MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13293/m_MTBLS13293_LC-MS_negative_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13293/a_MTBLS13293_LC-MS_negative_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13293/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13293/s_MTBLS13293.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13293<p>Relative intensities and the fraction of 13C isotopologues of each metabolite was analyzed using the Metabolomic Analysis and Visualization Engine (MAVEN) software (2011.6.17)&nbsp;</p>MetaboLightsPublicLiquid Chromatography MS - negative - reverse phase<p>UHPLC (Thermo Fisher Scientific Dionex UltiMate 3000) was used. Acquity UPLC BEH C18 Column (particle size 1.7 um, 2.1 mm x 100 mm) maintained at 25 C with an injection volume of 10 uL. The eluents consisted of 97:3 (v/v) water:methanol with 15 mM acetic acid and 10 mM tributylamine (eluent A) and methanol (eluent B) (CITATION). The flow rate was 0.18 mL min-1.</p>A phosphoketolase bypasses reductive stress in P. aeruginosa under anaerobic survival.Nanqing ZhouUniversity of DelawareCell Pelletmass spectrometry assay<p>Metabolites extraction using MeOH:ACN:H2O = 40:40:20, and incubated at 4 C for 30 min</p>Pseudomonas aeruginosahttps://www.ebi.ac.uk/metabolights/MTBLS13293Ludmilla Aristilde. Northwestern University. ludmilla.aristilde@northwestern.edu.<p>A GNPS vendor conversion program was used to convert the raw MS data into mzXML files.</p>Intracellular metabolite quantificationnqzhou@udel.edu<p>Cell pellets of Pseudomonas aeruginosa collected before and after anaerobic survival.</p>MetabolomicsPseudomonas aeruginosaStresstargeted metabolitesPseudomonas aeruginosatargeted metabolitesStress<p>High-resolution mass spectrometry (Thermo Fisher Scientific Q Exactive quadrupole-Orbitrap) with electrospray ionization operating in negative mode was applied to collect MS data. m/z range: 80-1200.</p>falseA phosphoketolase bypasses reductive stress in P. aeruginosa under anaerobic survival<p>Across diverse contexts, bacteria experience loss of electron acceptors due to fluctuating environmental conditions, requiring strategies to avoid metabolic arrest due to reductive stress. Yet, microbial metabolism has been primarily studied with cells growing aerobically under nutrient replete conditions. To study how cells metabolically preserve redox homeostasis and viability during growth-arrest, we explored how the opportunistic pathogen&nbsp;<em>Pseudomonas aeruginosa&nbsp;</em>remodels its metabolism during anaerobic survival and oxidant limitation. During anaerobic survival on glucose,&nbsp;<em>P. aeruginosa&nbsp;</em>relies on flux through the pentose phosphate pathway (PPP) and a previously undescribed phosphoketolase (here-in termed&nbsp;<em>xfp).&nbsp;</em>This re-routing bypasses&nbsp;<em>P. aeruginosa’</em>s<em>&nbsp;</em>canonical glucose-catabolizing Entner-Doudoroff pathway (EDP) avoiding reducing equivalent generation. Moreover, anaerobic survival on diverse carbon sources triggers PPP metabolite accumulation and ribonucleotide salvage, with this phosphoketolase being essential to mediating ribonucleotide homeostasis and avoiding lethal metabolic dysregulation. This study expands our understanding of&nbsp;<em>P. aeruginosa</em>’s anaerobic survival strategies and suggests that phosphoketolases play a previously unappreciated role in avoiding reductive stress and mediating ribonucleotide homeostasis. Our work serves as a reminder that large gaps remain in our understanding of growth arrest physiology even in well-studied model organisms, highlighting the potential for basic discovery in the realm of non-growth metabolism.</p>2026-06-032025-11-10MTBLS13293