基于酶触发的点击化学反应测定β-葡萄糖苷酶活性
收稿日期: 2018-03-09
修回日期: 2018-04-21
网络出版日期: 2018-06-01
基金资助
国家自然科学基金（No.21402056）资助项目.
Determination of β-Glucosidase Activity Based on Enzyme-Triggered Click Chemistry
Received date: 2018-03-09
Revised date: 2018-04-21
Online published: 2018-06-01
Supported by
Project supported by the National Natural Science Foundation of China (No. 21402056).
建立了一种新的β-葡萄糖苷酶活性定量检测的方法，其原理是通过β-葡萄糖苷酶对2-O-β-D-葡萄糖基-L-抗坏血酸的特异性水解，释放出抗坏血酸来还原Cu（Ⅱ），原位生成的Cu（I）催化荧光较弱的香豆素和苄基叠氮进行环加成点击反应，产生高荧光强度的三氮唑，从而利用荧光光谱检测体系荧光强度的变化，反映出β-葡萄糖苷酶的活性.实验结果显示，在1～40 U/L范围内，荧光强度增强程度与β-葡萄糖苷酶活性呈线性关系，可实现定量检测，其最低检测限为0.456 U/L.
关键词： β-葡萄糖苷酶活性; 测定; 2-O-β-D-葡萄糖基-L-抗坏血酸; 点击化学
王龙文 , 马济美 , 程鑫 , 李子龙 , 孙林皓 , 曾贞 , 江洪 . 基于酶触发的点击化学反应测定β-葡萄糖苷酶活性[J]. 有机化学, 2018 , 38(10) : 2775 -2779 . DOI: 10.6023/cjoc201803011
A novel method to quantify β-glucosidase activity was developed by coupling the cleavage of 2-O-(β-glucopyra-nosyl)ascorbic acid with the reduction of Cu (Ⅱ). The in situ generated Cu (I) catalyzed the cycloaddition between weakly fluorescent coumarin and benzyl azide to yield a highly fluorescent triazole product. The fluorescence intensity was dependently enhanced on the increase of the activity of β-glucosidase and a linear relationship was found between 1～40 U/L. This cascade allows detection of β-glucosidase with a limit of detection of 0.456 U/L.
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