Modulation of the Keap1–Nrf2 pathway using a peptide derived from edible mushrooms: In silico and in vitro studies

In this study, the peptide WV-9, obtained from Hericium erinaceus (Lion’s mane mushroom), was shown to modulate the Keap1–Nrf2 antioxidant pathway through direct molecular interaction. Molecular docking and molecular dynamics simulations demonstrated that WV-9 binds stably and specifically to the Keap1 Kelch domain by mimicking the ETGE motif of Nrf2. This interaction suggests a competitive mechanism that may disrupt Keap1-mediated degradation of Nrf2, promoting its nuclear translocation and transcriptional activation. In vitro analysis using RAW264.7 macrophage cells revealed that WV-9 enhanced the expression of key antioxidant genes, including superoxide dismutase 2 ( Sod-1 ), NAD(P)H quinone dehydrogenase 1 ( Nqo-1 ), and heme oxygenase 1 ( HO-1 ). Importantly, protein-level validation by Western blot analysis demonstrated a significant upregulation of HO-1 expression in WV-9-treated cells, whereas other antioxidant enzymes, such as SOD-1 and CAT, showed increasing trends that did not reach statistical significance, indicating a selective activation of the Nrf2 antioxidant response. In silico ADMET analysis predicted a favorable safety profile, with low cytochrome P450 interaction and minimal toxicity. Unlike conventional Nrf2 activators such as sulforaphane, which act via redox-based mechanisms, WV-9 operates through direct interference with the Keap1–Nrf2 interface. These findings support the potential application of WV-9 as a natural therapeutic candidate or a bioactive ingredient in functional foods aimed at combating oxidative stress and immune dysregulation. WV-9, a peptide derived from Hericium erinaceus , interacts with the Keap1 Kelch domain as predicted by molecular docking and molecular dynamics simulations. This interaction is associated with modulation of the Keap1–Nrf2 pathway and increased expression of Nrf2-regulated antioxidant and immunomodulatory genes in macrophage cells. • WV-9, a peptide from Hericium erinaceus , shows potential modulation of the Keap1–Nrf2 pathway. • Docking and molecular dynamics simulations suggest stable interaction with the Keap1 Kelch domain. • WV-9 enhances antioxidant-related gene expression (SOD-1, NQO-1, HO-1) in macrophage cells. • In silico ADMET analysis predicts a favorable safety profile with low toxicity. • WV-9 is a promising mushroom-derived bioactive for oxidative stress modulation.A novel peptide (WV-9) was identified from H. erinaceus with potential Keap1–Nrf2 binding activity.