School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 401331, China.
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.
Molecules. 2023 Jul 15;28(14):5429. doi: 10.3390/molecules28145429.
A biomimetic mineralization method was used in the facile and rapid preparation of nanoflowers for immobilizing alcohol dehydrogenase (ADH). The method mainly uses ADH as an organic component and zinc phosphate as an inorganic component to prepare flower-like ADH/Zn(PO) organic-inorganic hybrid nanoflowers (HNFs) with the high specific surface area through a self-assembly process. The synthesis conditions of the ADH HNFs were optimized and its morphology was characterized. Under the optimum enzymatic reaction conditions, the Michaelis-Menten constant () of ADH HNFs (β-NAD as substrate) was measured to be 3.54 mM, and the half-maximal inhibitory concentration (IC) of the positive control ranitidine (0.2-0.8 mM) was determined to be 0.49 mM. Subsequently, the inhibitory activity of natural medicine Pursh and nine small-molecule compounds on ADH was evaluated using ADH HNFs. The inhibition percentage of the aqueous extract of is 57.9%. The vanillic acid, protocatechuic acid, gallic acid, and naringenin have obvious inhibitory effects on ADH, and their percentages of inhibition are 55.1%, 68.3%, 61.9%, and 75.5%, respectively. Moreover, molecular docking analysis was applied to explore the binding modes and sites of the four most active small-molecule compounds to ADH. The results of this study can broaden the application of immobilized enzymes through biomimetic mineralization, and provide a reference for the discovery of ADH inhibitors from natural products.
一种仿生矿化方法被用于简便快速地制备固定化醇脱氢酶（ADH）的纳米花。该方法主要以 ADH 为有机组分，以磷酸锌为无机组分，通过自组装过程制备具有高比表面积的花状 ADH/Zn(PO)有机-无机杂化纳米花（HNFs）。优化了 ADH HNFs 的合成条件，并对其形态进行了表征。在最佳酶反应条件下，测定了 ADH HNFs（以β-NAD 为底物）的米氏常数（Km）为 3.54 mM，阳性对照雷尼替丁（0.2-0.8 mM）的半最大抑制浓度（IC）为 0.49 mM。随后，采用 ADH HNFs 评价了天然药物 Pursh 和九种小分子化合物对 ADH 的抑制活性。的水提物对 ADH 的抑制率为 57.9%。香草酸、原儿茶酸、没食子酸和柚皮苷对 ADH 有明显的抑制作用，其抑制率分别为 55.1%、68.3%、61.9%和 75.5%。此外，还应用分子对接分析探讨了四种最活跃的小分子化合物与 ADH 的结合模式和结合位点。该研究结果可以通过仿生矿化拓宽固定化酶的应用，并为从天然产物中发现 ADH 抑制剂提供参考。
