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.
- 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
- High-throughput enzyme screening
- High-specificity enzyme engineering
Intellectual Property & Development Status:
Patent pending. Available for licensing and/or research collaboration.