1992 Fiscal Year Final Research Report Summary
Fabrication and Evaluation of Functional Lipid Bilayer Membranes which oxidize NADH.
| Project/Area Number |
03805071
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
工業物理化学・複合材料
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
MATSUE Tomokazu Tohoku University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (70173797)
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| 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)
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| Project Period (FY) |
1991 – 1992
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| Keywords | NADH / Diaphorase / Monolayer / Bilayer Lipid Membrane / BLM / Ion Channel / Electron Transfer / Impedance |
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| 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.
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