Combinations of Legume Protein Hydrolysates Synergistically Inhibit Biological Markers Associated with Adipogenesis

The objective was to investigate the anti-adipogenesis potential of selected legume protein hydrolysates (LPH) and combinations using biochemical assays and in silico predictions. Black bean, green pea, chickpea, lentil and fava bean protein isolates were hydrolyzed using alcalase (A) or pepsin/pancreatin (PP). The degree of hydrolysis ranged from 15.5% to 35.5% for A-LPH and PP-LPH, respectively. Antioxidant capacities ranged for ABTS^•+ IC₅₀ from 0.3 to 0.9 Trolox equivalents (TE) mg/mL, DPPH^• IC₅₀ from 0.7 to 13.5 TE mg/mL and nitric oxide (NO) inhibition IC₅₀ from 0.3 to 1.3 mg/mL. LPH from PP-green pea, A-green pea and A-black bean inhibited pancreatic lipase (PL) (IC₅₀ = 0.9 mg/mL, 2.2 mg/mL and 1.2 mg/mL, respectively) (p < 0.05). For HMG-CoA reductase (HMGR) inhibition, the LPH from A-chickpea (0.15 mg/mL), PP-lentil (1.2 mg/mL), A-green pea (1.4 mg/mL) and PP-green pea (1.5 mg/mL) were potent inhibitors. Combinations of PP-green pea + A-black bean (IC₅₀ = 0.4 mg/mL), A-green pea + PP-green pea (IC₅₀ = 0.9 mg/mL) and A-black bean + A-green pea (IC₅₀ = 0.6 mg/mL) presented synergistic effects to inhibit PL. A-chickpea + PP-lentil (IC₅₀ = 0.8 mg/mL) and PP-lentil + A-green pea (IC₅₀ = 1.3 mg/mL) interacted additively to inhibit HMGR and synergistically in the combination of A-chickpea + PP-black bean (IC₅₀ = 1.3 mg/mL) to block HMGR. Peptides FEDGLV and PYGVPVGVR inhibited PL and HMGR in silico, showing predicted binding energy interactions of -7.6 and -8.8 kcal/mol, respectively. Combinations of LPH from different legume protein sources could increase synergistically their anti-adipogenic potential.