#METABOLOMICS WORKBENCH ariesm_20200320_145353 DATATRACK_ID:1948 STUDY_ID:ST001351 ANALYSIS_ID:AN002247 PROJECT_ID:PR000921
VERSION             	1
CREATED_ON             	April 7, 2020, 5:33 pm
#PROJECT
PR:PROJECT_TITLE                 	NMR Metabolomic Analysis of Bacterial Resistance Pathways Using Multivalent
PR:PROJECT_TITLE                 	Quaternary Ammonium Functionalized Macromolecules
PR:PROJECT_TYPE                  	NMR Hydrophilic Metabolomic Analysis
PR:PROJECT_SUMMARY               	Multivalent antimicrobial dendrimers are an exciting new system that is being
PR:PROJECT_SUMMARY               	developed to address the growing problem of drug resistant bacteria. Nuclear
PR:PROJECT_SUMMARY               	Magnetic Resonance (NMR) metabolomics is a quantitative and reproducible method
PR:PROJECT_SUMMARY               	for the determination of bacterial response to environmental stressors and for
PR:PROJECT_SUMMARY               	visualization of perturbations to biochemical pathways. NMR metabolomics is used
PR:PROJECT_SUMMARY               	to elucidate metabolite differences between wild type and antimicrobially
PR:PROJECT_SUMMARY               	mutated Escherichia coli (E. coli) samples. Proton (1H) NMR hydrophilic
PR:PROJECT_SUMMARY               	metabolite analysis was conducted on samples of E. coli after 33 growth cycles
PR:PROJECT_SUMMARY               	of a minimum inhibitory challenge to E. coli by poly(amidoamine) dendrimers
PR:PROJECT_SUMMARY               	functionalized with mannose and with C16-DABCO quaternary ammonium endgroups and
PR:PROJECT_SUMMARY               	compared to the metabolic profile of wild type E. coli. The wild type and
PR:PROJECT_SUMMARY               	mutated E. coli samples were separated into distinct sample sets by hierarchical
PR:PROJECT_SUMMARY               	clustering, principal component analysis (PCA) and sparse partial least squares
PR:PROJECT_SUMMARY               	discriminate analysis (sPLS-DA). Metabolite components of membrane fortification
PR:PROJECT_SUMMARY               	and energy related pathways had a significant p-value and fold change between
PR:PROJECT_SUMMARY               	the wild type and mutated E. coli. Amino acids commonly associated with membrane
PR:PROJECT_SUMMARY               	fortification from cationic antimicrobials, such as lysine, were found to have a
PR:PROJECT_SUMMARY               	higher concentration in the mutated E. coli than the wild type E. coli.
PR:PROJECT_SUMMARY               	N-acetylglucosamine, a major component of peptidoglycan synthesis, was found to
PR:PROJECT_SUMMARY               	have a 25 fold higher concentration in the mid log phase of the mutated E. coli
PR:PROJECT_SUMMARY               	than the mid log phase of the wild type. The metabolic profile suggests that E.
PR:PROJECT_SUMMARY               	coli change their peptidoglycan composition in order to garner protection from
PR:PROJECT_SUMMARY               	the highly positively charged and multivalent C16 DABCO and mannose
PR:PROJECT_SUMMARY               	functionalized dendrimer.
