Detection of glucose via enzyme-coupling reaction based on a DT-diaphorase fluorescence probe

Abstract

Enzyme-coupling reactions play an important role in the assay of analytes. In this manuscript, we developed a new fluorescent probe for the detection of glucose through the enzyme-coupling reaction of DT-diaphorase (DTD). The probe was synthesized through a mild and simple synthetic procedure, and showed good fluorescence response to DTD. The reactions for the detection of glucose proceed as follows: glucose dehydrogenase oxidizes glucose to gluconolactone with NAD(+) as the electron acceptor to yield NADH, and NADH can be utilized by DTD to further react with the probe releasing resorufin. As a result of these tandem reactions, fluorescence off-on response will occur. The method showed high selectivity for glucose with a detection limit of 0.2 µM, which may provide a potential way for fluorescence detection of glucose through enzyme-coupling reactions. Furthermore, the applicability of the method has been demonstrated by detecting glucose in human urine samples.