Oligosaccharide synthesis using activated glycoside derivative, glycosyl transferase and catalytic amount of nucleotide phosphate

US5952203A · kind A · utility

Assignee

• University of British Columbia · CA

Inventors

• Stephen Withers · Vancouver, CA
• Brenda Lougheed · Vancouver, CA

Key dates

• Filing date: Apr 11, 1997
• Grant date: Sep 14, 1999
• Expiry date: Apr 11, 2017

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P19/18
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Oligosaccharides are prepared using glycosyl transferase and activated glycosyl derivatives as donor sugars without the use of sugar nucleotides as donor sugars. A reaction mixture composition containing an activated glycoside derivative such as glycosyl fluoride or glycosyl mesylate, an acceptor substrate such as lactose or other oligosaccharide, a glycosyl transferase and a catalytic amount of a nucleotide phosphate or nucleotide phosphate analog is reacted to produce a glycosylated acceptor. In addition to an oligosaccharide, the acceptor substrate may be a monosaccharide, a fluorescent-labeled saccharide or a saccharide derivative such as an aminoglycoside antibiotic. The glycosyl transferase may be immobilized by removing its membrane-binding domain and attaching in its place a cellulose-binding domain. In a preferred embodiment, galactosyl transferase is the glycosyl transferase, .alpha.-D-galactosyl fluoride is the activated glycoside derivative and lactose is the acceptor substrate.