#METABOLOMICS WORKBENCH katherine_heal_20251104_165618 DATATRACK_ID:6626 STUDY_ID:ST004337 ANALYSIS_ID:AN007242 PROJECT_ID:PR002738 VERSION 1 CREATED_ON November 7, 2025, 6:24 pm #PROJECT PR:PROJECT_TITLE Conserved pathway for homarine catabolism in environmental bacteria PR:PROJECT_SUMMARY Homarine (N-methylpicolinic acid) is a ubiquitous marine metabolite produced by PR:PROJECT_SUMMARY phytoplankton and noted for its bioactivity in marine animals, yet its microbial PR:PROJECT_SUMMARY degradation pathways are uncharacterized. Here, we identify a conserved operon PR:PROJECT_SUMMARY (homABCDER) that mediates homarine catabolism in bacteria using comparative PR:PROJECT_SUMMARY transcriptomics, mutagenesis, and targeted knockouts. Phylogenetic and genomic PR:PROJECT_SUMMARY analyses show this operon distributed across abundant bacterial clades, PR:PROJECT_SUMMARY including coastal copiotrophs (e.g., Rhodobacterales) and open-ocean oligotrophs PR:PROJECT_SUMMARY (e.g., SAR11, SAR116). High-resolution mass spectrometry revealed PR:PROJECT_SUMMARY N-methylglutamic acid and glutamic acid as key metabolic products of homarine in PR:PROJECT_SUMMARY both model and natural systems, with N-methylglutamate dehydrogenase catalyzing PR:PROJECT_SUMMARY their conversion. Metatranscriptomics showed responsive and in situ expression PR:PROJECT_SUMMARY of hom genes aligned with homarine availability. These findings uncover the PR:PROJECT_SUMMARY genetic and metabolic basis of homarine degradation, establish its ecological PR:PROJECT_SUMMARY relevance, and highlight homarine as a versatile growth substrate that feeds PR:PROJECT_SUMMARY into central metabolism via glutamic acid in diverse marine bacteria. PR:INSTITUTE University of Washington, School of Oceanography PR:LAST_NAME Heal PR:FIRST_NAME Katherine PR:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle PR:EMAIL katherine.heal@pnnl.gov PR:PHONE 612-616-4840 PR:PUBLICATIONS DOI 10.21203/rs.3.rs-7359689/v1 #STUDY ST:STUDY_TITLE Homarine catabolism: Ruegeria pomeroyi DSS-3 comparative metabolomics under ST:STUDY_TITLE homarine and glucose supported growth ST:STUDY_SUMMARY Cultures of Ruegeria pomeroyi DSS-3 were revived from cryostocks onto ½ YTSS ST:STUDY_SUMMARY agar plates and incubated at 30 °C for 6 days. Single colonies were ST:STUDY_SUMMARY inoculated into 11 mL of glucose minimal media (GMM) and grown overnight at ST:STUDY_SUMMARY 30 °C with shaking at 200 rpm. GMM was prepared using a modified L1 minimal ST:STUDY_SUMMARY medium with glucose 12 mM C as the sole carbon source. All cultures were ST:STUDY_SUMMARY maintained in sterile 15 mL assay tubes. Overnight cultures were diluted to an ST:STUDY_SUMMARY optical density of 0.1 at 600 nm (OD₆₀₀) in fresh GMM, incubated for ST:STUDY_SUMMARY 11–12 h, and amended with glucose (4 mM C, 0.8 mM NH4, control), homarine ST:STUDY_SUMMARY (500 nM C, no additional NH4), or glucose + homarine (1 mM C, 2 mM C, ST:STUDY_SUMMARY respectively with 0.8 mM NH4). Homarine additions were staggered across time ST:STUDY_SUMMARY points to ensure consistent incubation durations. At each time point, samples ST:STUDY_SUMMARY were collected for cell counts and particulate metabolites. For cell counts, ST:STUDY_SUMMARY 1 mL of culture was fixed with glutaraldehyde (final concentration 1%) in ST:STUDY_SUMMARY labeled cryovials, held at 4 °C for 20 minutes, and then stored at ST:STUDY_SUMMARY –80 °C. Particulate metabolites were collected by filtering cultures ST:STUDY_SUMMARY through combusted glass fiber filters using a vacuum manifold set to 8 psi; ST:STUDY_SUMMARY filters were wrapped in combusted foil and flash-frozen in liquid nitrogen. The ST:STUDY_SUMMARY experiment cultures were sampled after two hours in biological triplicate, as ST:STUDY_SUMMARY well as the three control conditions: glucose-only controls, glucose plus ST:STUDY_SUMMARY homarine controls (uninoculated), and a filter blank. ST:INSTITUTE University of Washington, School of Oceanography ST:LAST_NAME Heal ST:FIRST_NAME Katherine ST:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle ST:EMAIL katherine.heal@pnnl.gov ST:PHONE 612-616-4840 ST:STUDY_COMMENTS Part of project 6620 ST:PUBLICATIONS DOI 10.21203/rs.3.rs-7359689/v1 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Ruegeria pomeroyi SU:TAXONOMY_ID 246200 SU:GENOTYPE_STRAIN DSS-3 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Gluc_A Sample source:Filtered bacterial cells | treatment:Glucose RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Gluc_A.