#METABOLOMICS WORKBENCH Lu_Group_20210907_071320 DATATRACK_ID:2827 STUDY_ID:ST001938 ANALYSIS_ID:AN003151 PROJECT_ID:PR001227 VERSION 1 CREATED_ON October 12, 2021, 6:37 pm #PROJECT PR:PROJECT_TITLE Metabolomics characterized concentration-dependent metabolic influence of PR:PROJECT_TITLE magnesium on biofilm formation in Escherichia coli PR:PROJECT_TYPE Targeted MS quantitative analysis PR:PROJECT_SUMMARY Metabolomics characterized concentration-dependent metabolic influence of PR:PROJECT_SUMMARY magnesium on biofilm formation in Escherichia coli PR:INSTITUTE Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University PR:DEPARTMENT Shanghai Center for Systems Biomedicine PR:LABORATORY Lu Group PR:LAST_NAME Lu PR:FIRST_NAME Haitao PR:ADDRESS 800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China PR:EMAIL haitao_lu@sjtu.edu.cn PR:PHONE 15221478139 #STUDY ST:STUDY_TITLE Metabolomics characterized concentration-dependent metabolic influence of ST:STUDY_TITLE magnesium on biofilm formation in Escherichia coli (Part1) ST:STUDY_SUMMARY Biofilms are broadly formed by a diversity of microorganisms that enable them to ST:STUDY_SUMMARY adapt stressful environments. Biofilms often impose harmful influences in many ST:STUDY_SUMMARY niches, as they can cause food contamination, antibiotics resistance, and ST:STUDY_SUMMARY environmental issues. However, eradicating biofilms remains difficult since the ST:STUDY_SUMMARY formation mechanism of biofilms are still incompletely clarified. In this study, ST:STUDY_SUMMARY we aimed at exploring the regulatory role of magnesium (Mg2+) on biofilm ST:STUDY_SUMMARY formation in Escherichia coli (E. coli) using phenotype visualization combined ST:STUDY_SUMMARY with targeted metabolomics method. We found that Mg2+ could exert significant ST:STUDY_SUMMARY influence on biofilm formation in a concentration-dependent manner by regulating ST:STUDY_SUMMARY phenotypic morphology and triggering metabolic modifications of biofilm. ST:STUDY_SUMMARY Phenotypic imaging revealed that increasing concentration of Mg2+ gradually ST:STUDY_SUMMARY inhibited biofilm formation, Mg2+ was observed to restore the microstructure of ST:STUDY_SUMMARY E. coli strain in biofilms to that in the relevant planktonic cells. In ST:STUDY_SUMMARY addition, our metabolomics analysis characterized 20 differential metabolites ST:STUDY_SUMMARY and associated 2 metabolic pathways including nucleotide metabolism and amino ST:STUDY_SUMMARY acid metabolism that were notably modified during biofilm formation under the ST:STUDY_SUMMARY treatments of different concentrations of Mg2+. Altogether, our work provides a ST:STUDY_SUMMARY novel insight into the influence of Mg2+ on biofilm formation at a metabolic ST:STUDY_SUMMARY level, which are implicated in the novel solution to disturb biofilm formation ST:STUDY_SUMMARY through the regulation of Mg2+ and functional metabolite interaction, then ST:STUDY_SUMMARY biofilms associated harmful impacts in different niches could be well tangled ST:STUDY_SUMMARY accordingly. ST:INSTITUTE Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University ST:DEPARTMENT Shanghai Center for Systems Biomedicine ST:LABORATORY Lu Group ST:LAST_NAME Lu ST:FIRST_NAME Haitao ST:ADDRESS 800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China ST:EMAIL haitao_lu@sjtu.edu.cn ST:PHONE 15221478139 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Escherichia coli SU:TAXONOMY_ID 562 #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 - 20210306-4-C-1 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210306-4-C-1 SUBJECT_SAMPLE_FACTORS - 20210306-4-C-2 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210306-4-C-2 