain (PPTX 49 kb)

    ESM 1

    CBM_4_9 superfamily domain (GIF 905 bytes)

    ESM 2

    tandem Big_2 domains (bacterial Ig-like domain) (GIF 1088 bytes)

    ESM 3

    glycosaminoglycan (GAG)-lyase superfamily domain (GIF 905 bytes)

    ESM 4

    PL family 8N-terminal alpha helical domain (GIF 1.32 kb)

    ESM 5

    PL family 8 super- sandwich domain (GIF 997 bytes)

    Table S1

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    Table S2

    (PPTX 51 kb)

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    Kurata, A., Matsumoto, M., Kobayashi, T. et al. Hyaluronate Lyase of a Deep-Sea Bacillus niacini . Mar Biotechnol 17, 277–284 (2015). https://doi.org/10.1007/s10126-015-9618-z

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    • DOI: https://doi.org/10.1007/s10126-015-9618-z

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