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Characterization of extracellular lipases produced by xerophilic molds

Research Project

Project/Area Number 20K05808
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38020:Applied microbiology-related
Research InstitutionKobe University

Principal Investigator

Takenaka Shinji  神戸大学, 農学研究科, 教授 (40314512)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
KeywordsAspergillus / 鰹節カビ / 好乾性糸状菌 / 脂質分解酵素 / 鰹節 / かび付け発酵熟成 / リパーゼ
Outline of Research at the Start

鰹節の製造には、かび付けと呼ばれる好乾性糸状菌が働く発酵熟成工程が含まれ、節の香味や色調そして保存性が高まる。申請者は、同工程には2種類の優占種が関わることを見出し、リパーゼなどの菌体外酵素の生産特性も明らかにした。しかし、鰹節カビとその酵素群が、低水分活性下で働くメカニズムには不明な点が多い。リパーゼは発酵工程で脂質を分解する鍵酵素として働いていることが考えられる。本研究では、優占種について、脂質分解に特化したリパーゼの特定とその性質および同遺伝子を明らかにする。さらに、優占種間の酵素特性や発現様式を比較し、鰹節カビとリパーゼ群の働きを含めた低水分活性下での発酵メカニズムを考察する。

Outline of Final Research Achievements

A mild-flavored soup stock made from katsuobushi is an important element of traditional Japanese cuisine and is the basic seasoning responsible for the taste. Fermented and ripened katsuobushi, known as karebushi, is manufactured by simmering skipjack tuna that is then smoke-dried, fermented, and ripened in a repeated molding process by five dominant Aspergillus species. Our aim was to characterize and identify the lipolytic enzymes secreted by the dominant Aspergillus species, especially A. chevalieri and A. pseudoglaucus, which are involved in hydrolyzing lipids during the molding process. Five and three lipolytic enzymes from A. chevalieri and A. pseudoglaucus were heterologously expressed. Recombinant enzymes, rcLip 1 to 5 and rpLip 1 to 3 were compared in the terms of catalytic properties.

Academic Significance and Societal Importance of the Research Achievements

鰹節の製造には、かび付けと呼ばれる好乾性糸状菌が働く発酵熟成工程が含まれる。鰹節カビとその酵素群が、低水分活性下で働く発酵メカニズムには不明な点が多いが、脂質分解酵素群は発酵工程で脂質を分解することで、香味や保存性を高める鍵酵素として働いていることが考えられる。そこで本研究では、2種の優占種が分泌するリパーゼ群について、脂質分解に特化したリパーゼの特定とその性質および同遺伝子の発現様式を明らかにした。これにより、高品質な鰹節製造につながる知見だけでなく、低水分活性下で働く加水分解酵素の耐性機構の解明や利活用にも展開できる。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (11 results)

All 2023 2022 2021 2020

All Journal Article (5 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 5 results,  Open Access: 1 results) Presentation (6 results)

  • [Journal Article] Improvement in salt-tolerance of Aspergillus oryzae γ-glutamyl transpeptidase via protein chimerization with Aspergillus sydowii homolog2023

    • Author(s)
      Senba Hironori、Nishikawa Arisa、Kimura Yukihiro、Tanaka Shinichi、Matsumoto Jun-ichi、Doi Mikiharu、Takenaka Shinji
    • Journal Title

      Enzyme and Microbial Technology

      Volume: 167 Pages: 110240-110240

    • DOI

      10.1016/j.enzmictec.2023.110240

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Isolation and characterization of a salt-tolerant γ-glutamyl transpeptidase from xerophilic Aspergillus sydowii2022

    • Author(s)
      Nishikawa Arisa、Senba Hironori、Kimura Yukihiro、Yokota Satoko、Doi Mikiharu、Takenaka Shinji
    • Journal Title

      3 Biotech

      Volume: 12 Issue: 10 Pages: 253-253

    • DOI

      10.1007/s13205-022-03259-3

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification and characterization of extracellular enzymes secreted by Aspergillus spp. involved in lipolysis and lipid-antioxidation during katsuobushi fermentation and ripening2021

    • Author(s)
      Takenaka Shinji、Ogawa Chiaki、Uemura Mariko、Umeki Tomoya、Kimura Yukihiro、Yokota Satoko、Doi Mikiharu
    • Journal Title

      International Journal of Food Microbiology

      Volume: 353 Pages: 109299-109299

    • DOI

      10.1016/j.ijfoodmicro.2021.109299

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Improvement of the halotolerance of a Bacillus serine protease by protein surface engineering2021

    • Author(s)
      Takenaka Shinji、Takada Airi、Kimura Yukihiro、Watanabe Masanori、Kuntiya Ampin
    • Journal Title

      Journal of Basic Microbiology

      Volume: 62 Issue: 2 Pages: 174-184

    • DOI

      10.1002/jobm.202100335

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Characterization of surface Aspergillus community involved in traditional fermentation and ripening of katsuobushi2020

    • Author(s)
      Takenaka Shinji、Nakabayashi Reina、Ogawa Chiaki、Kimura Yukihiro、Yokota Satoko、Doi Mikiharu
    • Journal Title

      International Journal of Food Microbiology

      Volume: 327 Pages: 108654-108654

    • DOI

      10.1016/j.ijfoodmicro.2020.108654

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] Aspergillus pseudoglaucus 由来カルボキシエステラーゼ/リパーゼの特性解析と異種発現2022

    • Author(s)
      豊嶋 瞭太, 仙波 弘雅, 上村 真理子, 木村 行宏, 土居 幹治, 竹中 慎治
    • Organizer
      日本生物工学会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 鰹節カビAspergillus属糸状菌による鰹節 (荒節) の燻煙香成分の代謝分解2022

    • Author(s)
      藤原 卓巳, 米子 響, 木村 行宏, 横田 仁子, 土居 幹治, 〇竹中 慎治
    • Organizer
      日本農芸化学会
    • Related Report
      2021 Research-status Report
  • [Presentation] Aspergillus sydowiiの生産する耐塩性γ-グルタミルトランスペプチダーゼの特性解析2021

    • Author(s)
      ○西川有咲,仙波弘雅,木村行宏,横田仁子,土居幹治,竹中慎治
    • Organizer
      日本生物工学会
    • Related Report
      2021 Research-status Report
  • [Presentation] Aspergillus chevalieri由来カルボキシエステラーゼ/リパーゼの特性解析2021

    • Author(s)
      〇梅木智哉,上村真理子,木村行宏,横田仁子,土居幹治,竹中慎治
    • Organizer
      日本生物工学会
    • Related Report
      2021 Research-status Report
  • [Presentation] 好乾性糸状菌が生産する耐塩性セルラーゼの探索と諸性質の検討2021

    • Author(s)
      〇斉藤大輔, 木村行宏, 横田仁子, 土居幹治, 竹中慎治
    • Organizer
      日本生物工学会
    • Related Report
      2021 Research-status Report
  • [Presentation] 鰹節カビAspergillus 属糸状菌が生産する脂質分解酵素の特性解析2021

    • Author(s)
      小川千晶、上村真理子、木村行宏、横田仁子 、土居幹治 、○竹中慎治
    • Organizer
      日本農芸化学会
    • Related Report
      2020 Research-status Report

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Published: 2020-04-28   Modified: 2024-01-30  

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