{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001980","ANALYSIS_ID":"AN003230","VERSION":"1","CREATED_ON":"September 8, 2021, 3:37 am"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic profiles in S. mutans, S. gordonii, and S. oralis cells treated with D-tagatose","PROJECT_SUMMARY":"Metabolomics data for Streptococcus mutans, Streptococcus gordonii, and Streptococcus oralis in the presence or absence of D-tagatose.","INSTITUTE":"Osaka University","LAST_NAME":"Mayumi","FIRST_NAME":"Shota","ADDRESS":"1-8, Yamadaoka","EMAIL":"mayumi@dent.osaka-u.ac.jp","PHONE":"+81-6-6879-2922"},

"STUDY":{"STUDY_TITLE":"Metabolomic profiles in S. mutans, S. gordonii, and S. oralis cells treated with D-tagatose","STUDY_SUMMARY":"Recent studies have shown phenotypic and metabolic heterogeneity in related species including Streptococcus oralis, a typical oral commensal bacterium, Streptococcus mutans, a cariogenic bacterium, and Streptococcus gordonii, which functions as an accessory pathogen in periodontopathic biofilm. In this study, metabolites characteristically contained in the saliva of individuals with good oral hygiene were determined, after which the effects of an identified prebiotic candidate, D-tagatose, on phenotype, gene expression, and metabolic profiles of those three key bacterial species were investigated. Examinations of the saliva metabolome of 18 systemically healthy volunteers identified salivary D-tagatose as associated with lower dental biofilm abundance in the oral cavity (Spearman’s correlation coefficient; r = -0.603, p = 0.008), then the effects of D-tagatose on oral streptococci were analyzed in vitro. In chemically defined medium (CDM) containing D-tagatose as the sole carbohydrate source, S. mutans and S. gordonii each showed negligible biofilm formation, whereas significant biofilms were formed in cultures of S. oralis. Furthermore, even in the presence of glucose, S. mutans and S. gordonii showed growth suppression and decreases in the final viable cell count in a D-tagatose concentration-dependent manner. In contrast, no inhibitory effects of D-tagatose on the growth of S. oralis were observed. To investigate species-specific inhibition by D-tagatose, the metabolomic profiles of D-tagatose-treated S. mutans, S. gordonii, and S. oralis cells were examined. The intracellular amounts of pyruvate-derived amino acids in S. mutans and S. gordonii, but not in S. oralis, such as branched-chain amino acids and alanine, tended to decrease in the presence of D-tagatose. This phenomenon indicates that D-tagatose inhibits growth of those bacteria by affecting glycolysis and its downstream metabolism. In conclusion, the present study provides evidence that D-tagatose is abundant in saliva of individuals with good oral health. Additionally, experimental results demonstrated that D-tagatose selectively inhibits growth of the oral pathogens S. mutans and S. gordonii. In contrast, the oral commensal S. oralis seemed to be negligibly affected, thus highlighting the potential of administration of D-tagatose as an oral prebiotic for its ability to manipulate the metabolism of those targeted oral streptococci.","INSTITUTE":"Osaka University","LAST_NAME":"Mayumi","FIRST_NAME":"Shota","ADDRESS":"1-8, Yamadaoka","EMAIL":"mayumi@dent.osaka-u.ac.jp","PHONE":"+81-6-6879-2922"},

