{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000258","ANALYSIS_ID":"AN000410","VERSION":"1","CREATED_ON":"01-03-2020"},

"PROJECT":{"PROJECT_TITLE":"Effects of synthetic large-scale genome reduction on metabolism and metabolic preferences in a nutritionally complex environment","PROJECT_SUMMARY":"The soil bacterium Sinorhizobium meliloti forms nodules on the roots of leguminous plants, where N2 is reduced to ammonia.  Its genome includes a 3.65 Mb chromosome, a 1.35 Mb pSymA megaplasmid, and a 1.68 Mb pSymB chromid.    pSymA and pSymB  constitute ~45% of the genome and here a non-targeted approach was used to identify the metabolic consequences of the removal of these replicons.  Polar and non-polar metabolites from wild-type, ∆pSymA, ∆pSymB, and ∆pSymAB cells and supernatants across a growth curve were analyzed by LC-HILIC-TOF-MS.  2008 metabolite features were identified in the extracellular metabolome of cells grown in LBmc containing yeast extract and casein hydrolysate.  1474 features were found from the intracellular metabolites of cells grown in minimal M9-sucrose medium.  Analysis revealed both time and genotype influenced the metabolome, with the removal of pSymB having a much greater effect than the loss of pSymA.  Strains lacking pSymB showed an increase in sugar, amino acid, and nucleotide metabolites in the intracellular metabolome, and the loss of pSymB clearly impaired the cell’s ability to catabolize exogenous amino acids.  We conclude that despite the ability of wild-type, ∆pSymA, ∆pSymB, and ∆pSymAB strains to grow in both M9-sucrose and LBmc media, the removal of pSymA, and particularly pSymB, had clear and dramatic effects on the S. meliloti metabolome.  The larger effect associated with the pSymB chromid is consistent with the large number of metabolic genes on this replicon and the greater genetic and metabolic integration of this replicon with the S. meliloti 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","DOI":"http://dx.doi.org/10.21228/M87W2S"},

"STUDY":{"STUDY_TITLE":"Metabolic contribution of pSymA and pSymB megaplasmid/chromid for multipartite Sinorhizobium 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. meliloti, the intracellular metabolome was analyzed at five time points (exponential and stationary growth phases) across the growth curve of strains with or without pSymA and/or pSymB 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","SUBMIT_DATE":"2015-09-17","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  tRNAarg and the smb20996-engA operon from pSymB were integrated to chromosome in strains 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":{"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":{"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 which reached ~4.8)"},

"SAMPLEPREP":{"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 ; L-phenylalanine-d8, diphenylalanine and glycine-phenylalanine as internal standards for peak area normalization","CELL_TYPE":"Gram negative bacterium"},

"CHROMATOGRAPHY":{"METHODS_FILENAME":"Opt_silicaHILIC.m","INSTRUMENT_NAME":"Agilent 1200 RR Series II","COLUMN_NAME":"Phenomenex Kinetex HILIC 100A (50 x 2.1 mm, 2.6um)","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; 14.0-24.0 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","CHROMATOGRAPHY_TYPE":"HILIC"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_COMMENTS":"NEGATIVE","INSTRUMENT_NAME":"Bruker MicrOTOF II Mass Spectrometer","SOFTWARE_VERSION":"Bruker Hystar","OPERATOR_NAME":"Fan Fei","DETECTOR_TYPE":"electron multiplier detector","DATA_FORMAT":".d","ANALYSIS_DISPLAY":"ESI/QTOF negative ion mode","NUM_FACTORS":"4","NUM_METABOLITES":"56"},

"MS":{"INSTRUMENT_NAME":"Bruker MicrOTOF II","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","CAPILLARY_VOLTAGE":"4500 V","DRY_GAS_FLOW":"8.0 L/min","DRY_GAS_TEMP":"250ºC","SCAN_RANGE_MOVERZ":"50-1000 m/z"},

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

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