This study evaluated the iron-chelating capacity (ICC) of Lupinus mutabilis protein hydrolysate (LMPH) and its peptide fractions obtained through ultrafiltration and purification by immobilized metal ion affinity chromatography (IMAC) and gel filtration chromatography (GFC). Peptides were identified by LC-MS/MS, and their interactions with Fe2+ were analysed using molecular docking. LMPH was produced by enzymatic hydrolysis with Alcalase and subsequently subjected to ultrafiltration to concentrate peptides smaller than 2 kDa. This fraction exhibited higher ICC (35.1 mg Fe2+·g−1) than the hydrolysate (22.75 mg Fe2+·g−1). Sequential purification by IMAC and GFC yielded peptide fractions with enhanced ICC values (45.20 and 13.51 mg Fe2+·g−1). A total of 176 peptides were identified by de novo LC-MS/MS sequencing, from which nine were selected based on favourable structure–ICC relationships and absence of predicted toxicity. Molecular docking analysis suggested spatial proximity between Fe2+ and the selected peptides. Although stable multi-site binding was not predicted under the applied computational model, the results support the potential of these sequences to interact with Fe2+. These findings provide molecular and chemical insights supporting the iron-binding potential of LMPH-derived peptides and highlight their future potential as functional ingredients for preventing and managing iron deficiency.
Vila-Santillán et al. (Fri,) studied this question.