{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001298","ANALYSIS_ID":"AN002162","VERSION":"1","CREATED_ON":"January 7, 2020, 2:46 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolome Profiling of Synechococcus elongatus PCC 11801 strains engineered for Succinate Production","PROJECT_TYPE":"Measurement of relative metabolite pools of wild type and engineered Synechococcus elongatus PCC 11801 strains","PROJECT_SUMMARY":"Experiments to measure relative metabolite pools of wild type Synechococcus elongatus PCC 11801 and its recombinants producing succinate. The wild type and the engineered strains producing succinate were cultivated at 1% CO2 and their metabolome data was collected in three biological and three technical replicates (n=9). The study aims to find metabolomics changes between the wild type and the engineered to identify potential rate-limiting steps that be used as targets for improved production.","INSTITUTE":"Indian Institute of Technology Bombay (IIT Bombay)","DEPARTMENT":"Department of Chemical Engineering","LABORATORY":"Bio systems Engineering Lab","LAST_NAME":"Wangikar","FIRST_NAME":"Pramod P","ADDRESS":"Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, Maharashtra, India - 400076","EMAIL":"wangikar@iitb.ac.in","PHONE":"+91 (22) 2576 7232","FUNDING_SOURCE":"Department of Biotechnology (DBT), PAN-IIT Centre for Bioenergy (Grant No: BT/EB/PAN IIT/2012)"},

"STUDY":{"STUDY_TITLE":"Metabolome Profiling of Synechococcus elongatus PCC 11801 strains engineered for Succinate Production","STUDY_TYPE":"Measurement of relative metabolite pools of wild type and engineered Synechococcus elongatus PCC 11801 strains using Isotopic Ratio Method","STUDY_SUMMARY":"Experiments to measure relative metabolite pools of wild type Synechococcus elongatus PCC 11801 and its recombinants producing succinate. The wild type and the engineered strains producing succinate were cultivated at 1% CO2 and their metabolome data was collected in three biological and three technical replicates (n=9). The study aims to find metabolomics changes between the wild type and the engineered to identify potential rate-limiting steps that be used as targets for improved production.","INSTITUTE":"Indian Institute of Technology Bombay (IIT Bombay)","DEPARTMENT":"Department of Chemical Engineering","LABORATORY":"Bio systems Engineering Lab","LAST_NAME":"Wangikar","FIRST_NAME":"Pramod P","ADDRESS":"Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, Maharashtra, India - 400076","EMAIL":"wangikar@iitb.ac.in","PHONE":"22254215"},

"SUBJECT":{"SUBJECT_TYPE":"Bacteria","SUBJECT_SPECIES":"Synechococcus elongatus PCC 11801","TAXONOMY_ID":"2219813"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"WT BR1-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR1 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR1-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR1 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR1-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR1 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR2-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR2 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR2-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR2 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR2-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR2 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR3-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR3 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR3-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR3 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT BR3-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 1PC KS BR3 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR1-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR1 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR1-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR1 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR1-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR1 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR2-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR2 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR2-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR2 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR2-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR2 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR3-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR3 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR3-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR3 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA2 BR3-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 2-GENE BR3 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR1-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR1 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR1-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR1 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR1-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR1 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR2-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR2 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR2-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR2 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR2-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR2 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR3-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR3 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR3-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR3 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA3 BR3-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SA2 3-GENE BR3 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR1-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR1 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR1-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR1 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR1-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR1 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR2-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR2 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR2-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR2 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR2-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR2 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR3-1",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR3 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR3-2",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR3 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"WT EP BR3-3",
"Factors":{"Genotype":"Wild type"},
"Additional sample data":{"RAW_FILE_NAME":"11801 EP 1PC KS BR3 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR1-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR1 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR1-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR1 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR1-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR1 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR2-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR2 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR2-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR2 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR2-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR2 IS IDA-3"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR3-1",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR3 IS IDA-1"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR3-2",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR3 IS IDA-2"}
},
{
"Subject ID":"-",
"Sample ID":"SA7 BR3-3",
"Factors":{"Genotype":"△DOP62_03525 PpsbaI OgdA PpsbaIII SsaD  Pcpcb300 PEPC SmR ,  DDOP62_03790 Prbcl400 gltA Prbcl400  SBPase/FBPase KanR"},
"Additional sample data":{"RAW_FILE_NAME":"11801 SBPase EP 1PC KS BR3 IS IDA-3"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"The experiment was conducted in a shaker maintained at a temperature of 38°C, 1% CO2, and 120 rpm. The cells were grown in 100 ml shake flasks with 20 ml culture volume and the samples for metabolomics analysis were collected at a metabolic steady-state (OD 730 nm = 0.6). Samples were quenched with methanol and extracted using the methanol-chloroform-water method. Extracts were stored at -80°C till LCMS analysis. LCMS analysis was done in the negative ion mode using information-dependent acquisition (IDA) method.","SAMPLE_TYPE":"Bacterial cells","COLLECTION_TUBE_TEMP":"4 degree centrigrade"},

