Yamada Yohei, Hayashi Tetsuya, Sakuraba Haruhiko, Yabutani Tomoki, Takayanagi Toshio
Department of Chemical Science and Technology, Institute of Science and Technology, The University of Tokushima, Tokushima, Japan.
Anal Sci. 2013;29(1):79-83. doi: 10.2116/analsci.29.79.
We fabricated a thermostable quinoprotein aldose sugar dehydrogenase (tPQQ-ASD derived from a hyperthermophilic archaeon Pyrobaculum aerophilum) immobilized electrode. The electrode was prepared by immobilizing agarose gel mixed with the enzyme and carbon nanofiber (CNF) on a carbon paste (CP) electrode containing p-benzoquinone (BQ) as an electron mediator. The electrocatalytic response was clearly observed by the addition of D-glucose at the electrode. The electrode properties such as pH, temperature dependency and substrate selectivity basically followed the enzyme properties. The current response against D-glucose increased with measurement temperatures up to 70°C, and response perturbation caused by dissolved oxygen level was not observed at the electrode. As for the results of long-term stability evaluation, the current response was stable for 30 days when the electrode was stored in HEPES buffer solution (pH 7.0) at 4°C.
我们制备了一种固定化电极，其固定有来自嗜热古菌嗜气栖热菌的热稳定醌蛋白醛糖脱氢酶（tPQQ-ASD）。该电极是通过将与酶和碳纳米纤维（CNF）混合的琼脂糖凝胶固定在含有对苯醌（BQ）作为电子媒介体的碳糊（CP）电极上制备而成。在电极上添加D-葡萄糖时，可清晰观察到电催化响应。电极的性质，如pH值、温度依赖性和底物选择性，基本遵循酶的性质。在高达70°C的测量温度下，对D-葡萄糖的电流响应随温度升高而增加，且在电极上未观察到由溶解氧水平引起的响应扰动。至于长期稳定性评估的结果，当电极在4°C下保存在HEPES缓冲溶液（pH 7.0）中时，电流响应在30天内保持稳定。