PR:INSTITUTE                     	Montana State University
PR:DEPARTMENT                    	Chemistry and Biochemistry
PR:LABORATORY                    	Cloninger
PR:LAST_NAME                     	Aries
PR:FIRST_NAME                    	Michelle
PR:ADDRESS                       	Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT,
PR:ADDRESS                       	59717 USA
PR:EMAIL                         	michelle.aries@montana.edu
PR:PHONE                         	4069943051
#STUDY
ST:STUDY_TITLE                   	NMR Metabolomic Analysis of Bacterial Resistance Pathways Using Multivalent
ST:STUDY_TITLE                   	Quaternary Ammonium Functionalized Macromolecules
ST:STUDY_TYPE                    	NMR Hydrophilic Metabolomics
ST:STUDY_SUMMARY                 	Multivalent antimicrobial dendrimers are an exciting new system that is being
ST:STUDY_SUMMARY                 	developed to address the growing problem of drug resistant bacteria. Nuclear
ST:STUDY_SUMMARY                 	Magnetic Resonance (NMR) metabolomics is a quantitative and reproducible method
ST:STUDY_SUMMARY                 	for the determination of bacterial response to environmental stressors and for
ST:STUDY_SUMMARY                 	visualization of perturbations to biochemical pathways. NMR metabolomics is used
ST:STUDY_SUMMARY                 	to elucidate metabolite differences between wild type and antimicrobially
ST:STUDY_SUMMARY                 	mutated Escherichia coli (E. coli) samples. Proton (1H) NMR hydrophilic
ST:STUDY_SUMMARY                 	metabolite analysis was conducted on samples of E. coli after 33 growth cycles
ST:STUDY_SUMMARY                 	of a minimum inhibitory challenge to E. coli by poly(amidoamine) dendrimers
ST:STUDY_SUMMARY                 	functionalized with mannose and with C16-DABCO quaternary ammonium endgroups and
ST:STUDY_SUMMARY                 	compared to the metabolic profile of wild type E. coli. The wild type and
ST:STUDY_SUMMARY                 	mutated E. coli samples were separated into distinct sample sets by hierarchical
ST:STUDY_SUMMARY                 	clustering, principal component analysis (PCA) and sparse partial least squares
ST:STUDY_SUMMARY                 	discriminate analysis (sPLS-DA). Metabolite components of membrane fortification
ST:STUDY_SUMMARY                 	and energy related pathways had a significant p-value and fold change between
ST:STUDY_SUMMARY                 	the wild type and mutated E. coli. Amino acids commonly associated with membrane
ST:STUDY_SUMMARY                 	fortification from cationic antimicrobials, such as lysine, were found to have a
ST:STUDY_SUMMARY                 	higher concentration in the mutated E. coli than the wild type E. coli.
ST:STUDY_SUMMARY                 	N-acetylglucosamine, a major component of peptidoglycan synthesis, was found to
ST:STUDY_SUMMARY                 	have a 25 fold higher concentration in the mid log phase of the mutated E. coli
ST:STUDY_SUMMARY                 	than the mid log phase of the wild type.The metabolic profile suggests that E.
ST:STUDY_SUMMARY                 	coli change their peptidoglycan composition in order to garner protection from
ST:STUDY_SUMMARY                 	the highly positively charged and multivalent C16-DABCO and mannose
ST:STUDY_SUMMARY                 	functionalized dendrimer.
ST:INSTITUTE                     	Montana State University
ST:DEPARTMENT                    	Chemistry and Biochemistry
ST:LABORATORY                    	Cloninger
ST:LAST_NAME                     	Aries
ST:FIRST_NAME                    	Michelle
ST:ADDRESS                       	Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT,
ST:ADDRESS                       	59717 USA
ST:EMAIL                         	michelle.aries@montana.edu
ST:PHONE                         	4069943051
#SUBJECT
SU:SUBJECT_TYPE                  	Bacteria
SU:SUBJECT_SPECIES               	Escherichia coli
SU:TAXONOMY_ID                   	562
SU:GENOTYPE_STRAIN               	ATCC 25922
#FACTORS
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	WT_S_1	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=fid - WT_2_1 - S
SUBJECT_SAMPLE_FACTORS           	-	WT_S_2	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=fid - WT_2_2 - S
SUBJECT_SAMPLE_FACTORS           	-	WT_S_5	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_S_6	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_S_7	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_S_9	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=fid - WT_2_9 - S
SUBJECT_SAMPLE_FACTORS           	-	WT_S_10	Genotype:Wild Type | Phase:Stationary	RAW_FILE_NAME=fid - WT_2_10 - S
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_1	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_2	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=fid - Mut_2_2 - S
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_3	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_7	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_8	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_9	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=fid - Mut_2_9 - S
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_10	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut_S_12	Genotype:Mutant | Phase:Stationary	RAW_FILE_NAME=fid - Mut_2_12 - S
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_1	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=fid - Mut_1_2 - ML
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_2	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_3	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_4	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=fid - Mut_1_4 - ML
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_6	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_7	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_8	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_9	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	Mut _ML_10	Genotype:Mutant | Phase:Mid Log	RAW_FILE_NAME=fid - Mut_1_10 - ML
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_1	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=fid - WT_1_1 - ML
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_2	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=fid - WT_1_2 - ML
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_3	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_6	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_7	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_8	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=fid - WT_1_8 - ML
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_9	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_10	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_11	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_12	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=-
SUBJECT_SAMPLE_FACTORS           	-	WT_ML_13	Genotype:Wild Type | Phase:Mid Log	RAW_FILE_NAME=fid - WT_1_13 -ML
#COLLECTION
CO:COLLECTION_SUMMARY            	Samples were collected in the mid log and stationary phases, washed with cold
CO:COLLECTION_SUMMARY            	PBS, frozen at -80 ºC. Please see attached PDF for detailed procedures.
