Enzyme-assisted hydrolysis is frequently used as a cost-effective and efficient method to obtain functional ingredientsfrom bioresources. This study involved the enzyme-assisted hydrolyzation and purification of fucoidan from Eckloniamaxima stipe and the investigation of its anti-inflammatory activity in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Fucoidans of Viscozyme-assisted hydrolysate from E. maxima (EMSFs) harvested in Jeju, Korea. Structural and chemicalcharacterizations were performed using fourier transform infrared spectroscopy, scanning electron microscope, andmonosaccharide analysis. Among fucoidans, EMSF6 was rich in fucose and sulfate and had a similar structural characterto commercial fucoidan. EMSF6 showed a strong inhibitory effect on nitric oxide generation in LPS-induced RAW 264.7cells and significantly decreased the production of LPS-induced pro-inflammatory cytokines, including interleukin-6,interleukin-1β, and tumor necrosis factor α. The anti-inflammatory potential of EMSF6 was mediated through the downregulationof inducible nitric oxide synthase and cyclooxygenase-2 expression. Thus, fucoidans from E. maxima stipe arepromising candidates for functional food products.