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Gluc_B Sample source:Filtered bacterial cells | treatment:Glucose RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Gluc_B.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Gluc_C Sample source:Filtered bacterial cells | treatment:Glucose RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Gluc_C.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Hom_A Sample source:Filtered bacterial cells | treatment:Homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Hom_A.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Hom_B Sample source:Filtered bacterial cells | treatment:Homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Hom_B.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Hom_C Sample source:Filtered bacterial cells | treatment:Homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Hom_C.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Mix_A Sample source:Filtered bacterial cells | treatment:Glucose + homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Mix_A.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Mix_B Sample source:Filtered bacterial cells | treatment:Glucose + homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Mix_B.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_Mix_C Sample source:Filtered bacterial cells | treatment:Glucose + homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_Mix_C.mzML; Raw_data_type=Experimental Sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_DDAneg20 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_DDAneg20.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_DDAneg35 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_DDAneg35.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_DDAneg50 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_DDAneg50.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Full1 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Full1.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Full2 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Full2.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Full3 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Full3.mzML; Raw_data_type=Pooled experimental sample SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Half1 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Half1.mzML; Raw_data_type=Pooled experimental sample (half strength) SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Half2 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Half2.mzML; Raw_data_type=Pooled experimental sample (half strength) SUBJECT_SAMPLE_FACTORS - 240304_Poo_EpicFate_Half3 Sample source:QC | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Poo_EpicFate_Half3.mzML; Raw_data_type=Pooled experimental sample (half strength) SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_BLK_Gluc_C Sample source:Blank | treatment:Glucose RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_BLK_Gluc_C.mzML; Raw_data_type=Media blank for glucose treatment SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_BLK_Hom_A Sample source:Blank | treatment:Homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_BLK_Hom_A.mzML; Raw_data_type=Media blank for homarine treatment SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_BLK_Hom_B Sample source:Blank | treatment:Homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_BLK_Hom_B.mzML; Raw_data_type=Media