文献类型：期刊文献

英文题名：alpha-Glucosidase immobilization on polydopamine-coated cellulose filter paper and enzyme inhibitor screening

作者：Li, Peng[1];Ma, Xiao-Hui[2];Dong, Yu-Ming[1];Jin, Ling[2];Chen, Juan[1]

第一作者：Li, Peng

通信作者：Chen, J[1];Jin, L[2]

机构：[1]Lanzhou Univ, Sch Pharm, Lanzhou 730000, Peoples R China;[2]Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China

第一机构：Lanzhou Univ, Sch Pharm, Lanzhou 730000, Peoples R China

通信机构：[1]corresponding author), Lanzhou Univ, Sch Pharm, Lanzhou 730000, Peoples R China;[2]corresponding author), Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China.|[1073501e14fb35863569f]甘肃中医药大学药学院（西北中藏药协同创新中心办公室）;[10735]甘肃中医药大学;

年份：2020

卷号：605

外文期刊名：ANALYTICAL BIOCHEMISTRY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000569074500008)】；

基金：This work was funded by Special Foundation for National Science and Technology Basic Resources of China (2018FY100701), the Collaborative and Innovative Science and Technology Team Supporting Program for Universities in Gansu Province, China (2016C-05), and National Survey of Traditional Chinese Medicine Resources, Special Subsidy for Public Health Services of Traditional Chinese Medicine, China (CS [2017]-66).

语种：英文

外文关键词：Cellulose filter paper; Polydopamine composite layer; Immobilized alpha-glucosidase; Capillary electrophoresis; Inhibitor screening

摘要：Immobilized enzyme has been gradually applied to the screening of enzyme inhibitors owing to its retained catalytic activity and reusability. In this work, the cheap and available cellulose filter paper (CFP) was used as a carrier for the immobilization of alpha-glucosidase (alpha-Glu). In virtue of the self-polymerization-adhesion behavior of dopamine, CFP was coated with a polydopamine composite layer and then alpha-glucosidase is covalently bound to the modified CFP through Schiff base reaction and Michael addition reaction. Combined with capillary electrophoresis (CE) analysis, enzyme reaction kinetics, inhibition kinetics and other performance of the prepared immobilized enzyme (CFP/Dopa/alpha-Glu) were examined and verified. Its Michaelis constant (K-m) was calculated to be 0.83 mM. And the inhibition constant (Ki) and half-maximal inhibitory concentration (IC50) for acarbose were determined to be 0.16 and 0.17 mu M, respectively. CFP/Dopa/alpha-Glu had the same optimum working pH value (7.0) as free alpha-Glu and slightly higher working temperature (65 degrees C) than free alpha-Glu. In addition, it exhibited good batch-to-batch reproducibility with an RSD value of 4.4% (n = 10), and excellent reusability with 71% of the initial enzyme activity after being recycled 11 times. Finally, the CFP/Dopa/alpha-Glu was applied to screen alpha-glucosidase inhibitors from 11 traditional Chinese medicines, and Terminalia chebula possessed the strongest inhibition effect on alpha-glucosidase.