"SUBJECT":{"SUBJECT_TYPE":"Bacteria","SUBJECT_SPECIES":"Streptococcus gordonii DL1 Challis / Streptococcus mutans UA159 / Streptococcus oralis ATCC9811"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"So_control_1",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"So_control_2",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"So_control_3",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"So_control_4",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"So_tagatose_1",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"So_tagatose_2",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"So_tagatose_3",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"So_tagatose_4",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_control_1",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_control_2",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_control_3",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_control_4",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_tagatose_1",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_tagatose_2",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_tagatose_3",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sm_tagatose_4",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_contorol_1",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_contorol_2",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_contorol_3",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_contorol_4",
"Factors":{"Hours":"3","D-tagatose (%)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_tagatose_1",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_tagatose_2",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_tagatose_3",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
},
{
"Subject ID":"-",
"Sample ID":"Sg_tagatose_4",
"Factors":{"Hours":"3","D-tagatose (%)":"5"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Cells were collected and washed with Milli-Q water by centrifugation and then flash-frozen in liquid N2.","SAMPLE_TYPE":"Bacterial cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"S. mutans, S. gordonii, and S. oralis cells were incubated aerobically at 37℃ in chemically defined medium containing 0.2% D-glucose without or with 5% D-tagatose."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Ribitol was added as an internal standard for normalization, then acetonitrile was added to remove protein. After thoroughly mixing the samples, centrifugation was performed. The upper 1.6 ml was evaporated in a centrifugal concentrator and freeze-dried overnight. Methoxyamine hydrochloride in pyridine and N-methyl-N-trimethylsilyltrifluoroacetamide were added to the samples as derivatizing agents, then they were thoroughly mixed and centrifuged, after which the upper 50 μl of each sample was transferred to a vial."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Shimadzu GC-MS/MS-TQ8040","COLUMN_NAME":"GL Sciences InertCap 5MS/NP capillary column (0.25 mm × 30 m, 0.25 μm)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Shimadzu GC-MS/MS-TQ8040","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"EI","ION_MODE":"POSITIVE","MS_COMMENTS":"The selected mass range was set to 85-500 m/z with electron impact ionization. Obtained GC/MS data were analyzed with an ABF converter (https://www.reifycs.com/AbfConverter/index.html) and the MS-DIAL software package, ver. 4.60 (http://prime.psc.riken.jp/Metabolomics_Software/MS-DIAL/index.html)."},

"MS_METABOLITE_DATA":{
"Units":"intensity",

