Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Talanta. 2014 Mar;120:456-61. doi: 10.1016/j.talanta.2013.12.032. Epub 2013 Dec 21.
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.
酶偶联反应在分析物的测定中起着重要作用。在本文中，我们通过 DT-二氢蝶啶还原酶（DTD）的酶偶联反应开发了一种用于检测葡萄糖的新型荧光探针。该探针通过温和简单的合成步骤合成，对 DTD 表现出良好的荧光响应。用于检测葡萄糖的反应如下：葡萄糖脱氢酶以 NAD(+)作为电子受体将葡萄糖氧化为葡萄糖酸内酯，生成 NADH，NADH 可被 DTD 进一步利用与探针反应，释放出 Resorufin。由于这些串联反应，将发生荧光关闭-开启响应。该方法对葡萄糖具有高选择性，检测限为 0.2µM，这可能为通过酶偶联反应进行荧光检测葡萄糖提供了一种潜在方法。此外，该方法的适用性已通过检测人尿液样本中的葡萄糖得到证明。
