The development of non‐hydrolysable oligosaccharide activity‐based inactivators for endoglycanases: A case study on a‐1,6 mannanases

DOI: 10.1002/chem.202101255

Authors: Hermen S. Overkleeft (Leiden University) , Sybrin Schröder (Leiden University) , Wendy Offen (University of York) , Alexandra Males (University of York) , Yi Jin (University of York) , Casper De Boer (Leiden University) , Jacopo Enotarpi (Leiden University) , Gijs Van Der Marel (Leiden University) , Bogdan Florea (Leiden University) , Jeroen Codée (Leiden University) , Gideon Davies (University of York)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Chemistry – A European Journal

State: Published (Approved)
Published: April 2021
Diamond Proposal Number(s): 13587 , 18598

Abstract: There is a vast genomic resource for enzymes active on carbohydrates. Lagging far behind, however, are functional chemical tools for the rapid characterization of carbohydrate‐active enzymes. Activity‐based probes (ABPs) offer one chemical solution to these issues with ABPs based upon cyclophellitol epoxide and aziridine covalent and irreversible inhibitors representing a potent and widespread approach. Such inhibitors for enzymes active on polysaccharides are potentially limited by the requirement for several glycosidic bonds, themselves substrates for the enzyme targets. Here we show that non‐hydrolysable trisaccharide can be synthesized and applied even to enzymes with challenging subsite requirements. We find that incorporation of carbasugar moieties, which we accomplished by cuprate‐assisted regioselective trans‐diaxial epoxide opening of carba‐mannal we synthesised for this purpose, yields inactivators that act as powerful activity‐based inhibitors for a‐1,6 endo‐mannanases. 3‐D structures at 1.35 – 1.47 Å resolutions confirm the design rationale and binding to the enzymatic nucleophile. Carbasugar oligosaccharide cyclophellitols offer a powerful new approach for the design of robust endoglycosidase inhibitors, while the synthesis procedures presented here should allow adaptation towards activity‐based endoglycosidase probes as well as configurational isosteres targeting other endoglycosidase families.

Journal Keywords: endoglycosidase; cyclophellitol; carbasugar; mechanism-based inhibitor; polysaccharides

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I04-Macromolecular Crystallography

Added On: 22/04/2021 08:34

Discipline Tags:

Biochemistry Chemistry Organic Chemistry Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)