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Evolutionary engineering of sesquiterpene synthases from basidiomycetous fungi

Research Project

Project/Area Number 22K19212
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 40:Forestry and forest products science, applied aquatic science, and related fields
Research InstitutionKyushu University

Principal Investigator

Ichinose Hirofumi  九州大学, 農学研究院, 准教授 (00432948)

Project Period (FY) 2022-06-30 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2023: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2022: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywordsセスキテルペン合成酵素 / 担子菌
Outline of Research at the Start

担子菌(キノコ)に由来するセスキテルペン合成酵素をひとつの実験例として、新奇な酵素機能を生み出す技術戦略を確立し、遺伝子工学を軸とするモノ創りバイオ技術に新概念を与える。「生物は不都合な酵素活性を封印しながら進化した」という作業仮説に基づき、「封印された酵素を遺伝子工学的に逆進化させることでユニークな機能が誕生する」ことを実証する。本挑戦が結実すれば、担子菌のSTSを例として「不活化された酵素が逆進化によってスーパー酵素に変貌する」という新たな研究概念が誕生する。

Outline of Final Research Achievements

Based on our original hypothesis that organisms evolved by quenching detrimental enzyme activities, we performed to synthesize novel natural products by introducing genetic engineering into basidiomycetous sesquiterpene synthases (STSs). A series of sesquiterpene compounds was produced when putative cDNAs of STSs, whose transcription were not confirmed in nature, was prepared from genomic DNA and applied for heterologous expression in yeast. In addition, chimerization of STS genes, that could not show enzymatic activity as native form, resulted in emerging active STSs that produced novel sesquiterpenes. The compounds and their derivatives obtained in this study may play important roles in the biological functions of basidiomycetes.

Academic Significance and Societal Importance of the Research Achievements

本研究では、担子菌の進化過程で活性を喪失したと考えられるセスキテルペン合成酵素に焦点をあて、その活性を復元することで新規化合物が獲得可能となることを示した。担子菌類には非常に多くのSTS遺伝子が見出される一方で、これらの中には酵素活性を示さない分子種も多く存在する。本研究は、不活性なSTSを人為的に改変することでユニークなセスキテルペン合成活性が発現することを明らかにした例であり、本研究を基軸として様々なセスキテルペンの合成が可能になると期待される。

Report

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

    (10 results)

All 2024 2023 2022 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (8 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results) Remarks (1 results)

  • [Journal Article] Conversion and synthesis of chemicals catalyzed by fungal cytochrome P450 monooxygenases: A review2023

    • Author(s)
      Permana Dani、Kitaoka Takuya、Ichinose Hirofumi
    • Journal Title

      Biotechnology and Bioengineering

      Volume: 120 Issue: 7 Pages: 1725-1745

    • DOI

      10.1002/bit.28411

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Presentation] Epicubenolおよびα-muurolol の代謝変換に関与する担子菌シトクロムP4502024

    • Author(s)
      陳シン、一瀬博文、北岡卓也
    • Organizer
      第74回日本木材学会大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] α-サンタレン合成酵素および糸状菌シトクロムP450モノオキシゲナーゼを利用した希少セスキテルペノイドの合成2024

    • Author(s)
      西村瑞樹、一瀬博文、北岡卓也
    • Organizer
      第74回日本木材学会大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 担子菌Resinicium bicolorが有するセスキテルペン合成酵素の機能解明2024

    • Author(s)
      袁暁悦、一瀬博文、北岡卓也
    • Organizer
      第74回日本木材学会大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Application of fungal cytochrome P450s for production of natural and pseudonatural sesquiterpenoids2023

    • Author(s)
      Ichinose Hirofumi
    • Organizer
      2023 International Joint Meeting of the 23rd International Conference on Cytochrome P450 and the 38th Annual Meeting of the Japanese Society for the Study of Xenobiotics
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Application of fungal cytochrome P450 monooxygenases for production of α-santalene derivatives2023

    • Author(s)
      Nishimura Mizuki, Kitaoka Takuya, Ichinose Hirofumi
    • Organizer
      2023 International Joint Meeting of the 23rd International Conference on Cytochrome P450 and the 38th Annual Meeting of the Japanese Society for the Study of Xenobiotics
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Identification of cytochrome P450 monooxygenases involved in fungal metabolism of sesquiterpene alcohols2023

    • Author(s)
      Chen Chen, Kitaoka Takuya, Ichinose Hirofumi
    • Organizer
      2023 International Joint Meeting of the 23rd International Conference on Cytochrome P450 and the 38th Annual Meeting of the Japanese Society for the Study of Xenobiotics
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 子嚢菌を宿主とした担子菌二次代謝関連遺伝子の転写と成熟mRNAの合成2023

    • Author(s)
      庄嶋菜月、一瀬博文、北岡卓也
    • Organizer
      第73回日本木材学会大会
    • Related Report
      2022 Research-status Report
  • [Presentation] 子嚢菌を利用した担子菌二次代謝関連遺伝子の転写と成熟mRNAの合成2022

    • Author(s)
      庄嶋菜月、一瀬博文、北岡卓也
    • Organizer
      令和4年度内外環境応答・代謝酵素研究会
    • Related Report
      2022 Research-status Report
  • [Remarks] 生物資源化学研究室

    • URL

      https://bm.wood.agr.kyushu-u.ac.jp

    • Related Report
      2023 Annual Research Report

URL: 

Published: 2022-07-05   Modified: 2025-01-30  

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