CO:SAMPLE_TYPE                   	Bacterial cells
#TREATMENT
TR:TREATMENT_SUMMARY             	The samples were not treated with anything. I collected the samples then
TR:TREATMENT_SUMMARY             	extracted the hydrophilic metabolites, placed them in an NMR buffer then ran
TR:TREATMENT_SUMMARY             	them on the NMR. All these procedures were uploaded.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The hydrophilic metabolites were extracted, the samples were added to an NMR
SP:SAMPLEPREP_SUMMARY            	buffer, run on the NMR and profiled, standardized to their concentrations and
SP:SAMPLEPREP_SUMMARY            	analyzed. See uploaded protocol for more details.
#ANALYSIS
AN:ANALYSIS_TYPE                 	NMR
#NMR
NM:INSTRUMENT_NAME               	Buker Avance III
NM:INSTRUMENT_TYPE               	FT-NMR
NM:NMR_EXPERIMENT_TYPE           	1D-1H
NM:SPECTROMETER_FREQUENCY        	600 MHz
#NMR_METABOLITE_DATA
NMR_METABOLITE_DATA:UNITS	mM
NMR_METABOLITE_DATA_START
Samples	WT_S_1	WT_S_2	WT_S_5	WT_S_6	WT_S_7	WT_S_9	WT_S_10	Mut_S_1	Mut_S_2	Mut_S_3	Mut_S_7	Mut_S_8	Mut_S_9	Mut_S_10	Mut_S_12	Mut _ML_1	Mut _ML_2	Mut _ML_3	Mut _ML_4	Mut _ML_6	Mut _ML_7	Mut _ML_8	Mut _ML_9	Mut _ML_10	WT_ML_1	WT_ML_2	WT_ML_3	WT_ML_6	WT_ML_7	WT_ML_8	WT_ML_9	WT_ML_10	WT_ML_11	WT_ML_12	WT_ML_13
Factors	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Wild Type | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Stationary	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Mutant | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log	Genotype:Wild Type | Phase:Mid Log
Acetate	5.004207	1.420349	3.673355	2.396972	2.696732	2.342503	3.050962	2.195773	0.663715	0.3064	1.279346	1.157738	1.651761	0.362918	0.892798	4.604748	3.940933	2.762371	3.306119	2.218623	4.01517	3.520154	3.1311	4.27284	3.696428	4.507632	6.053971	4.605458	5.477735	5.041596	4.587081	4.10203	5.280802	5.514488	4.901498
Acetoacetate	0.24309	0.123455	0.283431	0.10069	0.306197	0.323773	0.288621	0.139559	0.066026	0.225876	0.14557	0.103174	0.140322	0.151581	0.11931	0.671179	0.081061	0.421237	0.272755	0.56972	0.296206	0.258009	0.278807	0.37612	0	0.159002	0.052112	0	0.084259	0.042129	0.042129	0.079501	0.105557	0	0.116406
Adenosine	0.61653	0.066666	0.497739	0.086758	0.477647	0.378948	0.576345	0.936612	0.878938	0.978256	0.967761	0.522493	0.166048	0.99891	0.681298	0.638963	0.890187	0.385446	0.393061	0.377831	0.604839	0.711331	0.335407	0.764575	0.707489	0.428092	0.272013	0.491446	0.291119	0.391282	0.499304	0.567791	0.350053	0.275403	0.310225
Alanine	3.095242	3.603872	3.238241	3.640412	3.201702	3.381241	3.022163	3.745169	2.799352	3.728143	3.126668	3.661522	4.523691	2.508166	3.27707	3.633135	4.2206	4.285751	4.944485	3.627018	4.11156	4.209	4.383348	3.926867	5.898304	5.433286	5.261134	5.754285	5.142107	5.448196	5.520206	5.665795	5.34721	5.610267	5.023081
AMP	0.667406	0.900793	0.625175	1.012276	0.513692	0.582944	0.444441	1.021736	0.967562	0.70847	0.855155	1.329448	1.691333	0.688573	1.115823	1.220935	0.583634	0.543861	0.529897	0.557824	0.884384	0.560594	0.696222	0.902284	0.267592	0.631895	0.274798	0.265641	0.448609	0.357125	0.3581	0.449743	0.453346	0.263691	0.62242