SUBJECT_SAMPLE_FACTORS - 20210306-4-C-4 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210306-4-C-4 SUBJECT_SAMPLE_FACTORS - 20210306-4-C-5 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210306-4-C-5 SUBJECT_SAMPLE_FACTORS - 20210306-4-C-6 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210306-4-C-6 SUBJECT_SAMPLE_FACTORS - 20210306-4-30-1 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210306-4-30-1 SUBJECT_SAMPLE_FACTORS - 20210306-4-30-2 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210306-4-30-2 SUBJECT_SAMPLE_FACTORS - 20210306-4-30-4 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210306-4-30-4 SUBJECT_SAMPLE_FACTORS - 20210306-4-30-5 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210306-4-30-5 SUBJECT_SAMPLE_FACTORS - 20210306-4-30-6 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210306-4-30-6 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-1 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-1 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-2 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-2 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-3 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-3 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-4 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-4 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-5 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-5 SUBJECT_SAMPLE_FACTORS - 20210306-4-50-6 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210306-4-50-6 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-1 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-1 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-2 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-2 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-3 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-3 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-4 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-4 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-5 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-5 SUBJECT_SAMPLE_FACTORS - 20210306-4-80-6 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210306-4-80-6 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-1 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-1 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-2 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-2 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-3 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-3 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-4 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-4 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-5 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-5 SUBJECT_SAMPLE_FACTORS - 20210306-4-100-6 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210306-4-100-6 #COLLECTION CO:COLLECTION_SUMMARY After 72 hours of culture, the planktonic cells were isolated, and the biofilms CO:COLLECTION_SUMMARY were washed with PBS solvent for three times. Then, planktonic cells and biofilm CO:COLLECTION_SUMMARY solutions were centrifuged at 2000 rpm and 4°C for 15 minutes, to collect the CO:COLLECTION_SUMMARY strain pellets. CO:SAMPLE_TYPE Biofilm #TREATMENT TR:TREATMENT_SUMMARY Colony-forming antigen (CFA) medium (1% casamino acids, 0.15% yeast extract, TR:TREATMENT_SUMMARY 0.005% magnesium sulfate, and 0.0005% manganese chloride) was used to form TR:TREATMENT_SUMMARY biofilms. The E. coli strain was incubated in LB-agar plate for 12 hours, one TR:TREATMENT_SUMMARY colony was isolated to LB broth for further 4 hours incubation, then diluted the TR:TREATMENT_SUMMARY solution into CFA medium at a ratio of 1:1000 and the cultures were incubated TR:TREATMENT_SUMMARY for another 72 h at 30°C to form mature biofilms. To test the influence of TR:TREATMENT_SUMMARY MgSO4 on biofilm formation, the purified compound of MgSO4 was added to the CFA TR:TREATMENT_SUMMARY medium at different concentrations for the preparation of treated culture TR:TREATMENT_SUMMARY medium; The left cultural procedures were kept as the same as the protocol TR:TREATMENT_SUMMARY listed above. