{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000258","ANALYSIS_ID":"AN000409","VERSION":"1","CREATED_ON":"2016-09-17"},

"PROJECT":{"PROJECT_TITLE":"Effects of synthetic large-scale genome reduction on metabolism and metabolic in a nutritionally complex environment","PROJECT_SUMMARY":"The soil bacterium Sinorhizobium meliloti forms nodules on the roots of plants, where N2 is reduced to ammonia. Its genome includes a 3.65 Mb a 1.35 Mb pSymA megaplasmid, and a 1.68 Mb pSymB chromid. pSymA and pSymB ~45% of the genome and here a non-targeted approach was used to identify the consequences of the removal of these replicons. Polar and non-polar metabolites wild-type, ?pSymA, ?pSymB, and ?pSymAB cells and supernatants across a growth were analyzed by LC-HILIC-TOF-MS. 2008 metabolite features were identified in extracellular metabolome of cells grown in LBmc containing yeast extract and hydrolysate. 1474 features were found from the intracellular metabolites of grown in minimal M9-sucrose medium. Analysis revealed both time and genotype the metabolome, with the removal of pSymB having a much greater effect than the of pSymA. Strains lacking pSymB showed an increase in sugar, amino acid, and metabolites in the intracellular metabolome, and the loss of pSymB clearly the cell’s ability to catabolize exogenous amino acids. We conclude that the ability of wild-type, ?pSymA, ?pSymB, and ?pSymAB strains to grow in both and LBmc media, the removal of pSymA, and particularly pSymB, had clear and effects on the S. meliloti metabolome. The larger effect associated with the chromid is consistent with the large number of metabolic genes on this replicon the greater genetic and metabolic integration of this replicon with the S. chromosome.","INSTITUTE":"McMaster University","DEPARTMENT":"Department of Biology","LAST_NAME":"Finan","FIRST_NAME":"Turlough","ADDRESS":"Department of Biology, McMaster University, Hamilton, Canada L8S4K1","EMAIL":"finan@mcmaster.ca","PHONE":"(+1)905-525-9140 ext 22932"},

"STUDY":{"STUDY_TITLE":"Metabolic contribution of pSymA and pSymB megaplasmid/chromid for multipartite meliloti cultured in minimal M9 medium","STUDY_TYPE":"megaplasmid deletion","STUDY_SUMMARY":"To understand the contribution of pSymA and pSymB to the metabolism of S. the intracellular metabolome was analyzed at five time points (exponential and growth phases) across the growth curve of strains with or without pSymA and/or grown in a defined, minimal medium (M9).","INSTITUTE":"McMaster University","DEPARTMENT":"Department of Biology","LAST_NAME":"Finan","FIRST_NAME":"Turlough","ADDRESS":"Department of Biology, McMaster University, Hamilton, Canada L8S4K1","EMAIL":"finan@mcmaster.ca","PHONE":"(+1)905-525-9140 ext 22932","NUM_GROUPS":"20","TOTAL_SUBJECTS":"115"},