Aspartate	6.022608	4.25488	5.513727	5.45505	4.313557	5.004846	3.622267	7.298124	5.442504	6.527577	7.484072	5.890425	6.338346	7.67002	6.483623	7.758444	12.54406	6.876525	9.075297	4.677754	10.43766	10.41235	8.943502	10.15125	5.573016	8.877851	5.391347	5.582218	6.955479	6.268848	6.264247	7.225434	7.134599	5.59142	8.519611
Betaine	0.409897	0.624221	0.417007	0.562086	0.479143	0.424118	0.534167	0.459575	0.767859	0.47037	0.599755	0.589995	0.412131	0.739935	0.639975	0.759292	0.694907	0.922818	0.982652	0.862985	0.825154	0.769228	0.972041	0.727099	0.45394	0.555435	0.345407	0.398049	0.394515	0.396282	0.424227	0.504687	0.450421	0.342158	0.443623
Cysteine	1.321165	2.693369	2.020742	2.719391	1.99472	2.72032	1.26912	2.033308	1.614617	2.668459	1.862824	2.209791	2.804966	1.692341	2.037308	2.183831	1.398319	2.489392	1.777957	3.200827	1.671801	1.869338	1.961323	1.791075	3.417311	2.66579	3.087591	3.04343	3.354397	3.198914	3.385854	3.04155	2.876691	2.669549	3.621203
Cytosine	0.462037	0.195059	0.280464	0.278467	0.197057	0.098891	0.295222	0.358148	0.608798	0.179239	0.341424	0.496371	0.383944	0.324701	0.439148	0	0.180636	0.099953	0	0.199906	0.060212	0.153741	0.049977	0.090318	0	0	0	0	0	0	0	0	0	0	0
Formate	0.417895	0.659218	0.483477	0.558975	0.583721	0.549059	0.618382	0.679903	0.694903	0.523275	0.527521	0.826128	0.957354	0.375138	0.675798	1.55034	0.886207	1.127311	1.394999	0.859623	1.044682	1.222226	0.912405	1.218274	0.443544	0.425953	0.384413	0.371388	0.344614	0.358001	0.394079	0.434749	0.405183	0.299232	0.304815
Fucose	0.144041	0.433328	0.358468	0.288685	0.503111	0.572895	0.433328	0	0.844496	0	0.220528	0.422248	0	0.441057	0.428518	0.505916	0.152094	0.495804	0.248877	0.742731	0.311514	0.28316	0.386168	0.329005	1.608868	0.093295	0	1.608868	0.046648	0.827758	0.827758	0.851082	0.046648	1.608868	0.093295
Fumarate	0.157433	0.046913	0.220996	0.048546	0.219364	0.28456	0.154167	0.183029	0.148548	0.134083	0.165692	0.149115	0.149682	0.148355	0.148862	0.590333	1.021356	1.725985	2.129692	1.322278	0.907359	1.248642	1.418187	0.805844	0.416222	1.694952	0.584747	0.502094	0.74279	0.622442	0.579506	1.055587	1.13985	0.587966	0.900833
Glutamate	8.661314	10.26138	10.03912	10.41403	9.886475	11.41693	8.356019	16.24401	9.487716	12.47825	14.51173	12.55416	15.6206	12.77946	12.62926	14.34389	14.04207	13.38274	12.54213	14.22334	13.92286	14.31771	13.38268	14.19298	6.938229	8.758527	9.502878	8.049554	9.145848	8.597701	8.042038	7.848378	9.130703	9.16088	8.788818
Glycine	0.192923	0.33749	0.172573	0.268287	0.241776	0.152223	0.33133	0.310767	0.477869	0.443713	0.40964	0.397878	0.317886	0.508513	0.434756	0.692909	0.71028	0.393948	0.397653	0.390244	0.574582	0.624741	0.357253	0.701595	1.327406	0.938182	1.085668	1.310741	0.895778	1.103259	1.111592	1.132794	1.011925	1.294077	0.705887
Histidine	1.570525	1.291414	1.626615	1.187657	1.730372	1.682705	1.778039	0.80834	1.338867	1.037878	0.868342	1.160881	0.982894	0.928344	1.083369	0.768991	0.037769	0.815597	0.933979	0.697215	0.424593	0.326349	0.641307	0.40338	0.44262	1.019327	1.097865	0.734506	0.987599	0.861053	0.71511	0.730974	1.058596	1.026392	0.877334
