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The consortium was diluted to OD_600= 0.02 in YCFA supplemented with 0.4 % (w/v) glucose and grown for 8 h at 37 °C in an anaerobic chamber (Don Whitley, Bingley, UK or Coy Laboratory Products, Grass Lake, MI, USA) to reach late exponential phase. Cultures were then diluted to a starting OD_600= 0.02 in fresh YCFA containing either 0.4 % (w/v) HMOs (supplemented with 20 % 2′-fucosyllactose), 0.4 % (w/v) fiber consisting of equal proportions of arabinoxylan and locust bean gum mannan, or a mixed condition containing 0.4 % (w/v) of HMOs and fiber (total carbohydrate concentration 0.8 % (w/v)). Each condition was prepared in technical triplicates. Cultures were incubated at 37 °C until exponential phase monitored by measuring OD_600 every 15 min in 200 µL volumes using a BioTek Epoch 2 microtiter plate reader (Agilent, Santa Clara, CA, USA). Cells were harvested by centrifugation, washed three times with sterile PBS, and stored at −20 °C overnight prior to submission for proteomic analysis. Sample Preparation: 50 µL of lysis buffer (6 M guanidinium hydrochloride, 10 mM TCEP, 40 mM CAA, 50 mM HEPES, pH 8.5) was added to each pellet, and samples were boiled for 5 minutes at 95 °C, followed by sonication on high for 5 cycles of 60 seconds of sonication with 30-second pauses using a Bioruptor Pico sonicator (Diagenode). Samples were centrifuged at 18,000 RCF for 10 minutes, and supernatants were transferred to clean Eppendorf Protein LoBind tubes. Protein concentrations of the supernatants were determined using the BCA Rapid Gold assay (Thermo Scientific), and 20 µg of protein was taken forward for digestion. Volumes were normalized to 20 µL with lysis buffer, and samples were diluted threefold in digestion buffer (10% acetonitrile in 50 mM HEPES). Four hundred nanograms of LysC protease (Wako) were added to each sample, followed by incubation for 4 hours at 37 °C with shaking at 750 RPM on a Thermomixer (Eppendorf). Samples were then diluted to a final tenfold dilution in digestion buffer, and 200 ng of trypsin (MS-grade, Thermo Scientific) was added to each sample before incubation for 18 hours and 30 minutes at 37 °C with shaking at 750 RPM. Samples were acidified by adding 2% TFA to a final concentration of 1%, and the resulting peptides were desalted using a SOLA SPE plate (HRP 10mg/2mL, Thermo). Between each application, solvents were passed through the plate by centrifugation at 350 RCF. For each sample, the filters were activated with 200 µL of 100% methanol, followed by 200 µL of 80% acetonitrile containing 0.1% formic acid. The filters were equilibrated twice with 200 µL of 1% TFA and 3% acetonitrile, after which the samples were loaded. Following two washes with 200 µL of 0.1% formic acid, peptides were eluted into clean Eppendorf Protein LoBind tubes using 40% acetonitrile containing 0.1% formic acid. Eluted peptides were concentrated using a Savant SpeedVac SPD120 (Thermo Scientific) and reconstituted in 12 µL of A* buffer (2% acetonitrile, 1% TFA), sonicated for 1 minute in a water bath sonicator and centrifuged for 10 minutes at 18,000 RCF. Peptide concentrations were determined by NanoDrop measurement using a DeNovix DS-11 FX+ spectrophotometer. MS Analysis: 500 ng of peptides were loaded onto an µPAC trapping column (Thermo Scientific, 3003438) connected in line with a 50 cm µPAC NEO column (Thermo Scientific, 7500467) using 100% solvent A (0.1% formic acid in water) at 750 bar on a Vanquish NEO UHPLC system (Thermo). The column oven was operated at 50 °C. Peptides were eluted over a 68-minute gradient ranging from 4% to 45% solvent B (80% acetonitrile, 0.1% formic acid) at a flow rate of 250 nL/min. The Orbitrap Exploris 480 mass spectrometer (Thermo Fisher Scientific) was operated using a DIA method. Full MS spectra were acquired at a resolution of 120,000 with a normalized AGC target of 300% or maximum injection time set to auto, over a scan range of 400–1000 m/z. MS2 spectra were acquired at a resolution of 30,000 with an AGC target of 800% or maximum injection time set to auto and a normalized collision energy of 27. Isolation windows were set to 12 m/z with window placement optimization enabled, and the MS2 scan range was set to 145–1450 m/z. MS performance was verified for consistency using the Pierce™ HeLa Protein Digest Standard (Thermo Scientific), and chromatography was monitored to ensure reproducibility."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Raw files were analyzed using Spectronaut™ (version 17.4). Spectra were searched against the UniProt reference database for the relevant taxa (taxon IDs: 391904, 367928, 536231, 585394 and 622312). Carbamidomethylation of cysteines was set as a fixed modification, while methionine oxidation and protein N-terminal acetylation were included as variable modifications. All identifications were filtered to achieve a 1% false discovery rate (FDR) at both the peptide and protein levels. Protein quantification was performed at the MS2 level using Spectronaut’s intensity-based algorithms, and protein groups were inferred using IDPicker to account for shared peptides. The resulting dataset provides robust, high-confidence protein identifications suitable for downstream metaproteomic analysis and functional interpretation."],"omics_type":["Proteomics"],"labhead":["Marie Vestergaard"],"instrument_platform":[""],"labhead_affiliation":["Proteomics Core, DTU Bioengineering, Technical University of Denmark (DTU), Lyngby, Denmark"],"submission_type":["PARTIAL"],"species":["Bifidobacterium Longum Subsp. Infantis (strain Atcc 15697 / Dsm 20088 / Jcm 1222 / Nctc 11817 / S12)","Roseburia Hominis A2-183","Bifidobacterium Adolescentis Atcc 15703","Roseburia Intestinalis L1-82","Roseburia Inulinivorans Dsm 16841","Bacteria"],"publication":["10.1038/s41467-026-73297-5"],"submitter_mail":["vasi@dtu.dk"],"submitter_affiliation":["DTU Proteomics Core"],"submitter_country":["Denmark"],"additional_accession":[]},"is_claimable":false,"name":"Human milk oligosaccharides’ impact on maturation of the gut microbiota","description":"This dataset provides a comprehensive proteomic characterization of a defined five-strain human gut bacterial consortium, including Bifidobacterium and Roseburia species. The consortium was grown on different carbohydrate sources, human milk oligosaccharides, dietary fibers, or a combination of both, to investigate how substrate availability influences microbial protein expression and metabolic activity. The resulting data captures functional responses and interactions within the community, offering insights into carbohydrate utilization, cross-feeding dynamics, and gut microbial ecology. This resource can support studies of microbiome function, microbial interactions, and dietary modulation of gut microbiota.","dates":{"publication":"2026-04-23","submission":"2026-02-06"},"accession":"PXD074160","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:1126","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1367493","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:342808","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:99802","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:1549675","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:96795","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:2830","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:1092586","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402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