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Visualizing cell wall polymers biosynthesis and hydrolysis

Biomolecular interactions involving carbohydrates are fundamental processes relevant to the recognition, biosynthesis, and degradation of nearly all biomass in nature as well as industrial biochemical unit operations. However, classical enzymology techniques and biochemical assays are not suitable for understanding CAZyme adsorption or activity at solid-liquid interfaces. We develop and apply advanced single-molecule biophysical characterization and real-time imaging techniques to probe CAZymes and proteins active on carbohydrate-based interfaces. We are studying cellulases binding and processive motility to cellulose using single-molecule force spectroscopy. We are visualizing polysaccharides biosynthesis in real-time using super-resolution fluorescence imaging to characterize how single cells synthesize and assemble their cell walls.

Force spectroscopy enabled enzyme & cellular engineering

Mechanical forces are increasingly being recognized to play a pivotal role in most biological processes. Nearly every cell-biomacromolecule, cell-cell, and cell-pathogen interaction often involves protein-glycan or glycan-glycan interactions under conditions far from equilibrium. Here, we develop and utilize a highly multiplexed single-molecule force spectroscopy technique (called Acoustic Force Spectroscopy) enabled bio-panning tool to facilitate rational structure-guided and directed evolution based engineering of glycan-binding proteins and cellular systems, under physiologically relevant conditions far from equilibrium.