"Data":[{"Metabolite":"1,6-Anhydroglucose","So_control_1":"324.9047759","So_control_2":"202.4163732","So_control_3":"590.6940631","So_control_4":"466.7317189","So_tagatose_1":"389.9228499","So_tagatose_2":"498.3733484","So_tagatose_3":"393.573945","So_tagatose_4":"325.0359617","Sm_control_1":"460.9172422","Sm_control_2":"263.4637744","Sm_control_3":"501.8001361","Sm_control_4":"521.6458519","Sm_tagatose_1":"369.2868923","Sm_tagatose_2":"303.2265742","Sm_tagatose_3":"268.0493597","Sm_tagatose_4":"438.6424057","Sg_contorol_1":"569.2978231","Sg_contorol_2":"544.1097837","Sg_contorol_3":"417.3394399","Sg_contorol_4":"611.2513071","Sg_tagatose_1":"492.1764242","Sg_tagatose_2":"506.6982997","Sg_tagatose_3":"732.836574","Sg_tagatose_4":"591.8705821"},{"Metabolite":"1-Hexadecanol","So_control_1":"160.3390019","So_control_2":"101.4542254","So_control_3":"161.0977394","So_control_4":"323.4887801","So_tagatose_1":"178.0311589","So_tagatose_2":"107.8123443","So_tagatose_3":"302.0354982","So_tagatose_4":"298.1740473","Sm_control_1":"200.6014253","Sm_control_2":"267.9896116","Sm_control_3":"125.6846642","Sm_control_4":"273.2804044","Sm_tagatose_1":"251.9057436","Sm_tagatose_2":"353.5721814","Sm_tagatose_3":"284.7173164","Sm_tagatose_4":"443.7253068","Sg_contorol_1":"197.6025856","Sg_contorol_2":"263.3222473","Sg_contorol_3":"211.6733989","Sg_contorol_4":"418.6245207","Sg_tagatose_1":"155.3938892","Sg_tagatose_2":"192.4337475","Sg_tagatose_3":"147.5392995","Sg_tagatose_4":"151.934463"},{"Metabolite":"2-Aminobutyric acid","So_control_1":"553.9197427","So_control_2":"414.9976526","So_control_3":"535.9924073","So_control_4":"746.0485618","So_tagatose_1":"470.424419","So_tagatose_2":"527.1041903","So_tagatose_3":"359.3516887","So_tagatose_4":"593.7338078","Sm_control_1":"1726.403669","Sm_control_2":"2085.156459","Sm_control_3":"2017.428539","Sm_control_4":"2043.584597","Sm_tagatose_1":"1339.385487","Sm_tagatose_2":"1334.218006","Sm_tagatose_3":"1387.403954","Sm_tagatose_4":"1085.771335","Sg_contorol_1":"505.8954656","Sg_contorol_2":"551.7466903","Sg_contorol_3":"595.1765148","Sg_contorol_4":"379.9417625","Sg_tagatose_1":"190.4835206","Sg_tagatose_2":"180.5738278","Sg_tagatose_3":"217.6635632","Sg_tagatose_4":"204.223452"},{"Metabolite":"2-Deoxyadenosine","So_control_1":"1528.149121","So_control_2":"1419.359448","So_control_3":"2285.713367","So_control_4":"2262.179432","So_tagatose_1":"2452.542405","So_tagatose_2":"2146.326916","So_tagatose_3":"1976.503025","So_tagatose_4":"1768.291158","Sm_control_1":"73.02772323","Sm_control_2":"85.10534762","Sm_control_3":"60.71689258","Sm_control_4":"43.96292001","Sm_tagatose_1":"98.34077872","Sm_tagatose_2":"101.0831123","Sm_tagatose_3":"64.99794188","Sm_tagatose_4":"88.14516906","Sg_contorol_1":"356.9328876","Sg_contorol_2":"564.5736196","Sg_contorol_3":"618.3788573","Sg_contorol_4":"538.8902928","Sg_tagatose_1":"415.171299","Sg_tagatose_2":"376.270421","Sg_tagatose_3":"325.2262123","Sg_tagatose_4":"308.5244577"},{"Metabolite":"2-Deoxyguanosine","So_control_1":"11807.54646","So_control_2":"8734.375","So_control_3":"12411.63349","So_control_4":"14201.06567","So_tagatose_1":"11185.81222","So_tagatose_2":"10844.88109","So_tagatose_3":"9909.35322","So_tagatose_4":"10159.71344","Sm_control_1":"82.90248122","Sm_control_2":"114.5384496","Sm_control_3":"111.9872249","Sm_control_4":"117.5904401","Sm_tagatose_1":"64.65737487","Sm_tagatose_2":"88.63967088","Sm_tagatose_3":"73.19214396","Sm_tagatose_4":"119.3808633","Sg_contorol_1":"4187.40296","Sg_contorol_2":"3700.897643","Sg_contorol_3":"3759.945942","Sg_contorol_4":"3852.413733","Sg_tagatose_1":"2688.622117","Sg_tagatose_2":"2536.082496","Sg_tagatose_3":"2709.561509","Sg_tagatose_4":"2681.506241"},{"Metabolite":"2-Deoxyribose 