Direct electrochemistry of thermophilic cytochrome P450

嗜热细胞色素 P450 的直接电化学

• 批准号: 17550149
• 负责人: NAKAMURA Nobuhumi
• 金额: $ 2.3万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17550149/
• 关键词: 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

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.

细胞色素P450催化惰性烃分子与还原性活性分子氧的选择性氧化。细胞色素P450在生物传感器和生物反应器等生物设备中具有潜在的有用应用。细胞色素P450催化反应的电化学控制由于系统易化而将使其应用得到推广。研究了PEO修饰的P450 st在PEO 200溶剂中的高温直接电子转移，并利用紫外-可见吸收光谱、共振拉曼光谱和电化学方法研究了嗜热细胞色素P450-DDAB膜的性质。拉曼光谱表明，载玻片表面DDAB膜中血红素铁的构象接近天然构象，而在塑料成型碳（PFC）电极表面，P450 st-DDAB膜的构象与血红素-DDAB膜的构象非常相似，表明PFC电极上DDAB膜中的P450 st释放了血红素。当NaBr以作为盐加入到铸膜液中时，拉曼光谱的结果表明，即使在PFC电极上，DDAB膜中的P450 st也具有近天然构象，但没有观察到具有近天然结构的P450 st的氧化还原电位。这项研究表明，当与血红素蛋白-DDAB膜一起工作时，在某些情况下，蛋白质中的血红素铁在电极表面上释放。

项目成果

The Electrochemical Properties of Thermophilic Cytochrome P450 CYP119A2 at Extremely High Temperature in Poly(ethylene oxide)

嗜热细胞色素 P450 CYP119A2 在聚环氧乙烷中极高温度下的电化学性能

• 发表时间: 2007
• 期刊: Electrochem.Commun. 9
• 作者: H.Matsumura;N.Nakamura;M.Yohda;H.Ohno
• 通讯作者: H.Ohno

Spectroscopic and Electrochemical Characterization of Cytochrome P450st-DDAB Films on a Plastic-Formed Carbon Electrode

塑料成型碳电极上细胞色素 P450st-DDAB 薄膜的光谱和电化学表征

• 发表时间: 2007
• 期刊: Electroanalysis 19
• 作者: H.Matsumura;K.Murata;S.Wiwatchaiwong
• 通讯作者: S.Wiwatchaiwong

A Novel Method for Direct Electrochemistry of a Thermoacidophilic Cytochrome P450

嗜热嗜酸细胞色素 P450 直接电化学的新方法

• 发表时间: 2006
• 期刊: Electrochem. Commun. 8
• 作者: S.Wiwatchaiwong;H.Matsumura;N.Nakamura;M.Yohda;H.Ohno;K.Murata;S.Wiwatchaiwong;K.Yokoyama;K.Yokoyama;松村洋寿;S.Wiwatchaiwong;H.Matsumura
• 通讯作者: H.Matsumura

Effect of Supporting Electrolytes on the Redox Potential for Thermophilic Cytochrome P450 in Poly(ethylene oxide)-With the Aim of the Utilization as a Catalyst for the Biocathode

支持电解质对聚环氧乙烷中嗜热细胞色素 P450 氧化还原电位的影响——目的是用作生物阴极催化剂

• 发表时间: 2006
• 期刊: Kobunshi Ronbunshu 63
• 作者: H.Matsumura;N.Nakamura;M.Yohda;H.Ohno
• 通讯作者: H.Ohno

pectrpscopic and Electrochemical Characterization of Cytochrome P450st-DDAB Films on a Plastic-formed Carbon Electrode

塑料成型碳电极上细胞色素 P450st-DDAB 薄膜的光谱和电化学表征

• 发表时间: 2007
• 期刊: Electroanalysis 19
• 作者: S.Wiwatchaiwong;H.Matsumura;N.Nakamura;M.Yohda;H.Ohno;K.Murata;S.Wiwatchaiwong
• 通讯作者: S.Wiwatchaiwong