biostudies-literatureZhu WProject of MOE Key Laboratory of Geriatric Diseases and Immunology, Soochow UniversityMOST | National Natural Science Foundation of China (NSFC)MOST | National Natural Science Foundation of Chinae0077224https://www.ebi.ac.uk/biostudies/studies/S-EPMC11656735biostudies-literatureUnknown9(12)Enzyme-IIA (EIIA^Glc, Crr) of the phosphotransferase system (PTS) connects the uptake of glucose-family sugars to the cAMP-Crp regulatory cascade; phosphorylated EIIA^Glc enhances cAMP-Crp activity, which then contributes to the antibiotic-mediated accumulation of reactive oxygen species (ROS) and cell death. Defects in PTS cause antibiotic and disinfectant tolerance. We report that mannitol, a carbon source whose uptake does not use EIIA^Glc, reduces antibiotic-mediated killing of <i>Escherichia coli</i> without affecting antibiotic minimal inhibitory concentration. Thus, mannitol promotes antibiotic tolerance. The tolerance pathway was defined by the loss of ciprofloxacin lethality from the deletion of <i>ptsI</i> (first gene in PTS), <i>mtlA</i> (mannitol-smSphereAntibiotic tolerance due to restriction of cAMP-Crp regulation by mannitol, a non-glucose-family PTS carbon source.PMC1165673582172316KJS2401Liao YDrlica KWang BChen MGuo WWang DSu JXue YZhao XHong YZhang XNiu JNong YZhu WfalseAntibiotic tolerance due to restriction of cAMP-Crp regulation by mannitol, a non-glucose-family PTS carbon source.Enzyme-IIA (EIIA^Glc, Crr) of the phosphotransferase system (PTS) connects the uptake of glucose-family sugars to the cAMP-Crp regulatory cascade; phosphorylated EIIA^Glc enhances cAMP-Crp activity, which then contributes to the antibiotic-mediated accumulation of reactive oxygen species (ROS) and cell death. Defects in PTS cause antibiotic and disinfectant tolerance. We report that mannitol, a carbon source whose uptake does not use EIIA^Glc, reduces antibiotic-mediated killing of <i>Escherichia coli</i> without affecting antibiotic minimal inhibitory concentration. Thus, mannitol promotes antibiotic tolerance. The tolerance pathway was defined by the loss of ciprofloxacin lethality from the deletion of <i>ptsI</i> (first gene in PTS), <i>mtlA</i> (mannitol-s2024-01-01T00:00:00Z2024 Dec2025-04-18T14:40:30.653Z2025-04-07T00:57:03.99ZS-EPMC116567353956512710.1128/msphere.00772-24