Ghelichi_2025_Mar.Drugs_23_392

This study investigates the anti-obesity and antidiabetic potential of P. palmata extracts produced through sequential enzymatic and alkaline treatments. Among the treatment groups, the extract treated solely with Alcalase((a)) (Alc) demonstrated the highest protein content (10.11 +/- 0.15%) and degree of hydrolysis (30.36 +/- 0.77%), significantly outperforming other treatments (p < 0.05). The Alc extract also exhibited superior inhibitory activity against porcine pancreatic lipase and alpha-amylase, achieving the lowest IC(50) for lipase (2.29 +/- 0.87 mg.mL(-1)) and showing significant enzyme inhibition across all tested concentrations (p < 0.05). Ultrafiltration of the Alc extract revealed that peptide fractions < 1 kDa and 1-3 kDa were most effective in enzyme inhibition, with IC(50) values of 3.25-3.55 mg.mL(-1) for both lipase and alpha-amylase. Peptides were identified via LC-MS/MS analysis and database searching using SequestHT, resulting in 536 sequences, of which bioinformatic screening yielded 51 non-toxic, non-allergenic candidates (PeptideRanker score > 0.6); four of these contained known inhibitory motifs for lipase and alpha-amylase. Molecular docking confirmed strong binding affinities between these peptides and their respective enzymes, supporting their potential as natural enzyme inhibitors. These findings indicate the functional food potential of Alcalase((a))-derived P. palmata peptides for managing obesity and type 2 diabetes.