blank for homarine treatment SUBJECT_SAMPLE_FACTORS - 240304_Smp_WT_BLK_Mix_A Sample source:Blank | treatment:Glucose + homarine RAW_FILE_NAME(raw_file_name)=240304_Smp_WT_BLK_Mix_A.mzML; Raw_data_type=Media blank for glucose+homarine treatment SUBJECT_SAMPLE_FACTORS - 240304_Std_4uMStdsMix1InH2O_1 Sample source:Standards | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Std_4uMStdsMix1InH2O_1.mzML; Raw_data_type=Standards run SUBJECT_SAMPLE_FACTORS - 240304_Std_4uMStdsMix1InH2O_2 Sample source:Standards | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Std_4uMStdsMix1InH2O_2.mzML; Raw_data_type=Standards run SUBJECT_SAMPLE_FACTORS - 240304_Std_4uMStdsMix2InH2O_1 Sample source:Standards | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Std_4uMStdsMix2InH2O_1.mzML; Raw_data_type=Standards run SUBJECT_SAMPLE_FACTORS - 240304_Std_4uMStdsMix2InH2O_2 Sample source:Standards | treatment:N/A RAW_FILE_NAME(raw_file_name)=240304_Std_4uMStdsMix2InH2O_2.mzML; Raw_data_type=Standards run #COLLECTION CO:COLLECTION_SUMMARY Cultures of Ruegeria pomeroyi DSS-3 were revived from cryostocks onto ½ YTSS CO:COLLECTION_SUMMARY agar plates and incubated at 30 °C for 6 days. Single colonies were CO:COLLECTION_SUMMARY inoculated into 11 mL of glucose minimal media (GMM) and grown overnight at CO:COLLECTION_SUMMARY 30 °C with shaking at 200 rpm. GMM was prepared using a modified L1 minimal CO:COLLECTION_SUMMARY medium with glucose 12 mM C as the sole carbon source. All cultures were CO:COLLECTION_SUMMARY maintained in sterile 15 mL assay tubes. Overnight cultures were diluted to an CO:COLLECTION_SUMMARY optical density of 0.1 at 600 nm (OD₆₀₀) in fresh GMM, incubated for CO:COLLECTION_SUMMARY 11–12 h, and amended with glucose (4 mM C, 0.8 mM NH4, control), homarine CO:COLLECTION_SUMMARY (500 nM C, no additional NH4), or glucose + homarine (1 mM C, 2 mM C, CO:COLLECTION_SUMMARY respectively with 0.8 mM NH4). Homarine additions were staggered across time CO:COLLECTION_SUMMARY points to ensure consistent incubation durations. At each time point, samples CO:COLLECTION_SUMMARY were collected for cell counts and particulate metabolites. For cell counts, CO:COLLECTION_SUMMARY 1 mL of culture was fixed with glutaraldehyde (final concentration 1%) in CO:COLLECTION_SUMMARY labeled cryovials, held at 4 °C for 20 minutes, and then stored at CO:COLLECTION_SUMMARY –80 °C. Particulate metabolites were collected by filtering cultures CO:COLLECTION_SUMMARY through combusted glass fiber filters using a vacuum manifold set to 8 psi; CO:COLLECTION_SUMMARY filters were wrapped in combusted foil and flash-frozen in liquid nitrogen. The CO:COLLECTION_SUMMARY experiment cultures were sampled after two hours in biological triplicate, as CO:COLLECTION_SUMMARY well as the three control conditions: glucose-only controls, glucose plus CO:COLLECTION_SUMMARY homarine controls (uninoculated), and a filter blank. CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY Cultures were amended with glucose (4 mM C, 0.8 mM NH4, control), homarine (500 TR:TREATMENT_SUMMARY nM C, no additional NH4), or glucose + homarine (1 mM C, 2 mM C, respectively TR:TREATMENT_SUMMARY with 0.8 mM NH4). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For metabolite extractions, a one phase extraction was performed with SP:SAMPLEPREP_SUMMARY 40:40:20:0.01 methanol:acetonitrile:water:formic acid solution as the extraction SP:SAMPLEPREP_SUMMARY solvent (Canelas et al 2009). Filters were placed in 15 mL Teflon tubes with SP:SAMPLEPREP_SUMMARY pre-chilled extraction solvent, incubated at -20 ºC for 10 minutes, bead beaten SP:SAMPLEPREP_SUMMARY with silica beads, and centrifuged. The solvent was then collected, transferred SP:SAMPLEPREP_SUMMARY into glass tubes, and the procedure was repeated three times while keeping SP:SAMPLEPREP_SUMMARY samples cold as much as possible. Samples were dried down under nitrogen gas, SP:SAMPLEPREP_SUMMARY reconstituted in 400 uL of