#METABOLOMICS WORKBENCH uchimiya_20231029_173020 DATATRACK_ID:4425 STUDY_ID:ST002957 ANALYSIS_ID:AN004857 PROJECT_ID:PR001837 VERSION 1 CREATED_ON November 1, 2023, 11:05 pm #PROJECT PR:PROJECT_TITLE Metabolite flux from temperature-acclimated diatom strains PR:PROJECT_SUMMARY The temperature increase occurring in the surface ocean has fundamental PR:PROJECT_SUMMARY implications for physiological rates and processes of marine microbes. Here we PR:PROJECT_SUMMARY asked whether the temperature at which a marine diatom strain is acclimated PR:PROJECT_SUMMARY affects carbon transfer to a co-cultured heterotrophic bacterium. Model systems PR:PROJECT_SUMMARY were established in which the diatom Thalassiosira pseudonana was acclimated for PR:PROJECT_SUMMARY three months at temperatures below (14°C), equal to (20°C), and above (28°C) PR:PROJECT_SUMMARY the temperature of optimal growth, and then inoculated with the heterotrophic PR:PROJECT_SUMMARY bacterium Ruegeria pomeroyi. This deposition is for the results of diatom PR:PROJECT_SUMMARY endometabolites obtained from the main experiment of this study. PR:INSTITUTE University of Georgia PR:LABORATORY Moran Lab, Edison Lab PR:LAST_NAME Uchimiya PR:FIRST_NAME Mario PR:ADDRESS 315 Riverbend Rd, Athens, GA, 30602, USA PR:EMAIL mario.uchimiya@uga.edu PR:PHONE (706) 542-8387 PR:FUNDING_SOURCE National Science Foundation, Joint Genome Institute, Swedish Research Council PR:PROJECT_COMMENTS See also Metabolomics Workbench_UGA_temp_Oct2023_main.docx for details about PR:PROJECT_COMMENTS this deposition. PR:CONTRIBUTORS Malin Olofsson #STUDY ST:STUDY_TITLE Metabolite flux from temperature-acclimated diatom strains (main experiment) ST:STUDY_SUMMARY The temperature increase occurring in the surface ocean has fundamental ST:STUDY_SUMMARY implications for physiological rates and processes of marine microbes. Here we ST:STUDY_SUMMARY asked whether the temperature at which a marine diatom strain is acclimated ST:STUDY_SUMMARY affects carbon transfer to a co-cultured heterotrophic bacterium. Model systems ST:STUDY_SUMMARY were established in which the diatom Thalassiosira pseudonana was acclimated for ST:STUDY_SUMMARY three months at temperatures below (14°C), equal to (20°C), and above (28°C) ST:STUDY_SUMMARY the temperature of optimal growth, and then inoculated with the heterotrophic ST:STUDY_SUMMARY bacterium Ruegeria pomeroyi. This deposition is for the results of diatom ST:STUDY_SUMMARY endometabolites obtained from the main experiment of this study. ST:INSTITUTE University of Georgia ST:LABORATORY Moran Lab, Edison Lab ST:LAST_NAME Uchimiya ST:FIRST_NAME Mario ST:ADDRESS 315 Riverbend Rd, Athens, GA, 30602, USA ST:EMAIL mario.uchimiya@uga.edu ST:PHONE (706) 542-8387 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Thalassiosira pseudonana SU:TAXONOMY_ID 296543 #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 Endometabolome 1 Temperature:14 | Bacteria presence:1 RAW_FILE_NAME=8; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=14°C Co-culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 2 Temperature:14 | Bacteria presence:1 RAW_FILE_NAME=12; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=14°C Co-culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 3 Temperature:14 | Bacteria presence:1 RAW_FILE_NAME=16; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=14°C Co-culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 4 Temperature:14 | Bacteria presence:1 RAW_FILE_NAME=20; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=14°C Co-culture; Biological replicate=4 SUBJECT_SAMPLE_FACTORS Endometabolome 5 Temperature:20 | Bacteria presence:1 RAW_FILE_NAME=24; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=20°C Co-culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 6 Temperature:20 | Bacteria presence:1 RAW_FILE_NAME=28; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=20°C Co-culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 7 Temperature:20 | Bacteria presence:1 RAW_FILE_NAME=32; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=20°C Co-culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 8 Temperature:20 | Bacteria presence:1 RAW_FILE_NAME=36; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=20°C Co-culture; Biological