Green layer, Enteromorpha prolifera, is regarded as one of important materials for food processing in Korea. The acidic water-soluble polysaccharide (CPC-PS) isolated from the alga with hot water and cetylpyridium chloride was mainly constituted of rhamnose, xylose, uronic acid and sulfate. To determine the glycosyl-linkages and positions of sulfate by methylation, the CPC-PS was reduced and/or sulfates. A marked increase of glucose content in the reduced polysaccharide indicated that glucuronic acid was a major sugar in the polymer and sulfation was deduced to occur on O-3 of rhamnose and O-2 of xylose. According to the methylation analysis of the native, reduced, desulfated and reduced-desulfated polymers, CPC-PS mainly composed of 1,4- and 1,2,3-linked rhamnose 3-sulfate, 1,4-linked xylose 2-sulfate, 1,4-linked xylose and 1,4-linked glucuronic acid. Minor 1,4-linked rhamnose and 1,4,6-linked galactose residues were also detected.