{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T10:48:06Z","timestamp":1782470886298,"version":"3.54.5"},"reference-count":78,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,6,14]],"date-time":"2026-06-14T00:00:00Z","timestamp":1781395200000},"content-version":"vor","delay-in-days":13,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Acetate is a major metabolic end\u2010product of\n                    <jats:italic>Haemophilus influenzae<\/jats:italic>\n                    and is produced via the phosphotransacetylase\u2013acetate kinase (Pta\u2013AckA) pathway, which also generates adenosine triphosphate (ATP). Despite its importance, the contribution of this pathway to\n                    <jats:italic>H. influenzae<\/jats:italic>\n                    physiology and virulence remains poorly understood. Here, we have investigated the individual roles of AckA and Pta for these processes by generating nonpolar single\u2010gene knockouts in a chronic obstructive pulmonary disease (COPD)\u2010isolate strain (Hi2019) and characterized their metabolic and infection phenotypes. Both mutants exhibited significant growth impairments under microaerobic and anaerobic conditions, with reductions in growth rate of up to 50% compared with the wild type. Loss of AckA or Pta decreased ATP levels to ~\u200950% of wild\u2010type values and caused marked overoxidation of the NAD\n                    <jats:sup>+<\/jats:sup>\n                    \/NADH pool. Metabolomic analyses revealed distinct perturbations at the pyruvate node: the\n                    <jats:italic>pta<\/jats:italic>\n                    mutant produced minimal acetate but accumulated pyruvate and\n                    <jats:sc>d<\/jats:sc>\n                    \u2010lactate, while the\n                    <jats:italic>ackA<\/jats:italic>\n                    mutant continued to produce acetate, likely via nonenzymatic breakdown of acetyl\u2010phosphate. Both mutations also increased sensitivity to oxidative stress and enhanced biofilm formation. In infection models, including bronchial epithelial cells, primary human nasal epithelia, and murine macrophages, intracellular survival of both mutants was significantly reduced during early infection stages, although attenuation diminished over time. These findings demonstrate that the Pta\u2013AckA pathway is critical for metabolic homeostasis, stress resistance, and early intracellular colonization, highlighting its potential as a target for management of early\u2010stage\n                    <jats:italic>H. influenzae<\/jats:italic>\n                    infections.\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70324","type":"journal-article","created":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T03:53:54Z","timestamp":1781495634000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Acetate\u2010Linked Energy Metabolism as a Determinant of Early\n                    <i>Haemophilus influenzae<\/i>\n                    Infection Fitness"],"prefix":"10.1002","volume":"15","author":[{"given":"Marufa","family":"Nasreen","sequence":"first","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Hosmer","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saurab Kishore","family":"Munshi","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3433-8136","authenticated-orcid":false,"given":"Riya","family":"Joshi","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5143-4608","authenticated-orcid":false,"given":"Ayaho","family":"Yamamoto","sequence":"additional","affiliation":[{"name":"Child Health Research Centre The University of Queensland South Brisbane Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horst J.","family":"Schirra","sequence":"additional","affiliation":[{"name":"School of Environment and Science, Griffith Institute for Drug Discovery Griffith University Nathan Queensland Australia"},{"name":"Centre for Advanced Imaging The University of Queensland Brisbane Queensland Australia"},{"name":"Institute for Biomedicine and Glycomics Griffith University Southport Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6305-2201","authenticated-orcid":false,"given":"Peter","family":"Sly","sequence":"additional","affiliation":[{"name":"Child Health Research Centre The University of Queensland South Brisbane Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alastair G.","family":"McEwan","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2642-1319","authenticated-orcid":false,"given":"Ulrike","family":"Kappler","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences The University of Queensland St. Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,14]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftx042"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8060846"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep43587"},{"key":"e_1_2_10_5_1","volume-title":"Current Protocols in Molecular Biology","author":"Ausubel F. M.","year":"1987"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1080\/03079457.2015.1062841"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-016-7832-x"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01540-07"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.55.4.882-887.1987"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.181.21.6656-6663.1999"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.01604"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.9.4408-4410.2003"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1136\/gut.28.10.1221"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1097\/CCE.0000000000001030"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-019-03380-2"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.686833"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.01743"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.01219"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.04002-22"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/1873-3468.12351"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2007.06.004"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002244"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.02005-14"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0100645"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.16.5951"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01040-06"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1042\/ETLS20220092"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010209"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2007.05890.x"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1012282"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.27515-0"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01160-15"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.74.4.2498-2502.2006"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1186\/2001-1326-1-10"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00564-07"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0094816"},{"issue":"127","key":"e_1_2_10_37_1","article-title":"Precise, High\u2010Throughput Analysis of Bacterial Growth","author":"Kurokawa M.","year":"2017","journal-title":"Journal of Visualized Experiments: JoVE"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid2110.150004"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmii.2024.10.002"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.9b00359"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.5b01089"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1996.tb02636.x"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftz015"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.0c00242"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1359513"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.637213"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.3390\/antiox11081557"},{"key":"e_1_2_10_48_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2014.00069"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13205-019-1578-y"},{"key":"e_1_2_10_50_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.00256.2010"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-321-6:57"},{"key":"e_1_2_10_52_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0179621"},{"key":"e_1_2_10_53_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00687-24"},{"key":"e_1_2_10_54_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00995-10"},{"issue":"55","key":"e_1_2_10_55_1","article-title":"Use of the EpiAirway Model for Characterizing Long\u2010Term Host\u2013Pathogen Interactions","author":"Ren D.","year":"2011","journal-title":"JoVE (Journal of Visualized Experiments)"},{"key":"e_1_2_10_56_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp045"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00042-13"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.3.1322-1328.2001"},{"key":"e_1_2_10_59_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.00233"},{"key":"e_1_2_10_60_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00808a019"},{"key":"e_1_2_10_61_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.compbiolchem.2023.107942"},{"key":"e_1_2_10_62_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2012.00097"},{"key":"e_1_2_10_63_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0960-9822(02)00478-5"},{"key":"e_1_2_10_64_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(82)90376-2"},{"key":"e_1_2_10_65_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-020518-115526"},{"key":"e_1_2_10_66_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(14)70734-0"},{"key":"e_1_2_10_67_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep31007"},{"key":"e_1_2_10_68_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jnutbio.2010.07.009"},{"key":"e_1_2_10_69_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2013.06.003"},{"key":"e_1_2_10_70_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0050588"},{"key":"e_1_2_10_71_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid2303.161552"},{"key":"e_1_2_10_72_1","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.69.1.12-50.2005"},{"key":"e_1_2_10_73_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00660-20"},{"key":"e_1_2_10_74_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2007.05747.x"},{"key":"e_1_2_10_75_1","doi-asserted-by":"publisher","DOI":"10.1038\/s12276-019-0276-5"},{"key":"e_1_2_10_76_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-38901-3"},{"key":"e_1_2_10_77_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12918-018-0677-4"},{"key":"e_1_2_10_78_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.02959-20"},{"key":"e_1_2_10_79_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi702334h"}],"container-title":["MicrobiologyOpen"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mbo3.70324","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/mbo3.70324","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mbo3.70324","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T09:54:43Z","timestamp":1782467683000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/mbo3.70324"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6]]},"references-count":78,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,6]]}},"alternative-id":["10.1002\/mbo3.70324"],"URL":"https:\/\/doi.org\/10.1002\/mbo3.70324","archive":["Portico"],"relation":{},"ISSN":["2045-8827","2045-8827"],"issn-type":[{"value":"2045-8827","type":"print"},{"value":"2045-8827","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6]]},"assertion":[{"value":"2026-02-22","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-14","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70324"}}