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Fabrication and Evaluation of Enzyme Switches

Research Project

Project/Area Number 01470076
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 工業物理化学
Research InstitutionTohoku University

Principal Investigator

UCHIDA Isamu  Tohoku University, Faculty of Engineering, Professor, 工学部 教授 (50005302)

Co-Investigator(Kenkyū-buntansha) NISHINA Tatsuo  Tohoku University, Faculty of Engineering, Research Associate, 工学部, 助手 (60172673)
MATSUE Tomokazu  Tohoku University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (70173797)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1989: ¥4,900,000 (Direct Cost: ¥4,900,000)
KeywordsEnzyme switch / Microarray electrode / Diaphorase / Mediator / Polypyrrole / NADH / Enzyme-immobilized / Conductive polymer
Research Abstract

The purpose pf the present project is to fabricate enbricate enzyme switches which operate based on the specific catalytic capability of enzymes. The following results are obtained throughout this project.
1. Diaphorase (Dp) and glucose oxidase (GOD) were immobilized on electrode surfaces by electropolymerization of pyrroles. Voltammetric behavior the enzyme-immobilized electrodes indicated that these enzymes remained their activity. We have proposed a new electrochemical method for determination of various kinetic parameters of the enzyme reactions. A detailed analysis of the parameters suggested that active center of Dp is hydrophobic.
2. The electrochemical behavior of interdigitated microarray electrodes (Au, Pt, or ITO ; eletrode and gap widths, 4-20mum) was investigated in stationary and flowing conditions. The array eletrodes have capabilities of magnifying the current responses due to redox cycling and their behavior can be predicted by the digital simulation. The array electrodes coated with redox polymers exhibit switch-like characteristics.
3. Based on the above results, an enzyme device was fabricated by the eletrochemical coating pyrrole/N-methylpyrrole copolymer containing Dp on the microarray electrodes. The addition of NADH, a substrate for Dp, resulted in reduction of the polymer on the array surface to insulating state through the enzyme reaction. Therefore, the array device exhibited the on-off switching characteristics upon of NADH. The treatment of array electrode with octadecyltriethoxysilane made the surface hydrophobic ; this process is essential to obtain this polymer formation at gap area and rapid switching responses.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (43 results)

All Other

All Publications (43 results)