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Each pellet sample was mixed with 2 mL of 80% ice-cold methanol, and the mixed SP:SAMPLEPREP_SUMMARY solution was homogenized on ice for 2 minutes and repeated the same procedure SP:SAMPLEPREP_SUMMARY for three times. Next, the collected samples were centrifuged at 12000 rpm and SP:SAMPLEPREP_SUMMARY 4°C for 15 minutes, to collect the supernatants. The supernatants were further SP:SAMPLEPREP_SUMMARY mixed with 800 μL of ice-cold acetonitrile for 20 minutes before further SP:SAMPLEPREP_SUMMARY centrifuge at 12000 rpm and 4°C for 15 minutes. At last, the isolated SP:SAMPLEPREP_SUMMARY supernatants were gone through a 0.22 μm filter membrane (nylon) before the SP:SAMPLEPREP_SUMMARY lyophilization. The lyophilized samples were stored at -80°C for metabolomics SP:SAMPLEPREP_SUMMARY analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The chromatographic separation of biological samples was implemented on a Waters CH:CHROMATOGRAPHY_SUMMARY ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 μm) with an optimized CH:CHROMATOGRAPHY_SUMMARY gradient-elution program (mobile phase A and B were 0.1% formic acid in water CH:CHROMATOGRAPHY_SUMMARY and acetonitrile (v/v)): 0-2 minutes, 98% A; 2-10 minutes, 98%-65% A; 10-12 CH:CHROMATOGRAPHY_SUMMARY minutes, 65%-20% A; 12-14 minutes, 20%-2% A; 14-30 minutes, 2% A. The column CH:CHROMATOGRAPHY_SUMMARY temperature was at 40°C and the flow rate was set at 0.3 mL/ minutes. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6495 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Agilent MassHunter Workstation Data Acquisition Agilent MassHunter MS:MS_COMMENTS QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ) #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS - MS_METABOLITE_DATA_START Samples 20210306-4-C-1 20210306-4-C-2 20210306-4-C-4 20210306-4-C-5 20210306-4-C-6 20210306-4-30-1 20210306-4-30-2 20210306-4-30-4 20210306-4-30-5 20210306-4-30-6 20210306-4-50-1 20210306-4-50-2 20210306-4-50-3 20210306-4-50-4 20210306-4-50-5 20210306-4-50-6 20210306-4-80-1 20210306-4-80-2 20210306-4-80-3 20210306-4-80-4 20210306-4-80-5 20210306-4-80-6 20210306-4-100-1 20210306-4-100-2 20210306-4-100-3 20210306-4-100-4 20210306-4-100-5 20210306-4-100-6 Factors Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.40% Concentration of MgSO4(w/v):0.50% Concentration of MgSO4(w/v):0.50% Concentration of MgSO4(w/v):0.50% Concentration of MgSO4(w/v):0.50% Concentration of MgSO4(w/v):0.50% Concentration of MgSO4(w/v):0.50% Citrate 2326.255528 1280.785882 4063.517177 1839.75724 3062.152221 3603.364042 2142.995903 1532.07668 1929.731169 3623.052209 2479.242136 2258.901048 2665.656664 2991.956826 2898.325035 1893.672754 3258.477249 3655.46426 5053.056551 3609.94553 2634.883083 1966.503815 2680.175269 3955.084539 2980.199756 3281.84596 6983.01619 2799.920457 Isocitric acid 2363.674418 977.3233325 3246.91494 1439.187121 2239.935192 3102.968352 2467.309266 1385.704266 1724.373602 3077.912221 2122.658304 1979.950073 3009.109463 2771.458397 2436.872019 1675.732229 2693.766891 3232.695035 4496.944736 3241.793694 2178.865465 2028.805872 2630.943088 3391.765592 2339.642917 3452.600213 