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Reversible utilization of enzyme by grafting and refolding

Research Project

Project/Area Number 18K04821
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 27020:Chemical reaction and process system engineering-related
Research InstitutionTokyo University of Agriculture and Technology

Principal Investigator

Ohashi Hidenori  東京農工大学, 工学(系)研究科(研究院), 准教授 (00541179)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords固定化酵素 / タンパク質 / リフォールディング / 大気圧プラズマ / グラフト重合 / 酵素 / グラフト / 可逆性
Outline of Final Research Achievements

Conventional immobilized enzyme is hard to recover its function once denatured. On the other hand, in the case of free enzyme as protein, after unfolded to a linear chain of amino acid sequences with denaturant, it can recover its function by removing denaturant to refold the active 3D structure. In the conventional design, immobilized enzyme is hard to refold, because the structure was stiffly anchored to mitigate the inactivation of protein. In the present study, novel type of immobilized enzyme was designed, which can recover its function reversibly by unfolding/refolding by temporary and subsequent covalent immobilization with terminal groups. The specific functional groups for both fixation were introduced to a model protein and to a material surface respectively, and they were combined in order. Further, atmospheric-pressure plasma graft polymerization technique was thoroughly investigated to facilitate the functionalization of material surface.

Academic Significance and Societal Importance of the Research Achievements

酵素は高い基質特異性を持ち、副生成物の少ない高選択的な反応を可能とする優れた生体触媒であり、分離操作を必要としない化学プロセスへの応用が見込まれる。一方で、酵素は化成品と比べると高価な上、失活しやすいため、失活を防ぐ目的で酵素の固定化法が開発されてきた。しかし従来の固定化法では一旦失活すると機能回復が極めて困難である。本課題では、失活しても容易に機能回復を可能とする固定化酵素設計により、高価故に見送られてきた機能性酵素を再度実用化に近づけることを可能とする。また、グラフト鎖導入を簡便に行う手法を開発したことで、様々な材料の表面に、種々の高機能を付与することが可能になった。

Report

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

    (20 results)

All 2020 2019 2018 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results) Presentation (10 results) (of which Invited: 1 results) Remarks (3 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Proton diffusion facilitated by indirect interactions between proton donors through several hydrogen bonds2019

    • Author(s)
      Takaya Ogawa, Hidenori Ohashi, Takanori Tamaki, and Takeo Yamaguchi
    • Journal Title

      Chemical Physics Letters

      Volume: 731 Pages: 136627-136627

    • DOI

      10.1016/j.cplett.2019.136627

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Autonomous shrinking/swelling phenomenon driven by macromolecular interchain cross-linking via β-cyclodextrin-triazole complexation2019

    • Author(s)
      Yuuki Sugawara, Toshiki Takei, Hidenori Ohashi, Hidenori Kuroki, Shoji Miyanishi, and Takeo Yamaguchi
    • Journal Title

      Macromolecules

      Volume: 52 Issue: 22 Pages: 8551-8562

    • DOI

      10.1021/acs.macromol.9b01627

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Inhibition of Agrobacterium tumefaciens biofilm formation by acylase I-immobilized polymer surface grafting of a zwitterionic group-containing polymer brush2019

    • Author(s)
      Qian Bao, Li Xie, Hidenori Ohashi, Masaaki Hosomi, Akihiko Terada
    • Journal Title

      Biochemical Engineering Journal

      Volume: 152 Pages: 107372-107372

    • DOI

      10.1016/j.bej.2019.107372

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Performance Analysis of a Water-Gas Shift Membrane Reactor for Integrated Coal Gasification Combined Cycle Plant2018

    • Author(s)
      Wataru Yonamine, Sivasakthivel Thangavel, Hidenori Ohashi, Chihiro Fushimi
    • Journal Title

      Energy Conversion and Management

      Volume: 174 Pages: 552-564

    • DOI

      10.1016/j.enconman.2018.08.022

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Alkali-resistant Anion Exchange Membranes with Grafted Polyelectrolyte for Fuel Cells2018

