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%.