Shuang Guo,Shuo Liu,Jing Meng,Dongyu Gu,Yi Wang,Dajun He,Yi Yang
Morus alba leaves are a natural product with great antidiabetic potential. However, the therapeutic efficacy of natural products is usually achieved through the interaction of active compounds with specific targets. Among them, active compounds with multi-target therapeutic functions are more effective than single-target enzymes. In this study, a bienzyme system was constructed by co-immobilizing α -amylase and α -glucosidase onto Fe 3 O 4 for affinity screening of dual-target active components in the complex extract from M. alba leaves. As a result, a potential active compound was selectively screened by ligand fishing, separated by high-speed countercurrent chromatography using a solvent system of ethyl acetate– n -butanol–water (3:2:5, v/v), and identified as rutin. In addition, the result of molecular docking showed that rutin could interact with the active center of α -amylase and α -glucosidase through multiple hydrogen bonds, van der Waals forces, etc. to play an inhibitory role. These results demonstrate the effectiveness of the polydopamine magnetically immobilized bienzyme system for dual-target affinity screening of active substances. This study not only reveals the chemical basis of the antidiabetic activity of M. alba leaves from a dual-target perspective, but also promotes the progress of multitarget affinity screening.
Shuang Guo,Jing Meng,Shuai Wang,Dongyu Gu,Tingting Chu,Yi Yang
Natural products are an important source of drug development and have good prospects in the treatment of many diseases. However, the efficacy of many natural drugs is achieved through multi-targets, which poses a huge obstacle to their development. In this study, a bienzyme microcapsule system based on α-amylase and α-glucosidase was constructed for the ligand fishing of natural compounds. The microcapsules were formed on the surface of calcium carbonate particles by self-polymerization of dopamine, which physically encapsulated iron oxide and α-glucosidase. Then, α-amylase was immobilized on the surface of magnetic material with glutaraldehyde as crosslinking agent, and calcium carbonate template was removed by EDTA to form a bienzyme system for affinity screening of active components from Toona sinensis . The results showed that two compounds, 1,2,3,4,6-penta-O-galloyl-β- D -glucose and quercitrin were affinity screened from the extracts of Toona sinensis . In vitro activity experiment proved that they were α-amylase and α-glucosidase inhibitors, respectively. Moreover, both of them were mixed type inhibitors that can interact with active centers of corresponding enzymes through multiple hydrogen bonds , van der Waals forces, etc. This study not only proved the effectiveness of the bienzyme system, but also provided a successful example for multi-target screening of natural products.
危险品化学品经营许可证（带存储）