Exploiting cyclodextrins as artificial chaperones to enhance enzyme protection through supramolecular engineering

We report a method of enzyme stabilisation exploiting the artificial protein chaperone properties of beta-cyclodextrin (beta-CD) covalently embedded in an ultrathin organosilica layer. Putative interaction points of this artificial chaperone system with the surface of the selected enzyme were studied in silico using a protein energy landscape exploration simulation algorithm. We show that this enzyme shielding method allows for drastic enhancement of enzyme stability under thermal and chemical stress conditions, along with broadening the optimal temperature range of the biocatalyst. The presence of the beta-CD macrocycle within the protective layer supports protein refolding after treatment with a surfactant. Protecting a surface-immobilised enzyme with an enzyme-thin organosilica layer produced using a protein chaperone building block allows drastic improvement in the enzyme's thermal stability.