{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002431","ANALYSIS_ID":"AN003957","VERSION":"1","CREATED_ON":"January 5, 2023, 5:29 pm"},

"PROJECT":{"PROJECT_TITLE":"Linking genetic and phenotypic changes in the long term evolution experiment using metabolomics","PROJECT_TYPE":"MS","PROJECT_SUMMARY":"The metabolome of a cell is the integration point of an organism's environment, genetics, and gene expression pattern. The metabolic phenotype can be under selection and is known to contribute to adaption. However, the metabolome's inherent networked and convoluted nature makes relating mutations, metabolic changes, and effects on fitness challenging. To overcome this challenge, we use the Long Term Evolution Experiment (LTEE) as a model to understand how mutations can transduce themselves through a cellular network, eventually affecting metabolism and perhaps fitness. We used mass-spectropscopy to broadly survey the metabolomes of both ancestors and all 12 evolved lines and combined this with genomic and expression data to suggest how mutations that alter specific reaction pathways, such as the biosynthesis of nicotinamide adenine dinucleotide, might increase fitness in the system. Our work brings the field closer to a complete genotype-phenotype map for the LTEE and a better understanding of how mutations might affect fitness through the metabolome.","INSTITUTE":"Rutgers University","DEPARTMENT":"Genetics","LABORATORY":"Shah","LAST_NAME":"Favate","FIRST_NAME":"John","ADDRESS":"145 Bevier Road, Piscataway, NJ, 08854","EMAIL":"john.favate@rutgers.edu","PHONE":"7325984642"},

"STUDY":{"STUDY_TITLE":"MS profiling of the Long Term Evolution Experiment","STUDY_SUMMARY":"The metabolome of a cell is the integration point of an organism's environment, genetics, and gene expression pattern. The metabolic phenotype can be under selection and is known to contribute to adaption. However, the metabolome's inherent networked and convoluted nature makes relating mutations, metabolic changes, and effects on fitness challenging. To overcome this challenge, we use the Long Term Evolution Experiment (LTEE) as a model to understand how mutations can transduce themselves through a cellular network, eventually affecting metabolism and perhaps fitness. We used mass-spectropscopy to broadly survey the metabolomes of both ancestors and all 12 evolved lines and combined this with genomic and expression data to suggest how mutations that alter specific reaction pathways, such as the biosynthesis of nicotinamide adenine dinucleotide, might increase fitness in the system. Our work brings the field closer to a complete genotype-phenotype map for the LTEE and a better understanding of how mutations might affect fitness through the metabolome. We used mass-spectroscopy to profile metabolic changes in the Long Term Evolution Experiment and link these change to upstream changes in gene expression and mutations.","INSTITUTE":"Rutgers University","DEPARTMENT":"Genetics","LABORATORY":"Shah","LAST_NAME":"Favate","FIRST_NAME":"John","ADDRESS":"145 Bevier Road, Piscataway, NJ, 08854","EMAIL":"john.favate@rutgers.edu","PHONE":"7325894642"},

