Methods and compositions for synthesis of oligosaccharides,...

Details Methods and compositions for synthesis of oligosaccharides,... Methods and compositions for synthesis of oligosaccharides,...

C - Chemistry, Metallurgy

12

N

C12N 9/42 (2006.01) C12N 9/24 (2006.01) C12N 9/26 (2006.01) C12P 19/04 (2006.01) C12P 19/14 (2006.01)

Patent

CA 2165041

Mutant glycosidase enzymes are formed in which the normal nucleophilic amino acid within the active site has been changed to a non-nucleophilic amino acid. These enzymes cannot hydrolyze disaccharide products, but which can still form them. Using this enzyme, oligosaccharides are synthesized by preparing a mixture of an .alpha.-glycosyl fluoride and a glycoside acceptor molecule; enzymatically coupling the .alpha.-glycosyl fluoride to the glycoside acceptor molecule to form a glycosyl glycoside product using the mutant glycosidase enzyme; and recovering the glycosyl glycoside product. Particular enzymes include a mutant form of Agrobacterium .beta.-Glucosidase in which the normal glutamic acid residue at position 358 is replaced with an alanine residue.

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