Rapid screening and identification of α-glucosidase inhibitors from mulberry leaves using enzyme-immobilized magnetic beads coupled with HPLC/MS and NMR.
α-Glucosidase plays important roles in the digestion and absorption of carbohydrates in the small intestine. The inhibition of α-glucosidase is regarded as a potential way to treat diabetes. We established an approach to screening α-glucosidase inhibitors from medicinal plants using enzyme-coated magnetic bead. Using 1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide and N-hydroxysuccinimide as reaction reagents, α-glucosidase was immobilized on the magnetic beads by covalent linkage. The conjugation of α-glucosidase to the magnetic beads was characterized using scanning electron microscope and X-ray diffractometer. The proposed approach was applied in fishing potential α-glucosidase inhibitors from extract of Morus alba, a Chinese medicinal plant. The structures of potential active compounds were identified via liquid chromatography-mass spectrometry and nuclear magnetic resonance. The results demonstrated that two flavonoids (isoquercitrin and astragalin) could bind to α-glucosidase, which was confirmed via conventional α-glucosidase inhibitory assay. Our findings suggested that enzyme-coated magnetic beads may be suitable for discovering active compounds from medicinal plants.
Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
Yi
Y
Tao
Yufeng
Y
Zhang
Yiyu
Y
Cheng
Yi
Y
Wang
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors* / Q000378:metabolism; D065089:Glycoside Hydrolase Inhibitors; D044949:Kaempferols / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D008280:Magnetics / Q000379:methods*; D059346:Magnets; D013058:Mass Spectrometry / Q000379:methods; D000432:Methanol / Q000737:chemistry; D031625:Morus / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D018515:Plant Leaves / Q000737:chemistry; D011794:Quercetin / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D000520:alpha-Glucosidases / Q000378:metabolism
D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D065089:Glycoside Hydrolase Inhibitors; D044949:Kaempferols; D010936:Plant Extracts; C016527:isoquercitrin; D011794:Quercetin; C001579:astragalin; D000520:alpha-Glucosidases; D000432:Methanol