Illuminating glycoscience: synthetic strategies for FRET-enabled carbohydrate active enzyme probes
Carbohydrates are an essential class of biomolecule and carbohydrate active enzymes (CAZys) catalyse their synthesis, refinement, and degradation, hence contributing an overall regulatory capacity to their underpinning physiological roles. As such, there is a considerable current requirement to be able to monitor, quantify and inhibit CAZy activity. It is here that Förster resonance energy transfer (FRET) is emerging as a powerful tool in enabling this, through synthetic conjugation of appropriate fluorogenic partners into a particular CAZy substrate. In this review we survey recent accomplishments in synthetic methodology for accessing defined carbohydrate structures, suitably equipped with FRET probe capability, followed by their utilisation in studying particular classes of CAZy. The majority of examples concern endo-acting glycosidases, but emergent probes for exo-glycosidases, glycosyl transferases and catalytic antibodies are also examined.