"SUBJECT":{"SUBJECT_TYPE":"Bacteria","SUBJECT_SPECIES":"Escherichia coli","TAXONOMY_ID":"562"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"am1_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am1_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am1_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am1_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am2_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am2_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am2_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am2_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am3_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am3_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am3_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am3_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am4_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am4_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am4_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am4_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am5_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am5_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am5_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am5_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am6_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am6_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am6_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"am6_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap1_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap1_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap1_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap1_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap2_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap2_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap2_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap2_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap3_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap3_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap3_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap3_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap4_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap4_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap4_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap4_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap5_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap5_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap5_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap5_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap6_e_r1",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap6_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap6_e_r2",
"Factors":{"age":"evolved","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"ap6_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel606_e_r1",
"Factors":{"age":"ancestral","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"rel606_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel606_e_r2",
"Factors":{"age":"ancestral","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"rel606_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel607_e_r1",
"Factors":{"age":"ancestral","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"rel607_e_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel607_e_r2",
"Factors":{"age":"ancestral","growth_phase":"exponential"},
"Additional sample data":{"RAW_FILE_NAME":"rel607_e_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am1_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am1_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am1_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am1_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am2_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am2_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am2_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am2_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am3_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am3_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am3_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am3_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am4_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am4_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am4_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am4_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am5_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am5_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am5_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am5_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am6_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am6_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"am6_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"am6_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap1_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap1_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap1_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap1_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap2_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap2_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap2_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap2_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap3_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap3_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap3_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap3_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap4_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap4_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap4_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap4_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap5_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap5_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap5_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap5_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap6_s_r1",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap6_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"ap6_s_r2",
"Factors":{"age":"evolved","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"ap6_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel606_s_r1",
"Factors":{"age":"ancestral","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"rel606_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel606_s_r2",
"Factors":{"age":"ancestral","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"rel606_s_r2.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel607_s_r1",
"Factors":{"age":"ancestral","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"rel607_s_r1.mzXML"}
},
{
"Subject ID":"-",
"Sample ID":"rel607_s_r2",
"Factors":{"age":"ancestral","growth_phase":"stationary"},
"Additional sample data":{"RAW_FILE_NAME":"rel607_s_r2.mzXML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Bacterial cells were grown in DM25 medium and collected via vacuum filtration at 2hr and 24hr timepoints.","SAMPLE_TYPE":"Bacterial cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"No treatment was applied to the samples. The samples consist of the ancestral strains of the LTEE, REL606 and REL607, and clones from each of the evolved lines, designated Ara-1:6 and Ara+1:6"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Samples were prepared using the 40:40:20 acetonitrile:methanol:water protocol."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Waters Acquity BEH Amide (150 x 2.1mm, 1.7um)","SOLVENT_A":"95% water/5% acetonitrile; 20 mM acetic acid; 40 mM ammonium hydroxide, pH 9.4","SOLVENT_B":"20% water/80% acetonitrile; 20 mM acetic acid; 40 mM ammonium hydroxide, pH 9.4","FLOW_GRADIENT":"0 min, 100% B; 3 min, 100% B; 3.2 min, 90% B; 6.2 min, 90% B; 6.5 min, 80% B; 10.5 min, 80% B; 10.7 min, 70% B; 13.5 min, 70% B; 13.7 min, 45% B; 16 min, 45% B; 16.5 min, 100% B; and 22 min, 100% B","FLOW_RATE":"0.3mL/min","COLUMN_TEMPERATURE":"25"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"EL-MAVEN"},

