Patent ID: 8771995
Filing Date: 2014-07-08
Classification: C12P

Abstract:
1. A method of preparing heparin from a first polysaccharide comprised of 1-4 glycosidically linked alternating polymer of uronic acid and glucosamine residues, wherein the uronic acid is selected from iduronic and glucuronic acid, wherein the glucosamine is partially N-sulfated; the method comprising (a) treating the first polysaccharide with a reaction mixture comprising recombinant 2-O-sulfotransferase (OST) enzyme comprising the amino acid sequence of SEQ ID NO: 1, to yield a second polysaccharide; (b) treating the second polysaccharide with a reaction mixture comprising recombinant 6-O-sulfotransferase (OST) enzyme to yield a third polysaccharide; (c) treating the third polysaccharide with a reaction mixture comprising recombinant 3-O-sulfotransferase (OST) enzyme isoform 1 to yield heparin; wherein in each of steps (a)-(c) the reaction mixture further comprises 3′-phosphoadenosine 5′-phosphosulfate (PAPS), an arylsulfotransferase and an aryl sulfate.