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Investigation for unrevealed lytic pathway of polysaccharides initiated with oxidation of a hydroxyl group

Research Project

Project/Area Number 21K19070
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 38:Agricultural chemistry and related fields
Research InstitutionNiigata University

Principal Investigator

Kitaoka Motomitsu  新潟大学, 自然科学系, 教授 (60353984)

Project Period (FY) 2021-07-09 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2023: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2021: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords水酸基の酸化 / 多糖分解 / 酵素反応 / 新規分解経路 / β脱離 / セルロース / 酸化 / セロビオースホスホリラーゼ / N-アセチルヘキソサミン1-キナーゼ / グルコシルマンノースホスホリラーゼ / ピラノースオキシダーゼ / 酸化酵素 / LPMO
Outline of Research at the Start

酵素によるグリコシル結合の分解は、通常グリコシド結合に直接作用する酵素により行われる。本研究では、1-デオキシ-2-ケトセロビオースに見出されたβ脱離によるグリコシド結合の自発分解が、セルロースのグリコシル結合に直接関与しない水酸基の酸化に起因した自発分解として普遍的に観測できる現象であるかを、モデル化合物を調製することにより検証する。さらに、水酸基の酵素酸化反応がセルロースなどの結晶性多糖分解へ寄与している実例について探索する。

Outline of Final Research Achievements

2-Keto-cellobiose, a model compound representing oxidized cellulose at the position 2 of the reducing-end glucose residue, has successfully been synthesized from cellobiose by the concerted actions of cellobiose phosphorylase and pyranose oxidase at 10 °C in the presence of a catalytic amount of phosphate. It was degraded by β-elimination reaction at pH 7.0 and 30°C with a half-life of 103 hours. Because 1-deoxy-2-ketocellobiose, a model compound representing oxidized cellulose at the position 2 of the internal glucose residue of cellulose, has already been shown to degrade with a half-life of 17 hours under the same conditions, it was proved that when the hydroxy group at position 2 of cellulose is oxidized, the bond is generally eliminated even at room temperature under neutral pH.

Academic Significance and Societal Importance of the Research Achievements

本研究により、セルロースなどの多糖類の水酸基が酸化されることにより結合が自動的に分解されることが証明された。このことから水酸基酸化酵素による未知の多糖分解経路の存在する可能性が示されたことになる。この成果をもとに、特に難分解性の結晶性多糖の新規な分解経路の発見が期待される。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (2 results)

All 2024 2023

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results)

  • [Journal Article] Molecular Basis of Absorption at 340 nm of 3-Ketoglucosides under Alkaline Conditions2024

    • Author(s)
      Kitaoka Motomitsu、Takano Ayu、Takahashi Mei、Yamakawa Yoshiki、Fushinobu Shinya、Yoshida Nobuyuki
    • Journal Title

      Journal of Applied Glycoscience

      Volume: 71 Issue: 1 Pages: 9-13

    • DOI

      10.5458/jag.jag.JAG-2023_0014

    • ISSN
      1344-7882, 1880-7291
    • Year and Date
      2024-03-20
    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Automatic Calculation of the Kinetic Parameters of Enzymatic Reactions with Their Standard Errors Using Microsoft Excel2023

    • Author(s)
      Kitaoka Motomitsu
    • Journal Title

      Journal of Applied Glycoscience

      Volume: 70 Issue: 1 Pages: 33-37

    • DOI

      10.5458/jag.jag.JAG-2022_0012

    • ISSN
      1344-7882, 1880-7291
    • Year and Date
      2023-03-03
    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access

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Published: 2021-07-13   Modified: 2025-01-30  

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