Peptidoglycan recycling is critical for cell division, cell wall integrity, and β-lactam resistance in Caulobacter...

Most bacteria possess a peptidoglycan (PG) sacculus, which is continuously remodeled during cell growth and division. The turnover products generated in this process are typically imported into the cell and reused for PG biosynthesis. While the underlying pathways have been studied intensively in gammaproteobacteria, knowledge of their presence and physiological roles in other bacterial lineages remains limited. Here, we comprehensively investigate PG recycling in the alphaproteobacterial model organism Caulobacter crescentus. Characterizing the activities of key enzymes in vitro and in vivo, we show that this species contains a functional PG recycling pathway, including the MurU shunt. Our results reveal that PG recycling is critical for C. crescentus cell morphology and division and is dynamically regulated to balance the flux of metabolic intermediates toward PG biosynthesis and central carbon metabolism. Importantly, defects in PG recycling strongly impair the intrinsic ampicillin resistance of C. crescentus without changing the activity of its β-lactamase BlaA, likely by limiting PG precursor biosynthesis and thereby decreasing the activity of the cell wall biosynthetic machinery in the presence of residual antibiotic. These findings underscore the central role of PG recycling in bacterial fitness and suggest that inhibiting this process could provide a promising strategy to combat β-lactam-resistant pathogens.