| Project/Area Number |
17550149
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
NAKAMURA Nobuhumi Tokyo University of Agriculture and Technology, Institute of Symbiotic Agriculture and Technology, Associate Professor, 大学院共生科学技術研究院, 助教授 (60313293)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Cytochrome P450 / Electrocatalytic reaction / Immobilization of protein / Electron transfer reaction / Ionic liquid / Resonance Raman spectroscopy / Redox reaction / site-directed mutagenesis / シトクロムP450 / 酸化還元電位 / 直接電気化学 / タンパク質固定化法 / サイクリックボルタンメトリー / 表面増強共鳴ラマンスペクトル / 酵素 / 触媒反応 / 非水溶媒 / 化学修飾 |
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| Research Abstract |
Cytochrome P450s catalyze the selective oxidation of inert hydrocarbon molecules with reductively activate molecular oxygen. Cytochrome P450s have a potentially useful application for biodevices such as biosensors and bioreactors. Electrochemical controlling of the cytochrome P450 catalytic reaction will cause the application to spread because of system facilitation. We investigated the direct electron transfer of P450st modified with PEO in PEO200 solvent at remarkably high temperatures.
We have also studied the characterization of thermophilic cytochrome P450-didodecyldimethylammoniumbromide (DDAB) films by using UV-vis absorption, resonance Raman spectroscopy, and electrochemical methods. The observed Raman spectrum indicated near-native conformation of the heme iron in DDAB film on the surface of a glass slide, while on the surface of a plastic-formed carbon (PFC) electrode, the conformation of P450st-DDAB was very similar to that of heme-DDAB film, suggesting the release of heme from P450st in DDAB films on PFC electrodes. When NaBr was added as salt to the casting solution, the result of Raman spectrum indicated near-native conformation of P450st in DDAB film even on the PFC electrode, but no redox potential of P450st which has near native structure was observed. This study suggests the essential experimental conditions when working with heme protein-DDAB films as, in some cases, heme iron from proteins is released on the surface of the electrode.
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