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α-Glucosidase immobilization on chitosan-modified cellulose filter paper: Preparation, property and application

Author:
Liu, Dong-Mei, Chen, Juan, Shi, Yan-Ping
Source:
International journal of biological macromolecules 2019 v.122 pp. 298-305
ISSN:
0141-8130
Subject:
Oriental traditional medicine, alpha-glucosidase, capillary electrophoresis, cellulose, enzymatic reactions, enzyme inhibitors, enzyme kinetics, glutaraldehyde, immobilized enzymes, pH, temperature
Abstract:
In the present study, α-glucosidase (α-Glu) was immobilized on the chitosan-modified cellulose filter paper (CFP/CS). For α-Glu immobilization, glutaraldehyde (GA) was used as a coupling agent. CFP, environmentally friendly and commercially available with low cost, will avoid the tedious procedure for synthesizing immobilization carrier. In addition, the CFP/CS-immobilized α-Glu can be directly taken out from the reaction mixture after an enzymatic reaction. This makes the instantaneous separation of immobilized enzyme comes true and is convenient to the subsequent study. Combined with capillary electrophoresis (CE), the CFP/CS-immobilized α-Glu was then used for enzyme kinetic and inhibition study. The CFP/CS-immobilized α-Glu exhibited enhanced pH and temperature tolerance. In addition, the performance of the CFP/CS-immobilized α-Glu was studied. Immobilized α-Glu exhibited excellent batch-to-batch reproducibility (RSD = 6.7%, n = 5) and improved reusability (71.0% of its initial activity after 10 repeated cycles). Finally, immobilized α-Glu was used to screen enzyme inhibitors from 11 traditional Chinese medicines (TCMs). The results showed that CFP/CS has potential development prospects as a novel enzyme immobilization carrier.