Novel peptides generated from solid-state fermented hempseed cake were characterized by peptidomics and validated for bioactivity. Fermentation with Aspergillus niger , Bacillus subtilis , and Lactobacillus rhamnosus markedly increased peptide numbers compared with the unfermented control and promoted the release of short peptides mainly from Cupin type-1 domain proteins, with leucine frequently occurring at cleavage termini. In silico prediction and molecular docking identified antioxidant, angiotensin-converting enzyme (ACE), and dipeptidyl peptidase-IV (DPP-IV) inhibitory candidates. Synthesized peptides confirmed that LDVSP and LVSPL showed potent ACE inhibition, with IC50 values of 107.47 ± 6.57 and 127.06 ± 12.11 μM, respectively, while LVSPL also inhibited DPP-IV (IC50 = 2.21 ± 0.04 mM). B. subtilis fermentation yielded the strongest antioxidant and ACE-inhibitory activities, whereas A. niger produced the highest DPP-IV inhibitory activity, highlighting fermented hempseed cake as a promising source of multifunctional bioactive peptides. • Solid-state fermentation greatly increased peptide numbers from hempseed protein. • Cupin-derived short peptides enriched, often with leucine at cleavage ends. • In silico tools predicted antioxidant, ACE- and DPP-IV-inhibitory hempseed peptides. • Synthesized peptides confirmed ACE inhibitory activity of LDVSP and LVSPL.
Feng et al. (Wed,) studied this question.