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Construction of a ribonucleopeptide receptor-based synthetic enzyme with high turnover number by controlling the affinity for the product

Research Project

Project/Area Number 18K14335
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 37010:Bio-related chemistry
Research InstitutionKyoto University

Principal Investigator

Nakano Shun  京都大学, エネルギー理工学研究所, 助教 (40650809)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsリセプター / RNA-ペプチド複合体 / in vitroセレクション / ライブラリー / mRNAディスプレイ / 蛍光センサー / 人工酵素 / ribonucleopeptide / receptor / fluorescent sensor / library / 触媒 / RNP / in vitro セレクション / セレクション / 回転数
Outline of Final Research Achievements

RNA-peptide complex has been used to construct a receptor that was used to develop the receptor-based fluorescent sensor and catalyst. In this study, I tried to develop a construction method of the receptor-based artificial enzyme with high turnover number by controlling the affinity for the substrate and product. The highly selective ATP receptor could be constructed by applying in vitro selection using RNP scaffold. However, the method for construction of the peptide library that can be used to SELEX could not be established. As the other related results, the reaction conditions was successfully optimized for the modification of cysteine-introduced Rev peptide and the formation of covalent linkage between RNA and peptide subunit of RNP scaffold.

Academic Significance and Societal Importance of the Research Achievements

リセプターに触媒分子を修飾する設計によって、触媒活性や反応回転数をどの程度向上できるのかについては、十分な検証がなされていない。リセプターの改変による人工酵素作製の試みでは、「生成物の解離」をどのように実現するかが大きな課題となる。触媒分子修飾リセプターの反応回転数の向上を行うために、「生成物の解離」を考慮したセレクションをおこない、触媒活性の向上が達成できるのかについて検討するために必要となる技術開発を行った。この成果は今後、高い触媒活性を有する酵素を目的に応じた様々な用途に合わせて任意に作製するための基礎的な技術を確立するために有用な知見となる。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (15 results)

All 2020 2019 2018 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 1 results) Presentation (8 results) (of which Int'l Joint Research: 4 results,  Invited: 1 results) Book (2 results) Remarks (2 results)

  • [Journal Article] Receptor-based fluorescent sensors constructed from ribonucleopeptide2020

    • Author(s)
      Nakano Shun、Konishi Hiroaki、Morii Takashi
    • Journal Title

      Methods in Enzymology

      Volume: 641 Pages: 183-223

    • DOI

      10.1016/bs.mie.2020.04.041

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] RNA-Peptide Conjugation through an Efficient Covalent Bond Formation2020

    • Author(s)
      Nakano Shun、Seko Taiki、Zhang Zhengxiao、Morii Takashi
    • Journal Title

      Applied Sciences

      Volume: 10 Issue: 24 Pages: 8920-8920

    • DOI

      10.3390/app10248920

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Highly selective dual sensing of ATP and ADP using fluorescent ribonucleopeptide sensors2019

    • Author(s)
      Nakano Shun、Shimizu Musashi、Dinh Huyen、Morii Takashi
    • Journal Title

      Chemical Communications

      Volume: 55 Issue: 11 Pages: 1611-1614

    • DOI

      10.1039/c8cc09934k

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] Highly selective detection of ATP and ADP by using fluorescent ribonucleopeptide sensors2019

    • Author(s)
      Shun Nakano, Musashi Shimizu, Huyen Dinh, Takashi Morii
    • Organizer
      CISNAC2019
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] Selective detection of ATP and ADP by using fluorescent ribonucleopeptide sensors2019

    • Author(s)
      Shun Nakano, Takashi Morii
    • Organizer
      The 2nd International Symposium of Biofunctional Chemistry
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] Construction of tailor-made receptors and sensors from RNA-peptide complex2019

    • Author(s)
      Shun Nakano, Takashi Morii
    • Organizer
      Lecture in Jinan University
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Fluorescent ribonucleopeptide sensors for selective detection of ATP and ADP2019

    • Author(s)
      NAKANO, S. ; Shimizu, M.; Dinh, H.; Morii, T.
    • Organizer
      日本化学会第99春季年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Dual sensing of ATP and ADP by fluorescent ribonucleopeptide sensors2018

    • Author(s)
      Nakano, S.; Shimizu, M.; Morii, T.
    • Organizer
      ISNAC2018/第45回国際核酸化学シンポジウム
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] ATP とADP を高選択的に検出する蛍光性リボヌクレオペプチドセンサー2018

    • Author(s)
      仲野 瞬、森井 孝
    • Organizer
      第12回バイオ関連化学シンポジウム
    • Related Report
      2018 Research-status Report
  • [Presentation] RNA-ペプチド複合体を用いた蛍光性センサーの作製2018

    • Author(s)
      仲野 瞬、森井 孝
    • Organizer
      生体機能関連化学部会若手の会第30回サマースクール
    • Related Report
      2018 Research-status Report
  • [Presentation] 酵素連続反応の効率化へ向けたナノ空間区画化技術の開発2018

    • Author(s)
      仲野 瞬、小西 宏明、Dinh Huyen、中田 栄司、森井 孝
    • Organizer
      2018年度微細構造解析プラットフォームシンポジウム
    • Related Report
      2018 Research-status Report
  • [Book] 核酸科学ハンドブック2020

    • Author(s)
      日本核酸化学会、杉本 直己
    • Total Pages
      576
    • Publisher
      講談社
    • ISBN
      9784065207864
    • Related Report
      2020 Annual Research Report
  • [Book] Kinetic Control in Synthesis and Self-Assembly (Chapter 8: Covalent Bond Formation by Modular Adaptors to Locate Multiple Enzymes on a DNA Scaffold)2019

    • Author(s)
      Nakata, E.; Nakano, S.; Rajendran, A.; Morii, T.
    • Total Pages
      324
    • Publisher
      Elsevier
    • ISBN
      9780128121269
    • Related Report
      2018 Research-status Report
  • [Remarks] 京都大学 エネルギー理工学研究所  生物機能化学研究分野 森井研究室 ホームページ

    • URL

      http://www.iae.kyoto-u.ac.jp/material/index.html

    • Related Report
      2019 Research-status Report
  • [Remarks] 京都大学 エネルギー理工学研究所 エネルギー利用過程研究部門 生物機能化学研究分野 森井研究室

    • URL

      http://www.iae.kyoto-u.ac.jp/material/index.html

    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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