MetaboLights

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Mobile phase A consisted of water/acetonitrile (95:5, v/v) containing 0.1% formic acid, and mobile phase B consisted of acetonitrile/isopropanol/water (47.5:47.5:5, v/v/v) containing 0.1% formic acid. The injection volume was 3 μL, and the column temperature was maintained at 40℃. Metabolite detection and analysis of bacterial strains.Miao JiangAntwerp universityPoolculture supernatantmass spectrometry assay200 μL of each sample was transferred into a 1.5 mL centrifuge tube. Then, 800 μL of extraction solvent consisting of methanol/acetonitrile (1:1, v/v) was added. The extraction solvent contained four internal standards, including L-2-chlorophenylalanine at a concentration of 0.02 mg/mL. The mixture was vortexed for 30 s and then subjected to low-temperature ultrasonic extraction for 30 min at 5℃ and 40 kHz. The supernatant was collected and dried under a gentle stream of nitrogen. Pseudarthrobacter oxydansmixed samplehttps://www.ebi.ac.uk/metabolights/MTBLS14926Miao Jiang. Antwerp university. jiangmiao0203@gmail.com.Li Xiangnan. Chinese Academy of Sciences. lixiangnan@iga.ac.cn. The raw data were preprocessed using Progenesis QI software, and a three-dimensional data matrix was exported in CSV format. The matrix contained sample information, metabolite names, and mass spectral response intensities. Internal standard peaks and known false-positive peaks, including noise peaks, column bleed peaks, and derivatization reagent-related peaks, were removed from the data matrix. Subsequently, peak de-redundancy and peak merging were performed to obtain the final processed dataset. Treatmentjiangmiao0203@gmail.comFor bacterial culture-supernatant metabolomics, the standardized bacterial suspension was inoculated into TSB medium without PEG 6000 or TSB medium supplemented with 10% (w/v) PEG 6000, representing the control and osmotic-stress treatments, respectively. Six independent biological replicates were prepared for each treatment. After incubation at 27 ℃ for 3 d, culture supernatants were collected for LC-MS/MS metabolomic profiling.Metabolomicspooled quality control sampleMetabolomicsosmotic stress treatment designThermo Scientific Orbitrap Exploris 240untargeted analysiscomparative designPseudarthrobacter oxydansmixed sampleculture supernatantexperimental sampleuntargeted metabolite profilingThermo Scientific Vanquish UHPLC SystemPoolpooled quality control sampleMetabolomicsosmotic stress treatment designThermo Scientific Orbitrap Exploris 240untargeted analysiscomparative designPseudarthrobacter oxydansmixed sampleculture supernatantexperimental sampleuntargeted metabolite profilingThermo Scientific Vanquish UHPLC SystemPoolMass spectrometric detection was performed using electrospray ionization (ESI) in both positive and negative ion modes. The scan range was set to m/z 70-1050. The sheath gas flow rate and auxiliary gas flow rate were set to 60 and 20 arb, respectively. The heater temperature was maintained at 350℃, and the capillary temperature was set to 320℃. falseMetabolite detection and analysis of bacterial strainsThis study investigated whether P. oxydans-secreted metabolites function as upstream metabolic signals that modulate host responses. We therefore compared the extracellular metabolomic profiles of P. oxydans cultured under control conditions and under 10% PEG 6000-induced osmotic stress.2026-07-032026-07-03MTBLS14926