Detection of glucose via enzyme-coupling reaction based on a DT-diaphorase fluorescence probe.
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.
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Xinghui
X
Gao
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. Electronic address: lixh@iccas.ac.cn.
Xiaohua
X
Li
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Qiongqiong
Q
Wan
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Zhao
Z
Li
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. Electronic address: mahm@iccas.ac.cn.
Huimin
H
Ma
D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism*; D005947:Glucose / Q000032:analysis* / Q000378:metabolism; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism; D006029:Glycosuria; D006801:Humans; D057230:Limit of Detection; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D010084:Oxidation-Reduction; D013050:Spectrometry, Fluorescence / Q000379:methods
D005456:Fluorescent Dyes; D042843:Glucose 1-Dehydrogenase; D016660:NAD(P)H Dehydrogenase (Quinone); D005947:Glucose
Enzyme-coupling reaction; Fluorescence probe; Glucose; Resorufin