Institute of Analytical Science / Shaanxi Provincial Key Laboratory of Electroanalytical Chemistry, Northwest University, Xi'an, Shaanxi 710069, China.
Bioelectrochemistry. 2012 Aug;86:46-53. doi: 10.1016/j.bioelechem.2012.01.009. Epub 2012 Feb 1.
We report on the direct electrochemistry of myoglobin (Mb) immobilized on a composite matrix based on chitosan (CHIT) and titanium carbide nanoparticles (TiC NPs) underlying on glassy carbon electrode (GCE). The cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the modified electrode. In deaerated buffer solutions, the cyclic voltammetry of the composite films of Mb-TiC NPs-CHIT showed a pair of well-behaved redox peaks that are assigned to the redox reaction of Mb, confirming the effective immobilization of Mb on the composite film. The electron transfer rate constant was estimated to be 3.8 (±0.2)·s(-1), suggested that the interaction between the protein and certain electrode surfaces may mimic some physiological situations and may elucidate the relationship between the protein structures and biological functions. The linear dynamic range for the detection of hydrogen peroxide was 0.5-50 μM with a correlation coefficient of 0.999 and the detection limit was estimated at about 0.2 μM (S/N=3). The calculated apparent Michaelis-Menten constant was 0.07 (±0.01) mM, which suggested a high affinity of the redox protein-substrate. The immobilized Mb in the TiC NPs-CHIT composite film retained its bioactivity. Furthermore, the method presented here can be easily extended to immobilize and obtain the direct electrochemistry of other redox enzymes or proteins.
我们报告了基于壳聚糖(CHIT)和碳化钛纳米粒子(TiC NPs)的复合基质固定化肌红蛋白(Mb)的直接电化学。循环伏安法和电化学阻抗谱用于表征修饰电极。在脱氧缓冲溶液中,Mb-TiC NPs-CHIT 复合膜的循环伏安法显示出一对行为良好的氧化还原峰,归因于 Mb 的氧化还原反应,证实了 Mb 在复合膜上的有效固定化。电子转移速率常数估计为 3.8(±0.2)·s(-1),表明蛋白质与某些电极表面之间的相互作用可能模拟某些生理情况,并阐明蛋白质结构与生物功能之间的关系。检测过氧化氢的线性动态范围为 0.5-50 μM,相关系数为 0.999,检测限约为 0.2 μM(S/N=3)。计算出的表观米氏常数为 0.07(±0.01)mM,表明氧化还原蛋白-底物具有高亲和力。固定在 TiC NPs-CHIT 复合膜中的 Mb 保留了其生物活性。此外,这里提出的方法可以很容易地扩展到固定化和获得其他氧化还原酶或蛋白质的直接电化学。