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Glycosylated cyclophellitol-derived activity-based probes and inhibitors for cellulases.

Casper de BoerNicholas G S McGregorEvert PeterseSybrin P SchröderBogdan I FloreaJianbing JiangJos ReijngoudArthur F J RamGilles P van WezelGijsbert A van der MarelJeroen D C CodéeHerman S OverkleeftGideon J Davies
Published in: RSC chemical biology (2020)
Cellulases and related β-1,4-glucanases are essential components of lignocellulose-degrading enzyme mixtures. The detection of β-1,4-glucanase activity typically relies on monitoring the breakdown of purified lignocellulose-derived substrates or synthetic chromogenic substrates, limiting the activities which can be detected and complicating the tracing of activity back to specific components within complex enzyme mixtures. As a tool for the rapid detection and identification of β-1,4-glucanases, a series of glycosylated cyclophellitol inhibitors mimicking β-1,4-glucan oligosaccharides have been synthesised. These compounds are highly efficient inhibitors of HiCel7B, a well-known GH7 endo-β-1,4-glucanase. An elaborated activity-based probe facilitated the direct detection and identification of β-1,4-glucanases within a complex fungal secretome without any detectable cross-reactivity with β-d-glucosidases. These probes and inhibitors add valuable new capacity to the growing toolbox of cyclophellitol-derived probes for the activity-based profiling of biomass-degrading enzymes.
Keyphrases
  • highly efficient
  • loop mediated isothermal amplification
  • small molecule
  • living cells
  • ionic liquid
  • label free
  • fluorescence imaging
  • quantum dots
  • fluorescent probe