Cyclophellitol, cyclophellitol aziridine and derivatives thereof are selective and potent, covalent mechanism based inhibitors of exo-acting, retaining glycosyl hydrolases. Activity Based Probes (ABPs) based on these inhibitors have contributed tremendously to our understanding of exo-acting enzyme activity in health and disease.[1,2] The construction of cyclophellitol containing carbohydrate oligomers allows the study of endo-acting glycosidases.
In this communication the design and synthesis of dedicated ABPs based on hemicellulose and cellulose motives (the main carbohydrate derived components of plant biomass) is presented. The main challenge is the incorporation of the cyclophellitol warhead in the oligomers and the discovery of reaction conditions that are compatible with the warhead in the late stage of the synthesis. Examples of strategies employed to synthesize different motives will be discussed.
The obtained probes are used to visualize active xylanases, cellulases and xyloglucanases. These probes can aid in the discovery of unknown enzymes acting on recalcitrant biomass leading to higher sugar yields for renewable fuel production.
- Jiang, J.; Kuo, C.-L.; Wu, L.; Franke, C.; Kallemeijn, W. W.; Florea, B. I.; van Meel, E.; van der Marel, G. A.; Codée, J. D. C.; Boot, R. G.; et al. Detection of Active Mammalian GH31 α-Glucosidases in Health and Disease Using In-Class, Broad-Spectrum Activity-Based Probes. ACS Cent. Sci. 2016, 2 (5), 351–358.
- Witte, M. D.; Kallemeijn, W. W.; Aten, J.; Li, K.-Y.; Strijland, A.; Donker-Koopman, W. E.; van den Nieuwendijk, A. M. C. H.; Bleijlevens, B.; Kramer, G.; Florea, B. I.; et al. Ultrasensitive in Situ Visualization of Active Glucocerebrosidase Molecules. Nat. Chem. Biol. 2010, 6 (12), 907–913.