Identification and in vitro Characterization of Novel Antidiabetic Peptides Released Enzymatically from Peanut Protein

Plant Foods Hum Nutr. 2024 Mar;79(1):66-72. doi: 10.1007/s11130-023-01118-x. Epub 2023 Nov 23.

Abstract

Bioactive peptides derived from proteins found in various foods provide significant health benefits, including regulating blood sugar levels by inhibiting carbohydrate-hydrolyzing enzymes. Hydrolysates of peanut protein were prepared using alcalase (AH) or trypsin (TH) to generate antidiabetic peptides with high activity against α-amylase (IC50 of 6.46 and 5.71 mg/mL) and α-glucosidase (IC50 of 6.30 and 5.57 mg/mL), as well as antiradical activity to scavenge DPPH (IC50 of 4.18 and 3.12 mg/mL) and ABTS•+ (IC50 of 2.87 and 2.56 mg/mL), respectively. The bioactivities of hydrolysates were greatest in the ultrafiltration-generated F3 fraction (< 3 kDa). The most active fraction was TH-F3, which was purified by gel filtration chromatography to generate sub-fractions (SF). With IC50 values of 1.05 and 0.69 mg/mL, the F3-SF8 fraction was the most effective at inhibiting the activity of α-amylase and α-glucosidase, respectively. This fraction was further purified using RP-HPLC to generate sub-subfractions (SSF), the most active of which were F3-SF8-SSF9 and SSF10. The peptide sequences F3-SF8-SSF9 and SSF10 were determined using LC-MS/MS. Two novel antidiabetic peptides with the potential to inhibit α-amylase and α-glucosidase were identified, with the sequences Asp-Trp-Arg (476.22 Da, IC50 of 0.78, and 0.35 mg/mL) and Phe-Tyr (329.15 Da, IC50 of 0.91, and 0.41 mg/mL). These results suggest that peptides derived from peanut protein are attractive natural ingredients for diabetes management applications.

Keywords: Antidiabetic peptides; Antioxidants; LC-MS/MS; Peanut protein hydrolysate; α-Amylase; α-Glucosidase.

MeSH terms

  • Arachis* / metabolism
  • Chromatography, Liquid
  • Hypoglycemic Agents* / chemistry
  • Hypoglycemic Agents* / pharmacology
  • Peptides / pharmacology
  • Tandem Mass Spectrometry
  • alpha-Amylases
  • alpha-Glucosidases / metabolism

Substances

  • Hypoglycemic Agents
  • alpha-Glucosidases
  • Peptides
  • alpha-Amylases