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This study investigated whether selenium-containing peptides derived from selenium-rich black beans exert inhibitory effects on α-amylase and evaluated their potential hypoglycemic activity in vitro. Selenium-rich black beans from Guangxi, China, were used as raw materials, and selenium-containing peptides were screened and synthesized through a combination of single-factor experiments, response surface methodology, and peptidomics analysis. Four peptides—Se-MetLP, Se-MetIP, TLTSe-MetLR, and Se-CPSFGQ—were identified and evaluated for their α-amylase inhibitory activity. Among the four peptides, Se-MetIP exhibited the highest inhibitory effect, achieving an inhibition rate of 70.58%±2.21% at a concentration of 4 mg/mL, whereas TLTSe-MetLR showed the lowest half-maximal inhibitory concentration (IC50) of 0.47 mg/mL, indicating superior inhibitory potency. Based on their strong α-amylase inhibitory activity, Se-MetIP and TLTSe-MetLR were selected for further investigation of inhibition mechanisms using circular dichroism spectroscopy, isothermal titration calorimetry, and molecular docking analyses, and their in vitro hypoglycemic effects were subsequently evaluated in a hyperglycemic HepG2 cell model. Circular dichroism analysis revealed that peptide binding induced conformational changes in α-amylase, characterized by increased β-sheet content and reduced β-turn and random coil structures, suggesting alterations in the enzyme spatial conformation. Isothermal titration calorimetry demonstrated that peptide-enzyme interactions were exothermic. Molecular docking results showed that the binding free energies of Se-MetIP and TLTSe-MetLR with α-amylase were −5.399 and −6.401 kcal/mol, respectively, with van der Waals forces and hydrogen bonding as the dominant stabilizing interactions, complemented by hydrophobic effects. Furthermore, cellular assays demonstrated that Se-MetIP and TLTSe-MetLR significantly enhanced glucose consumption and promoted glycogen synthesis in insulin-resistant HepG2 cells, thereby improving glucose utilization. Overall, selenium-containing peptides from black beans effectively inhibit α-amylase activity and exhibit in vitro hypoglycemic potential, providing a theoretical basis for the development of hypoglycemic peptides from selenium-rich black beans.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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