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Although cellulose has been
identified as the foremost candidate for the replacement of fossil fuels, its
recalcitrant nature prevents the full deployment of technologies based on its
saccharification. We recently reported a possible strategy to resolve this
conundrum: using cellulases under “RAging” - a solvent-free process that
utilizes enzymes under mechanochemical conditions - to achieve fast, efficient
hydrolytic depolymerization of cellulosic materials into glucose.
β-Glucosidases catalyze the last and often limiting step of this process, i.e. the formation of glucose from
cellobiose. Here, we reveal the high sensitivity of β-glucosidases to ball
milling, as well as an unexpected stabilization effect of inert surfaces,
enabling the protection of β-glucosidases under mechanochemical treatment. This
approach provides an unexpected strategy to control the reactivity of enzymes
under mechanochemical conditions. Finally, our results also provide the very
first demonstration of enzymatic equilibrium under mechanochemical conditions.