Chin. J. Org. Chem. ›› 2018, Vol. 38 ›› Issue (10): 2775-2779.DOI: 10.6023/cjoc201803011 Previous Articles     Next Articles

Notes

基于酶触发的点击化学反应测定β-葡萄糖苷酶活性

王龙文, 马济美, 程鑫, 李子龙, 孙林皓, 曾贞, 江洪   

  1. 华中农业大学理学院 武汉 430070
  • 收稿日期:2018-03-09 修回日期:2018-04-21 发布日期:2018-06-01
  • 通讯作者: 马济美,E-mail:majimei@mail.hzau.edu.cn;江洪,E-mail:jianghong0066@126.com E-mail:majimei@mail.hzau.edu.cn;jianghong0066@126.com
  • 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

Wang Longwen, Ma Jimei, Cheng Xin, Li Zilong, Sun Linhao, Zeng Zhen, Jiang Hong   

  1. Department of Chemistry, College of Science, Huazhong Agricultural University, Wuhan 430070
  • Received:2018-03-09 Revised:2018-04-21 Published:2018-06-01
  • Contact: 10.6023/cjoc201803011 E-mail:majimei@mail.hzau.edu.cn;jianghong0066@126.com
  • Supported by:

    Project supported by the National Natural Science Foundation of China (No. 21402056).

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.

Key words: β-glucosidase activity, determination, 2-O-(β-glucopyranosyl)ascorbic acid, click chemistry