Zhang Ke, Zhou Hao, Hu Ping, Lu Qing
Institute of Environmental Medicine, State Key Laboratory of Environment Health (Incubation), Key Laboratory of Environment and Health, Ministry of Education, Key Laboratory of Environment and Health (Wuhan), Ministry of Environmental Protection, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology #13 Hangkong Road Wuhan Hubei 430030 China
Department of Clinical Laboratory, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology #1277 Jiefang Avenue Wuhan Hubei 430022 China.
RSC Adv. 2019 Nov 14;9(64):37207-37213. doi: 10.1039/c9ra07082f. eCollection 2019 Nov 13.
It is rather difficult to achieve the direct electrochemistry of nitrate reductase (NR) as it is a complex homodimeric enzyme. However, in this study, the direct electron transfer between NR's redox centers and the electrode surface was achieved with the aid of aminated graphene sheets (am-GSs) which could immobilize NR stably and control the orientation of the enzyme molecules on the surface of the modified electrode through electrostatic attractions. Moreover, when the gold nanoparticles (AuNPs) which could act as electronic wire were introduced to the modified electrode, the NR-based enzymatic reduction of nitrate was promoted and a sensitive electrochemical response regarding the electrochemical reduction of nitrate could be obtained at the NR/AuNPs/am-GSs/GC electrode. Under optimized conditions, a wide linear range from 1.0 × 10 mol L to 2.0 × 10 mol L was acquired with a low detection limit of 7 × 10 mol L (S/N = 3). The biosensor was successfully employed to determine the total nitrogen in environmental water samples and the results were in good accordance with those obtained by ultraviolet-visible spectrophotometry.
由于硝酸还原酶(NR)是一种复杂的同二聚体酶,实现其直接电化学较为困难。然而,在本研究中,借助胺化石墨烯片(am-GSs)实现了NR氧化还原中心与电极表面之间的直接电子转移,胺化石墨烯片能够稳定固定NR,并通过静电引力控制酶分子在修饰电极表面的取向。此外,当将可作为电子导线的金纳米颗粒(AuNPs)引入修饰电极时,促进了基于NR的硝酸盐酶促还原,并且在NR/AuNPs/am-GSs/GC电极上可获得关于硝酸盐电化学还原的灵敏电化学响应。在优化条件下,获得了1.0×10⁻⁵ mol/L至2.0×10⁻³ mol/L的宽线性范围,检测限低至7×10⁻⁶ mol/L(S/N = 3)。该生物传感器成功用于测定环境水样中的总氮,结果与紫外可见分光光度法所得结果高度一致。