Sequential Degradation of Heparin in Flavobacterium heparinum

Five enzymes from Flavobacterium heparinum, capable of degrading heparin to its basic constituents, have been purified and characterized. The purified heparinase degrades heparin to trisulfated disaccharide with a yield of 70 to 80%. The trisulfated disaccharide is desulfated by a disaccharide sulfoesterase yielding the disulfated disaccharide. This product is then a substrate for the glycuronidase forming glucosamine-2,6-disulfate and α,β-keto acid. Glucosamine-2,6-disulfate is desulfated by a sulfoesterase and a sulfamidase yielding free glucosamine and inorganic sulfate. The substrate specificity of these enzymes led to the formulation of a pathway for the sequential degradation of heparin. It was shown, by the action of these purified enzymes, that the trisulfated disaccharide is one of the fundamental structural units of heparin.