Structural highlights
FunctionMANA_ASPNC Endo-1,4-mannanase, a crucial enzyme for depolymerization of seed galactomannans and wood galactoglucomannans. Publication Abstract from PubMedbeta-Mannanases are endo-acting glycoside hydrolases (GHs) that cleave beta-1,4 glycosidic linkages in mannan-rich plant cell wall polysaccharides. They find application in the food and paper industries. Activity-based probes (ABPs) are powerful tools for GH profiling in complex biological samples, yet to date, bespoke ABPs reporting on mannanases have not been reported. Here, we describe the synthesis of cyclophellitol-inspired ABPs based on mannobiose, mannotriose, and glucomannose, and their use in reporting mannanase activities in secretomes from saprophytic bacteria and fungi grown on mannan-containing biomass polysaccharides. In addition to mannanases, our ABPs also labelled cellulases in secretomes from both Aspergillus niger and Cellvibrio japonicus, which may indicate broader ("negative-subsite") substrate specificity in these enzymes. Mechanistic proof of active-site nucleophile labelling by our ABPs was obtained for both AnManA and CjMan26C by X-ray crystallography and for both AnManA and AnMan26A by mass spectrometry. Together, our results establish mannanase-targeted ABPs that may find use alongside existing reagents that report on retaining GHs that process other bulk polysaccharides. The development of activity-based mannanase probes.,Tedeschi M, Lit VAJ, McGregor NGS, Gote T, Kooloth Valappil P, Arentshorst M, Florea BI, Gagestein B, Armstrong Z, Codee JDC, Nin-Hill A, Rovira C, Ram AFJ, Davies GJ, Overkleeft HS Chem Sci. 2026 Jul 13. doi: 10.1039/d6sc04720c. PMID:42445515[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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