Preparation of fucosylated oligosaccharides
AssigneeNeose Technologies, Inc.
InventorEric R. Sjoberg
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Abstract
This invention provides methods for practical enzymatic conversion of GDP-mannose to GDP-fucose. These methods are useful for efficient synthesis of reactants used in the synthesis of fucosylated oligosaccharides.
Claims11
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;
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- d) incubating the reaction mixture for a sufficient time to convert the GDP-4-keto-6-deoxymannose to GDP-fucose.
Dependencies are inferred from the claim text.