Activity-Based Profiling of Carbohydrate-Active Enzymes

Activity-based profiling exploits chemically reactive probes that covalently bind the active sites of carbohydrate-active enzymes (CAZymes), enabling direct detection, characterisation and quantification of enzyme function in complex biological settings. By targeting catalytic nucleophiles or cofactors within glycoside hydrolases, carbohydrate esterases and auxiliary activities, activity-based probes (ABPs) illuminate the distribution and dynamics of CAZymes in proteomes, microbial secretomes and tissue specimens. This approach overcomes limitations of sequence-based annotation by reporting only on catalytically competent species, and it has been instrumental in identifying enzyme defects in metabolic disorders, mapping glycan-processing pathways in gut microbiota and evaluating biomass-degrading machineries in environmental and industrial contexts. Key chemical scaffolds, such as cyclophellitol derivatives and mechanism-based inactivators, confer both selectivity and irreversible labelling, facilitating downstream enrichment, imaging or mass-spectrometric analysis. The ability to multiplex distinct probe warheads and reporter tags has recently expanded the throughput of enzyme-function screening, accelerating discovery of novel CAZymes and enabling rational design of enzyme cocktails for sustainable biotechnologies. This integrated chemical-biological strategy continues to redefine our understanding of carbohydrate metabolism across health, ecology and biotechnology.

Research from Nature Portfolio

No recent Nature Portfolio content available.

Activity-Based Profiling of Carbohydrate-Active Enzymes publication trend

The graph below shows the total number of articles in activity-based profiling of carbohydrate-active enzymes across all publications each year (not limited to Nature Index journals).

Technical terms

Activity-based probe (ABP): A molecule bearing a reactive “warhead” and a reporter tag that irreversibly binds an enzyme active site to enable detection and isolation of catalytically active enzymes.

Activity-based protein profiling (ABPP): A chemical proteomic technique that uses ABPs to survey active enzymes in complex proteomes rather than relying on sequence or abundance alone.

Carbohydrate-Active Enzymes (CAZymes): A broad family of enzymes, including glycoside hydrolases, glycosyltransferases, carbohydrate esterases and auxiliary activities, that assemble or break down complex carbohydrates.

Glycoside hydrolases (GHs): Enzymes that catalyse the hydrolysis of glycosidic bonds in oligo- and polysaccharides, classified into families based on sequence and mechanism.

Cyclophellitol: A natural epoxide-bearing inhibitor scaffold used to design retaining glycosidase ABPs, exploiting its mimicry of sugar substrates to achieve covalent enzyme labelling.

References

1. From Mechanism-Based Retaining Glycosidase Inhibitors to Activity-Based Glycosidase Profiling. Journal of the American Chemical Society (2024).
2. A Multiplexing Activity-Based Protein-Profiling Platform for Dissection of a Native Bacterial Xyloglucan-Degrading System. ACS Central Science (2023).
3. Rational Design of Mechanism-Based Inhibitors and Activity-Based Probes for the Identification of Retaining α‑l‑Arabinofuranosidases. Journal of the American Chemical Society (2020).