Invention Summary:
The ability to synthesize specific glycosidic linkages provides a way to produce designer glycans. Glycosytransferases that synthesize oligosaccharides are expressed poorly, have narrow substrate specificity and use expensive nucleotide sugars, limiting their utility in scaling-up glycan synthesis. Glycosynthases (GSs) offer an alternative biosynthetic approach to producing glycans in a facile manner. However, to date, only a limited number of GSs have been created from wild-type GHs using an inefficient empirical strategy.
Rutgers researchers have implemented a directed evolution-based protein engineering approach to create GS variants possessing enhanced activity for making specific glycosidic linkages. This is a fluorescence-activated cell sorting (FACS)-based high-throughput screening (HTS) method for rapid identification of GSs with high glycosynthase activity and/or substrate specificity. This technique employs click-chemistry-enabled detection of glycosyl azides as donor sugar reactants for rapid detection of GS activity. Pilot runs have led to the successful identification of GSs capable of synthesizing fucosylated glycans.
Market Applications:
- A versatile HTS method for engineering new glycosynthases
- Identification of GSs for the synthesis of oligosaccharide prebiotics and glycoconjugates for diverse pharmaceutical and biomedical applications
Advantages:
- High-throughput enzyme screening
- High-specificity enzyme engineering
Intellectual Property & Development Status:
Patent pending. Available for licensing and/or research collaboration.