H2O, and stored at -80 ºC until analysis by LC-MS. SP:SAMPLEPREP_SUMMARY Isotopically-labeled internal standards were added for normalization purposes, SP:SAMPLEPREP_SUMMARY as reported in Table S17 of https://doi.org/10.21203/rs.3.rs-7359689/v1 #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY For HILIC chromatography, a SeQuant ZIC-pHILIC column (5 um particle size, 2.1 CH:CHROMATOGRAPHY_SUMMARY mm x 150 mm, from Millipore) was used with 10 mM ammonium carbonate in 85:15 CH:CHROMATOGRAPHY_SUMMARY acetonitrile to water (Solvent A) and 10 mM ammonium carbonate in 85:15 water to CH:CHROMATOGRAPHY_SUMMARY acetonitrile (Solvent B) at a flow rate of 0.15 mL/min. This column was compared CH:CHROMATOGRAPHY_SUMMARY with a Waters UPLC BEH amide and a Millipore cHILIC column; the pHILIC showed CH:CHROMATOGRAPHY_SUMMARY superior reproducibility and peak shapes. The column was held at 100% A for 2 CH:CHROMATOGRAPHY_SUMMARY minutes, ramped to 64% B over 18 minutes, ramped to 100% B over 1 minute, held CH:CHROMATOGRAPHY_SUMMARY at 100% B for 5 minutes, and equilibrated at 100% A for 25 minutes (50 minutes CH:CHROMATOGRAPHY_SUMMARY total). The column was maintained at 30 C. The injection volume was 2 µL for CH:CHROMATOGRAPHY_SUMMARY samples and standard mixes. When starting a batch, the column was equilibrated CH:CHROMATOGRAPHY_SUMMARY at the starting conditions for at least 30 minutes. To improve the performance CH:CHROMATOGRAPHY_SUMMARY of the HILIC column, we maintained the same injection volume, kept the CH:CHROMATOGRAPHY_SUMMARY instrument running water blanks between samples as necessary, and injected CH:CHROMATOGRAPHY_SUMMARY standards in a representative matrix in addition to standards in water. After CH:CHROMATOGRAPHY_SUMMARY each batch, the column was flushed with 10 mM ammonium carbonate in 85:15 water CH:CHROMATOGRAPHY_SUMMARY to acetonitrile for 20 to 30 minutes. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm,5um) CH:SOLVENT_A 85% acetonitrile/15% water; 10 mM ammonium carbonate CH:SOLVENT_B 15% acetonitrile/85% water; 10 mM ammonium carbonate CH:FLOW_GRADIENT The column was held at 100% A for 2 minutes, ramped to 64% B over 18 minutes, CH:FLOW_GRADIENT ramped to 100% B over 1 minute, held at 100% B for 5 minutes, and equilibrated CH:FLOW_GRADIENT at 100% A for 25 minutes CH:FLOW_RATE 0.15 mL/min CH:COLUMN_TEMPERATURE 30 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS MS acquisition Comments: Polarity switching was used with a scan range of 60 to MS:MS_COMMENTS 900 m/z and a resolution of 60,000. MS parameters were as follows: capillary MS:MS_COMMENTS temperature was 320 ∞C, the H-ESI spray voltage was 3.3 kV, and the auxiliary MS:MS_COMMENTS gas heater temperature was 100 ∞C. The S-lens RF level was 65. Sheath gas, MS:MS_COMMENTS auxiliary gas, and sweep gas flow rates were maintained at 16, 3, and 1, MS:MS_COMMENTS respectively. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS normalized peak area MS_METABOLITE_DATA_START Samples 240304_Smp_WT_Gluc_A 240304_Smp_WT_Gluc_B 240304_Smp_WT_Gluc_C 240304_Smp_WT_Hom_A 240304_Smp_WT_Hom_B 240304_Smp_WT_Hom_C 240304_Smp_WT_Mix_A 240304_Smp_WT_Mix_B 240304_Smp_WT_Mix_C Factors Sample source:Filtered bacterial cells | treatment:Glucose Sample source:Filtered bacterial cells | treatment:Glucose Sample source:Filtered bacterial cells | treatment:Glucose Sample source:Filtered bacterial cells | treatment:Homarine Sample source:Filtered bacterial cells | treatment:Homarine Sample source:Filtered bacterial cells | treatment:Homarine Sample source:Filtered bacterial cells | treatment:Glucose + homarine Sample source:Filtered bacterial cells | treatment:Glucose + homarine Sample source:Filtered bacterial cells | treatment:Glucose + homarine Adenine 263062649.32287663 296156678.66630286 307420589.8659045 239526376.85705036 286409168.2175333 381732819.4789112 379444601.3877231 282075612.97467273 241313340.31544444 Adenosine 397184504.27751666 1542419272 