Patent ID: 7247456
Filing Date: 2007-07-24
Classification: C12N,C12P

Abstract:
1. A method for the preparation of a fucosylated oligosaccharide, the method comprising contacting an acceptor saccharide with a fucosylation reaction mixture that comprises GDP-fucose and a fucosyltransferase which transfers fucose from the GDP-fucose to provide said fucosylated oligosaccharide, wherein the efficiency of said fucosylation is enhanced by forming said GDP-fucose by enzymatic conversion of GDP-mannose to GDP-fucose by: a) providing a reaction mixture that comprises GDP-mannose, GDP-mannose 4,6-dehydratase, and NADP+; b) incubating the reaction mixture for a sufficient time to convert at least about 90% of the GDP-mannose to GDP-4-keto-6-deoxymannose; c) adding to the product of step b) a polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO:1, wherein the polypeptide has GDP-4-keto-6-deoxymannose 3,5-epimerase and GDP-4-keto-6-galactose reductase activities; and d) incubating the reaction mixture for a sufficient time to convert the GDP-4-keto-6-deoxymannose to GDP-fucose.