5-phosphate","So_control_1":"580.7098714","So_control_2":"277.4084507","So_control_3":"305.0673473","So_control_4":"329.0086207","So_tagatose_1":"546.7199059","So_tagatose_2":"305.0536051","So_tagatose_3":"199.6444436","So_tagatose_4":"410.7286489","Sm_control_1":"44.60782402","Sm_control_2":"359.1361327","Sm_control_3":"155.0209846","Sm_control_4":"131.7355337","Sm_tagatose_1":"248.4793131","Sm_tagatose_2":"146.0891303","Sm_tagatose_3":"316.0149873","Sm_tagatose_4":"348.0585551","Sg_contorol_1":"350.6266041","Sg_contorol_2":"235.4129803","Sg_contorol_3":"265.552519","Sg_contorol_4":"72.07188916","Sg_tagatose_1":"292.0723438","Sg_tagatose_2":"363.1362511","Sg_tagatose_3":"253.3365001","Sg_tagatose_4":"317.3989175"},{"Metabolite":"2-Hydroxypyridine (also found in blank)","So_control_1":"9047.53006","So_control_2":"5824.137324","So_control_3":"7661.929478","So_control_4":"8068.285392","So_tagatose_1":"7276.845625","So_tagatose_2":"7126.615704","So_tagatose_3":"6339.323734","So_tagatose_4":"6475.32761","Sm_control_1":"7568.787675","Sm_control_2":"7198.627553","Sm_control_3":"7142.299512","Sm_control_4":"6586.580434","Sm_tagatose_1":"6087.638032","Sm_tagatose_2":"5913.421393","Sm_tagatose_3":"5548.657824","Sm_tagatose_4":"9629.121673","Sg_contorol_1":"7369.298774","Sg_contorol_2":"7699.352599","Sg_contorol_3":"6830.263533","Sg_contorol_4":"6679.811491","Sg_tagatose_1":"5309.227282","Sg_tagatose_2":"5348.065597","Sg_tagatose_3":"5224.350286","Sg_tagatose_4":"5154.225068"},{"Metabolite":"3-Hydroxy butyrate","So_control_1":"2350.498192","So_control_2":"2980.08216","So_control_3":"2758.49348","So_control_4":"3923.54228","So_tagatose_1":"2248.288874","So_tagatose_2":"1777.063609","So_tagatose_3":"718.5660948","So_tagatose_4":"2354.7108","Sm_control_1":"64.94102462","Sm_control_2":"135.6392506","Sm_control_3":"36.03618421","Sm_control_4":"79.35582813","Sm_tagatose_1":"177.5058619","Sm_tagatose_2":"119.3270264","Sm_tagatose_3":"287.5668449","Sm_tagatose_4":"153.2854916","Sg_contorol_1":"2924.018505","Sg_contorol_2":"3849.444624","Sg_contorol_3":"1766.991516","Sg_contorol_4":"1847.824445","Sg_tagatose_1":"1953.232998","Sg_tagatose_2":"599.7592142","Sg_tagatose_3":"1115.705238","Sg_tagatose_4":"888.0126833"},{"Metabolite":"3-Phenyllactic acid","So_control_1":"1595.549273","So_control_2":"1218.88659","So_control_3":"1731.61419","So_control_4":"2466.214694","So_tagatose_1":"4549.108944","So_tagatose_2":"3809.514911","So_tagatose_3":"3936.001762","So_tagatose_4":"3944.73377","Sm_control_1":"163.7528664","Sm_control_2":"99.94390563","Sm_control_3":"32.55151146","Sm_control_4":"101.1330657","Sm_tagatose_1":"73.72997634","Sm_tagatose_2":"88.33996328","Sm_tagatose_3":"99.95628694","Sm_tagatose_4":"89.20995947","Sg_contorol_1":"246.0250642","Sg_contorol_2":"419.6209235","Sg_contorol_3":"472.3100833","Sg_contorol_4":"364.8231091","Sg_tagatose_1":"350.5310467","Sg_tagatose_2":"391.1337953","Sg_tagatose_3":"362.7130703","Sg_tagatose_4":"311.6595306"},{"Metabolite":"3-Phosphoglyceric acid","So_control_1":"236385.0584","So_control_2":"174056.4847","So_control_3":"248823.3528","So_control_4":"265745.3214","So_tagatose_1":"27372.67429","So_tagatose_2":"26046.47037","So_tagatose_3":"34198.05468","So_tagatose_4":"30481.68022","Sm_control_1":"67009.28227","Sm_control_2":"86386.57958","Sm_control_3":"78300.44805","Sm_control_4":"74710.00595","Sm_tagatose_1":"51631.16664","Sm_tagatose_2":"62382.10594","Sm_tagatose_3":"64949.72558","Sm_tagatose_4":"47485.55962","Sg_contorol_1":"20750.37232","Sg_contorol_2":"19324.41072","Sg_contorol_3":"17758.135","Sg_contorol_4":"20505.7947","Sg_tagatose_1":"4731.509955","Sg_tagatose_2":"3176.897463","Sg_tagatose_3":"3381.34144","Sg_tagatose_4":"3763.45478"},{"Metabolite":"4-Aminobutyric 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orol_4":"59330.23992","Sg_tagatose_1":"9625.231618","Sg_tagatose_2":"9247.339285","Sg_tagatose_3":"15573.82144","Sg_tagatose_4":"9076.414751"},{"Metabolite":"Icosanoic 