Unravelling mechanistic insights of sprouting and autoclaving to enhance the nutritional and structural profile of horse gram (Macrotyloma uniflorum L.)

Horse gram is an underutilized legume with solid nutritional potential, but its consumption is limited due to high anti-nutritional factors and poor digestibility. The systematic elucidation to evaluate the effects of sprouting, autoclaving, and their combination on the physicochemical, structural, and nutritional properties of horse gram (Macrotyloma uniflorum). The research sought to determine how these treatments influence Amino Acid composition, anti-nutritional factors, Texture profile and structural perspectives. Horse gram (HG) were subjected to sprouting of 36 h, autoclaving (121 °C, 15 psi for 15 min), and combined sprouting-autoclaving treatments. Instrumental analysis was conducted using Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Texture profile analysis (TPA), proximate composition, anti-nutritional factor (ANF) quantification, and amino acid profiling were performed following standard protocols. Sprouted Horse Gram (SHG) exhibited the highest protein content (23.68%) and total essential amino acid content (60.1 g/100 g). All treatments significantly (p ≤ 0.05) reduced trypsin inhibitors, phytic acid, tannins, and oxalates. FTIR revealed substantial changes in amide I and II bands and hydroxyl regions, indicating protein denaturation and hydrogen bonding alterations SEM analyses confirmed surface disruption and porosity increase in treated samples. Sprouting enhanced protein quality and reduced anti-nutritional factors, while autoclaving improved digestibility and textural properties. The findings provide novel insights into how traditional treatments alters the nutritional, and structural properties of horse gram, making a sustainable and systematically validated approach for enhancing its functionality in plant-based food systems. This research bridges biochemical and structural evidence with practical food applications in future, underscoring its significance for developing nutritionally superior legume based diets for health-conscious consumers and the growing vegan population.