Methods and compositions for synthesis of oligosaccharides using mutant glycosidase enzymes
US6284494A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Sep 29, 1998 |
| Grant date | Sep 4, 2001 |
| Priority date | — |
| Expiry date | Sep 29, 2018 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/02
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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 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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.