Methods and compositions for synthesis of oligosaccharides, and

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

1995-12-12

1998-02-10

Wax, Robert A.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435100, 4351721, 435200, 435201, 435209, 536 41, C12P 1944, C12P 1912, C12N 1500, C12N 924

Patent

active

057168121

ABSTRACT:
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 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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