Enzymatic synthesis of sulfated polysaccharides
AssigneesThe University of North Carolina at Chapel Hill, Rensselaer Polytechnic Institute
InventorsJian Liu, Robert J. Linhardt, Fikri Y. Avci, Ava M. Munoz, Jinghua Chen
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Abstract
Heparin is synthesized from a polysaccharide comprised of a 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; by a series of selective reactions catalyzed by recombinant enzymes.
Claims6
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.
Dependencies are inferred from the claim text.