  • [Publications] 末永 智一: "Determination of Catechlamines by an Electrochemical FlowーThrough Detector with IndiumーTin Oxide Microelectrode Arrays Coated with Nafion" Chemistry Letters. 133-136 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 張 憲彰: "Effect of Viologen Structure on Electroreduction of NAD^+ Catalyzed by Diaphorase Immobilized on Electrodes" Chemistry Letters. 1119-1122 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "Multichannel Electrochemical Detection System for Flow Analysis" Analytical Chemistry. 62. 407-409 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "Electrooxidation of NADH with Co(phen)^<3+>_3 Catalyzed by NADH Dehydrogenase Purified from Bacillus Stearothermophilus" Bioelectrochemistry and Bioenergetics. 24. 347-354 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "Electron Transferase Activity of Diaphorase(NADH:Acceptor Oxidoreductase)from Bacillus Stearoーthermophilus" Biochimica et Biophysica Acta. 1038. 29-38 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 張 憲彰: "Enzyme Sensors for LーAmino Acid Based on Electrochemical Detection of NADH" Denki Kagaku. 58. 1211-1212 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 青木 純: "Electrochemical Response at Microarray Electrodes in Flowing Streams and Application to Determination of Catecholamrnes" Analytical Chemistry. 62. 2206-2210 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "ElectronーTransfer from NADH to Polypyrroie and its Applicabitity to Electrochemical Oxidation of NADH" J.Electroanalytical Chemistry.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 青木 純: "Multichannel Electrochemical Detection with Microelectrode Array in Flowing Streams" Analytical Chemistry.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 張 憲彰: "A Novel Polyviologen Electrode Fabricated by Electrochemical Crosslinking" Chemical Communications.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 張 憲彰: "Enzymeーimmobilized for NADH sensing and Application to the Kinetic Studies of NADH Dependent Dehydrogenases" The Analyst.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "An Enzyme Switch Sensitive to NADH" Chemical Communications.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "Ultramicroelectrodes as Sensing Devices for Biological Moleculues,Chemical Sensor Technolgy vol.3" Kodansha / Elsevier, 17 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Matsue,: "Determination of Catecholamines by an Electrochemical Flow-Through Detector with Indium-Tin Oxide Microelectrode Arrays Coated with Nafion." Chem. Lett.,. 133-136 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Chang,: "Effect of Viologen Structure on Electroreduction of NAD^+ Catalyzed by Diaphorase Immobilized on Electrodes." Chem. Lett.,. 1119-1122 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Matsue,: "Multichannel Electrochemical Detection System for Flow Analysis." Anal. Chem.62. 407-409 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Maysue,: "Electron Transferase Activity of Diaphorase (NADH : Acceptor Oxidoreductase) from Bacillus Stearothermophius." Biochim. Biophys. Acta,. 1038. 29-38 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Chang,: "Enzyme Sensors for L-Amion Acid Based on Electrochemical Detection of NADH." Denki Kagaku. 58. 1211-1212 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] A. Aoki,: "Electrochemical Response at Microarray Electrodes in Flowing Streams and Application to Determination of Catecholamines." Anal. Chem.62. 2206-2210 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Matsue,: "Electron-Transfer from NADH to Polypyrrole and Its Applicability to Electrochemical Oxidation of NADH." J. Electroanal. Chem.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] A. Aoki,: "Multichannel Electrochemical Detection with Microelctrode Array in Flowing Strems."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Chang,: "A Novel Polyviologen Electrode Fabricated by Electrichemical Crosslinking."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Chang,: "Enzyme-Immobilized Electrode for NADH Sensing and Application to the Kinetic Studies of NADH Dependent Dehydrogenases."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Matsue,: "An Enzyme Switch Sensitive to NADH."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Matsue,: Kodansha/Elsevier. Ultramicroelectrodes as Sensing Devices for Biological Molecules, in "Chemical Sensor Technology. Vol. 3", (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 末永 智一: "Determination of Catecholamines by an Electrochemical FlowーThrough Detector with IndiumーTin Oxide Microelectrode Arrays Coated with Nafion" Chemistry Letters. 133-136 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 張 憲彰: "Effect of Viologen Structure on Electroreduction of NAD^+ Catalyzed by Diaphorase Immobilized on Electrodes" Chemistry Letters. 1119-1122 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "Multichannel Electrochemical Detection System for Flow Analysis" Analytical Chemistry. 62. 407-409 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "Electron Transferase Activity of Diaphorase(NADH:Acceptor Oxidoreductase) from Bacillus Stearoーthermophilus" Biochimica et Biophysica Acta. 1038. 29-38 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "Electrooxidation of NADH with Co(Phen)^<3+>_3 Catalyzed by NADH Dehydrogenase Purified from Bacillus Stearothermophilus" Bioelectrochemistry and Bioenergetics. 24. 347-354 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 張 憲彰: "Enzyme Sensors for LーAmino Acid Based on Electrochemical Detection of NADH" Denki kagaku. 58. 1211-1212 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 青木 純: "Electrochemical Response at Microarray Electrodes in Flowing Streams and Application to Determination of Catecholamines" Analytical Chemistry. 62. 2206-2210 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "ElectronーTransfer from NADH to polypyrrole and its Applicability to Electrochemical Oxidation of NADH" J.Electroanalytical Chemistry.

    • Related Report
      1990 Annual Research Report
  • [Publications] 青木 純: "Multichannel Electrochemical Detection with Microelectrode Array in Flowing Streams" Analytical Chemistry.

    • Related Report
      1990 Annual Research Report
  • [Publications] 張 憲彰: "A Novel Polyviologen Electrode Fabricated by Electrochemical Cresslinking" Chemical Communications.

    • Related Report
      1990 Annual Research Report
  • [Publications] 張 憲彰: "Enzymeーimmobilized Electrode for NADH sensing and Application to the Kinetic Studies of NADH Dependent Dehydroqenases" The Analyst.

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "An Enzyme Switch Sensitive to NADH" Chemical Communications.

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永 智一: "Ultramicroelectrodes as Sensing Devices for Biological Moleculues,Chemical Sensor Technology Vol.3" Kodansha/Elsevier, 17 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 末永智一: "A Multichannel Electrochemical Detection System for Flow Analysis" Analytical Chemistry. 62. 407-409 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 末永智一: "Electron Transferase Activity of Diaphorase(NADH:Acceptor)(Oxidoreductase)Purified from Bacillus Stearothermophilus" Biochimica et Biophysica Acta.

    • Related Report
      1989 Annual Research Report
  • [Publications] 青木純: "Electrochemical Response at Microarray Electrodes in Flowing Streams and Application to Determination of Catecholamines" Analytical Chemistry.

    • Related Report
      1989 Annual Research Report
  • [Publications] 末永智一: "Electrooxidation of NADH with Co(phen)_3^<3+> Catalyzed by NADH Dehydrogenase Purified from Bacillus Stearothermophilus" Bioelectrochemistry and Bioenergetics.

    • Related Report
      1989 Annual Research Report
  • [Publications] 末永智一: "Ultramicroelectrodes as Sensing Devices for Biological Molecules" Kodansha/Elsevier,

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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