Aldehyde‐Based Inhibitors of the Peptidoglycan O‐Acetylesterase Ape

The O‐acetylation of the muramic acid residues in peptidoglycan (PG) is a modification that protects the bacteria from lysis due to the action of lysozyme. In Gram‐negative bacteria, deacetylation is required to allow lytic transglycosylases to promote PG cleavage during cell growth and division. This deacetylation is catalyzed by O‐acetylpeptidoglycan esterase (Ape) which is a serine esterase and employs covalent catalysis via a serine‐linked acyl enzyme intermediate. Loss of Ape activity affects the size and shape of bacteria and dramatically reduces virulence. In this work, we report the first rationally designed aldehyde‐based inhibitors of Ape from Campylobacter jejuni. The most potent of these acts as a competitive inhibitor with a Ki value of 13 μM. We suspect that the inhibitors are forming adducts with the active site serine that closely mimic the tetrahedral intermediate of the normal catalytic cycle. Support for this notion is found in the observation that reduction of the aldehyde to an alcohol effectively abolishes the inhibition. Aldehyde‐based inhibitors of the peptidoglycan deacetylase Ape were shown to act as competitive inhibitors with Ki values in the low micromolar range. Reduction of the aldehyde to an alcohol abolishes inhibition, suggesting that the binding may involve the formation of a tetrahedral adduct with the active site serine nucleophile.