“Turn-On” Fluorescence Determination of β-Glucosidase Activity Using Fluorescent Polymer Nanoparticles Formed from Polyethylenimine Cross-Linked with Hydroquinone AITranslate
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β-Glucosidase is associated with many diseases, so it is important to be able to accurately determine β-glucosidase activity. Here, we describe a turn-on fluorescence method for sensitively determining β-glucosidase activity. The method involves the formation of water-soluble fluorescent polymer nanoparticles. β-Glucosidase initially hydrolyzes the β-arbutin substrate to release hydroquinone (1,4-dihydroxybenzene). The hydroquinone then forms cross-links between branched polyethylenimine molecules to give fluorescent water-soluble polymer nanoparticles. The polymer nanoparticles were found to fluoresce with excitation and emission peaks at 373 and 510 nm, respectively, and the fluorescence intensity was related to the β-glucosidase activity. The fluorescence intensity could therefore be used to determine the β-glucosidase activity. The lowest β-glucosidase activity that could be determined was 0.4 U L–1, and the dynamic linear range was 1.0–36.0 U L–1. The method was very selective for β-glucosidase activity and was not sensitive to other proteins. The method was successfully used to determine β-glucosidase in spiked samples containing human serum albumin at a concentration of 0.5%, and the recoveries were 95.2%–104.5%.
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DOI:https://doi.org/10.1021/acsapm.9b00728
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β-Glucosidase is associated with many diseases, so it is important to be able to accurately determine β-glucosidase activity. Here, we describe a turn-on fluorescence method for sensitively determining β-glucosidase activity. The method involves the formation of water-soluble fluorescent polymer nanoparticles. β-Glucosidase initially hydrolyzes the β-arbutin substrate to release hydroquinone (1,4-dihydroxybenzene). The hydroquinone then forms cross-links between branched polyethylenimine molecules to give fluorescent water-soluble polymer nanoparticles. The polymer nanoparticles were found to fluoresce with excitation and emission peaks at 373 and 510 nm, respectively, and the fluorescence intensity was related to the β-glucosidase activity. The fluorescence intensity could therefore be used to determine the β-glucosidase activity. The lowest β-glucosidase activity that could be determined was 0.4 U L–1, and the dynamic linear range was 1.0–36.0 U L–1. The method was very selective for β-glucosidase activity and was not sensitive to other proteins. The method was successfully used to determine β-glucosidase in spiked samples containing human serum albumin at a concentration of 0.5%, and the recoveries were 95.2%–104.5%.
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| GB/T 7714-2015 | [1] Jinshui Liu, Huijuan Bao, Chenfu Liu, et al. ACS Applied Polymer Materials, 2019(1). DOI:10.1021/acsapm.9b00728. |
| MLA | [1] Jinshui Liu, et al., ACS Applied Polymer Materials, no. 1, 2019, https://doi.org/10.1021/acsapm.9b00728. |
| APA | [1] Jinshui Liu, Huijuan Bao, Chenfu Liu, Fangfei Wu, & Feng Gao. (2019). ACS Applied Polymer Materials(1). https://doi.org/10.1021/acsapm.9b00728 |
| IEEE | [1] Jinshui Liu, Huijuan Bao, Chenfu Liu, Fangfei Wu, and Feng Gao, ACS Applied Polymer Materials, no. 1, 2019, doi: 10.1021/acsapm.9b00728. keywords: {fluorescence;nonconjugated fluorescent polymer;βglucosidase activity;βarbutin;polyethylenimine;hydroquinone} |
