α-Glucosidase immobilization on cellulose filter paper coated with a blend of chitosan and polyvinyl alcohol for inhibitor screening from traditional Chinese medicines.

چکیده اصلی

In this work, α-glucosidase was successfully immobilized on commercially available cellulose filter paper and applied to screen inhibitors from traditional Chinese medicines combined with capillary electrophoresis analysis. For the immobilization of α-glucosidase, cellulose filter paper was employed as the carrier. Which was first coated with a chitosan-polyvinyl alcohol blend solution and subsequently activated with polyethyleneimine to introduce a positively charged surface. Finally, α-glucosidase was immobilized onto the modified cellulose filter paper via electrostatic adsorption. The carrier was characterized, and several parameters affecting the catalytic activity of immobilized enzymes were optimized. The results showed that the tensile strength of chitosan-polyvinyl alcohol-modified cellulose filter paper was 1.8 times that of the original cellulose filter paper, indicating that the mechanical properties of the carrier were enhanced. Moreover, the immobilized α-glucosidase showed good storage stability (retained 51.7 ± 0.43% of its initial activity after storing at 4℃ for 30 days), excellent reusability (retained 79.5 ± 1.18% of its initial activity after reusing 10 times), improved environmental tolerance (70°C and pH 8.0). In addition, the reliability of the immobilized enzyme for enzyme inhibitor screening was verified using acarbose as a model inhibitor, and its half-maximal inhibitory concentration was 0.85 ± 0.02 μM. Eventually, the immobilized α-glucosidase was used to screen inhibitors from 12 traditional Chinese medicine extracts combined with capillary electrophoresis analysis, and Sanguisorbae Radix exhibited the strongest inhibitory effect. This established platform provides a rapid and reliable strategy for screening α-glucosidase inhibitors from traditional Chinese medicine.

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