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Fabrication and Evaluation of Functional Lipid Bilayer Membranes which oxidize NADH.

Research Project

Project/Area Number 03805071
Research Category

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

Allocation TypeSingle-year Grants
Research Field 工業物理化学・複合材料
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

MATSUE Tomokazu  Tohoku University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (70173797)

Co-Investigator(Kenkyū-buntansha) NISHINA Tatsuo  Tohoku University, Faculty of Engineering, Research Associate, 工学部, 助手 (60172673)
UCHIDA Isamu  Tohoku University, Faculty of Engineering, Professor, 工学部, 教授 (50005302)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsNADH / Diaphorase / Monolayer / Bilayer Lipid Membrane / BLM / Ion Channel / Electron Transfer / Impedance / ボルタンメトリ- / 膜透過係数 / シミュレ-ション / ジアフォラ-ゼ / 自己集積膜 / 固定化酵素
Research Abstract

1.Regeneration of NAD(H) at Diaphorase-Immobilized Surfaces
Diaphorase was immobilized onto an Au electrode by step-by-step reaction with aminoethanethiol and glutaraldehyde. The surface concentration was found to be 6x10^<-12> mol/cm^2. Cyclic voltammetry with ferrocenylmethanol (FMA) as mediator showed a remarkable increase in the oxidation peak for FMA in the presence of NADH in solution. From the intercept and slope of the 1/i vs. exp[F(E-E)/RT] plot at the onset current region, the Michaelis constant and the molecular activity have been determined, respectively, to be 8x10^<-7> M and k_2=9s^<-1>.
2.Ion-and Electron-Transport Across a Planer Bilayer Lipid Membrane (BLM)
The electron-transport across the BLM containing tetracyanoquinodimethane(TCNQ) and the ion transport through the BLM with alamethicin ion channels were investigated by electrochemical methods. From the impedance spectroscopy of the BLM, heterogeneous electron-transfer rates between redox species and the BLM were determined. The results demonstrated that hydrophobic and electrostatic interactions have important roles in the electron-transfer. The selective determination of I^- transported through alamethicin channels was also investigated by measuring the oxidation current of I^- at a microdisk electrode located near the BLM. From the analysis of the oxidation current upon application of membrane potential, the flux of I^- through the BLM was determined.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] T.MATSUE: "Electron-Transfer from NADH dehydrogenase to Polypyrrole and its Applicability to Electrochemical Oxidation of NADH" J.Electroanal.Chem.300. 111-118 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Chang: "Enzyme-Immobilized Electrode for NADH Sensing and Application to the Kinetic Studies of NADH Dependent Dehydrogenases" Analyst. 116. 793-796 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.MATSUE: "An Enzyme Switch Sensitive to NADH" J.Chem.Soc.,Chem.Commun. 1029-1031 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.YAMADA: "A Microvoltammetric Study of Permeation of Ferrocene Derivatives Through a Planer Bilayer Lipid Membrane" Biochem.Biophys.Res.Commun. 180. 1330-1334 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.MATSUE: "An Ultramicroelectrode for Determination of Intracellular Oxygen. Light-Irradiation-Induced Change in Oxygen Concentration in an Algae Protoplast" Biochim.Biophys.Acta. 1101. 69-72 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 山田 弘: "平面脂質二分子膜に近接させた電極による膜透過の評価" 電気化学. 60. 664-665 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 樺澤 明裕: "微分容量測定およびIn situ表面増強ラマン散乱によるチオール単分子膜の安定性の検討" 電気化学. 60. 989-991 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N.MATSUDA: "Surface-Enhanced Infrared and Raman Studies of Electrochemical Reuction of Self-Assembled Monolayers Formed from p-Nitrothiophenol at Silver" Chem.Lett.1385-1388 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.SAWAGUCHI: "Catalytic Capability of DiAphorase Bound to a Self-Assembled Thiol Monolayer at a Gold Electrode." Bioelectrochem.Bioenerg.29. 127-133 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 末永 智一: "マイクロ電極を用いた生体物質の極微量分析" BME. 5. 15-22 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 末永 智一: "マイクロバイオセンサ" 表面技術. 43. 759-763 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Matsue: "Electron-Transfer from NADH dehydrogenase to Polypyrrole and its Applicability to Electrochemical Oxidation of NADH" J. Electroanal. Chem.300. 111-118 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Chang: "Enzyme-Immobilized Electrode for NADH Sensing and Application to the Kinetic Studies of NADH Dependent Dehydrogenases" Analyst. 116. 793-796 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Matsue: "An Enzyme Switch Sensitive to NADH" J. Chem. Soc., Chem. Commun.1029-1031 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Yamada: "A Microvoltammetric Study of Permeation of Ferrocene Derivatives Through a Planer Bilayer Lipid Membrane." Biochem. Biophys. Res. Commun.180. 1330-1334 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Matsue: "An Ultramicroelectrode for Determination of Intracellular Oxygen. Light-Irradiation-Induced Change in Oxygen Concentration in an Algae Protoplast." Biochem. Biophys. Acta. 1101. 69-72 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Yamada: "Study of Permeation Through Planer Bilayer Lipid Membrane with a Microelectrode Located Near the Membrane." Denki Kagaku. 60. 664-665 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] A. Kabasawa: "Study on Electrochemical Stability of Thiol Monolayer on Au and Ag Electrodes by AC Impedance and In situ Surface-Enhanced Raman Scattering" Denkikagaku. 60. 989-991 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N. Matsuda: "Surface-Enhanced Infrared and Raman Studies of Electrochemical Reuction of Self-Assembled Monolayers Formed from p-Nitrothiophenol at Silver" Chem. Lett.1385-1388 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Sawaguchi: "Catalytic Capability of Diaphorase Bound to a Self-Assembled Thiol Monolayer at a Gold Electrode" Bioelectrochem. Bioenerg.29. 127-133 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H. Yamada: "Determination of Active Diaphorase Immobilized at Electrode Surfaces" Bioelectrochem. Bioenerg.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Matsue: "Trace Analysis of Biological Molecules Using Microelectrodes" BME. 5. 15-22 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Matsue: "Microbiosensor" Hyoumen Gijutsu. 43. 759-763 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.Sawaguchi: "Catalytic Capability of Diaphorase Bound to a Self-Assembled Thiol Monolayer at a Gold Electrode" Bioelectrochem.Bioenerg.29. 127-133 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 末永 智一: "マイクロ電極を用いた生体物質の極微量分析" BME. 5. 15-22 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 末永 智一: "マイクロバイオセンサ" 表面技術. 43. 759-763 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Matsuda: "Surface-Enhanced Infrared and Raman Studies of Electrochemical Reuction of Self-Assembled Monolayers Formed from p-Mitrothiophenol at Silver" Chem.Lett.1385-1388 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 樺澤 明裕: "微分容量測定およびIn situ表面増強ラマン散乱によるチオール単分子膜の安定性の検討" 電気化学. 60. 989-991 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 山田 弘: "A Microvoltammetric Study of Permeation of Ferrocene Derivatives Through a Planer Bilayer Lipid Membrane," Biochem.Biophys.Res.Commun.,. 180. 1330-1334 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 山田 弘: "平面脂質二分子膜に近接させた電極による膜透過の評価," 電気化学.

    • Related Report
      1991 Annual Research Report
  • [Publications] 末永 智一: "An Ultramicroelectrode for Determination of lntracellular Oxygen.Lightーlrradiationーlnduced Change in Oxygen Concentration in an Algae Protoplast," Biochim.Biophys.Acta.

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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