Homocysteine	0.628704	0	0.314352	0.314352	0	0	0	0	0	1.120523	0.560261	0	0	1.120523	0.373508	4.425639	1.335668	5.675852	1.72561	9.626093	2.65326	2.748859	3.937162	2.880654	0.959736	0.948677	0.930711	0.945224	0.939694	0.942459	0.949715	0.954207	0.939694	0.930711	0.948677
Isocitrate	5.194404	3.943877	4.150518	4.664472	3.429922	3.106631	3.753213	4.936221	11.23881	5.616277	7.823816	9.053472	6.868138	10.71141	9.606118	3.579189	3.606699	4.074702	3.127956	5.021448	3.41451	3.807606	3.566173	3.592944	4.179185	3.093981	3.116632	4.440589	2.765644	3.603117	3.472415	3.636583	3.105307	4.701994	2.414656
Isoleucine	8.032809	8.434944	7.724972	7.73371	8.426207	7.417136	9.435278	4.50923	6.231235	5.172992	4.729615	4.996316	3.761398	4.95	4.980877	4.956644	4.184869	4.997374	4.601934	5.392813	4.288808	4.456005	4.361142	4.570756	6.256907	6.905562	6.555357	6.018087	6.548617	6.283352	6.402762	6.581234	6.73046	5.779267	6.541877
Lactate	0.570848	0.820036	0.661036	0.854604	0.626467	0.751223	0.50171	0.700565	0.828274	0.710102	0.688568	0.908725	0.989176	0.676571	0.83134	0.483578	0.601071	1.541712	0.619897	2.463527	0.583411	0.905283	1.103648	0.542324	0.375553	0.398455	0.439171	0.382887	0.433438	0.408163	0.404496	0.387004	0.418813	0.39022	0.427706
Leucine	12.5155	13.01003	14.28715	11.93343	15.36375	16.05879	14.6687	8.13346	11.34785	9.304006	8.512498	9.601222	7.854594	8.891535	9.36466	8.671739	9.097455	9.490957	9.461929	9.519985	8.895977	9.174604	9.204847	8.884597	10.69344	11.14734	10.84135	10.33886	10.04382	10.19134	10.36863	10.92039	10.99435	9.984277	9.246281
Lysine	7.314858	7.456419	7.361195	7.04127	7.776344	7.407532	8.145155	9.290829	9.022589	11.95526	11.53247	7.149046	5.275503	13.77411	9.357401	5.150635	4.490196	4.777008	5.019791	4.534225	4.756544	4.56483	4.702905	4.820415	3.148868	4.560288	5.128069	3.948319	5.106084	4.527202	4.127476	3.854578	4.844179	4.74777	5.0841
Methionine	3.075653	3.539427	3.296146	3.502394	3.333179	3.516638	3.14972	2.361894	2.163787	2.585489	2.015332	2.411333	2.658879	1.66877	2.163812	1.889732	2.571249	2.909464	3.836935	1.981994	2.550808	2.603098	3.050453	2.230491	2.812813	3.258294	2.580328	2.562455	2.652872	2.607663	2.732843	3.035554	2.919311	2.312096	2.725416
N-Acetylglucosamine	0	0.241355	0.190812	0.120678	0.311489	0.381623	0.241355	0	0.401441	0	0.20072	0.20072	0	0.401441	0	0.250356	0	1.407231	0	2.814462	0.083452	0.469077	0.703616	0.125178	0	0	0.078995	0	0.078995	0	0	0	0.039498	0	0.078995
NAD+	1.31022	0.619562	0.960043	0.93589	0.643715	0.609867	0.677564	2.101301	1.909536	1.697186	1.932121	2.098615	2.287694	1.762941	1.986724	2.778196	1.467438	1.339164	1.590611	1.087717	2.002867	1.649928	1.551066	2.122817	0.823942	1.183063	1.096535	0.934168	1.247749	1.090958	1.035845	1.003502	1.139799	1.044394	1.398962
Phenylalanine	3.798833	4.693122	4.205861	4.398568	4.500416	4.612889	4.387942	5.322827	2.446779	2.786141	3.727654	2.614432	2.782085	2.132481	2.453782	3.308325	2.233033	2.89435	2.770711	3.017989	2.689841	2.445924	2.711258	2.770679	4.424605	4.38187	3.222525	4.169726	3.323858	3.746792	3.874232	4.403238	3.802198	3.914847	3.425191
