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Establishment of a general heterologous expression system for difficult-to-express molybdenum enzymes and their functional analysis.

Research Project

Project/Area Number 20K15432
Research Category

Grant-in-Aid for Early-Career Scientists

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

Principal Investigator

Takeuchi Michiki  京都大学, 農学研究科, 特定助教 (40766193)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2022: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2021: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywordsモリブデン酵素 / 異種発現
Outline of Research at the Start

モリブデン酵素は、様々な反応を触媒するユニークな酸化還元酵素であり、幅広い分野に応用可能な酵素である。しかしながら、取り扱いの難しさから、研究があまり進んでこなかった。本研究では、モリブデン酵素の研究において重要な「モリブデン酵素の異種発現」を確立し、それを利用してモリブデン酵素の学術的な知見を深める。さらに、環境分野や、健康分野といった様々な分野へ応用する。

Outline of Final Research Achievements

Molybdenum enzymes are unique redox enzymes that catalyze a variety of reactions. By using Rhodococcus erythropolis as a heterologous expression, I succeeded in heterologous (partially homologous) expression of uracil/thymine dehydrogenase, xanthine oxidase, and polyphenol dehydroxylase. After examining the conditions for heterologous expression in the active form, I found that the addition of molybdic acid and optimization of codon frequency were important for heterologous expression in the active form.

Academic Significance and Societal Importance of the Research Achievements

モリブデン酵素は、様々な反応を触媒するユニークな酸化還元酵素であり、その反応の多
様性から、幅広い分野に応用可能な酵素である。例えば、ギ酸脱水素酵素による二酸化炭素が還元や、腸内細菌のポリフェノール脱水素酵素による腸内代謝など、環境分野や健康分野といった様々な分野に関与する重要な酵素である。モリブデン酵素の活性型での異種発現に関する知見は、モリブデン酵素の学術的知見を深めるうえで必要不可欠であり、その応用は幅広い分野に役立ち、社会的意義が高いものである。

Report

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

    (7 results)

All 2022 2021 2020

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 2 results) Presentation (1 results)

  • [Journal Article] Quantification of leuco-indigo in indigo-dye-fermenting suspension by normal pulse voltammetry.2022

    • Author(s)
      Kikuchi, M., K. Sowa, M. Takeuchi, K. Nakagawa, M. Matsunaga, A. Ando, K. Kano, J. Ogawa, E. Sakuradani.
    • Journal Title

      J Biosci Bioeng

      Volume: 134 Issue: 1 Pages: 84-88

    • DOI

      10.1016/j.jbiosc.2022.04.009

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Indigo-Mediated Semi-Microbial Biofuel Cell Using an Indigo-Dye Fermenting Suspension2021

    • Author(s)
      Kikuchi Mayu、Sowa Keisei、Nakagawa Kasumi、Matsunaga Momoka、Ando Akinori、Kano Kenji、Takeuchi Michiki、Sakuradani Eiji
    • Journal Title

      Catalysts

      Volume: 11 Issue: 9 Pages: 1080-1080

    • DOI

      10.3390/catal11091080

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Isolation and characterization of indigo-reducing bacteria and analysis of microbiota from indigo fermentation suspensions2021

    • Author(s)
      Nakagawa Kasumi、Takeuchi Michiki、Tada Manami、Matsunaga Momoka、Kugo Masami、Kiyofuji Suzuna、Kikuchi Mayu、Yomota Kazuya、Sakamoto Takaiku、Kano Kenji、Ogawa Jun、Sakuradani Eiji
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 86 Issue: 2 Pages: 273-281

    • DOI

      10.1093/bbb/zbab209

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Mechanistic Insights into Indigo Reduction in Indigo Fermentation: A Voltammetric Study.2021

    • Author(s)
      Nakagawa, K., M. Takeuchi, M. Kikuchi, S. Kiyofuji, M. Kugo, T. Sakamoto, K. Kano, J. Ogawa, E. Sakuradani
    • Journal Title

      Electrochemistry

      Volume: 89 Issue: 1 Pages: 25-30

    • DOI

      10.5796/electrochemistry.20-00123

    • NAID

      130007965922

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Voltammetric in-situ monitoring of leuco-indigo in indigo-fermenting suspensions2021

    • Author(s)
      Nakagawa, K., M. Takeuchi, M. Kikuchi, M. Tada, T. Sakamoto, K. Kano, J. Ogawa, E. Sakuradani
    • Journal Title

      J Biosci Bioeng

      Volume: - Issue: 5 Pages: 565-571

    • DOI

      10.1016/j.jbiosc.2021.01.005

    • NAID

      40022568972

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Characterization of xanthine oxidase from Cellulosimicrobium funkei possessing hypoxanthine‐metabolizing activity2020

    • Author(s)
      Kozono, I., M. Takeuchi, S. Kozono, A. Satomura, W. Aoki, M. Hibi, J. Ogawa
    • Journal Title

      Journal of Applied Microbiology

      Volume: - Issue: 6 Pages: 2132-2140

    • DOI

      10.1111/jam.14891

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] 藍染め染色液の電気化学的解析と微生物燃料電池への応用2022

    • Author(s)
      竹内道樹,中川香澄,菊地真由,宋和慶盛,松永桃花,阪本鷹行, 安藤晃規,小川 順,加納健司,櫻谷英治
    • Organizer
      電気化学会
    • Related Report
      2021 Research-status Report

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

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