A fluorescence signal amplification strategy for modification-free ratiometric determination of tyrosinase in situ based on the use of dual-templated copper nanoclusters
文献类型： 外文期刊
作者： Huang, Xuemin 1 ; Zhao, Huanan 1 ; Qiu, Wen 1 ; Wang, Jian 1 ; Guo, Longhua 1 ; Lin, Zhenyu 1 ; Pan, Wei 2 ; Wu, Yong 4 ; Qi 1 ;
作者机构： 1.Fuzhou Univ, Key Lab Analyt Sci Food Safety & Biol, Fujian Prov Key Lab Anal & Detect Food Safety Eel, Minist Educ, Fuzhou 350108, Fujian, Peoples R China
2.Fujian Acad Agr Sci, Inst Agr Qual Stand & Testing Technol Res, Fuzhou 350003, Fujian, Peoples R China
3.Fuzhou Univ, Dept Chem, Fuzhou 350108, Fujian, Peoples R China
4.Xiamen Anxinya Biol Technol Co Ltd, Xiamen 361005, Fujian, Peoples R China
关键词： Dual-templated copper nanoclusters; Fluorescence resonance energy transfer; Ratiometric detection; Tyrosinase
期刊名称：MICROCHIMICA ACTA （ 影响因子：5.833； 五年影响因子：5.357 ）
ISSN： 0026-3672
年卷期： 2020 年 187 卷 4 期
页码：
收录情况： SCI
摘要： A fluorescence resonance energy transfer (FRET)-based in situ fluorescence signal amplification strategy is described for the determination of tyrosinase (TYR). In this assay, a dual-templated copper nanocluster (CuNCs) stabilized by bovine serum albumin (BSA) and glycylglycine (Gly-Gly) was used as an energy donor. Metyrosine was employed as a TYR substrate because its enzyme catalytic product (methyldopa) was able to function as a monomer molecule to form fluorescent polymethyldopa (PMeDP) with the assistance of BSA/Gly-Gly CuNCs. In this process, PMeDP can combine with BSA/Gly-Gly CuNCs without extra modification and then acts as an energy receptor, which leads to a remarkable FRET from BSA/Gly-Gly CuNCs to PMeDP. Interestingly, the fluorescence intensity of PMeDP was strengthened greatly in the FRET-based sensor compared to the separate excitation, which provided good sensitivity for TYR sensing. Illuminated under a UV light source, the fluorescence signal change is observed from dark violet to bright green. Therefore, the present sensing system affords a reliable ratiometric assay for TYR determination. Also, the ratio of fluorescence intensity between PMeDP (lambda(em) at 505 nm, F505) and BSA/Gly-Gly CuNCs (lambda(em) at 415 nm, F415) was used for quantitative determination of TYR. The sensing system was easily operated in aqueous media with an exciting detection limit of 44.0 U L-1. This sensing strategy has been applied to the screening of inhibitors. Schematic representation of the strategy for the determination of tyrosinase.
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