A novel acidic polysaccharide named SSPA50-1 was isolated from Scapharca subcrenata using a simulated gastric fluid extraction method. SSPA50-1 is a heteropolysaccharide with an average molecular weight of 44.7 kDa that is composed of galacturonic acid, glucose, galactose, mannose, ribose, rhamnose, fucose, xylose and arabinose at a molar ratio of 1.00:5.40:9.04:3.10:1.59:4.01:2.10:2.21:2.28. The structural characterization based on the methylation and 1D/2D NMR analyses indicated that SSPA50-1 is composed of →3)-β-L-Rhap-(1→, →3)-β-L-2-O-Me-Fucp-(1→, →2)-α-D-Xylp-(1→, →5)-α-L-Araf-(1→, →3)-β-D-Galp-(1→, →6)-α-D-Glcp-(1→, →3,4)-β-D-Manp-(1→, →3,4)-β-D-Galp-(1→, β-D-Ribf-(1→, α-D-Glcp-(1→, and α-D-GalAp6Me-(1→. Furthermore, SSPA50-1 possessed potent immunoregulatory activity by enhancing the phagocytosis and NO, iNOS, TNF-α and IL-6 secretion capacity of RAW264.7 cells. Otherwise, SSPA50-1 significantly promoted the proliferation of splenic lymphocytes and RAW264.7 macrophages. These results indicated that SSPA50-1 could be developed as a potential ingredient for immunostimulatory agents.