"SUBJECT":{"SUBJECT_TYPE":"Bacterial cells","SUBJECT_SPECIES":"Sinorhizobium meliloti","TAXONOMY_ID":"382","GENOTYPE_STRAIN":"Rm2011 (SU47 str-3)|SmA818|RmP3009|RmP2917","HUMAN_ETHNICITY":"wild type|Rm2011 lack pSymA|Rm2011 lack pSymB|Rm2011 lack pSymA and pSymB | a and the smb20996-engA operon from pSymB were integrated to chromosome in RmP3009 and RmP2917","SPECIES_GROUP":"Microorganism"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"M1A",
"Sample ID":"A_M1a",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1A",
"Sample ID":"A_M1b",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1A",
"Sample ID":"A_M1c",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1A",
"Sample ID":"A_M1d",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1A",
"Sample ID":"A_M1e",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1A",
"Sample ID":"A_M1f",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2a",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2b",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2c",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2d",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2e",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M2A",
"Sample ID":"A_M2f",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3a",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3b",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3c",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3d",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3e",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M3A",
"Sample ID":"A_M3f",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4a",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4b",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4c",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4d",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4e",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M4A",
"Sample ID":"A_M4f",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5a",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5b",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5c",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5d",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5e",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M5A",
"Sample ID":"A_M5f",
"Factors":{"strains":"ΔpSymA"}
},
{
"Subject ID":"M1AB",
"Sample ID":"AB_M1a",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M1AB",
"Sample ID":"AB_M1b",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M1AB",
"Sample ID":"AB_M1c",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M1AB",
"Sample ID":"AB_M1d",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M1AB",
"Sample ID":"AB_M1e",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2a",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2b",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2c",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2d",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2e",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M2AB",
"Sample ID":"AB_M2f",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M3AB",
"Sample ID":"AB_M3a",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M3AB",
"Sample ID":"AB_M3b",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M3AB",
"Sample ID":"AB_M3c",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M3AB",
"Sample ID":"AB_M3d",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M3AB",
"Sample ID":"AB_M3e",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M4AB",
"Sample ID":"AB_M4a",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M4AB",
"Sample ID":"AB_M4b",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M4AB",
"Sample ID":"AB_M4c",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M4AB",
"Sample ID":"AB_M4d",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M4AB",
"Sample ID":"AB_M4e",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M5AB",
"Sample ID":"AB_M5a",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M5AB",
"Sample ID":"AB_M5b",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M5AB",
"Sample ID":"AB_M5c",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M5AB",
"Sample ID":"AB_M5d",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M5AB",
"Sample ID":"AB_M5e",
"Factors":{"strains":"ΔpSymAB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1a",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1b",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1c",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1d",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1e",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1B",
"Sample ID":"B_M1f",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2a",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2b",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2c",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2d",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2e",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M2B",
"Sample ID":"B_M2f",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3a",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3b",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3c",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3d",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3e",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M3B",
"Sample ID":"B_M3f",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4a",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4b",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4c",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4d",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4e",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M4B",
"Sample ID":"B_M4f",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M5B",
"Sample ID":"B_M5a",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M5B",
"Sample ID":"B_M5b",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M5B",
"Sample ID":"B_M5c",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M5B",
"Sample ID":"B_M5d",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M5B",
"Sample ID":"B_M5e",
"Factors":{"strains":"ΔpSymB"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1a",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1b",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1c",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1d",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1e",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M1WT",
"Sample ID":"WT_M1f",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2a",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2b",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2c",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2d",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2e",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M2WT",
"Sample ID":"WT_M2f",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3a",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3b",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3c",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3d",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3e",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M3WT",
"Sample ID":"WT_M3f",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4a",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4b",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4c",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4d",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4e",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M4WT",
"Sample ID":"WT_M4f",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5a",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5b",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5c",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5d",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5e",
"Factors":{"strains":"wild type"}
},
{
"Subject ID":"M5WT",
"Sample ID":"WT_M5f",
"Factors":{"strains":"wild type"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"-","SAMPLE_TYPE":"cells","VOLUMEORAMOUNT_COLLECTED":"6 x 108 cells","STORAGE_CONDITIONS":"-80ºC freezer","COLLECTION_VIALS":"eppendorf tube","TISSUE_CELL_IDENTIFICATION":"6 x 108 cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"-","ANIMAL_VET_TREATMENTS":"-80ºC freezer","ANIMAL_ANESTHESIA":"test tube","ANIMAL_ENDP_CLINICAL_SIGNS":"n/a","HUMAN_FASTING":"30ºC incubator","HUMAN_ENDP_CLINICAL_SIGNS":"OD600 values of approximately 0.3, 0.5, 0.9, 1.6, and 3.5 (except for Rm2011 reached ~4.8)"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"-","SAMPLEPREP_PROTOCOL_FILENAME":"Fei,_F.,_Bowdish,_D._M._E.,_&_McCarry,_B._E._(2014)._Analytical_and_Bioanalytical_Chemistry,_406,_3723?3733.","EXTRACTION_METHOD":"cell pellet extracted with 2:2:1 MeOH/EtOH/H2O with beadbeating","EXTRACT_STORAGE":"-80ºC freezer","SAMPLE_RESUSPENSION":"cell extracts suspended 60%v/v acetonitrile/water","SAMPLE_SPIKING":"L-methionine-d3, L-tryptophan-d5 as standards for recovery determination ; diphenylalanine and glycine-phenylalanine as internal standards for peak area","CELL_TYPE":"Gram negative bacterium"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Agilent 1200 RR Series II","COLUMN_NAME":"Phenomenex Kinetex HILIC 100A (50 x 2.1 mm, 2.6um)","METHODS_FILENAME":"Opt_silicaHILIC.m","COLUMN_TEMPERATURE":"40C","FLOW_GRADIENT":"0-0.5 min, 95%A; 0.5-12.5 min, 35%A; 12.5-13.0 min, 35%A; 13.0-14.0 min, 95%A; min, 95%A","FLOW_RATE":"200 ?L/min","INTERNAL_STANDARD":"L-phenylalanine-d8, diphenylalanine and glycine-phenylalanine","SAMPLE_INJECTION":"2 ?L","SOLVENT_A":"HPLC grade acetonitrile","SOLVENT_B":"10 mM ammonium acetate in HPLC grade water adjusted to pH 3 with formic acid","ANALYTICAL_TIME":"24 min"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","SOFTWARE_VERSION":"Bruker Hystar","OPERATOR_NAME":"Fan Fei","DETECTOR_TYPE":"electron multiplier detector","DATA_FORMAT":".d"},

"MS":{"INSTRUMENT_NAME":"Bruker MicrOTOF II","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","CAPILLARY_VOLTAGE":"-3800 V","DRY_GAS_FLOW":"8.0 L/min","DRY_GAS_TEMP":"250ºC","SCAN_RANGE_MOVERZ":"50-1000 m/z","MS_COMMENTS":"POSITIVE","MS_RESULTS_FILE":"ST000258_AN000409_Results.txt UNITS:normalized peak area Has m/z:Yes Has RT:Yes RT units:Minutes"},

"MS_METABOLITE_DATA":{
"Units":"normalized peak area",

"Data":[{"Metabolite":"2-amino-caprylic 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