In the present investigation, a metal-chelating bioactive peptide was derived from Citrullus lanatus seed milk fermented with Lactococcus lactis. The cationic fermented milk peptide (FMP) thus obtained was purified using the HiTrap-chelating column followed by rpHPLC. The FMP possessed the ability to chelate multiple divalent cations like Cu 2+ , Ca 2+ , and Fe 2+ with 86.81%, 61.04%, and 24.32% of chelation respectively and further it exhibited 78.03% of DPPH free radical scavenging activity. Interaction of FMP with metal ions was assessed by change in the absorption spectra and was analyzed by ultraviolet-visible and fluorescence spectroscopy. The FMP-metal complexes were found stable at simulated gastric conditions. In vitro analysis using intestinal Caco-2 cell lines revealed that there was an increase in metal bioavailability in the presence of the FMP and was least influenced by the addition of a dietary inhibitor, phytic acid. By LC-MS analysis the molecular mass of FMP was found to be 11.6 kD and it contains oxygen-rich and nitrogen-rich amino acids that favor the metal chelation. In our study, we have found that the fermented C. lanatus seed milk can serve as a potential functional food with bioenhancer peptides that increase metal bioavailability and enhance human health.
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