replicate=4 SUBJECT_SAMPLE_FACTORS Endometabolome 9 Temperature:28 | Bacteria presence:1 RAW_FILE_NAME=40; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=28°C Co-culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 10 Temperature:28 | Bacteria presence:1 RAW_FILE_NAME=44; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=28°C Co-culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 11 Temperature:28 | Bacteria presence:1 RAW_FILE_NAME=48; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=28°C Co-culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 12 Temperature:28 | Bacteria presence:1 RAW_FILE_NAME=52; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335 co-cultured with the heterotrophc bacteria Ruegeria pomeroyi DSS-3; Experiment name=Main experiment; Sample descritpion=28°C Co-culture; Biological replicate=4 SUBJECT_SAMPLE_FACTORS Endometabolome 13 Temperature:14 | Bacteria presence:0 RAW_FILE_NAME=56; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=14°C Axenic culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 14 Temperature:14 | Bacteria presence:0 RAW_FILE_NAME=60; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=14°C Axenic culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 15 Temperature:14 | Bacteria presence:0 RAW_FILE_NAME=64; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=14°C Axenic culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 16 Temperature:14 | Bacteria presence:0 RAW_FILE_NAME=68; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=14°C Axenic culture; Biological replicate=4 SUBJECT_SAMPLE_FACTORS Endometabolome 17 Temperature:20 | Bacteria presence:0 RAW_FILE_NAME=72; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=20°C Axenic culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 18 Temperature:20 | Bacteria presence:0 RAW_FILE_NAME=76; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=20°C Axenic culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 19 Temperature:20 | Bacteria presence:0 RAW_FILE_NAME=80; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=20°C Axenic culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 20 Temperature:20 | Bacteria presence:0 RAW_FILE_NAME=84; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=20°C Axenic culture; Biological replicate=4 SUBJECT_SAMPLE_FACTORS Endometabolome 21 Temperature:28 | Bacteria presence:0 RAW_FILE_NAME=88; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=28°C Axenic culture; Biological replicate=1 SUBJECT_SAMPLE_FACTORS Endometabolome 22 Temperature:28 | Bacteria presence:0 RAW_FILE_NAME=92; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=28°C Axenic culture; Biological replicate=2 SUBJECT_SAMPLE_FACTORS Endometabolome 23 Temperature:28 | Bacteria presence:0 RAW_FILE_NAME=96; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=28°C Axenic culture; Biological replicate=3 SUBJECT_SAMPLE_FACTORS Endometabolome 24 Temperature:28 | Bacteria presence:0 RAW_FILE_NAME=100; Organism=Phytoplankton, marine diatom, Thalassiosira pseudonana CCMP1335, axenic; Experiment name=Main experiment; Sample descritpion=28°C Axenic culture; Biological replicate=4 #COLLECTION CO:COLLECTION_SUMMARY Based on pre-experimental axenic growth curves, harvests occurred at days 3 CO:COLLECTION_SUMMARY (28°C), 4 (20°C), and 6 (14°C), starting 7 h into the light cycle. At each CO:COLLECTION_SUMMARY harvest, subsamples of 600 mL were filtered onto 2.0-µm Isopore filters CO:COLLECTION_SUMMARY (Millipore, Burlington, MA) for diatom endometabolite analysis and stored in CO:COLLECTION_SUMMARY 50-mL centrifuge tubes at -80°C. CO:COLLECTION_PROTOCOL_FILENAME 2_Collection protocol_UGA_temp_Oct2023_main.docx CO:SAMPLE_TYPE Algae CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Six treatment combinations of a marine diatom strain Thalassiosira pseudonana TR:TREATMENT_SUMMARY CCMP1335 were prepared: treatments incubated axenically at either 14, 20, or 28 TR:TREATMENT_SUMMARY oC, and treatments co-cultured with a bacterial strain Ruegeria pomeroyi DSS-3 TR:TREATMENT_SUMMARY at the corresponding temperatures (four replicates for each). L1 media was used TR:TREATMENT_SUMMARY with NaH13CO3 as a source of bicarbonate and a salinity of 35 ppt. The diatom TR:TREATMENT_SUMMARY