Ertapenem Exposure Is Associated with Downregulation of Purine Biosynthesis Proteins and Inhibition of Biofilm Formation in Staphylococcus aureus

Using LC-MS/MS analysis of biofilm-derived adherent S. aureus cells, we quantified 1,706 proteins and identified 59 proteins with statistically significant expression changes in ertapenem-treated versus untreated S. aureus cells (49 downregulated, 10 upregulated). Proteins associated with purine biosynthesis were predominantly downregulated, particularly FGAM synthase (PurQ) with a −2.29 log₂ fold change. Molecular docking supported by 100 ns molecular dynamics simulations demonstrated stable ertapenem binding to the PurQ active site (docking score: −9.615 kcal/mol) through interactions with key residues: Gly53, Asp54, Tyr55, Gln89, His141, Gly142, Glu143, and Gly144.

Collectively, these findings implicate a novel aspect of ertapenem’s activity, connected to the disruption of purine biosynthesis in S. aureus through targeted downregulation of FGAM synthase. This work provides a strong foundation for drug repurposing strategies aimed at targeting purine biosynthesis in drug-resistant bacterial pathogens.