Fractionation of sunflower protein hydrolysates by low pressure preparative gradient chromatography: An integrated in vitro and in silico assessment of bioactivities.

Bioactive peptides from plant proteins offer significant benefits. In this study, the structural and bioactive properties of sunflower protein isolate and its enzymatic hydrolysates were systematically investigated. The hydrolysates were fractionated using low-pressure preparative gradient chromatography (LP-PGC), and their antioxidant capacity, along with angiotensin-converting enzyme and dipeptidyl peptidase-IV inhibitory activities relevant to metabolic health, were assessed. Structural analyses via sodium dodecyl sulfate polyacrylamide gel electrophoresis and scanning electron microscopy revealed significant alterations in molecular weight distribution and surface morphology after hydrolysis and fractionation, confirming the generation of low-molecular-weight peptides linked to improved bioactivity. The results demonstrated a clear fraction-dependent bioactivity profile. MTT-based cell viability analysis of HepG2 and Caco-2 cells indicates that the non-toxicity of the hydrolysates depends on both their concentration and the cell type. Molecular docking studies were performed with the three highest-scoring peptides from each hydrolysate according to PeptideRanker. LP-PGC effectively reveals in vitro/in silico bioactivity of sunflower protein hydrolysates for multifunctional food use.