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Development of Biosensing system based on the enzyme swiching mechanism and the application of the system

Research Project

Project/Area Number 19750065
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Analytical chemistry
Research InstitutionKanagawa Institute of Technology

Principal Investigator

IIDA Yasuhiro  Kanagawa Institute of Technology, 応用バイオ科学部, 准教授 (40329305)

Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥3,370,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥270,000)
Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥2,200,000 (Direct Cost: ¥2,200,000)
Keywords生物・生体工学 / バイオセンサ / 酵素阻害剤 / 固定化酵素 / FIA / スクリーニング / ウレアーゼ / 固定化単体 / 固定化担体
Research Abstract

本研究では、酵素を1分子のスイッチと見立て、その活性を可逆的にON/OFF制御できるシステムの構築と、そのシステムを用いた分析、更には酵素の阻害剤のスクリーニングに関する研究を展開してきた。具体的には、1)新規酵素固定化担体の開発、2)酸性ウレアーゼ阻害剤のスクリーニング、3)金属酵素を用いたコファクターセンシングである。
1)に関しては、骨格にアミノ基を有する新規なモノリスシリカの作成方法を開発し、固定化担体として、酵素を高密度で集積できること、微量の試料を迅速かつ高感度に分析できること等の有用性を例証した。2)に関しては、1)で開発した固定化担体に酸性ウレアーゼを固定化、フローシステムに組込んだ。活性を評価した後、阻害剤と作用させ活性を消失させ、再度基質を注入して活性の回復を評価することにより、阻害剤の評価を行った。このように、酵素活性のON/OFFを評価して、薬剤の持続性を容易に評価できるシステムを用いて、市販生薬を対象にスクリーニングを行い、ダイオウ中に従来の阻害剤以上に持続性のある(不可逆的に活性を阻害する)ことを見出した。また、阻害剤評価のハイスループット化を実現するため、磁気ビーズを用いてバッチ式で薬剤の持続性を評価できるシステムを構築した。3)に関しては、亜鉛(II)イオンを活性中心に持つ金属酵素の一種である、アルカリファスファターゼを用いて、補因子(コファクター)である亜鉛(II)イオンを除去、付与することにより活性を制御、付与する金属溶液の濃度と活性の回復の間の相関を用いてZn(II)イオンおよびMg(II)イオンを計測できるシステムを構築し、当該イオンの計測を行った。
本研究により、酵素を分子スイッチとして制御できる系を構築し、その活性を指標として新たな金属分析用バイオセンシングシステムを開発し、また、酵素活性の不可逆的阻害を容易に評価できる薬剤評価法を開発することが達成され、その意義は極めて大きいと思われる。

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (40 results)

All 2009 2008 2007

All Journal Article (19 results) (of which Peer Reviewed: 8 results) Presentation (19 results) Book (2 results)

  • [Journal Article] Calorimetric biosensing for copper (II) ions using immobilized ASOD-preparations2009

    • Author(s)
      I. Satoh, Y. Kobayashi, Y. Iida
    • Journal Title

      Chemical Sensor 25-A

      Pages: 82-84

    • Related Report
      2008 Final Research Report
  • [Journal Article] Calorimetric biosensing for copper(II)ions using immobilized ASOD-preparations2009

    • Author(s)
      I. Satoh, Y. Kobayashi, Y. Iida
    • Journal Title

      Chemical Sensor 25, A

      Pages: 82-84

    • Related Report
      2008 Annual Research Report
  • [Journal Article] Antibacterial activity of ZnO powder with crystallographic orientation2008

    • Author(s)
      T. Ohira, O. Yama , Z. Nakagawa
    • Journal Title

      J. Mater. Sci : Mater Med 19

      Pages: 1407-1412

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Cofactor Sensing with use of Enzyme Switching Mechanism2008

    • Author(s)
      Y. Iida, D. Nojima, K. Yoshino, K. Matsumoto, I. Satoh
    • Journal Title

      Chemical Sensor 24-A

      Pages: 121-123

    • NAID

      40015986464

    • Related Report
      2008 Final Research Report
  • [Journal Article] リアクター型センサ2008

    • Author(s)
      飯田泰広
    • Journal Title

      マテリアルインテグレーション 21

      Pages: 299-306

    • Related Report
      2008 Final Research Report
  • [Journal Article] Antibacterial Characteristics of MgO-ZnO Solid Solution with Different Chemical Compositions2008

    • Author(s)
      T. Ohira, O. Yamamoto, M. Kawamura, Y. Iida, Z. Nakagawa
    • Journal Title

      Trans. MRS-J. 33

      Pages: 1273-1276

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Influence of the mixing ratio on antibacterial characteristics of MgO-ZnO solid solution in two phase coexistence region2008

    • Author(s)
      T. Ohira, M. Kawamura, Y. I , M. Fukuda, O. Yamamoto
    • Journal Title

      J. Ceramic Society of Japan 116

      Pages: 1234-1237

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Influence of the mixing ratio on antibacterial characteristics of MgO-ZnO solid solution in two phase coexistence region2008