    • Author(s)
      Hidenori Ohashi, Jung Hyangmi, Xueqin Chi, Shoji Miyanishi, Takeo Yamaguchi
    • Journal Title

      Chemistry Letters

      Volume: 47 Issue: 7 Pages: 857-859

    • DOI

      10.1246/cl.180321

    • NAID

      130007402793

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Immobilization of Azospira sp. strain I13 by gel entrapment for mitigation of N 2 O from biological wastewater treatment plants: Biokinetic characterization and modeling2018

    • Author(s)
      Suenaga Toshikazu、Aoyagi Ryo、Sakamoto Nozomi、Riya Shohei、Ohashi Hidenori、Hosomi Masaaki、Tokuyama Hideaki、Terada Akihiko
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 印刷中 Issue: 2 Pages: 213-219

    • DOI

      10.1016/j.jbiosc.2018.02.014

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] 大気圧プラズマグラフト重合法の基礎的検討とその応用2020

    • Author(s)
      橋本颯人,長澤寛規,大橋秀伯
    • Organizer
      化学工学会第86年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] グラファイトの表面微量酸官能基の定量2020

    • Author(s)
      福島悠歩,清水雄介,須黒雅博,野木栄信,大橋秀伯
    • Organizer
      化学工学会第86年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 高分子溶液の粘弾性測定を用いた分子拡散性予測2020

    • Author(s)
      松井凌我,大橋秀伯
    • Organizer
      膜シンポジウム2020
    • Related Report
      2020 Annual Research Report
  • [Presentation] バナジウムレドックスフロー電池用の価数選択型電解質膜の開発2020

    • Author(s)
      杉浦響也,大橋秀伯
    • Organizer
      膜学会第42年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 高分子中の分子拡散性予測における自由体積の加成性に関する議論2020

    • Author(s)
      松井凌我,大橋秀伯
    • Organizer
      膜学会第42年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] マイクロ流路型高比表面積圧透析によるタンパク質高速濃縮2020

    • Author(s)
      加藤葵、大橋秀伯
    • Organizer
      化学工学会 第85年会
    • Related Report
      2019 Research-status Report
  • [Presentation] グラファイトの気相修飾によるLIBの高温保存性能向上2020

    • Author(s)
      大橋秀伯・藤井一洋、清水雄介、野木秀信、須黒雅博、張涵
    • Organizer
      化学工学会 第85年会
    • Related Report
      2019 Research-status Report
  • [Presentation] マイクロ流路透析を用いた高濃度タンパク質リフォールディング法の開発2019

    • Author(s)
      加藤葵、大橋秀伯
    • Organizer
      膜シンポジウム2019
    • Related Report
      2019 Research-status Report
  • [Presentation] マイクロ流路透析による高濃度タンパク質リフォールディング2019

    • Author(s)
      大橋 秀伯, 加藤 葵
    • Organizer
      化学工学会 第84年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 移動物性を基盤としたデバイス・システム設計2018

    • Author(s)
      大橋 秀伯
    • Organizer
      化学工学会 第50回秋季大会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Remarks] 大橋研究室へようこそ

    • URL

      http://web.tuat.ac.jp/~ohashilab/

    • Related Report
      2020 Annual Research Report
  • [Remarks] Welcome to Ohashi Lab 大橋研究室へようこそ

    • URL

      http://web.tuat.ac.jp/~ohashilab

    • Related Report
      2019 Research-status Report
  • [Remarks] Welcome to Ohashi Lab 大橋研究室へようこそ

    • URL

      http://web.tuat.ac.jp/~ohashilab/link.html

    • Related Report
      2018 Research-status Report
  • [Patent(Industrial Property Rights)] リチウム二次電池、及びリチウム二次電池の製造方法2020

    • Inventor(s)
      清水雄介、張涵、野木秀信、須黒雅博、大橋秀伯・藤井一洋
    • Industrial Property Rights Holder
      清水雄介、張涵、野木秀信、須黒雅博、大橋秀伯・藤井一洋
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2020-032227
    • Filing Date
      2020
    • Related Report
      2019 Research-status Report

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

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