Li Zihong, Ren Mingguang, Zhao Yuping, Song Wenhui, Cheng Jie, Lin Weiying
Guangxi Key Laboratory of Electrochemical Energy Materials, Institute of Optical Materials and Chemical Biology, School of Chemistry and Chemical Engineering, Guangxi University, Nanning, Guangxi 530004, PR China; Institute of Fluorescent Probes for Biological Imaging, School of Materials Science and Engineering, School of Chemistry and Chemical Engineering, University of Jinan, Jinan, Shandong 250022, PR China.
Institute of Fluorescent Probes for Biological Imaging, School of Materials Science and Engineering, School of Chemistry and Chemical Engineering, University of Jinan, Jinan, Shandong 250022, PR China; Institute of State Key Laboratory of Biobased Material and Green Papermaking, Qilu University of Technology, Jinan 250353, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Apr 15;251:119446. doi: 10.1016/j.saa.2021.119446. Epub 2021 Jan 6.
β-galactosidase is of great significance to living organisms, which is an important marker of primary ovarian cancer and cellular senescence. To detect the activity of β-galactosidase, a novel fluorescent probe ESIPT-GAL which based on excited state intramolecular proton transfer (ESIPT) mechanism for detecting β-galactosidase is developed in this work with low background fluorescence and high sensitivity (Φ = 0.0045-0.2409). The fluorescence intensity at 552 nm of this probe increased by ~ 55 times with β-galactosidase addition (0-4 U/mL), and its detection limit is very low (3.9 × 10 U/mL). In addition, the spectral data (pseudo-first-order rate: 1.303 min) and enzyme kinetic parameter (V = 69.5 μΜ•S) both show that the probe can achieve rapid response to β-galactosidase. Moreover, the probe has good water solubility, which ensures that it has good biocompatibility and can be easily applied to detect β-galactosidase in living cells and tissues. Importantly, the probe ESIPT-GAL can monitor β-galactosidase in deep mouse tissue sections (90 μm).
β-半乳糖苷酶对生物体具有重要意义，它是原发性卵巢癌和细胞衰老的重要标志物。为了检测β-半乳糖苷酶的活性，本研究开发了一种基于激发态分子内质子转移（ESIPT）机制的新型荧光探针ESIPT-GAL用于检测β-半乳糖苷酶，该探针具有低背景荧光和高灵敏度（Φ = 0.0045 - 0.2409）。加入β-半乳糖苷酶（0 - 4 U/mL）后，该探针在552 nm处的荧光强度增加了约55倍，其检测限非常低（3.9×10 U/mL）。此外，光谱数据（伪一级反应速率：1.303 min）和酶动力学参数（V = 69.5 μΜ•S）均表明该探针能够对β-半乳糖苷酶实现快速响应。而且，该探针具有良好的水溶性，这确保了它具有良好的生物相容性，并且能够轻松应用于检测活细胞和组织中的β-半乳糖苷酶。重要的是，探针ESIPT-GAL能够监测深度为90μm的小鼠组织切片中的β-半乳糖苷酶。
