基于-氨基酚催化和 NADH 介导的银纳米粒子合成的新型光控比色检测硝还原酶法。

A novel light-controlled colorimetric detection assay for nitroreductase based on -aminophenol-catalyzed and NADH-mediated synthesis of silver nanoparticles.

机构信息

College of Chemistry, Nanchang University, Nanchang 330031, China.

出版信息

Anal Methods. 2021 May 20;13(19):2223-2228. doi: 10.1039/d1ay00231g.

Abstract

A novel and efficient light-controlled colorimetric assay for the quantification and detection of nitroreductase (NTR) was constructed based on p-aminophenol (pAP)-catalyzed and nicotinamide adenine dinucleotide (NADH)-mediated generation of AgNPs. Due to the hydrolysis of p-nitrophenol by NTR in the presence of NADH, the hydrolysis product can be used as a catalyst to catalyze the reduction of Ag+ by NADH under the light. As the concentration of NTR increases, the value of absorbance at ca. 400 nm (A400) decreases and the color of the solution turns from brown to bright yellow. A linear correlation was obtained between A400 and the NTR concentration in the range from 1-50 μg mL-1 and the limit of detection (LOD) is 0.27 μg mL-1. The detection system does not respond to other common biological molecules due to the specificity of enzymes and the effect of the nitroreductase inhibitor on the NTR activity was also tested. Finally, we applied the assay to determine NTR in human serum samples by spiking different concentrations of NTR with a recovery of 85.2%-92.5%.

摘要

基于对氨基苯酚（pAP）催化和烟酰胺腺嘌呤二核苷酸（NADH）介导的银纳米颗粒（AgNPs）生成，构建了一种新颖、高效的光控比色法来定量和检测硝基还原酶（NTR）。由于在 NADH 存在下 NTR 对 p-硝基苯酚的水解，水解产物可用作催化剂，在光照下催化 NADH 还原 Ag+。随着 NTR 浓度的增加，在约 400nm（A400）处的吸光度值降低，溶液颜色从棕色变为亮黄色。在 1-50μg mL-1 的范围内，A400 与 NTR 浓度之间存在线性关系，检测限（LOD）为 0.27μg mL-1。由于酶的特异性以及硝基还原酶抑制剂对 NTR 活性的影响，该检测系统不会对其他常见生物分子产生响应。最后，我们通过向人血清样品中加入不同浓度的 NTR 来测定 NTR，回收率为 85.2%-92.5%。