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Fabrication and Evaluation of Enzyme Switches

酶开关的制作和评估

基本信息

批准号：
01470076
负责人：
UCHIDA Isamu
金额：
$ 4.1万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
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/en/grant/KAKENHI-PROJECT-01470076/
关键词：
Enzyme switch Microarray electrode Diaphorase Mediator Polypyrrole NADH Enzyme-immobilized Conductive polymer

项目摘要

The purpose pf the present project is to fabricate enbricate enzyme switches which operate based on the specific catalytic capability of enzymes. The following results are obtained throughout this project.1. Diaphorase (Dp) and glucose oxidase (GOD) were immobilized on electrode surfaces by electropolymerization of pyrroles. Voltammetric behavior the enzyme-immobilized electrodes indicated that these enzymes remained their activity. We have proposed a new electrochemical method for determination of various kinetic parameters of the enzyme reactions. A detailed analysis of the parameters suggested that active center of Dp is hydrophobic.2. The electrochemical behavior of interdigitated microarray electrodes (Au, Pt, or ITO ; eletrode and gap widths, 4-20mum) was investigated in stationary and flowing conditions. The array eletrodes have capabilities of magnifying the current responses due to redox cycling and their behavior can be predicted by the digital simulation. The array electrodes coated with redox polymers exhibit switch-like characteristics.3. Based on the above results, an enzyme device was fabricated by the eletrochemical coating pyrrole/N-methylpyrrole copolymer containing Dp on the microarray electrodes. The addition of NADH, a substrate for Dp, resulted in reduction of the polymer on the array surface to insulating state through the enzyme reaction. Therefore, the array device exhibited the on-off switching characteristics upon of NADH. The treatment of array electrode with octadecyltriethoxysilane made the surface hydrophobic ; this process is essential to obtain this polymer formation at gap area and rapid switching responses.

本项目的目的是制造基于酶的特定催化能力运行的覆盖酶开关。本项目取得了以下成果： 1．通过吡咯的电聚合将心肌黄酶（Dp）和葡萄糖氧化酶（GOD）固定在电极表面。酶固定电极的伏安行为表明这些酶仍保持其活性。我们提出了一种新的电化学方法来测定酶反应的各种动力学参数。详细参数分析表明Dp活性中心具有疏水性。 2．在静止和流动条件下研究了叉指微阵列电极（Au、Pt 或 ITO；电极和间隙宽度，4-20mum）的电化学行为。阵列电极具有放大氧化还原循环引起的电流响应的能力，并且可以通过数字模拟来预测它们的行为。涂有氧化还原聚合物的阵列电极表现出类似开关的特性。 3．基于上述结果，通过在微阵列电极上电化学涂覆含Dp的吡咯/N-甲基吡咯共聚物制备了酶装置。添加 Dp 的底物 NADH，通过酶反应将阵列表面的聚合物还原至绝缘状态。因此，该阵列器件表现出基于NADH的开关特性。阵列电极采用十八烷基三乙氧基硅烷处理，使其表面具有疏水性；该过程对于在间隙区域形成聚合物和快速切换响应至关重要。