Assessment of molecular interactions and anti-inflammatory activity of fermented camel milk-derived peptides in RAW macrophages

Fermented camel milk is gaining attention as a functional food due to the formation of bioactive peptides during microbial fermentation. This study aimed to investigate the generation of bioactive peptides and their associated antidiabetic, antioxidant, and anti-inflammatory activities in camel milk fermented using Lacticaseibacillus rhamnosus M9 and Saccharomyces cerevisiae WBS2A. The in vitro fermentation process was evaluated in relation to proteolytic activity and peptide formation under varying fermentation times and inoculum concentrations. Antidiabetic activity was assessed using α-amylase and α-glucosidase inhibition assays, while antioxidant potential was determined using the ABTS assay. Peptide profiles and metabolite changes were characterized using RP-HPLC, SDS-PAGE, and 2D electrophoresis, and structural modifications were analysed through FTIR and CLSM. Fermentation significantly enhanced proteolytic activity, leading to the release of low-molecular-weight peptides (3-10 kDa) associated with improved bioactivity. Fermented samples exhibited notable α-amylase (70.68%) and α-glucosidase (59.18%) inhibition, along with maximum antioxidant activity (54.97%) after 48 h. In addition, fermented camel milk reduced intracellular ROS, pro-inflammatory cytokines, and apoptosis in LPS-stimulated RAW 264.7 macrophages. Molecular docking analysis further suggested potential interactions between identified peptides and target enzymes, supporting the observed in vitro activities.These findings demonstrate that fermentation-derived peptides contribute to the functional properties of camel milk; however, further in vivo studies are required to confirm their bioavailability and therapeutic relevance.