Basic Research for Development of Biocatalyst-modified Electrodes

生物催化剂修饰电极开发的基础研究

• 批准号: 01560150
• 负责人: IKEDA Tokuji
• 金额: $ 1.28万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01560150/
• 关键词: Biocatalyst Electrode; Enzyme electrode; Bioelectrocatalysis; Bioreactor; Biosensor; Biocatalyst; Electron Transfer; Modified Electrode; 酵素固定化電極; 電気化学リアクタ; エレクトロカタリシス; 接触電解酸化; 酸化還元酵素; NAD@NADH電解; 細胞膜タンパク質

This study was carried out for aiming at the development of enzyme-modified electrodes functioning as biocatalyst electrodes which are applicable to a wide variety of field such as those in biosensor and bioreactor technologies. Two kinds of enzymes, ie. diaphorase and membrane-bound enzymes were used. Diaphoraso was immobilized on the surface of a carbon paste electrode by the method developed by the authos. The electrode functioned as an exellent NADH sensor. A combination of the diaphorase reaction with a dehydrogenase reaction allowed for the detection of NADH and NAD as low as picomolar order, based on a cycling effect. A quantitative method has been establised for evaluating the capability of a variety of redox compounds for electron transfer mediation between the enzyme and the electrode. A quinoprotein and flavoprotein, both of which contain heme C in their molecules, were examined as candidates suitable for immobilizing on the surface of electrodes of a variety of materials. They were found to be adsorbed on these electrode surface keeping with their enzyme activity and allowing the direct electron transfer at the electrodes. Thus we have found a suitable system for constructiang the biocat alyst electrode. Some applications of this type of electrode were also studied ; for example, a possibility for microtype fructose sensor based on the direct electron transfer between a carbon paste electrode and fructose dehydrogenase adsorbed on it was suggested based on the present study.

本研究的目的是开发可应用于生物传感器和生物反应器技术等广泛领域的用作生物催化剂电极的酶修饰电极。两种酶，即。使用心肌黄酶和膜结合酶。用自行研制的固定化方法将硅藻土固定在碳糊电极表面。该电极作为一个优秀的NADH传感器。黄递酶反应与脱氢酶反应的组合允许检测低至皮摩尔级的NADH和NAD，基于循环效应。建立了一种定量方法，用于评价各种氧化还原化合物在酶和电极之间进行电子传递介导的能力。醌蛋白和黄素蛋白，这两者都含有血红素C在其分子中，被检查作为候选人适合于固定在各种材料的电极表面上。发现它们被吸附在这些电极表面上，保持它们的酶活性，并允许在电极处的直接电子转移。因此，我们找到了一个合适的体系，用于构建生物催化剂电极。还研究了这种电极的一些应用，例如，基于本研究提出了基于碳糊电极与吸附在其上的果糖脱氢酶之间的直接电子转移的微型果糖传感器的可能性。

项目成果

Tokuji Ikeda: "DーFructose dehydrogenaseーmodified carbon paste electrode containing pーbenzoquinone as a mediated amperometic Fructose seneei" Agric,Biol,Chem.,. 54. 2919-2924 (1990)

Tokuji Ikeda：“含有对苯醌作为介导的安培果糖塞尼酮的 D-果糖脱氢酶修饰碳糊电极”Agric,Biol,Chem.,. 54. 2919-2924 (1990)

池田篤治: "Amperometric response to reducing carbohydrates of an enzyme electrode based on oligosaccharide dehydrogenase." Analytica Chimica Acta. (1990)

Atsuji Ikeda：“基于寡糖脱氢酶的酶电极对碳水化合物的还原反应”（1990 年）。

S. Miyaoka, F. Matsushita, T. Ikeda and M. Senda: "Bioelectrocatalysis at Enzyme-Modified Electrodes. Effect of Various Electrode Materials." Rev. Polarogr (Kyoto). 36. 37 (1990)

S. Miyaoka、F. Matsushita、T. Ikeda 和 M. Senda：“酶修饰电极的生物电催化。各种电极材料的影响。”

T. Ikeda, K. Isono and M. Senda: "Nucleoside Oxidase Electrode. Cathodic Current Response Due to the Oxidation of Nucleosides." Rev. Polarogr (Kyoto). 36. 36 (1990)

T. Ikeda、K. Isono 和 M. Senda：“核苷氧化酶电极。核苷氧化引起的阴极电流响应。”

T. Ikeda, T. Shibata, S. Todoriki, M. Senda and H. Kinoshita: "Amperometric Response to Reducing Carbohydrates of an Enzyme Electrode Based on Oligosaccharide Dehydrogenase. Detection of Lactose and -Amylase." Analytica Chimica Acta. 230. 75-82 (1990)

T. Ikeda、T. Shibata、S. Todoriki、M. Senda 和 H. Kinoshita：“基于寡糖脱氢酶的酶电极还原碳水化合物的电流响应。乳糖和淀粉酶的检测。”