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

“Assessment of molecular interactions and anti-inflammatory activity of fermented camel milk-derived peptides in RAW macrophages. Padhiyar PN(1), Basaiawmoit B(2), Mankad PM(3), Sakure AA(4), Bhattacharya A(5), Kondepudi KK(5), Tiwary BK(6), Sarkar P(7), Koringa P(3), Adil S(8), Hati S(9). Author information: (1)Department of Dairy Microbiology, SMC College of Dairy Science, Kamdhenu University, Anand, Gujarat, 388110, India. (2)Department of Rural Development and Agricultural Production, North-Eastern Hill University, Chasingre, Meghalaya, India. (3)Department of Veterinary Biotechnology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Anand, Gujarat, 388110, India. (4)Department of Agriculture Biotechnology, Anand Agricultural University, Anand, Gujarat, 388110, India. (5)Healthy Gut Research Group, Food & Nutritional Biotechnology Division, National Agri-Food Biotechnology Institute, Mohali, Punjab, 140306, India. (6)Department of Microbiology, North Bengal St. Xavier's College, North Bengal University, Siliguri, West Bengal, India. (7)Department of Food Process Engineering, National Institute of Technology, Rourkela, India. (8)Department of Dairy Technology, Parul Institute of Technology, Parul University, Vadodara, Gujarat, 391 760, India. shaikh.adil23773@paruluniversity.ac.in. (9)Department of Dairy Microbiology, SMC College of Dairy Science, Kamdhenu University, Anand, Gujarat, 388110, India. subrota.hati@kamdhenuuni.edu.in. 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. © 2026. The Author(s). Conflict of interest statement: Declarations. Conflict of interest: The authors declare that they have no conflicts of interest or competing interests. Ethical approval and consent to participate: The study was approved by the PG Research Committee, Kamdhenu University, and Advisory Committee Members from the SMC College of Dairy Science, Anand, Gujarat, India. All committee members were duly informed of their rights and responsibilities and provided explicit written consent. The study was conducted in accordance with the guidelines governing research involving human participants, as outlined by the Ethics Committee of the SMC College of Dairy Science, Anand, Gujarat, India. There is no animal study in involved in the present work. We do not directly involve in procuring the milk from the animals. We have purchased the camel milk from the GCMMF Ltd., Anand, India available at the retailer shop at Anand, Gujarat, India. Consent for publication: Not applicable.”