MetaboLights

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identification was conducted by matching the detected features against NovoMetDB, an in-house high-quality MS/MS spectral database of Novogene, with a mass tolerance of 10 ppm and considering adduct information. The identified metabolites were then normalized to obtain relative peak areas. Compounds with a coefficient of variation (CV) > 30% in the quality control (QC) samples were removed. Finally, the identification and relative quantification results of metabolites were obtained.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseChromatographic separation was performed on a Hypersil Gold C18 column (100 × 2.1 mm, 1.9 μm; Thermo Fisher Scientific) maintained at 40°C. The mobile phase consisted of (A) 0.1% formic acid in water and (B) methanol. The gradient elution program was as follows: 0–1.5 min, 98% A / 2% B; 3 min, 15% A / 85% B; 10 min, 0% A / 100% B; 10.1 min, 98% A / 2% B; 11–12 min, 98% A / 2% B (re-equilibration). The flow rate was 0.2 mL/min.Metabolomics analysis of Streptomyces sp. 30177 in response to Sphingomonas sp. 06703 supernatant.qin xiaoSouth China Normal Universitycell supernatantmass spectrometry assayIn this study, solvent blank and pooled quality control (QC) samples were prepared and processed alongside the biological samples using the same procedure. The extraction protocol was as follows: Freeze-dry 1 mL of sample in a lyophilizer, then add 100 μL of 80% methanol aqueous solution. Vortex thoroughly, incubate on ice for 5 min, and centrifuge at 15,000 × g and 4°C for 15 min. Take an aliquot of the supernatant and dilute with MS-grade water to a final methanol concentration of 53%. Centrifuge again at 15,000 × g and 4°C for 15 min, collect the supernatant, and analyze by LC-MS.Streptomyces sp. 30177https://www.ebi.ac.uk/metabolights/MTBLS15203Dan Wang. Institute of Agricultural Resources and Environment, Guangdong Academy of Agricultural Sciences. No. 66, Jinying Road, Tianhe District, Guangzhou, Guangdong Province, China. wangdanzh@gdaas.cn.Wenjie Gu. Institute of Agricultural Resources and Environment, Guangdong Academy of Agricultural Sciences. No. 66, Jinying Road, Tianhe District, Guangzhou, Guangdong Province, China. guwenjie0818@163.com.Raw data files were converted to mzXML format using ProteoWizard. Peak detection and quantification were performed using XCMS, and peak alignment across different samples was carried out based on retention time and m/z ratio.Treatment19854815383@163.comSphingomonas sp. 06703 was activated on NA plates. A single colony was inoculated into NB medium and cultured at 30°C, 180 rpm until OD600 reached 1.0. Cells were collected by centrifugation at 5,000 g, washed twice with PBS buffer, and resuspended to OD600 = 1.0. The cell suspension was then transferred (1% inoculum) into M9 glucose medium and cultured at 30°C, 180 rpm. Glucose consumption was monitored daily using a glucose assay kit (glucose oxidase method). After complete glucose depletion on day 6, the culture was centrifuged at 10,000 g for 10 min, and the supernatant was filtered through a 0.22 μm aqueous membrane to obtain the primary metabolite. Spores of Streptomyces sp. 30177 were collected by washing with sterile water, and the mycelium was filtered through a 5 μm filter to obtain a spore suspension. The spores were inoculated (1% inoculum) into the primary metabolite of Sphingomonas sp. 06703 and cultured at 30°C, 180 rpm for 6 days. The control group was inoculated into M9 medium. After incubation, the growth of Streptomyces sp. 30177 was assessed by measuring wet cell weight. The culture was centrifuged at 10,000 g for 10 min at 4°C, and the supernatant was filtered through a 0.22 μm aqueous membrane to obtain the secondary metabolite. Primary and secondary metabolites from both strains were collected.Metabolomicscell supernatantMetabolomicsVanquishQ Exactive HF-Xuntargeted analysisStreptomycesSphingomonasexperimental blankStreptomyces sp. 30177cell supernatantMetabolomicsVanquishQ Exactive HF-Xuntargeted analysisStreptomycesSphingomonasexperimental blankStreptomyces sp. 30177The mass spectra were acquired in full-scan mode over the m/z range of 100–1500. The ESI source parameters were set as follows: spray voltage, 3.5 kV; sheath gas flow rate, 35 psi; auxiliary gas flow rate, 10 L/min; capillary temperature, 320°C; S-lens RF level, 60; auxiliary gas heater temperature, 350°C. Mass spectra were acquired in both positive and negative ion modes. MS/MS fragmentation was performed using data-dependent acquisition (DDA) mode.falseMetabolomics analysis of Streptomyces sp. 30177 in response to Sphingomonas sp. 06703 supernatantThis study investigated the metabolomic response of Streptomyces sp. 30177 to cell-free supernatant of Sphingomonas sp. 06703. Untargeted LC-MS metabolomics was performed to identify differentially regulated metabolites.2026-07-302026-07-30MTBLS15203