5437.795245 2656.892993 Methylmalonic acid 2926.428312 8150.115441 1012.942125 5513.15038 6155.366251 2793.235585 5007.874268 3386.216981 4332.459064 6080.478854 1740.726768 3780.173302 2681.150377 2379.626905 4755.901578 1944.229911 1314.720304 1613.028121 3416.610655 1531.017025 1157.048244 1935.058023 629.1302272 1748.992993 2087.466195 1779.099056 2170.881705 2826.916187 3-Hydroxybenzoic acid 18.76240298 30.79886472 29.35188962 41.41673102 30.99202664 15.10462575 22.13970915 23.38463559 21.92432696 36.25491714 9.255997122 28.79412661 17.75897871 17.03680109 23.74617928 12.32026423 14.66703969 27.63990409 41.78024559 24.71013153 20.93464966 16.78210231 26.99752722 23.21767019 34.45204222 17.06517425 23.15365437 18.06970494 4-Hydroxybenzoic acid 23.97068518 37.28448714 35.80039583 47.57974898 40.6241692 17.80980685 28.30878892 29.33949763 27.15919256 47.34984522 11.63402516 32.87402399 20.75054316 21.45699885 30.24104854 14.58768105 15.31053362 30.29997575 52.21065753 29.40070846 26.07177797 18.45998862 32.61508751 29.59493325 39.22712295 20.33972259 28.75106059 23.01199679 Azelaic acid 21.97977978 38.84546483 21.9193774 41.73451978 14.44504189 16.94088258 20.95855382 20.30574829 25.7573133 34.77674194 14.86363567 18.38195523 22.90784587 29.44807428 21.27124601 10.82718653 19.85409845 20.61193267 39.24820295 17.60966047 18.5894389 20.52981363 27.64584774 23.9040124 27.3327934 13.84091431 26.19881862 26.08334096 Maleic acid 377.7946379 532.0387637 1649.623714 1356.460457 735.7244064 50.73052084 188.5843367 239.0247669 132.9212231 183.4763432 58.30458619 236.9671009 60.79903257 47.01126716 68.60007347 77.00266851 55.05072746 804.6002578 211.3623364 213.083865 352.4346377 132.6420958 234.3103389 308.8754442 321.1345056 140.1653686 123.8412433 182.0579385 Lithocholic acid 14.62862639 26.72666879 26.78397051 39.78090039 59.09546783 42.44119268 13.25786603 16.12947682 21.39862224 54.3685932 24.27458238 60.62202308 18.18297997 11.88301476 45.60564907 11.87003553 17.59298168 12.25082028 23.35277328 29.61195876 19.97498714 12.52422285 46.47636274 11.95021885 12.230981 13.80416509 20.01538259 16.9440584 Sebacic acid 9.147112768 27.3429413 7.181066014 20.10144109 5.037050181 10.77675475 7.465198413 9.507902127 15.05911126 15.16077128 8.743767066 9.613698535 18.73850014 16.08668256 13.93187289 6.319475102 8.628506929 7.375747324 14.36263407 6.710786054 5.981328473 14.83860465 40.86245496 11.598111 13.22441259 10.94546312 13.91957719 19.57789542 N-Acetylglutamic acid 12858.9571 18709.71249 28697.33753 32311.96861 29251.1189 21728.28262 16042.70397 22219.3047 23220.71037 33463.16104 13401.73749 21069.31382 16521.33278 15610.52759 20491.07333 11354.33924 15871.79009 13098.99396 28858.58126 19700.49078 17014.51726 14134.70057 12439.50834 15498.57771 9996.565412 17249.53732 25787.17101 15049.87668 Beta-Glycerophosphate 9.21465312 14.08570173 9.554955772 21.80499719 13.35600711 13.71225313 8.655624986 4.660776935 8.553238037 17.42799572 5.529523461 8.734097426 15.45445334 12.7696676 18.02015023 4.666376358 4.996854755 5.740372532 4.160445213 3.293863815 3.024735095 4.345336855 3.813220378 2.702472465 3.09388881 3.144959008 4.619777047 5.263276995 Cis-Aconitic acid 21.81399892 11.76822185 24.18239324 20.89330815 25.29732633 47.85155488 33.52293147 21.71164236 24.56837772 45.50643326 31.34976004 31.50978829 50.7878917 49.95308796 41.80294095 24.76664674 37.16977336 56.99591667 58.3788491 53.40120308 32.1685688 34.9230334 35.28872479 45.70610177 31.59360289 37.31979339 92.34351025 47.65530165 D-Glucose 6-phosphate 29.94493638 32.61077972 13.52661787 35.96743534 24.99281952 48.90473298 31.85225493 23.14859446 36.60635045 64.96390495 