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

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CoA"},{"metabolite_name":"NADH"},{"metabolite_name":"Glutathione"},{"metabolite_name":"Aspartate"},{"metabolite_name":"Glutamate"},{"metabolite_name":"N-Acetylaspartate"},{"metabolite_name":"FAD"},{"metabolite_name":"Butyryl CoA"},{"metabolite_name":"N-acetyl-glutamate"},{"metabolite_name":"Glycerophosphocholine"},{"metabolite_name":"Acetyllysine"},{"metabolite_name":"Itaconic acid"},{"metabolite_name":"N-acetyl-L-ornithine"},{"metabolite_name":"4-Aminobutyrate"},{"metabolite_name":"Asparagine"},{"metabolite_name":"Homocitrulline"},{"metabolite_name":"Serine"},{"metabolite_name":"S-adenosyl-L-homocysteine"},{"metabolite_name":"Glutamine"},{"metabolite_name":"Histidine"},{"metabolite_name":"Acetyl-arginine"},{"metabolite_name":"Carnitine"},{"metabolite_name":"Threonine"},{"metabolite_name":"4-Guanidinobutanoic acid"},{"metabolite_name":"Alanine"},{"metabolite_name":"Thiamine"},{"metabolite_name":"Proline"},{"metabolite_name":"N-Acetylputrescine"},{"metabolite_name":"Pipecolic acid"},{"metabolite_name":"Dimethylglycine"},{"metabolite_name":"Pyridoxamine"},{"metabolite_name":"Methylnicotinamide"},{"metabolite_name":"Valine"},{"metabolite_name":"Tyrosine"},{"metabolite_name":"Xanthosine"},{"metabolite_name":"Betaine"},{"metabolite_name":"1-Methylimidazole acetic acid"},{"metabolite_name":"Homocysteine"},{"metabolite_name":"Urocanic acid"},{"metabolite_name":"Cyclic AMP"},{"metabolite_name":"Methionine"},{"metabolite_name":"Pantothenate"},{"metabolite_name":"Isoleucine"},{"metabolite_name":"Acetyl proline"},{"metabolite_name":"Leucine"},{"metabolite_name":"Pterin"},{"metabolite_name":"Phenylalanine"},{"metabolite_name":"Guanine"},{"metabolite_name":"Tryptophan"},{"metabolite_name":"Nicotinate"},{"metabolite_name":"1 3-Dimethyluric acid"},{"metabolite_name":"5-Aminoimidazole-4-carboxamide ribonucleoside (AICAr)"},{"metabolite_name":"N-Acetyltyrosine"},{"metabolite_name":"Deoxyinosine"},{"metabolite_name":"Hydroxyindoleacetic acid"},{"metabolite_name":"5-Methylcytosine"},{"metabolite_name":"Creatinine"},{"metabolite_name":"Hypoxanthine"},{"metabolite_name":"Acetyl-Methionine"},{"metabolite_name":"Uridine"},{"metabolite_name":"Hydroxyphenyllactic acid"},{"metabolite_name":"Adenosine"},{"metabolite_name":"Hydroxymandelic acid"},{"metabolite_name":"Adenine"},{"metabolite_name":"Phenylethanolamine"},{"metabolite_name":"Tryptamine"},{"metabolite_name":"Leucic acid"},{"metabolite_name":"Acetyl-Phenylalanine"},{"metabolite_name":"2 2-dimethylsuccinic acid"},{"metabolite_name":"Uracil"},{"metabolite_name":"Mandelic acid"},{"metabolite_name":"Hydroquinone"},{"metabolite_name":"Phenylacetylglycine"},{"metabolite_name":"Deoxyadenosine"},{"metabolite_name":"Phenyllactic acid"},{"metabolite_name":"Thymidine"},{"metabolite_name":"5- Methylthioadenosine"},{"metabolite_name":"3-Phenylbutyric acid"},{"metabolite_name":"Dihydroxybenzeneacetic acid"},{"metabolite_name":"Androstenedione"},{"metabolite_name":"11a-Hydroxyprogesterone"},{"metabolite_name":"Indole"},{"metabolite_name":"Tetrahydrocortisone"},{"metabolite_name":"3-Cresotinic acid"},{"metabolite_name":"Spermine"},{"metabolite_name":"3-Phosphoglycerate"},{"metabolite_name":"3-Phosphoserine"},{"metabolite_name":"6-Phosphogluconate"},{"metabolite_name":"CDP"},{"metabolite_name":"Citrulline"},{"metabolite_name":"Cytosine"},{"metabolite_name":"D-sedoheptulose-1 7-phosphate"},{"metabolite_name":"Dihydroorotate"},{"metabolite_name":"GDP"},{"metabolite_name":"Guanidinoacetate"},{"metabolite_name":"IMP"},{"metabolite_name":"Guanosine"},{"metabolite_name":"Inosine"},{"metabolite_name":"L-Octanoylcarnitine"},{"metabolite_name":"L-Palmitoylcarnitine"},{"metabolite_name":"Mannose-6-phosphate"},{"metabolite_name":"N-acetyl-glutamine"},{"metabolite_name":"O-Butanoylcarnitine"},{"metabolite_name":"O-Decanoyl-L-carnitine"},{"metabolite_name":"O-Propanoylcarnitine"},{"metabolite_name":"Phosphoenolpyruvate"},{"metabolite_name":"Ribulose-5-phosphate"},{"metabolite_name":"UDP"},{"metabolite_name":"Xylose-5-phosphate"},{"metabolite_name":"PosIS_Alanine-2H4-15N"},{"metabolite_name":"PosIS_Glutamine-2H5-15N2"},{"metabolite_name":"PosIS_Inosine-15N4"},{"metabolite_name":"PosIS_Lysine-2H8"},{"metabolite_name":"PosIS_Serine-13C3-15N1"}]
}

}