Zhou Guohua, Liu Zhicheng, Huang Yuning, Yang Rufei, Guo Xin, Jia Yongmei, Liu Peilian, Li Zhiguo, Cui Wenjuan
Key Laboratory of Clean Energy Materials Chemistry of Guangdong Higher Education Institutes, , Lingnan Normal University, Zhanjiang, 524048, Guangdong, China.
Mikrochim Acta. 2025 Apr 28;192(5):326. doi: 10.1007/s00604-025-07155-4.
A colorimetric method is presented for detecting tyrosinase activity and screening inhibitors, leveraging the in-situ formation of gold nanoparticles mediated by catechol. The active tyrosinase converses catechol into o-quinone leading to changes in both the size of AuNPs and their corresponding UV-Vis absorption spectrum as well as visual color. This enzyme-mediated alteration can be modulated by tyrosinase inhibitors through chemical regulation of enzyme activity. Based on these, facile detection of tyrosinase and high-throughput screening for its inhibitors can be achieved simply by visually observing color changes in solution. The proposed method is characterized by its sensitivity, simplicity, rapidity, and accuracy, allowing for quantitative determination of tyrosinase within the range 0.5 to 25 U/mL, with a detection limit of 0.038 U/mL. Furthermore, visual or instrumental observation of the color changes in gold nanoparticles enables high-throughput screening of inhibitors and calculation of their IC50 values. We anticipate that this approach will significantly contribute to tyrosinase research and facilitate the development, discovery, and high-throughput screening of effective inhibitors.
本文提出了一种比色法，用于检测酪氨酸酶活性和筛选抑制剂，该方法利用儿茶酚介导的金纳米颗粒原位形成。活性酪氨酸酶将儿茶酚转化为邻苯醌，导致金纳米颗粒的大小及其相应的紫外-可见吸收光谱以及视觉颜色发生变化。这种酶介导的改变可以通过酪氨酸酶抑制剂对酶活性的化学调节来调控。基于此，通过目视观察溶液中的颜色变化，即可简单地实现酪氨酸酶的简便检测及其抑制剂的高通量筛选。所提出的方法具有灵敏度高、操作简单、快速、准确的特点，能够在0.5至25 U/mL范围内定量测定酪氨酸酶，检测限为0.038 U/mL。此外，通过目视或仪器观察金纳米颗粒的颜色变化，可以实现抑制剂的高通量筛选并计算其IC50值。我们预计这种方法将对酪氨酸酶研究做出重大贡献，并促进有效抑制剂的开发、发现和高通量筛选。
