(387p) Targeting Endogenous Metabolism to Disrupt Bacterial Persistence: A New Approach to Combat Antibiotic Tolerance

Bacterial persisters, a subpopulation of phenotypic variants, pose a significant challenge in the treatment of chronic and relapsing infections due to their high tolerance to antibiotics. These non-heritable survivors are increasingly recognized as precursors to antimicrobial resistance, yet the metabolic mechanisms enabling their persistence remain poorly defined. In this study, we investigated the metabolic state of Escherichia coli persister cells arising in the late stationary phase, a physiologically relevant condition marked by nutrient depletion and high cell density.

Using a combination of genetic and metabolic assays, we identified the Crp/cAMP regulatory axis as a key driver of metabolic reprogramming during the transition to persistence. This shift redirects cellular metabolism from anabolic growth to oxidative phosphorylation, establishing a dependence on tricarboxylic acid (TCA) cycle–mediated energy production. Contrary to the prevailing notion of metabolic dormancy, our data reveal that persister viability relies on selective activation of internal carbon metabolism, most notably through lipid-derived glycerol generated via phospholipid recycling.

Targeted disruption of glycerol catabolism (via tpiA and gloA deletions) or TCA cycle function significantly impairs persister formation and alters stress-response dynamics. These findings demonstrate that energy homeostasis in nutrient-limited environments is sustained by endogenous carbon flux, providing a novel mechanistic framework for persistence. From a process development perspective, this work highlights metabolic nodes that may serve as tractable targets for combination therapies or antimicrobial adjuvants aimed at eradicating persistent infections.

Leading this project allowed me to develop a strong foundation in microbial metabolism, genetic engineering, and systems-level data integration. Through dissecting the metabolic adaptations that sustain persister survival, I gained practical expertise in identifying functional targets within complex biological networks, skills directly applicable to antimicrobial development, metabolic optimization, and industrial biotechnology.