18.99733222 38.30961726 55.20849744 34.51039536 48.16112751 16.46860636 82.5058757 26.28192198 117.6613383 142.2760479 55.40678705 107.8770769 18.5017891 22.18458217 24.14183331 56.26691421 108.0814229 63.53307377 Flavin adenine dinucleotide 137.4630379 170.7849819 104.8273287 157.6701087 179.3820024 153.0311737 239.9414158 188.6361984 215.0553771 396.2302031 125.6295437 240.1994215 213.710376 177.5060329 262.7212449 158.1075714 166.8195782 261.707579 319.2933524 204.8245938 227.0349505 172.0756634 188.9443392 186.1113428 231.4840173 190.4827786 255.2543203 261.3324756 Fumarate 385.7275593 524.7615817 1649.623714 1342.100052 735.7244064 50.73052084 200.1622549 232.6081752 138.595285 200.1083984 55.20047205 236.9671009 56.18998184 43.813717 73.4205796 77.00266851 55.33869988 804.6002578 204.2565797 213.083865 342.4007699 123.7684094 234.3103389 303.5616541 319.1827534 138.739886 139.2012489 175.3172504 Malic acid 288.0427192 573.9379142 194.9203058 172.9656461 274.1935818 269.9841929 430.2872871 304.1803993 220.4876593 569.972642 181.8083751 460.8277182 339.0734153 363.8300617 431.1314231 202.097085 219.2998852 555.4022609 607.5051876 593.0985331 337.1841429 314.1939299 630.6935014 392.6628147 1109.603188 487.3274519 633.8843461 621.5613543 Succinic acid 2732.065597 7137.470837 1012.363997 5327.504442 6150.371493 2641.150806 4795.594509 3285.533018 3967.768602 6159.900622 1652.488738 3828.43821 2715.423812 2304.496609 4750.248772 1848.206436 1261.614279 1466.125922 3196.125563 1580.773648 1107.989993 1729.473221 619.781628 1684.4804 2024.572268 1662.981212 2210.66327 2701.837509 p-Hydroxyphenylpropionic acid 130.9131586 136.0080228 121.735601 159.0194085 113.145427 106.9404991 115.8774576 114.1093193 108.1021929 195.0846701 59.45838439 109.8604706 87.86837229 90.61778739 159.6209921 59.89533409 88.34353881 97.58426294 146.9294073 86.53878569 75.98937116 54.98316843 151.1353826 89.46041787 164.0752436 84.86747316 135.8959312 110.1968391 Phenyllactic acid 110.2673008 232.6834671 340.7591044 517.0240686 409.6504766 113.9609884 149.0480822 143.3701365 182.9319349 263.6871602 110.7953612 181.2917666 99.54214774 81.02140365 129.5044225 118.0046706 157.1440605 247.7634213 247.1316793 165.8396851 176.6403653 146.1720787 193.3857913 199.6666015 311.2662809 120.6631413 147.1920665 143.7837575 Citramalic acid 15.27946979 13.53769892 16.02524078 24.13892116 21.68130794 8.963529643 10.40232566 8.439505459 12.37513356 16.51490381 10.81445707 9.18884266 10.18191915 12.09581144 7.471174301 9.651438692 8.809822897 6.406943017 8.876027667 3.665090061 5.104950282 5.300889872 5.887846024 5.948299182 8.880989242 8.421372399 16.99760292 6.665938115 Glutaric acid 383.0182702 807.8262494 1466.33725 1692.673144 1542.288939 494.1478105 233.3236082 692.5405174 716.8505177 519.6285445 516.0128755 345.3779376 329.2252008 493.667771 341.3162412 374.099365 350.9388329 266.628194 349.9245918 296.7923069 198.1315057 333.8200171 416.2217703 280.21172 422.4170246 350.0846128 449.6327403 261.0290787 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z RT Citrate 191.11 1.54 Isocitric acid 191.02 1.52 Methylmalonic acid 117.02 1.88 3-Hydroxybenzoic acid 137.02 6.26 4-Hydroxybenzoic acid 137.02 6.25 Azelaic acid 187.1 9.56 Maleic acid 115 1.77 Lithocholic acid 375.3 15.21 Sebacic acid 201.1 10.89 N-Acetylglutamic acid 188.05 1.7 Beta-Glycerophosphate 171 1.05 Cis-Aconitic acid 173 1.7 D-Glucose 6-phosphate 259.02 1.03 Flavin adenine dinucleotide 784.15 6.41 Fumarate 115 1.78 Malic acid 133.01 1.11 Succinic acid 117.02 1.88 p-Hydroxyphenylpropionic acid 165.05 6.8 Phenyllactic acid 165.05 8.29 Citramalic acid 147.03 2.01 Glutaric acid 131.03 3.45 METABOLITES_END #END