Antioxidant and immunomodulatory potency of Lacticaseibacillus rhamnosus NCDC24 fermented milk-derived peptides: A computationally guided in-vitro and ex-vivo investigation.

Infections of microbial and non-microbial origins have been associated with significant immunological manifestations, thereby underscoring the need for a thorough understanding and investigation of novel immunomodulatory and antioxidant molecules that could prevent these incidences. To this end, we herein aim to identify fermented milk peptides with antioxidant and immunomodulatory properties that could be exploited for specific future applications. Our computational prediction models indicate that these peptides are non-toxic and possess considerable hydrophobicity (19.82-38.96 %) and functionality. Further analyses reveal that two of the four peptides, i.e., Pep 1 (AGWNIPM) and Pep 4 (YLGYLEQLLR), possess higher in-vitro antioxidant activity. The immunomodulatory potential of these two peptides (Pep 1 and Pep 4) is further demonstrated by using a combination of molecular simulation trajectory and ex-vivo approaches. Both peptides demonstrate ability to control the production of pro- inflammatory (TNF-α, IL-1, and IL-6) and anti-inflammatory (IL-10) cytokines as well as nitric oxide release in LPS-stimulated murine peritoneal macrophages. Similarly, peptide interferences also lead to significant (P < 0.05) improvement in macrophage phagocytic capacity. Taken together, these findings highlight the antioxidant and immunomodulatory properties of fermented milk peptides (Pep 1 and Pep 4) within the cellular environment and should facilitate prospective studies exploring such bioactive peptides and related functional molecules mediating the benefits of fermented milk products on human health.