Molecular and functional characterization of an iron fortified plant based meat (IFPBM) for enhanced nutritional efficacy and cellular iron uptake

• Fe 3 O 4 nanoparticle fortification significantly enhanced iron bioavailability in HTC-16 cells. • Iron demonstrated binding interactions with lysine, cysteine, and glutamine amino acids. • Iron fortification contributed to stabilization of the β-sheet structure within IFPBM • Peptides derived from IFPBM exhibited strong binding affinity to the Dcytb enzyme. • Iron ion chelation decreases the digestibility of IFPBM protein. Growing consumer demand for nutrient-dense foods with low environmental footprints is driving innovation in plant based products. This study employed response surface methodology to optimize a plant based meat (PBM) formulation composed of pea protein isolate (80±0.04%), wheat gluten (18.5%±0.21%), and seasoning (1.5±0.11%), which exhibited superior water holding capacity of 14.7 ± 0.13 %, protein content of 38±0.64 %, and porosity of 23.64±0.88% ensuring desirable structural and functional characteristics. The optimized PBM was subsequently fortified with Fe 3 O 4 nanoparticles to address iron and protein deficiencies. The strategic incorporation of Fe-NP increased iron content to 16.9±0.04 mg/100g while cellular iron uptake increased by 79.32±5.71 % compared with non-fortified analogs. The MTT assay performed on HCT116 intestinal epithelial cells indicated good cellular compatibility, with an IC 50 ranging from (0.5±0.12mg/ml). Molecular interactions between Fe-NP and amino acid residues, particularly histidine, cysteine, and methionine, modulate protein digestibility, stabilize β-sheet structures, and significantly (p < 0.05) improve the colour and texture attributes of IFPBM. Additionally, in silico digestion and molecular docking of key peptides (YPTSPQQPGQW, PYPQPQP, QQPGQGQPGYY) revealed strong binding affinities of more than 7 kcal/mol with AMPK, ACE, DPP-IV, and Dcytb, suggesting multifunctional bioactivities with potential antihypertensive, antidiabetic, and iron transport–modulating effects. These findings provide evidence for exploring nanotechnology in improving the nutritional profile and functional properties of PBM, thereby improving consumer acceptance. Further studies should investigate the cellular and molecular pathways underlying the enhanced iron uptake using appropriate animal models.