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The gram-negative pathogen\n                      <jats:italic toggle=\"yes\">Vibrio cholerae<\/jats:italic>\n                      , the causative agent of cholera, is highly tolerant to \u03b2-lactam antibiotics. We previously found that the disruption of glycolysis, via deletion of\n                      <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                      (\n                      <jats:italic toggle=\"yes\">vc0374<\/jats:italic>\n                      , glucose-6-phosphate isomerase), resulted in significant cell wall damage and increased sensitivity toward \u03b2-lactam antibiotics. Here, we uncover the mechanism of this resulting damage. We find that glucose causes growth inhibition, partial lysis, and a damaged cell envelope in \u2206\n                      <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                      . Supplementation with N-acetylglucosamine, but not other carbon sources (either from upper glycolysis, TCA cycle intermediates, or cell wall precursors) restored growth, re-established wild-type \u03b2-lactam resistance, and recovered cellular morphology of a\n                      <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                      mutant exposed to glucose. Targeted metabolomics revealed the cell wall precursor synthetase enzyme GlmU (\n                      <jats:italic toggle=\"yes\">vc2762<\/jats:italic>\n                      , coding for the bifunctional enzyme that converts glucosamine-1P to UDP-GlcNAc) as a critical bottleneck and mediator of glucose toxicity in \u2206\n                      <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                      .\n                      <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                      assays of GlmU revealed that sugar phosphates (primarily glucose-1-phosphate) inhibit the acetyltransferase activity of GlmU (likely competitively), resulting in compromised peptidoglycan and lipopolysaccharide biosynthesis. These findings identify the molecular mechanism of \u0394\n                      <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                      glucose toxicity in\n                      <jats:italic toggle=\"yes\">V. cholerae<\/jats:italic>\n                      .\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Sugar-phosphate toxicity is a well-characterized phenomenon that is seen within diverse bacterial species, and yet the molecular underpinnings often remain elusive. We previously discovered that disrupting\n                        <jats:italic toggle=\"yes\">Vibrio cholerae\u2019s<\/jats:italic>\n                        ability to eat glucose (by disrupting the\n                        <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                        gene) resulted in a damaged cell envelope and enabled us to kill\n                        <jats:italic toggle=\"yes\">V. cholerae<\/jats:italic>\n                        more easily using antibiotics like penicillin. Upon deletion of\n                        <jats:italic toggle=\"yes\">pgi<\/jats:italic>\n                        , glucose-phosphate levels rapidly build up and inhibit the enzymatic activity of GlmU, a key step of bacterial peptidoglycan precursor synthesis. GlmU inhibition causes enhanced killing by antibiotics and a pronounced cell envelope defect. Thus, GlmU serves as a prime target for novel drug development. This research opens new routes through which central metabolism and sugar-phosphate toxicity modulate antibiotic susceptibility.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.01729-25","type":"journal-article","created":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T13:00:20Z","timestamp":1753707620000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Sugar phosphate-mediated inhibition of peptidoglycan precursor synthesis"],"prefix":"10.1128","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0675-7413","authenticated-orcid":false,"given":"Megan R.","family":"Keller","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05bnh6r87","id-type":"ROR","asserted-by":"publisher"}],"name":"Weill Institute for Cell and Molecular Biology, Cornell University","place":["Ithaca, USA"]},{"name":"Cornell Institute of Host-Microbe Interactions and Disease, Cornell University","place":["Ithaca, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3395-7429","authenticated-orcid":true,"given":"Vijay","family":"Soni","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02r109517","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, Division of Infectious Diseases, Weill Cornell Medicine","place":["New York, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Megan","family":"Brown","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02r109517","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, Division of Infectious Diseases, Weill Cornell Medicine","place":["New York, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelly M.","family":"Rosch","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05bnh6r87","id-type":"ROR","asserted-by":"publisher"}],"name":"Weill Institute for Cell and Molecular Biology, Cornell University","place":["Ithaca, USA"]},{"name":"Cornell Institute of Host-Microbe Interactions and Disease, Cornell University","place":["Ithaca, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anas","family":"Saleh","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02r109517","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, Division of Infectious Diseases, Weill Cornell Medicine","place":["New York, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4582-2895","authenticated-orcid":false,"given":"Kyu","family":"Rhee","sequence":"additional","affiliation":[{"name":"Cornell Institute of Host-Microbe Interactions and Disease, Cornell University","place":["Ithaca, USA"]},{"id":[{"id":"https:\/\/ror.org\/02r109517","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, Division of Infectious Diseases, Weill Cornell Medicine","place":["New York, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3283-9161","authenticated-orcid":false,"given":"Tobias","family":"D\u00f6rr","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05bnh6r87","id-type":"ROR","asserted-by":"publisher"}],"name":"Weill Institute for Cell and Molecular Biology, Cornell University","place":["Ithaca, USA"]},{"name":"Cornell Institute of Host-Microbe Interactions and Disease, Cornell University","place":["Ithaca, USA"]},{"id":[{"id":"https:\/\/ror.org\/05bnh6r87","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology, Cornell University","place":["Ithaca, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202051034"},{"key":"e_1_3_2_3_2","unstructured":"CDC. 2022. 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