Zheng Jia-Jia, Zhu Feiyan, Song Ningning, Deng Fang, Chen Qi, Chen Chen, He Jiuyang, Gao Xingfa, Liang Minmin
Experimental Center of Advanced Materials, School of Materials Science & Engineering, Beijing Institute of Technology, Beijing, China.
Laboratory of Theoretical and Computational Nanoscience, National Center for Nanoscience and Technology of China, Beijing, China.
Nat Protoc. 2024 Dec;19(12):3470-3488. doi: 10.1038/s41596-024-01034-7. Epub 2024 Aug 15.
Nanozymes are nanomaterials with enzyme-like catalytic properties. They are attractive reagents because they do not have the same limitations of natural enzymes (e.g., high cost, low stability and difficult storage). To test, optimize and compare nanozymes, it is important to establish fundamental principles and systematic standards to fully characterize their catalytic performance. Our 2018 protocol describes how to characterize the catalytic activity and kinetics of peroxidase nanozymes, the most widely used type of nanozyme. This approach was based on Michaelis-Menten enzyme kinetics and is now updated to take into account the unique physicochemical properties of nanomaterials that determine the catalytic kinetics of nanozymes. The updated procedure describes how to determine the number of active sites as well as other physicochemical properties such as surface area, shape and size. It also outlines how to calculate the hydroxyl adsorption energy from the crystal structure using the density functional theory method. The calculations now incorporate these measurements and computations to better characterize the catalytic kinetics of peroxidase nanozymes that have different shapes, sizes and compositions. This updated protocol better describes the catalytic performance of nanozymes and benefits the development of nanozyme research since further nanozyme development requires precise control of activity by engineering the electronic, geometric structure and atomic configuration of the catalytic sites of nanozymes. The characterization of the catalytic activity of peroxidase nanozymes and the evaluation of their kinetics can be performed in 4 h. The procedure is suitable for users with expertise in nano- and materials technology.
纳米酶是具有类酶催化特性的纳米材料。它们是有吸引力的试剂，因为它们没有天然酶的相同局限性（例如，成本高、稳定性低和储存困难）。为了测试、优化和比较纳米酶，建立基本原理和系统标准以全面表征其催化性能很重要。我们2018年的方案描述了如何表征过氧化物酶纳米酶（使用最广泛的纳米酶类型）的催化活性和动力学。这种方法基于米氏酶动力学，现在进行了更新，以考虑决定纳米酶催化动力学的纳米材料的独特物理化学性质。更新后的程序描述了如何确定活性位点的数量以及其他物理化学性质，如表面积、形状和尺寸。它还概述了如何使用密度泛函理论方法从晶体结构计算羟基吸附能。现在的计算纳入了这些测量和计算，以更好地表征具有不同形状、尺寸和组成的过氧化物酶纳米酶的催化动力学。这个更新后的方案更好地描述了纳米酶的催化性能，并有利于纳米酶研究的发展，因为进一步的纳米酶开发需要通过设计纳米酶催化位点的电子、几何结构和原子构型来精确控制活性。过氧化物酶纳米酶催化活性的表征及其动力学评估可以在4小时内完成。该程序适用于具有纳米和材料技术专业知识的用户。
