Colorimetric determination of tyrosinase activity and high-throughput screening of inhibitors based on in-situ formation of gold nanoparticles

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2025-04-28 DOI:10.1007/s00604-025-07155-4

Guohua Zhou, Zhicheng Liu, Yuning Huang, Rufei Yang, Xin Guo, Yongmei Jia, Peilian Liu, Zhiguo Li, Wenjuan Cui

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Abstract

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.

Graphical Abstract

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酪氨酸酶活性的比色测定及基于原位形成金纳米颗粒的抑制剂的高通量筛选

提出了一种检测酪氨酸酶活性和筛选抑制剂的比色法，利用由儿茶酚介导的金纳米颗粒的原位形成。活性酪氨酸酶将儿茶酚转化为邻醌，导致aunp的大小及其相应的紫外-可见吸收光谱和视觉颜色的变化。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节。在此基础上，酪氨酸酶的快速检测和其抑制剂的高通量筛选可以简单地通过视觉观察溶液的颜色变化来实现。该方法灵敏、简便、快速、准确，可在0.5 ~ 25 U/mL范围内定量测定酪氨酸酶，检出限为0.038 U/mL。此外，通过视觉或仪器观察金纳米颗粒的颜色变化，可以高通量筛选抑制剂并计算其IC50值。我们预计这种方法将对酪氨酸酶的研究做出重大贡献，并促进有效抑制剂的开发、发现和高通量筛选。图形抽象

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.