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Outline
In order to investigate the optimal extraction process and anti-inflammatory activity of white kidney bean polypeptides (WKBPs), this study employed white kidney bean protein as raw material and hydrolyzed white kidney bean protein by ultrasound-assisted enzymatic hydrolysis, utilizing the inhibition rate of bovine serum protein degeneration as the index. We identified the optimal extraction process with a single-factor combined response surface method and separated the prepared white kidney bean crude peptides with ultrafiltration tubes of different molecular weights. This study determined the anti-inflammatory activity of various components in vitro, detected the optimal components by liquid chromatography-mass spectrometry (LC-MS/MS), and examined the anti-inflammatory peptide segments by virtual screening and molecular docking techniques. The results indicated that the inhibition rate of WKBPs on bovine serum protein denaturation was 86.57% under the conditions of 166 W ultrasonic power, 18000 U/g enzyme addition, 128 min enzymatic hydrolysis time, and 46 ℃ enzymatic hydrolysis temperature. After ultrafiltration, the components with Mw<1 kDa revealed good anti-inflammatory activity (bovine serum protein denaturation inhibition, hyaluronidase inhibition, and NO inhibition), and the IC50 values were 1.181, 9.244 and 10.887 mg/mL, respectively. We detected 3306 peptides by LC-MS/MS, of which 73% were peptide sequences ranging from 3 to 10. Virtual screening Vina score with type and induced nitric oxide synthase (inducible nitric oxide synthase, iNOS) binding energy minimum peptide sequence of 4 peptide FFFR, molecular docking visualization findings demonstrated that 4-peptide FFFR could bind to the active site of iNOS via hydrogen bonding and hydrophobic interaction, thus exerting inhibitory effects. This study serves as a reference for further research on the anti-inflammatory activity of WKBPs.
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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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