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Diversiry and origin of the TCA cycles

Research Project

Project/Area Number 19H00988
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionJapan Agency for Marine-Earth Science and Technology

Principal Investigator

Nunoura Takuro  国立研究開発法人海洋研究開発機構, 海洋機能利用部門(生命理工学センター), センター長代理 (60359164)

Co-Investigator(Kenkyū-buntansha) 力石 嘉人  北海道大学, 低温科学研究所, 教授 (50455490)
千葉 洋子  国立研究開発法人理化学研究所, 環境資源科学研究センター, 上級研究員 (70638981)
Project Period (FY) 2019-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥45,110,000 (Direct Cost: ¥34,700,000、Indirect Cost: ¥10,410,000)
Fiscal Year 2022: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
Fiscal Year 2021: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000)
Fiscal Year 2020: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000)
Fiscal Year 2019: ¥11,830,000 (Direct Cost: ¥9,100,000、Indirect Cost: ¥2,730,000)
Keywordsクエン酸回路 / TCA回路 / メタボローム / 好熱菌 / アーキア / 炭酸固定 / CO2固定 / 炭素固定 / 不完全型TCA回路 / 超好熱菌 / 代謝解析 / 炭素中央代謝 / アイソトポマー / 中央代謝 / 水素酸化菌 / メタン菌 / アミノ酸 / アミノ酸生合成
Outline of Research at the Start

TCA回路は最も研究された代謝系の一つであるが、ゲノム解析から非モデル微生物には不完全型等多様なTCA回路の存在が示唆され、実際、研究代表者らが始原的な可逆的TCA回路を見出したように、未だ重要な知見の発見が相次ぐ。本研究では、培養が困難な環境微生物へ適用可能な独自の微量メタボローム解析技術を駆使し、(1) (超)好熱性アーキア及びバクテリアに分布する「不完全な」TCA回路の役割の解明、(2) TCA回路ともう一つの始型炭素固定経路(Wood-Ljungdahl経路)との関係の解明、(3) (1)/(2)の過程で明らかにされる未知アミノ酸生合成経路の解明を実施する。

Outline of Final Research Achievements

In order to understand the function of the putatively incomplete TCA cycle (citric acid cycle) in (hyper)thermophilic archaea and bacteria, we conducted multi-omics analyses including tracer-based metabolomics using stable isotope tracers. Moreover, we have developed a novel tracer-based method of metabolomics for amino acids in combination with capillary electrophoresis and Orbitrap Fusion Mass Spectrometry (CE-Orbitrap Fusion MS). The novel method has advantages in data quality and sample preparation; only 1x105 cells are required, comparing to the methods for the tracer-based metabolomics.

Academic Significance and Societal Importance of the Research Achievements

本研究において確立したCE-Orbirap Fusion MSによる代謝解析手法は、理論的にあらゆる単離微生物に対して僅か試験管1本で中央代謝経路及びアミノ産生合成経路を検証することを可能とする。つまり、本技術は、新規代謝経路の探索を容易とするだけでなく、ポストゲノム情報に不可欠な代謝マップ等の精度の向上に大きく貢献するポテンシャルを有すことを意味する。また、この技術を用いることにより、従属栄養性の超好熱バクテリア等におけるCO2固定能が従来の想定より相当程度大きいことを明らかにした。今後、これらの知見は微生物を用いる物質生産システムの構築にも貢献するであろう。

Report

(6 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • 2019 Comments on the Screening Results   Annual Research Report
  • Research Products

    (11 results)

All 2022 2021 2020

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 3 results) Presentation (8 results) (of which Int'l Joint Research: 4 results,  Invited: 1 results)

  • [Journal Article] Stable isotope analysis -1: MassWorks on conventional gas chromatograph-mass spectrometer2021

    • Author(s)
      力石嘉人、滝沢侑子、布浦拓郎
    • Journal Title

      低温科学

      Volume: 79 Pages: 23-29

    • DOI

      10.14943/lowtemsci.79.23

    • ISSN
      1880-7593
    • Year and Date
      2021-03-20
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Analysis of amino acids by Orbitrap mass spectrometry.2021

    • Author(s)
      島村繁、澄田智美、布浦拓郎
    • Journal Title

      低温科学

      Volume: 79 Pages: 37-42

    • DOI

      10.14943/lowtemsci.79.37

    • ISSN
      1880-7593
    • Year and Date
      2021-03-20
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Detection of tracing of stable isotope labelled amino acids using Orbitrap-Fusion CE-MS and Quadrupole GC-MS2021

    • Author(s)
      澄田智美、島村繁、布浦拓郎
    • Journal Title

      低温科学

      Volume: 79 Pages: 43-49

    • DOI

      10.14943/lowtemsci.79.43

    • ISSN
      1880-7593
    • Year and Date
      2021-03-20
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] The rediscovery of obligately chemolithoautotrophs as chemolithomixotrophs provides insights into the ancestral carbon fixation metabolism .2022

    • Author(s)
      Yoko Chiba, Tomomi Sumida, Masafumi Kameya, Yoshito Chikaraishi, Takuro Nunoura
    • Organizer
      Gordon Research Conference: Molecular Basis of Microbial One-Carbon Metabolism
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Variation of TCA cycle and Its Contribution of Carbon Fixation in (Hyper)thermophilic Archaea and Bacteria2022

    • Author(s)
      Takuro Nunoura
    • Organizer
      Gordon Research Conference: Molecular Basis of Microbial One-Carbon Metabolism
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Visualization of CO2 Fixation Pathways in Thermophilic Bacteria2022

    • Author(s)
      千葉洋子, 澄田智美, 亀谷将史, 力石嘉人, 布浦拓郎
    • Organizer
      Emergence in Biological Systems: Challenges to Bridging Hierarchies
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 微量メタボローム解析を用いたMethanothermobacter thermautotrophicusにおけるアミノ酸生合成系 の再構築2022

    • Author(s)
      福山宥斗、澄田智美、島村繁、酒井早苗、跡見晴幸、布浦拓郎
    • Organizer
      第34回日本Archaea研究会講演会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 微量メタボローム解析による従属栄養性超好熱バクテリアThermotogaのCO2再利用能の評価2022

    • Author(s)
      福山 宥斗, 島村 繁, 澄田 智美, 力石 嘉人,千葉 洋子, 跡見 晴幸, 布浦 拓郎
    • Organizer
      The 35th annual JSME conference
    • Related Report
      2022 Annual Research Report
  • [Presentation] Reassimilation of CO2 and acetate in Thermotoga maritima revealed by 13C isotope tracing experiments using microfluidic capillary electrophoresis-mass spectrometry2022

    • Author(s)
      Yuto Fukuyama, Tomomi Sumida, Shigeru Shimamura, Yoko Chiba, Hisato Chikaraishi, Haruyuki Atomi, Takuro Nunoura
    • Organizer
      Gordon Research Conference: Molecular Basis of Microbial One-Carbon Metabolism
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 分子内安定同位体プロービングを用いた好熱性独立栄養性細菌の炭酸固定経路の検証2020

    • Author(s)
      千葉 洋子、澄田 智美、力石 嘉人、布浦 拓郎
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Identification of CO2 fixation pathway of a thermophilic bacterium using a position-specific stable isotope probing2020

    • Author(s)
      千葉 洋子、澄田 智美、力石 嘉人、布浦 拓郎
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report

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Published: 2019-04-18   Modified: 2024-01-30  

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