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Creation of Apocarotenoid Diversity through Evolutionary Molecular Engineering of Carotenoid Oxidative Cleavage Enzymes

Research Project

Project/Area Number 19K23670
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeMulti-year Fund
Review Section 0601:Agricultural chemistry and related fields
Research InstitutionNational Institute of Advanced Industrial Science and Technology

Principal Investigator

Furubayashi Maiko  国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (90849895)

Project Period (FY) 2019-08-30 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsアポカロテノイド / カロテノイド / 生合成 / レチノイド / 生合成工学 / 合成生物学 / 天然化合物 / レチナール / 進化分子工学
Outline of Research at the Start

アポカロテノイドは多様な生物活性をもつ天然化合物の一群であり、その多くは植物ホルモン・香料・ビタミンなど多彩な機能をもつことから更なる新規分子構造の探索が望まれている。アポカロテノイド生合成の最初のステップは、カロテノイドを切断する「カロテノイド酸化開裂酵素(Carotenoid Cleavage Oxygenase, CCO)」と呼ばれ、様々な生物種から多様な基質特異性を示すものがみつかっている。本研究では、各種カロテノイドを生合成する組替大腸菌に、異種由来の様々な特異性をもつCCO遺伝子を掛け合わせて発現・進化工学し、天然・非天然アポカロテノイド多様性を生み出すことを目指す。

Outline of Final Research Achievements

Apocarotenoids are the general term for the oxidative cleavage products of carotenoids, and are known for various biological functions, including plant hormones and fragrance molecules. In this study, we aimed to create novel apocarotenoids in the laboratory by expressing a variety of carotenoid cleavage enzymes (CCOs) from diverse biological origins in recombinant E. coli that produce various carotenoids. By expressing a variety of CCO genes from microorganisms and plants in carotenoid-producing E. coli, we succeeded in the cleavage of carotenoids that are not the original substrates, in other words, the production of novel apocarotenoids. Furthermore, by introducing amino acid mutations into the CCO enzyme, we discovered the possibility of efficient production of new apocarotenoids.

Academic Significance and Societal Importance of the Research Achievements

本研究では、自然界に存在するカロテノイド開裂酵素(CCO)遺伝子をカロテノイド生産大腸菌に導入して調べることにより、その基質選択性を詳細に調べた。その結果、本来の基質以外のカロテノイドを切断することができることを見出した。この成果は、自然界のアポカロテノイド経路の進化のさらなる理解につながるだろう。また、アポカロテノイドは多様な生理活性や健康・美容効果が知られているが、本研究で生み出されたアポカロテノイド、およびこの方法を用いて生み出しうる新たなアポカロテノイド化合物の生物活性や物性を調べることにより、より機能性が高く価値のある化合物の発見に繋げられると考えられる。

Report

(5 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (8 results)

All 2023 2022 2021 2019

All Journal Article (5 results) (of which Peer Reviewed: 3 results,  Open Access: 1 results) Presentation (3 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Journal Article] Reconstruction of the Native Biosynthetic System of Carotenoids in <i>E. coli</i>─Biosynthesis of a Series of Carotenoids Specific to Paprika Fruit2023

    • Author(s)
      Hattan Jun-ichiro、Furubayashi Maiko、Maoka Takashi、Takemura Miho、Misawa Norihiko
    • Journal Title

      ACS Synthetic Biology

      Volume: 12 Issue: 4 Pages: 1072-1080

    • DOI

      10.1021/acssynbio.2c00578

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 彩りと香りをもたらすアポカロテノイドの魅力2022

    • Author(s)
      古林 真衣子
    • Journal Title

      Seibutsu-kogaku Kaishi

      Volume: 100 Issue: 7 Pages: 382-382

    • DOI

      10.34565/seibutsukogaku.100.7_382

    • ISSN
      0919-3758, 2435-8630
    • Year and Date
      2022-07-25
    • Related Report
      2022 Annual Research Report
    • Open Access
  • [Journal Article] Use of directed enzyme evolution to create novel biosynthetic pathways for production of rare or non-natural carotenoids2022

    • Author(s)
      Furubayashi Maiko、Umeno Daisuke
    • Journal Title

      Methods in Enzymology, Carotenoids: Carotenoid and apocarotenoid biosynthesis metabolic engineering and synthetic biology

      Volume: 671 Pages: 351-382

    • DOI

      10.1016/bs.mie.2022.03.008

    • ISBN
      9780323913539
    • Related Report
      2022 Annual Research Report
  • [Journal Article] Promiscuous activity of β‐carotene hydroxylase CrtZ on epoxycarotenoids leads to the formation of rare carotenoids with 6‐hydroxy‐3‐keto‐ε‐ends2022

    • Author(s)
      Furubayashi Maiko、Maoka Takashi、Mitani Yasuo
    • Journal Title

      FEBS Letters

      Volume: - Issue: 15 Pages: 1921-1931

    • DOI

      10.1002/1873-3468.14342

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Capsanthin Production in Escherichia coli by Overexpression of Capsanthin/Capsorubin Synthase from Capsicum annuum2021

    • Author(s)
      Furubayashi Maiko、Kubo Akiko、Takemura Miho、Otani Yuko、Maoka Takashi、Terada Yoshinobu、Yaoi Katsuro、Ohdan Kohji、Misawa Norihiko、Mitani Yasuo
    • Journal Title

      Journal of Agricultural and Food Chemistry

      Volume: 69 Issue: 17 Pages: 5076-5085

    • DOI

      10.1021/acs.jafc.1c00083

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] Engineering Natural and Non-natural Carotenoid/Apocarotenoid Pathways in Escherichia coli2023

    • Author(s)
      Maiko Furubayashi
    • Organizer
      Gordon Research Conference: Biology and Applications of Carotenoids, Retinoids and Other Apocarotenoids
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] カロテノイド生合成経路の構築方法2019

    • Author(s)
      古林真衣子
    • Organizer
      第33回カロテノイド若手の会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] カロテノイド生合成工学と磁性細菌の合成生物学2019

    • Author(s)
      古林真衣子
    • Organizer
      第13回ゲノム微生物学会若手の会
    • Related Report
      2019 Research-status Report

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Published: 2019-09-03   Modified: 2024-01-30  

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