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Study on new biosensors with molecular wiring function of enzyme information

具有酶信息分子接线功能的新型生物传感器研究

基本信息

批准号：
07640814
负责人：
YAO Toshio
金额：
$ 1.22万
依托单位：
Osaka Prefecture University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07640814/
关键词：
Biosensor Enzyme electrode Molecular wire Conducting polymer 五酸化バナジウムゲル

项目摘要

Oxidoreductases such as glucose oxidase does not directly transfer electrons to conventional electrodes because the distance between their redox centers and the electrode surface exceeds, even on closest approach, the distance across which electrons are transferred at sufficient rates. We showed here the direct electrical communication between the redox center of a large enzyme molecule and a simple metal electrode (glassy carbon, platinum, etc.) established through vanadium (V) oxide and redox polymer [Os(bpy)_2(PVP)_<10>C1]C1 (bpy=bipyridyl, PVP=poly-4-vinylpyridine). As enzymes, Flavo enzymes (glucose oxidase, lactate oxidase, glutamate oxidase), quino enzymes (fructose dehydrogenase, galactose oxidase), and peroxidases were selected in this work. The enzyme was codeposited with the redox polymer and cross-linking reagent on the surface of a electrode. The electron transfer from the reduced active center (ex. FADH_2 of glucose oxidase) in the core of the enzyme to the electrode surface is facilitated via the redox polymer matrix. The characterisation of the sensor in terms of linearity, reproducibility, pH dependence, stability and the response to interfering substances was also carried out. The prepared enzyme sensors responded specifically to each substrate.

氧化还原酶如葡萄糖氧化酶不直接将电子转移到常规电极，因为它们的氧化还原中心和电极表面之间的距离甚至在最接近时也超过电子以足够的速率转移的距离。我们在这里展示了一个大的酶分子的氧化还原中心和一个简单的金属电极（玻璃碳，铂等）之间的直接电通信。通过氧化钒（V）和氧化还原聚合物[Os（bpy）_2（PVP）_<10>C1]C1（bpy=联吡啶，PVP=聚-4-乙烯基吡啶）建立。作为酶，在本工作中选择了Flavo酶（葡萄糖氧化酶、乳酸氧化酶、谷氨酸氧化酶）、quino酶（果糖脱氢酶、半乳糖氧化酶）和过氧化物酶。将酶与氧化还原聚合物和交联剂共沉积在电极表面上。电子从还原的活性中心（ex.通过氧化还原聚合物基质促进酶核心中的葡萄糖氧化酶的FADH_2）到电极表面。还对传感器的线性、重现性、pH依赖性、稳定性和对干扰物质的响应进行了表征。所制备的酶传感器对每种底物都有特异性反应。