    • Author(s)
      T. Ohira, M. Kawamura, Y. Iida, M. Fukuda, O. Yamamoto
    • Journal Title

      J. Ceramic Society of Japan 116

      Pages: 1234-1237

    • NAID

      110006987523

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Antibacterial Characteristics of MgO-ZnO Solid Solution with Different Chemical Compositions2008

    • Author(s)
      T. Ohira, O. Yamamoto, M. Kawamura, Y. Iida, Z. Nakagawa
    • Journal Title

      Trans. MRS-J 33

      Pages: 1273-1276

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] リアクター型センサ2008

    • Author(s)
      飯田泰広
    • Journal Title

      マテグアルインテグレーション 21

      Pages: 299-306

    • Related Report
      2008 Annual Research Report
  • [Journal Article] Antibacterial activity of ZnO powder with crystallographic orientation2008

    • Author(s)
      Toshiaki Ohira, Osamu Yamamoto, Yasuhiro Iida, Zenbe-e Nakagawa
    • Journal Title

      J. Mater. Sci: Mater Med 19

      Pages: 1407-1412

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cofactor Sensing with use of Enzyme Switching Mechanism2008

    • Author(s)
      Yasuhiro IIDA, Daisuke Nojima, Kakeru Yoshino, Kunio Matsumoto, Ikuo SATOH
    • Journal Title

      Chemical Sensor 24-A

      Pages: 121-123

    • NAID

      40015986464

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Development of Nocel Silica Monolith and the Application to the Support for Immobilized Enzyme2007

    • Author(s)
      Y.Tsukada, N. Murayama, J. Yamaguchi, T. Saito, I. Satoh
    • Journal Title

      The papers of Technical Meeting on Chemical Sensor IEE Japan CHS-07

      Pages: 29-32

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Screening of Enzyme Inhibitors with Use of an Immobilized Enzyme as a Recognition Element2007

    • Author(s)
      Y. Iida, Y.Takahara, and I. Satoh
    • Journal Title

      Chemical Sensor 23-A

      Pages: 25-27

    • NAID

      40015522416

    • Related Report
      2008 Final Research Report
  • [Journal Article] Development of novel supporting materials for immobilizing enymes and the application of the preparation to micro biosensing2007

    • Author(s)
      Y. Iida, S. Aki, N. Murayama, Y. Tsukada, I. Satoh, and O. Yamamoto
    • Journal Title

      Chemical Sensor 23-B

      Pages: 100-102

    • NAID

      40015136605

    • Related Report
      2008 Final Research Report
  • [Journal Article] アルコール飲料中の尿素分析用FIAシステム2007

    • Author(s)
      飯田泰広・佐藤生男
    • Journal Title

      Flow Injection Anal. 24

      Pages: 90-101

    • NAID

      40015893956

    • Related Report
      2008 Final Research Report 2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Development of Nocel Silica Monolith and the Application to the Support for Immobilized Enzyme2007

    • Author(s)
      Y. Iida, Y. Tsukada, N. Murayama, J. Yamaguchi, T. Saito, I. Satoh
    • Journal Title

      The papers of Technical Meeting on Chemical Sensor IEE Japan CHS-07

      Pages: 29-32

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Screening of Enzyme Inhibitors with Use of an Immobilized Enzyme as a Recognition Element2007

    • Author(s)
      Y. Iida, Y. Takahara, and I. Satoh
    • Journal Title

      Chemical Sensor 23-A

      Pages: 25-27

    • NAID

      40015522416

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Evelopemnt of novel supporting materials for immobilizing enymes and the application of the preparation to micro biosensing2007

    • Author(s)
      Yasuhiro IIDA, Sho Aki, Naoko Murayama, Yuka Tsukada, Ikuo SATOH, and Osamu Yamamoto
    • Journal Title

      Chemical Sensor 23-B

      Pages: 100-102

    • Related Report
      2007 Annual Research Report
  • [Presentation] 酵素スイッチング機構を用いたコファクターセンシング2008

    • Author(s)
      飯田泰広、野島大佑、吉野翔、松本邦男、佐藤生男
    • Organizer
      電気化学会第74回大会
    • Related Report
      2008 Final Research Report
  • [Presentation] 持続的阻害効果を指標とする新規薬剤評価法の開発とウレアーゼ阻害剤探索への適用2008

    • Author(s)
      管博臣・安藝翔・佐藤生男・飯田泰広
    • Organizer
      防菌防黴学会第35回年次大会
    • Related Report
      2008 Final Research Report
  • [Presentation] Determination of zinc (II) ions with use of enzyme switching mechanism2008

    • Author(s)
      D. Nojima, I. Satoh, Y. Iida
    • Organizer
      The 15th International Conference on Flow Injection Analysis
    • Related Report
      2008 Final Research Report
  • [Presentation] Development of novel silica monolith as a support for immobilized enzyme based on the FIA system and application of the system for determination of urea2008

    • Author(s)
      Y. Iida, H. Kan, I. Satoh
    • Organizer
      The 15th International Conference on Flow Injection Analysis
    • Related Report
      2008 Final Research Report
  • [Presentation] Evaluation of properties of catechin as a tyrosinase inhibition2008