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ol_3":"487.1801186","Sg_contorol_4":"534.9480074","Sg_tagatose_1":"4779.162231","Sg_tagatose_2":"2101.169591","Sg_tagatose_3":"2127.164482","Sg_tagatose_4":"1793.672093"},{"Metabolite":"Putrescine_3TMS_2018","So_control_1":"12729.79484","So_control_2":"8602.637911","So_control_3":"12952.76253","So_control_4":"14525.47219","So_tagatose_1":"11007.10977","So_tagatose_2":"11440.66818","So_tagatose_3":"11568.89025","So_tagatose_4":"11540.73279","Sm_control_1":"899.3573812","Sm_control_2":"248.8298307","Sm_control_3":"200.538226","Sm_control_4":"191.6709783","Sm_tagatose_1":"82.69150653","Sm_tagatose_2":"147.2037604","Sm_tagatose_3":"97.53155784","Sm_tagatose_4":"476.0578749","Sg_contorol_1":"2329.444849","Sg_contorol_2":"1922.74782","Sg_contorol_3":"2018.592987","Sg_contorol_4":"1884.026083","Sg_tagatose_1":"2351.636113","Sg_tagatose_2":"2662.727102","Sg_tagatose_3":"2169.444972","Sg_tagatose_4":"2340.515814"},{"Metabolite":"Putrescine_4TMS_2018","So_control_1":"25.99997898","So_control_2":"5.699019953","So_control_3":"20.8247512","So_control_4":"1157.243329","So_tagatose_1":"944.9970111","So_tagatose_2":"827.9103435","So_tagatose_3":"22.97200735","So_tagatose_4":"45.45733544","Sm_control_1":"1139.789913","Sm_control_2":"1077.038048","Sm_control_3":"1090.798123","Sm_control_4":"966.7949747","Sm_tagatose_1":"1586.146745","Sm_tagatose_2":"1452.013974","Sm_tagatose_3":"1651.830319","Sm_tagatose_4":"2297.935266","Sg_contorol_1":"13.80067493","Sg_contorol_2":"10.13881014","Sg_contorol_3":"31.28469104","Sg_contorol_4":"18.11094168","Sg_tagatose_1":"1212.392864","Sg_tagatose_2":"1351.733783","Sg_tagatose_3":"4.134612705","Sg_tagatose_4":"23.2160803"},{"Metabolite":"Pyroglutamic 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acid(18_0) (also found in blank)","Rt(min)":"14.27","Quant mass":"117.0995"},{"Metabolite":"Succinic acid(or anhydride)","Rt(min)":"7.488","Quant mass":"147.15"},{"Metabolite":"Sucrose","Rt(min)":"16.727","Quant mass":"361.3068"},{"Metabolite":"Tagatose_1_minor","Rt(min)":"11.984","Quant mass":"103.1469"},{"Metabolite":"Tagatose_2_major","Rt(min)":"12.123","Quant mass":"217.2195"},{"Metabolite":"Threonine_2TMS_minor","Rt(min)":"7.321","Quant mass":"147.15"},{"Metabolite":"Threonine_3TMS_major","Rt(min)":"8.193","Quant mass":"218.2324"},{"Metabolite":"Thymidine_3","Rt(min)":"15.733","Quant mass":"103.1273"},{"Metabolite":"Thymine","Rt(min)":"8.302","Quant mass":"255.201"},{"Metabolite":"Trehalose","Rt(min)":"17.268","Quant mass":"361.3064"},{"Metabolite":"Tryptophan_1TMS","Rt(min)":"13.931","Quant mass":"130.15"},{"Metabolite":"Tryptophan_2TMS","Rt(min)":"14.108","Quant mass":"218.2028"},{"Metabolite":"Tryptophan_3TMS","Rt(min)":"14.293","Quant mass":"202.2"},{"Metabolite":"Turanose_1","Rt(min)":"17.312","Quant mass":"144.1333"},{"Metabolite":"Tyrosine","Rt(min)":"12.487","Quant mass":"218.2013"},{"Metabolite":"Uracil","Rt(min)":"7.714","Quant mass":"241.19"},{"Metabolite":"Urea","Rt(min)":"6.677","Quant mass":"147.1652"},{"Metabolite":"Uridine 5'-diphosphate_1","Rt(min)":"17.64","Quant mass":"169.1427"},{"Metabolite":"Uridine 5'-diphosphate_2","Rt(min)":"17.783","Quant mass":"169.1428"},{"Metabolite":"Uridine_4TMS","Rt(min)":"15.577","Quant mass":"217.2"},{"Metabolite":"Valine_1TMS_minor","Rt(min)":"5.113","Quant mass":"156.1714"},{"Metabolite":"Valine_2TMS","Rt(min)":"6.513","Quant mass":"144.2"},{"Metabolite":"Xanthine","Rt(min)":"13.022","Quant mass":"353.2625"},{"Metabolite":"Xanthosine","Rt(min)":"16.782","Quant mass":"325.2482"},{"Metabolite":"Xylitol","Rt(min)":"10.903","Quant mass":"103.15"}]
}

}