Methods and compositions for synthesis of oligosaccharides using mutant glycosidase enzymes
US7078194B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 17, 2001 |
| Grant date | Jul 18, 2006 |
| Priority date | — |
| Expiry date | Apr 17, 2021 |
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 nonnucleophilic 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 &agr;glycosyl fluoride and a glycoside acceptor molecule; enzymatically coupling the &agr.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 &bgr.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.