Proline	1.634986	0.570686	1.50528	0.92282	1.153146	1.375573	0.930719	1.244095	1.480111	1.013347	1.316515	1.345825	1.211539	1.388935	1.360195	6.551735	6.084411	4.123363	4.983056	3.26367	5.932722	5.212312	4.642692	6.318073	9.043043	7.556447	7.292855	8.658348	6.919372	7.78886	7.981208	8.299745	7.424651	8.273653	6.54589
Pyroglutamate	7.056526	9.122495	7.261623	8.161865	8.222253	7.466719	8.977787	4.278205	7.057067	3.789291	4.52454	5.824026	4.590985	4.770875	5.472976	3.944545	4.576674	3.509245	0.178163	0	4.032484	4.241556	3.542738	4.26061	5.281217	4.94951	3.846756	4.884861	4.146433	4.515647	4.713825	5.115363	4.398133	4.488506	4.44611
Succinate	2.794128	3.662377	3.407701	4.200158	2.86992	4.021274	1.718566	4.608149	6.528634	6.425008	3.633121	8.747076	10.96552	2.658093	6.717415	5.428326	6.800578	4.854486	6.046524	3.662448	7.286272	5.922236	7.242199	6.114452	7.157614	2.999096	8.387215	7.063175	9.254248	8.158711	8.205931	5.078355	5.693155	6.968736	10.12128
Tyrosine	4.47862	4.558224	4.410347	4.325739	4.642832	4.342073	4.94359	5.764929	3.282707	4.983996	4.835299	3.636291	3.989876	3.90567	3.726084	2.592496	2.926665	3.450796	4.482546	2.419046	2.950318	3.07141	3.391294	2.75958	3.790603	4.342158	3.485258	3.693336	3.434798	3.564067	3.612701	4.066381	3.913708	3.596068	3.384339
UDP-glucose	0.547829	0.462892	0.273914	0.542365	0.194442	0	0.388884	1.029476	0.648644	0.517398	0.956103	0.927882	1.20712	0.88273	0.912831	0.29204	0.672094	0.353843	0.269081	0.438604	0.485894	0.553017	0.423696	0.482067	0.125674	0.189888	0	0.125674	0	0.062837	0.062837	0.157781	0.094944	0.125674	0
Valine	10.36675	9.569699	8.70507	8.830361	9.444407	7.043386	11.84543	5.655956	7.02659	7.783004	5.76363	6.648744	6.270897	5.871304	6.389597	4.748054	6.343726	6.476718	7.190809	5.762627	5.850928	6.222979	6.468861	5.54589	8.299831	6.664932	8.223501	7.854432	7.332665	7.593548	7.816248	7.482381	7.444217	7.409034	6.441828
NMR_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	KEGG	PubChem	CAS #
Acetate	C00033	175	71-50-1
Acetoacetate	C00164	6971017	541-50-4
Adenosine	C00212	60961	58-61-7
Alanine	C01401	5950	56-41-7
AMP	C00020	6083	61-19-8
Aspartate	C00049	5960	56-84-8
Betaine	C00719	247	107-43-7
Cysteine	C00097	5862	54-90-4
Cytosine	C00380	597	71-30-7
Formate	C00058	283	141-53-7
Fucose	C02095	17106	2438-80-4
Fumarate	C00122	5460307	142-42-7
Glutamate	C00302	33032	56-86-0
Glycine	C00037	750	56-40-6
Histidine	C00135	6274	71-00-1
Homocysteine	C05330	91552	6027-13-0
Isocitrate	C00311	1198	320-77-4
Isoleucine	C16434	6306	73-32-5
Lactate	C00186	612	50-21-5
Leucine	C00123	6106	61-90-5
Lysine	C00047	5962	56-87-1
Methionine	C00073	6137	59-51-8
N-Acetylglucosamine	C00043	24139	7512-17-6
NAD+	C00003	5892	53-84-9
Phenylalanine	C00079	6140	63-91-2
Proline	C00148	145742	609-36-9
Pyroglutamate	C01879	7405	28874-51-3
Succinate	C00042	160419	110-15-6
Tyrosine	C00082	6057	60-18-4
UDP-glucose	C00029	8629	133-89-1
Valine	C00183	6287	72-18-4
METABOLITES_END
#END