Amperometric inhibitive biosensor based on horseradish peroxidase-nanoporous gold for sulfide determination.
Amperometric inhibitive biosensor based on horseradish peroxidase-nanoporous gold for sulfide determination.
复制标题
基于辣根过氧化物酶-纳米孔金的电流抑制生物传感器用于硫化物测定
DOI：
10.1038/srep30905
复制
发表时间：
2016-08-12
影响因子：
4.6
通讯作者：
Wang X
中科院分区：
文献类型：
--
作者：
Sun H;Liu Z;Wu C;Xu P;Wang X
硫化物作为一种众所周知的有毒污染物，对人体健康有害。在本研究中，开发了一种简单而灵敏的电流抑制生物传感器，用于测定环境中的硫化物。通过将纳米孔金（NPG）固定在玻碳电极（GCE）上，并将辣根过氧化物酶（HRP）封装在NPG上，以邻苯二胺（OPD）为底物，基于硫化物对HRP活性的抑制作用，成功构建了用于硫化物检测的HRP/NPG/GCE生物电极。由此产生的HRP/NPG/GCE生物电极在硫化物检测中实现了0.1–40μM的宽线性范围，具有1720μA mM−1cm−2的高灵敏度和0.027μM的低检测限。另外，通过Michaelis-Menten分析，硫化物对HRP的抑制是与OPD作为底物的竞争性抑制。值得注意的是，通过使用差分脉冲伏安法 (DPV) 技术在脱气 PBS（50 mM，pH 7.0）中清洗 HRP/NPG/GCE 生物电极仅 60 秒，即可快速恢复 HRP 活性。此外，还实现了HRP/NPG/GCE生物电极对硫化物的真实样品分析。基于以上结果，与之前报道的抑制性生物传感器相比，HRP/NPG/GCE生物电极可能是真正测定硫化物的更好选择。
As a well-known toxic pollutant, sulfide is harmful to human health. In this study, a simple and sensitive amperometric inhibitive biosensor was developed for the determination of sulfide in the environment. By immobilizing nanoporous gold (NPG) on glassy carbon electrode (GCE), and encapsulating horseradish peroxidase (HRP) onto NPG, a HRP/NPG/GCE bioelectrode for sulfide detection was successfully constructed based on the inhibition of sulfide on HRP activity witho-Phenylenediamine (OPD) as a substrate. The resulted HRP/NPG/GCE bioelectrode achieved a wide linear range of 0.1–40 μM in sulfide detection with a high sensitivity of 1720 μA mM−1cm−2and a low detection limit of 0.027 μM. Additionally, the inhibition of sulfide on HRP is competitive inhibition with OPD as a substrate by Michaelis-Menten analysis. Notably, the recovery of HRP activity was quickly achieved by washing the HRP/NPG/GCE bioelectrode using differential pulse voltammetry (DPV) technique in deaerated PBS (50 mM, pH 7.0) for only 60 s. Furthermore, the real sample analysis of sulfide by the HRP/NPG/GCE bioelectrode was achieved. Based on above results, the HRP/NPG/GCE bioelectrode could be a better choice for the real determination of sulfide compared to inhibitive biosensors previously reported.