Reduced glutathione levels in Enterococcus faecalis trigger metabolic and transcriptional compensatory adjustments during iron exposure.

Enterococcus faecalis, a facultative anaerobic pathogen and common constituent of the gastrointestinal microbiota, must navigate varying iron levels within the host. This study explores its response to iron supplementation in a glutathione-deficient mutant strain ({Delta}gsh). We examined the transcriptomic and metabolic responses of a glutathione synthetase mutant strain ({Delta}gsh) exposed to iron supplementation, integrating these data into a genome-scale metabolic model (GSMM). Our results show that under glutathione deficiency, E. faecalis reduces intracellular iron levels and shifts its transcriptional response to prioritize energy production genes. Notably, basal metabolites, including arginine, increase. The GSMM highlights the importance of arginine metabolism, particularly the arc operon (anaerobic arginine catabolism), as a compensatory mechanism for reduced glutathione during iron exposure. These findings provide insights into how E. faecalis adjusts metal homeostasis and transcriptional/metabolic processes to mitigate the effects of oxidative stress caused by iron. IMPORTANCEIron is essential for bacterial survival, yet its excess can be harmful through increase of oxidative stress. Enterococcus faecalis, a bacterium common member of the human gut, must carefully balance its iron levels in order to survive in changing environments. This study studies how E. faecalis compensates the reduced levels of glutathione --a key antioxidant-- when exposed to high iron concentrations. We discovered that E. faecalis lowers its intracellular iron levels under glutathione decrease and reprograms its metabolism to prioritize energy production. These findings provide valuable insights into bacterial adaptation mechanisms under oxidative stress conditions, which could influence the development of new strategies to combat bacterial infections.