School of Materials Science and Engineering, University of Jinan, Jinan 250022, China.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China.
Anal Chem. 2022 Mar 1;94(8):3485-3493. doi: 10.1021/acs.analchem.1c04018. Epub 2022 Feb 16.
Distinguished by the coupled catalysis-facilitated high turnover and admirable specificity, enzyme cascades have sparked tremendous attention in bioanalysis. However, three-enzyme cascade-based versatile platforms have rarely been explored without resorting to tedious immobilization procedures. Herein, we have demonstrated that formamide-converted transition metal-nitrogen-carbon (f-MNC, M = Fe, Cu, Mn, Co, Zn) with a high loading of atomically dispersed active sites possesses intrinsic peroxidase-mimetic activity following the activity order of f-FeNC > f-CuNC > f-MnNC > f-CoNC > f-ZnNC. Ulteriorly, benefitting from the greatest catalytic performance and explicit catalytic mechanism of f-FeNC, versatile enzyme cascade-based colorimetric bioassays for ultrasensitive detection of diabetes-related glucose and α-glucosidase (α-Glu) have been unprecedentedly devised using f-FeNC-triggered chromogenic reaction of 3,3',5,5'-tetramethylbenzidine as an amplifier. Notably, several types of α-Glu substrates can be effectively utilized in this three-enzyme cascade-based α-Glu assay, and it can be further employed for screening α-Glu inhibitors that are used as antidiabetic and antiviral drugs. These versatile assays can also be extended to detect other HO-generating or -consuming biomolecules and other bioenzymes that are capable of catalyzing glucose generation procedures. These nanozyme-involved multienzyme cascades without intricate enzyme-engineering techniques may provide a concept to facilitate the deployment of nanozymes in celestial versatile bioassay fabrication, disease diagnosis, and biomedicine.
酶级联因其耦合催化促进的高转化率和令人钦佩的特异性而在生物分析中引起了极大的关注。然而，在不借助繁琐的固定化程序的情况下，很少有人探索基于三酶级联的通用平台。在此，我们证明了具有原子分散活性位点的高负载量的甲酰胺转化的过渡金属-氮-碳（f-MNC，M=Fe、Cu、Mn、Co、Zn）具有固有的过氧化物酶模拟活性，其活性顺序为 f-FeNC>f-CuNC>f-MnNC>f-CoNC>f-ZnNC。此外，由于 f-FeNC 具有最大的催化性能和明确的催化机制，我们设计了基于多种酶级联的比色生物分析方法，以 f-FeNC 触发的 3,3',5,5'-四甲基联苯胺显色反应为放大器，用于超灵敏检测与糖尿病相关的葡萄糖和α-葡萄糖苷酶（α-Glu）。值得注意的是，该三酶级联的α-Glu 测定法可有效利用多种类型的α-Glu 底物，还可用于筛选用作抗糖尿病和抗病毒药物的α-Glu 抑制剂。这些通用测定法还可扩展到检测其他产生或消耗 HO 的生物分子和其他能够催化葡萄糖生成步骤的生物酶。这些无需复杂酶工程技术的纳米酶级联反应可能为在多功能生物分析、疾病诊断和生物医学中应用纳米酶提供了一个概念。
