MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/m_MTBLS14722_LC-MS_negative_normal-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/m_MTBLS14722_LC-MS_positive_normal-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/s_MTBLS14722.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/a_MTBLS14722_LC-MS_positive_normal-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722/a_MTBLS14722_LC-MS_negative_normal-phase.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14722The metabolites were identified by searching database, and the main databases (HMDB, Metlin, NIST, MassBank, Lipidblast, GNPS, etc)and the self-compiled Majorbio Database (MJDB) of Majorbio Biotechnology Co., Ltd. (Shanghai,China).MetaboLightsPublicLiquid Chromatography MS - positive - normal-phaseLiquid Chromatography MS - negative - normal-phaseExploris 240 systemequipped with an ACQUITY HSS T3 column (100 mm X 2.1 mm i.d., 1.8 um; Waters, USA)atMajorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). The mobile phases consisted of 0.1%formic acid in water:acetonitrile (95:5, v/v) (solvent A) and 0.1% formic acid in acetonitrile (solventB). The elution gradient conditions are as follows: 0-1.5 min, mobile phase B was increased from 0%to 5%; 1.5-2 min, mobile phase B was increased from 5% to 10%; 2-4.5 min, mobile phase B wasincreased from 10% to 30%; 5-6.3 min, mobile phase B was maintained at 100%; 6.3-6.4 min.mobile phase B was decreased from 100% to 0%; 6.4-8 min, mobile phase B was maintained at 0%.The flow rate was 0.40 mL/min and the column temperature was 40°C. The injection volume was 3uLMS conditions:The UPLC system was coupled to a UHPLC-Orbitrap Exploris 240 system Mass Spectrometerequipped with an electrospray ionization (ESI) source operating in positive mode and negative mode.The optimal conditions were set as followed: source temperature at 350°C; sheath gas flow rate at 60arb; Aux gas flow rate at 20 arb; ion-spray voltage floating (ISVF) at -2800V in negative mode and3400V in positive mode, respectively; Normalized collision energy, 20-40-60V rolling for MS/MS.Data acquisition was performed with the Data Dependent Acquisition (DDA) mode. The detectionwas carried out over a mass range of 70-1050 m/z.Microbial metabolism driven by nanobiohybrids via electromagnetic induction.Chaohui YangFujian Agriculture and Forestry UniversityPooled SampleWhole Organismmass spectrometry assayMetabolite Extraction:100 uL liquid sample was added to a 1.5 mL centrifuge tube with 400 uL solution (acetonitrile:methanol = 1:1(v:v))containing four internal standards (0.02 mg/mL L-2-chlorophenylalanine, etc.) to extract metabolites. The samples were mixed by vortex for 30 s and low-temperature sonicated for 30 min (5°C, 40 KHz). The samples were placed at -20°C for 30 min to precipitate the proteins. Thenthe samples were centrifuged for 15 min (4°C, 13000 g). The supernatant was removed and blowndry under nitrogen. The sample was then re-solubilized with 100 uL solution (acetonitrile: water =1:1)and extracted by low-temperature ultrasonication for 5 min (5°C, 40 KHz), followed bycentrifugation at 13000 g and 4°C for 10 min. The supernatant was transferred to sample vials forLC-MS/MS analysis.Pooled SampleThiobacillus denitrificanshttps://www.ebi.ac.uk/metabolights/MTBLS14722Chaohui Yang. Fujian Agriculture and Forestry University. yangchaohui@fafu.edu.cn.The pretreatment of LC/MS raw data was performed by Progenesis QI (Waters Corporation,Milford, USA) software, and a three-dimensional data matrix in CSV format was exported. Theinformation in this three-dimensional matrix included: sample information, metabolite name andmass spectral response intensity. Internal standard peaks, as well as any known false positive peaks(including noise, column bleed, and derivatized reagent peaks), were removed from the data matrix,deredundant and peak pooled. With or without DMFyangchaohui@fafu.edu.cnSample preparation:Samples were obtained from a constructed Thiobacillus denitrificans–graphite nanosheets (T. d-GNS) biohybrid system. The biohybrid was employed for biological denitrification under conditions either with or without a dynamic magnetic field (DMF), which were designated as the experimental group and control group, respectively. For metabolomic analysis, 5 mL of each sample was collected and subjected to cell disruption using a cell disruptor (200 W; 1 s on, 1 s off, for 10 min). The resulting lysate was filtered through a 0.22 μm membrane filter and immediately stored at −80 °C until further analysis.MetabolomicsExploris240（ThermoScientific）pooled quality control sampleMetabolomicsmagnetic fielduntargeted analysisPooled SampleVanquishUHPLC（ThermoScientific）Thiobacillus denitrificansWhole Organismexperimental sampleuntargeted metabolite profilingmicroorganismexperimental blankExploris240（ThermoScientific）pooled quality control sampleMetabolomicsmagnetic fielduntargeted analysisPooled SampleVanquishUHPLC（ThermoScientific）Thiobacillus denitrificansWhole Organismexperimental sampleuntargeted metabolite profilingmicroorganismexperimental blankExploris 240 systemequipped with an ACQUITY HSS T3 column (100 mm X 2.1 mm i.d., 1.8 um; Waters, USA)atMajorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). The mobile phases consisted of 0.1%formic acid in water:acetonitrile (95:5, v/v) (solvent A) and 0.1% formic acid in acetonitrile (solventB). The elution gradient conditions are as follows: 0-1.5 min, mobile phase B was increased from 0%to 5%; 1.5-2 min, mobile phase B was increased from 5% to 10%; 2-4.5 min, mobile phase B wasincreased from 10% to 30%; 5-6.3 min, mobile phase B was maintained at 100%; 6.3-6.4 min.mobile phase B was decreased from 100% to 0%; 6.4-8 min, mobile phase B was maintained at 0%.The flow rate was 0.40 mL/min and the column temperature was 40°C. The injection volume was 3uLMS conditions:The UPLC system was coupled to a UHPLC-Orbitrap Exploris 240 system Mass Spectrometerequipped with an electrospray ionization (ESI) source operating in positive mode and negative mode.The optimal conditions were set as followed: source temperature at 350°C; sheath gas flow rate at 60arb; Aux gas flow rate at 20 arb; ion-spray voltage floating (ISVF) at -2800V in negative mode and3400V in positive mode, respectively; Normalized collision energy, 20-40-60V rolling for MS/MS.Data acquisition was performed with the Data Dependent Acquisition (DDA) mode. The detectionwas carried out over a mass range of 70-1050 m/z.falseMicrobial metabolism driven by nanobiohybrids via electromagnetic inductionMicroorganisms sustain biogeochemical cycles via metabolically driven redox reactions that require essential external energy. Beyond conventional chemotrophy and phototrophy, many microorganisms remain metabolically active under extreme energy limitation, suggesting the existence of alternative energy acquisition strategies. Magnetic fields are pervasive in both natural and engineered environments, yet their potential as direct drivers of microbial metabolism remains unresolved. Here we demonstrate that microbial activity can be driven via electromagnetic induction as an energy conversion pathway under a dynamic magnetic field (DMF). Nanobiohybrids comprising Thiobacillus denitrificans and graphite nanosheets generated an electromotive force under DMF, enabling bio-denitrification with water as the electron donor. Comparable induction-driven processes were also observed in carbon and sulphur cycling, as well as pollutant degradation. These findings identify electromagnetic induction as a previously unrecognized mechanism enabling microbial energy conversion and reveal a mode of magnetic control over metabolism, with broad implications for biogeochemistry, bioenergy, and environmental technologies.2026-06-092026-06-09MTBLS14722