Pyropia yezoensis (P. yezoensis) is an important marine algae. Its high protein content serves as a good source of biologically active peptides. Potent inhibitory effects on the production of inflammatory mediators were observed in a bioactive peptide derived from P. yezoensis (peptide from P. yezoensis; PPY1), as demonstrated in lipopolysaccharide (LPS)-stimulated macrophages. The present study showed that peptide concentrations ranging from 250 to 1,000 ng/ml had no significant cytotoxicity in the cell viability assay when applied to the RAW 264.7 cells for 24 h. PPY1 completely inhibited LPS‑stimulated nitric oxide (NO) release in a dose-dependent manner. Fluorescence intensity, corresponding to intracellular reactive oxygen species (ROS) produced by 10 ng/ml LPS-stimulated cells, significantly shifted, indicating that the peptide reduced the level of ROS. Furthermore, PPY1 exerted potent inhibitory activity to reduce the release of pro-inflammatory cytokines (inducible NO synthase, cyclooxygenase-2, interleukin-1β and tumor necrosis factor-α) in LPS-stimulated macrophages in a dose-dependent manner. These results also showed that the anti-inflammatory activity of PPY1 was associated with downregulation of extracellular signal-regulated kinase, protein 38, and c-jun NH2-terminal kinase phosphorylation in the mitogen-activated protein kinase pathways. In conclusion, PPY1 can have a significant role as an anti-inflammatory agent, with a potential for use in marine products.