BioModelsCailean CarterNon-curatedconstraint-based model*Manuscript Acceptedhttps://www.ebi.ac.uk/biomodels/MODEL2507240001falseBioModelsModelsOtherCarter2025 Trimethoprim induced metabolic perturbations and counteractions2026MODEL2507240001Cailean CarterCailean Carternull, Trimethoprim is a clinically important antibiotic for treating urinary tract infections yet its mechanism of killing remains elusive due to a cascade of affected metabolic reactions. Metabolites available in growth media can selectively counteract trimethoprim which affects the interpretation of results. We sought to understand the full scope of trimethoprim’s impact on Escherichia coli metabolism and how metabolite availability affects trimethoprim outcomes. We applied flux balance analysis on a genome-scale metabolic model of E. coli to simulate trimethoprim activity under bacteriostatic and bactericidal conditions. Our results suggested that in the absence of environmental purines or nucleosides, trimethoprim induces salvage of ATP to repair DNA. We experimentally validated the result with ATP bioluminescence screening of 96 clinical E. coli isolates. Therefore, the choice of growth media composition significantly changes the outcome of trimethoprim challenge and opens opportunities for personalised medicine.. null, null. Quadram Institutecailean.carter@quadram.ac.ukQuadram InstitutefalseCarter2025 - Trimethoprim induced metabolic perturbations and counteractionsTrimethoprim is a clinically important antibiotic for treating urinary tract infections yet its mechanism of killing remains elusive due to a cascade of effected metabolic reactions. Metabolites available in growth media can selectively counteract trimethoprim which affects the interpretation of results. We sought to understand the full scope of trimethoprim’s impact on Escherichia coli metabolism and how metabolite availability affects trimethoprim outcomes. We applied flux balance analysis on a genome-scale metabolic model of E. coli to simulate trimethoprim activity under bacteriostatic and bactericidal conditions.2026-03-102026-03-112025-07-24MODEL2507240001MODEL2507240001