碳纳米管促进氧化还原蛋白质和酶的直接电子转移

Direct Electron Transfer of Redox Proteins and Enzymes Promoted by Carbon Nanotube

将血红蛋白(Hb)、辣根过氧化物酶(HRP)和葡萄糖氧化酶(GOx)分别固定在经碳纳米管修饰的玻碳电极(CNT/GC)上,制成Hb CNT/GC、HRP CNT/GC和GOx CNT/GC电极.Hb、HRP和GOx在CNT/GC电极表面均能发生有效和稳定的直接电子转移反应,其相应的循环伏安曲线均显示出一对几近对称的氧化还原峰;在60mV/s下,其式量电位E0'分别为-0.343V、-0.319V和-0.456V(vs.SCE,pH6.9),且不随扫速而变;以上三者在CNT/GC电极表面直接电子转移的表观速率常数ks依次为1.25±0.25、2.07±0.56和1.74±0.42s-1;根据式量电位E0'随缓冲溶液pH值的变化关系,确知在CNT/GC电极上,Hb或HRP发生的直接电化学遵从(1e+1H+)电极过程机理,而GOx发生的直接电化学反应则遵从(2e+2H+)机理.此外,固定在CNT/GC电极表面的Hb、HRP和GOx也同时表现出对各自底物的生物电催化活性.由本文制备的碳纳米管修饰电极及其固定生物蛋白质(酶)的方法具有简单、易于操作等优点,并可用于对其它生物氧化还原蛋白质和酶的直接电子转移测试.

The redox proteins (enzymes), such as hemoglobin (Hb), horseradish peroxidase (HRP) and glucose oxidase (GOx) were immobilized on the surface of the carbon nanotube modified with glassy carbon (CNT/GC) electrode, respectively. The experimental results showed that the redox proteins (enzymes) underwent effective and stable direct electron transfer reaction at the surface of CNT/GC electrode with a pair of nearly symmetrical redox peaks in phosphate buffer solution. The formal redox potential, E^(0'), is almost independent on the scan rates, the average value of E^(0') for Hb, HRP and GOx is -0.343±0.001, -0.319±0.002 and -0.456±0.001 V (vs.SCE,pH 6.9), respectively. The dependence of E^(0') on the pH of the buffer solution indicates that the direct electron transfer of Hb and HRP is a one-electron-transfer reaction process coupled with one-proton-transfer, while the GOx is a two-electron-transfer reaction process coupled with two-proton-transfer. The experimental results also demonstrated that the immobilized Hb and HRP retained their bioelectrocatalytic activity to the reduction of H_(2)O_(2), and immobilized GOx can catalyze the oxidation of glucose in the presence of ferrocene monocarboxylic acid. The method presented here can be easily extended to immobilize and obtain the direct electrochemistry of other redox enzymes or proteins.