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基于石墨烯量子点分散血红素的纳米酶用于比色检测谷胱甘肽。

Nanozyme Based on Dispersion of Hemin by Graphene Quantum Dots for Colorimetric Detection of Glutathione.

机构信息

Guangxi Medical University Cancer Hospital, Guangxi Medical University, Nanning 530021, China.

Key Laboratory of Surface & Interface Science of Polymer Materials of Zhejiang Province, Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou 310018, China.

出版信息

Molecules. 2022 Oct 11;27(20):6779. doi: 10.3390/molecules27206779.

Abstract

Compared with natural enzymes, nanozymes have the advantages of good catalytic performance, high stability, low cost, and can be used under extreme conditions. Preparation of highly active nanozymes through simple methods and their application in bioanalysis is highly desirable. In this work, a nanozyme based on dispersion of hemin by graphene quantum dot (GQD) is demonstrated, which enables colorimetric detection of glutathione (GSH). GQD was prepared by a one-step hydrothermal synthesis method. Hemin, the catalytic center of heme protein but with low solubility and easy aggregation that limits its catalytic activity, can be dispersed with GQD by simple sonication. The as-prepared Hemin/GQD nanocomplex had excellent peroxidase-like activity and can be applied as a nanozyme. In comparison with natural horseradish peroxidase (HRP), Hemin/GQD nanozyme exhibited a clearly reduced Michaelis-Menten constant () when tetramethylbenzidine (TMB) was used as the substrate. With HO being the substrate, Hemin/GQD nanozyme exhibited a higher maximum reaction rate () than HRP. The mechanisms underlying the nanozyme activity were investigated through a free radical trapping experiment. A colorimetric platform capable of sensitive detection of GSH was developed as the proof-of-concept demonstration. The linear detection range was from 1 μM to 50 μM with a low limit of detection of 200 nM (S/N = 3). Determination of GSH in serum samples was also achieved.

摘要

与天然酶相比,纳米酶具有催化性能好、稳定性高、成本低等优点,并且可以在极端条件下使用。通过简单的方法制备高活性的纳米酶并将其应用于生物分析是非常理想的。在这项工作中,展示了一种基于石墨烯量子点(GQD)分散血红素的纳米酶,可用于谷胱甘肽(GSH)的比色检测。GQD 通过一步水热合成法制备。血红素是血红素蛋白的催化中心,但由于溶解度低且容易聚集,限制了其催化活性,可通过简单的超声处理与 GQD 分散。所制备的血红素/GQD 纳米复合物具有优异的过氧化物酶样活性,可作为纳米酶使用。与天然辣根过氧化物酶(HRP)相比,当以四甲基联苯胺(TMB)为底物时,血红素/GQD 纳米酶的米氏常数(Km)明显降低。以 HO 为底物时,血红素/GQD 纳米酶的最大反应速率(Vmax)高于 HRP。通过自由基捕获实验研究了纳米酶活性的机制。作为概念验证的演示,开发了一种能够灵敏检测 GSH 的比色平台。线性检测范围从 1 μM 到 50 μM,检测限低至 200 nM(S/N = 3)。还实现了血清样品中 GSH 的测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0576/9608629/3f11ca91f068/molecules-27-06779-g001.jpg

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