"TREATMENT":{"TREATMENT_SUMMARY":"The metabolites were extracted using a methanol-chloroform-water method described in the Metabolite Extraction Protocol file of the collection data."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"One aliquot of the metabolite extract of each sample were reconstituted in 100µL 50:50 methanol-water and filtered using nylon syringe filters to remove any particulate matter. The metabolite extract of each test sample was mixed with equal volume of an extract of the PCC 11801 WT biomass that is fully labeled with 13C isotopic carbon by growing for ~5 generations in the presence of NaH13CO3 in modified BG-11 medium. 13C-labeled biomass of PCC 11801 that acted as an internal standard. The injection volume was 6 µL. The peak areas corresponding to the 12C and 13C monoisotopic peak for the metabolites of interest were quantified using MultiQuant 3.0.1 (SCIEX, Framingham, MA). The relative quantification of metabolites was done using isotopic ratio method by normalizing area under the peak for monoisotopic m/z of a particular metabolite by its respective highest possible isotopologue present in the internal standard giving area ratio.","PROCESSING_STORAGE_CONDITIONS":"On ice","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Shimadzu 20AD","COLUMN_NAME":"Phenomenex Synergi Hydro RP 100 A (100 x 2mm, 2.5um)","FLOW_GRADIENT":"The gradient method used is as follows: 0% B (0.01 min), 0% B (2 min), 35% B (8 min), 35% B (10.5 min), 90% B (15.50 min), 90% B (20.5 min), 0% B (22 min), and 0% B (30 min)","FLOW_RATE":"0.3 mL/minute","SOLVENT_A":"10 mM tributylamine + 15mM acetic acid in water","SOLVENT_B":"100% Methanol"},

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

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5600+ TripleTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"The peak areas corresponding to the 12C and 13C monoisotopic peak for the metabolites of interest were quantified using MultiQuant 3.0.1 (SCIEX, Framingham, MA). The relative quantification of metabolites was done using isotopic ratio method by normalizing area under the peak for monoisotopic m/z of a particular metabolite by its respective highest possible isotopologue present in the internal standard giving area ratio."},

"MS_METABOLITE_DATA":{
"Units":"Area Ratio",

"Data":[{"Metabolite":"RuBP","WT BR1-1":"1.36","WT BR1-2":"1.3","WT BR1-3":"1.36","WT BR2-1":"1.34","WT BR2-2":"1.45","WT BR2-3":"1.32","WT BR3-1":"1.61","WT BR3-2":"1.62","WT BR3-3":"1.51","SA2 BR1-1":"1.61","SA2 BR1-2":"1.55","SA2 BR1-3":"1.34","SA2 BR2-1":"1.02","SA2 BR2-2":"1.06","SA2 BR2-3":"1.02","SA2 BR3-1":"0.8","SA2 BR3-2":"0.77","SA2 BR3-3":"0.84","SA3 BR1-1":"1.29","SA3 BR1-2":"1.27","SA3 BR1-3":"1.29","SA3 BR2-1":"1.05","SA3 BR2-2":"0.95","SA3 BR2-3":"0.93","SA3 BR3-1":"1.63","SA3 BR3-2":"1.53","SA3 BR3-3":"1.57","WT EP BR1-1":"0.8107","WT EP BR1-2":"0.8499","WT EP BR1-3":"0.8469","WT EP BR2-1":"0.8244","WT EP BR2-2":"0.8613","WT EP BR2-3":"0.8341","WT EP BR3-1":"0.8637","WT EP BR3-2":"0.8384","WT EP BR3-3":"0.7833","SA7 BR1-1":"1.423","SA7 BR1-2":"1.426","SA7 BR1-3":"1.456","SA7 BR2-1":"1.502","SA7 BR2-2":"1.47","SA7 BR2-3":"1.471","SA7 BR3-1":"1.717","SA7 BR3-2":"1.768","SA7 BR3-3":"1.775"},{"Metabolite":"3PGA","WT BR1-1":"1.23","WT BR1-2":"1.21","WT BR1-3":"1.24","WT BR2-1":"1.22","WT BR2-2":"1.18","WT BR2-3":"1.22","WT BR3-1":"1.35","WT BR3-2":"1.38","WT BR3-3":"1.35","SA2 BR1-1":"1.36","SA2 BR1-2":"1.37","SA2 BR1-3":"1.37","SA2 BR2-1":"1.14","SA2 BR2-2":"1.1","SA2 BR2-3":"1.12","SA2 BR3-1":"1.1","SA2 BR3-2":"1.09","SA2 BR3-3":"1.14","SA3 BR1-1":"1.24","SA3 BR1-2":"1.21","SA3 BR1-3":"1.22","SA3 BR2-1":"0.92","SA3 BR2-2":"0.91","SA3 BR2-3":"0.93","SA3 BR3-1":"1.23","SA3 BR3-2":"1.23","SA3 BR3-3":"1.22","WT EP BR1-1":"0.7514","WT EP BR1-2":"0.731","WT EP BR1-3":"0.7729","WT EP BR2-1":"0.7506","WT EP BR2-2":"0.7395","WT EP BR2-3":"0.734","WT EP BR3-1":"0.7072","WT EP BR3-2":"0.7102","WT EP BR3-3":"0.6949","SA7 BR1-1":"0.5813","SA7 BR1-2":"0.5863","SA7 BR1-3":"0.5786","SA7 BR2-1":"0.366","SA7 BR2-2":"0.3653","SA7 BR2-3":"0.3647","SA7 BR3-1":"0.4387","SA7 BR3-2":"0.4482","SA7 BR3-3":"0.4454"},{"Metabolite":"FBP","WT BR1-1":"1.66","WT BR1-2":"1.73","WT BR1-3":"1.25","WT BR2-1":"1.63","WT BR2-2":"1.22","WT BR2-3":"1.79","WT BR3-1":"2.09","WT 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}

}