    • Author(s)
      T. Satoh, N. Maezumi, Y. Iida
    • Organizer
      The 15th International Conference on Flow Injection Analysis
    • Related Report
      2008 Final Research Report
  • [Presentation] Development of a microfluidic system and application of the system with use of immobilized acid urease to screening of urease inhibitors from herbal medicines2008

    • Author(s)
      H. Kan, S. Aki, Y. Iida
    • Organizer
      The 15th International Conference on Flow Injection Analysis
    • Related Report
      2008 Annual Research Report 2008 Final Research Report
  • [Presentation] Flow-injection analysis of Zn (II) and Mg (II) ions with an ALP-column based on an apoenzyme reactivation method2008

    • Author(s)
      D. Nojima, T. Satoh, I. Satoh, Y. Iida
    • Organizer
      214th Meeting of The Electrochemical Society
    • Related Report
      2008 Final Research Report
  • [Presentation] Development of novel screening method with use of an immobilized enzyme as a recognition element and the investigation of the properties of inhibitors2008

    • Author(s)
      Y. Iida, I. Satoh, N. Maezumi
    • Organizer
      214th Meeting of The Electrochemical Society
    • Related Report
      2008 Annual Research Report 2008 Final Research Report
  • [Presentation] Flow-injection analysis of Zn(II) and Mg(II) ions with an ALP-column based on an apoenzyme reactivation method2008

    • Author(s)
      D. Nojima, T. Satoh, I. Satoh, Y. Iida
    • Organizer
      214th Meeting of The Electrochemical Society
    • Place of Presentation
      Hawaii, USA
    • Related Report
      2008 Annual Research Report
  • [Presentation] Evaluation of properties of catechin as a tyrosinase inhibition2008

    • Author(s)
      T. Satoh, N. Maezumi, Y. Iida
    • Organizer
      The 15th International Conference on Flow Injecti on Analysis
    • Place of Presentation
      Aichi, Japan
    • Related Report
      2008 Annual Research Report
  • [Presentation] Development of novel silica monolith as a support for immobilized enzyme based on the FIA system and application of the system for determination of urea2008

    • Author(s)
      Y. Iida, H. Kan, I. Satoh
    • Organizer
      The 15th International Conference on Flow Injecti on Analysis
    • Place of Presentation
      Aichi, Japan
    • Related Report
      2008 Annual Research Report
  • [Presentation] Determination of zinc(II)ions with use of enzyme switching mechanism2008

    • Author(s)
      D. Nojima, I. Satoh, Y. Iida
    • Organizer
      The 15th International Conference on Flow Injecti on Analysis
    • Place of Presentation
      Aichi, Japan
    • Related Report
      2008 Annual Research Report
  • [Presentation] 持続的阻害効果を指標とする新規薬剤評価法の開発とウレアーゼ阻害剤探索への適用2008

    • Author(s)
      管博臣・安藝翔・佐藤生男・飯田泰広
    • Organizer
      防菌防〓学会第35回年次大会
    • Place of Presentation
      静岡(浜松)
    • Related Report
      2008 Annual Research Report
  • [Presentation] 酵素スイッチング機構を用いたコファクターセンシング2008

    • Author(s)
      飯田泰広、野島大佑、吉野 翔、松本邦男、佐藤生男
    • Organizer
      電気化学会第74回大会
    • Place of Presentation
      山梨大学
    • Related Report
      2007 Annual Research Report
  • [Presentation] 金メッシュ固定化アルカリホスファターゼを用いた微量亜鉛(II)イオンの計測2007

    • Author(s)
      野島大佑・佐藤生男・飯田泰広
    • Organizer
      Separation Science 2007
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Presentation] アミノ基導入型新規モノリスカラムの開発2007

    • Author(s)
      安藝翔・佐藤生男・飯田泰広
    • Organizer
      Separation Science 2007
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Presentation] 新規モノリスカラムの作成と微量バイオセンシングへの適用2007

    • Author(s)
      飯田泰広・安藝翔・佐藤生男
    • Organizer
      Separation Science 2007
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Presentation] 酵素固定化用モノリスカラムの開発と微量計測への応用2007

    • Author(s)
      飯田泰広・安藝翔・村山尚子・塚田結香・佐藤生男・山本修
    • Organizer
      2007年電気化学会秋季大会
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Presentation] 新規モノリスシリカカラムの開発と酵素固定化担体への適用2007

    • Author(s)
      飯田泰広・塚田結香・村山尚子・山口淳一・斎藤貴・佐藤生男
    • Organizer
      平成19年度電気学会センサ・マイクロマシン部門総合研究会
    • Related Report
      2008 Final Research Report 2007 Annual Research Report
  • [Book] 先進化学センサ2008

    • Author(s)
      飯田泰広(分担)
    • Publisher
      (株)ティー・アイ・シィー
    • Related Report
      2008 Final Research Report
  • [Book] 先進化学センサ2008

    • Author(s)
      飯田泰広(分担)
    • Total Pages
      365
    • Publisher
      (株)テイー・アイ・シイー
    • Related Report
      2008 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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