954409863.5453106 1941950169.192544 2769343803.0601354 3773390095.923965 1147807614.7729814 357447956.8 519012675.6 Adenosine monophosphate 22234698.086137865 12235037.45185508 11183046.423580948 2737593.8261854355 4908268.761553557 6739852.732517135 34279271.16329464 17082834.379946586 14951122.881968755 L-Arginine 400928552.6281133 394876177.50675446 399140950.2817123 34200828.869831614 77014177.34496465 50302266.18824117 263804548.39271075 184536313.35673425 239464942.18987983 L-Asparagine 5002063.037289706 5632248.304 3613564.187720337 3289955.8256143723 4536649.705135552 3673776.113534178 6857931.546976386 4256751.1226436095 4309802.850307174 Citrulline 3855410.5444433033 32055666.22108508 5165246.448418219 11552004.351396332 17205198.504498396 18230869.051319953 8434087.278 17423505.27232003 15794178.639418188 Cytosine 42323422.99947487 29065668.75201696 26839008.755992953 11495636.622125486 14699351.61578952 15850946.633819578 42131484.23418347 36930955.93455931 38351323.51537908 Deoxyadenosine 8189221.405169141 26677977.527551893 17043351.91239258 17947747.024750873 38056035.57778971 46961175.12624361 21973715.43195867 7792733.339679408 13328244.374116035 L-Glutamic acid 17875256051.02259 12446040965.56043 15993840570.110704 5504889851.984282 7338453736.819024 8162298435.401269 16407818257.411428 13664232242.529709 13279007453.28972 Glutathione disulfide 57165067.82270419 25020354.582286857 34649410.129118524 11155085.888867078 11048760.960568655 12011457.75775163 14867524.094230324 17214461.694795758 Guanosine 4125151.1183555713 12598533.158804534 9759638.149088187 12424104.354881668 21506381.223990597 30419352.19553123 7766039.024 1584934.443782368 2348547.7367270594 L-Homoserine 4314114.184547655 2967359.0909994747 3490068.994820535 4867476.780948039 6198330.5395367555 6440367.945 4149965.433648589 L-Lysine 48061320.67356071 39055583.47941644 31155554.408299457 2637391.6336665987 7243342.517599266 3682881.236219349 24230717.78766854 20278417.99001114 25843261.129678413 L-Phenylalanine 71119628.95597236 136872424.1321154 165253677.6617225 65256448.76 177650794.82994786 123798367.32158992 219501265.90115315 232803162.9 381568550.1062452 L-Proline 178917792 149035216 178752144 27283098 89747168 71818504 331793568 279292416 303104448 L-Serine 18307219.970552824 19111872.662378613 11097126.304723352 21801563.204020225 27549098.298876707 27319699.96059308 20777493.40006555 13500917.563330162 8929284.345043056 L-Tryptophan 7062583.203853015 14011073.54612236 13843862.032921376 2537768.0345535967 9652940.286260637 5842547.259 15105296.35157753 9215261.282271558 13886294.282648174 L-Tyrosine 12518414.75 24986956.423377685 31655763.639100533 2680816.314707432 8221782.490675614 6033904.651 31326714.763738994 30993082.53317589 28523719.71384552 Homarine 2050645605.8132436 161946639.93391758 29940116.55972373 2611197125.5880527 10600266.071187047 5199096.25 204544803.0096527 14372575.767537331 N-Methyl-L-glutamic acid 6353242.522416147 10671345.249319533 6233413.299054862 2810245773.6917963 882093225.3188734 940113264.9529332 43551919.55989248 32875830.25606022 21907452.52263856 n-methyl glutamine 45348802.193521336 42903847.15151643 38721720.74814429 7076868638.371584 9651132188.766151 10623367339.26326 22362651.739176255 11383247.187181402 4579333.754744482 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name rt mz Adenine 5.25 136.0623 Adenosine 5.04 268.1046 Adenosine monophosphate 11.59 346.055 L-Arginine 19.78 175.1195 L-Asparagine 11.66 133.0613 Citrulline 12.43 176.1035 Cytosine 7.57 112.0511 Deoxyadenosine 3.85 252.1097 L-Glutamic acid 12.18 148.061 Glutathione disulfide 14.71 613.1598 Guanosine 9.65 284.0995 L-Homoserine 11.58 120.0661 L-Lysine 18.9 147.1134 L-Phenylalanine 5.94 166.0868 L-Proline 9.58 116.0712 L-Serine 12 106.0504 L-Tryptophan 8.19 205.0977 L-Tyrosine 9.83 182.0817 Homarine 6.25 138.0555 N-Methyl-L-glutamic acid 10.65 162.0766 n-methyl glutamine 9.85 161.0921 METABOLITES_END #END