AbstractThe effect of a 2,5B boat conformation on xyloside reactivity has been investigated by studying the hydrolysis and glycosylation of a series of synthetic xyloside analogues based on a 2‐oxabicyclo[2.2.2]octane framework, which forces the xylose analogue to adopt a 2,5B conformation. The locked β‐xylosides were found to hydrolyze 100–1200 times faster than methyl β‐D‐xylopyranoside, whereas the locked α‐xylosides hydrolyzed up to 2×104 times faster than methyl α‐D‐xylopyranoside. A significant rate enhancement was also observed for the glycosylation reaction. The high reactivity of these conformers can be related to the imposition of a 2,5B conformation, which approximates a transition state (TS) boat conformation. In this way, the energy penalty required to go from the chair to the TS conformation is already paid. These results parallel and support the observation that the GH‐11 xylanase family force their substrate to adopt a 2,5B conformation to achieve highly efficient enzymatic glycosidic bond hydrolysis.