used for the co-cultured treatments was B12 stressed to emphasize the known TR:TREATMENT_SUMMARY co-existing system. The light cycle consisted of 16 h light (120 µmol photons TR:TREATMENT_SUMMARY m-2 s-1) and 8 h of dark. TR:TREATMENT_PROTOCOL_FILENAME 3_Treatment protocol_UGA_temp_Oct2023_main.docx #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Filter samples were transferred into 15-mL of ultrapure MilliQ water in 50-mL SP:SAMPLEPREP_SUMMARY tubes, and diatom cells were removed from the filters by sonication in an SP:SAMPLEPREP_SUMMARY ice-water bath for 7 min (cycle: 50 s on and 10 s off). The liquid fraction was SP:SAMPLEPREP_SUMMARY subsequently collected in new tubes and the procedure repeated three times, SP:SAMPLEPREP_SUMMARY after which fractions were combined and stored at -80°C until further SP:SAMPLEPREP_SUMMARY processing. Samples were lyophilized (Labconco, Kansas City, MO, USA) and SP:SAMPLEPREP_SUMMARY pellets mixed with 600 μL of phosphate buffer (30 mmol L-1 phosphate in SP:SAMPLEPREP_SUMMARY deuterated water, pH 7.4) and 1 mmol L-1 internal standard SP:SAMPLEPREP_SUMMARY 2,2-dimethyl-2-silapentane-5-sulfonate (DSS). Samples were vortexed for 5 min, SP:SAMPLEPREP_SUMMARY centrifuged at 20,800 relative centrifugal force (RCF) for 10 min, and SP:SAMPLEPREP_SUMMARY supernatants were transferred to 5-mm NMR tubes (Bruker, Billerica, MA, USA). SP:SAMPLEPREP_SUMMARY Extraction and buffer blank controls were also prepared. Additionally, one SP:SAMPLEPREP_SUMMARY pooled control sample was prepared by combining aliquots of all the samples and SP:SAMPLEPREP_SUMMARY used for annotation. SP:SAMPLEPREP_PROTOCOL_FILENAME 4_Sample preparation protocol_UGA_temp_Oct2023_main.docx #ANALYSIS AN:ANALYSIS_PROTOCOL_FILE 5_Analysis protocol_UGA_temp_Oct2023_main.docx #NMR NM:INSTRUMENT_NAME AVANCE III HD instrument (Bruker) NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 2D-1H-13C NM:SPECTROMETER_FREQUENCY 600 MHz NM:NMR_PROBE TCI cryoprobe NM:NMR_TUBE_SIZE 5-mm #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS Intensity NMR_METABOLITE_DATA_START Samples 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Factors Temperature:14 | Bacteria presence:1 Temperature:14 | Bacteria presence:1 Temperature:14 | Bacteria presence:1 Temperature:14 | Bacteria presence:1 Temperature:20 | Bacteria presence:1 Temperature:20 | Bacteria presence:1 Temperature:20 | Bacteria presence:1 Temperature:20 | Bacteria presence:1 Temperature:28 | Bacteria presence:1 Temperature:28 | Bacteria presence:1 Temperature:28 | Bacteria presence:1 Temperature:28 | Bacteria presence:1 Temperature:14 | Bacteria presence:0 Temperature:14 | Bacteria presence:0 Temperature:14 | Bacteria presence:0 Temperature:14 | Bacteria presence:0 Temperature:20 | Bacteria presence:0 Temperature:20 | Bacteria presence:0 Temperature:20 | Bacteria presence:0 Temperature:20 | Bacteria presence:0 Temperature:28 | Bacteria presence:0 Temperature:28 | Bacteria presence:0 Temperature:28 | Bacteria presence:0 Temperature:28 | Bacteria presence:0 Glycine-betaine 3.58E-11 2.68E-11 5.89E-11 3.57E-11 1.00E-10 1.04E-10 7.54E-11 6.69E-11 8.69E-10 1.00E-09 1.88E-09 8.25E-10 6.59E-09 7.69E-09 6.04E-09 6.07E-09 8.82E-09 9.96E-09 1.04E-08 9.37E-09 1.76E-08 1.39E-08 1.63E-08 1.16E-08 Proline 5.44E-10 4.72E-10 5.94E-10 6.30E-10 7.29E-10 6.96E-10 6.82E-10 5.85E-10 5.73E-10 4.98E-10 4.30E-10 3.61E-10 4.40E-10 5.71E-10 4.65E-10 4.43E-10 4.17E-10 4.77E-10 4.58E-10 4.91E-10 1.21E-10 8.43E-11 1.22E-10 1.62E-10 DHPS 2.14E-10 1.89E-10 2.15E-10 2.35E-10 2.56E-10 2.52E-10 2.39E-10 2.12E-10 1.45E-10 1.39E-10 1.32E-10 1.06E-10 2.12E-10 2.55E-10 1.99E-10 2.12E-10 1.69E-10 1.77E-10 1.96E-10 1.74E-10 1.24E-10 9.59E-11 1.21E-10 8.98E-11 DMSP 2.72E-11 2.13E-11 1.84E-11 2.48E-11 2.52E-11 2.67E-11 2.67E-11 1.43E-11 1.43E-11 1.60E-11 1.31E-11 9.89E-12 4.97E-10 5.01E-10 4.43E-10 5.97E-10 2.53E-10 3.24E-10 3.99E-10 3.18E-10 1.65E-10 1.89E-10 2.35E-10 1.01E-10 Homarine 8.88E-11 8.77E-11 1.08E-10 1.14E-10 1.01E-10 1.03E-10 9.39E-11 7.83E-11 2.05E-12 3.98E-12 3.23E-12 2.99E-12 1.73E-10 2.33E-10 1.83E-10 1.94E-10 1.55E-10 1.54E-10 1.64E-10 1.58E-10 3.73E-12 3.17E-12 4.08E-12 2.64E-12 beta-1,3-glucan 2.84E-09 3.11E-09 3.46E-09 4.05E-09 3.68E-09 3.94E-09 4.11E-09 3.78E-09 4.03E-09 4.05E-09 3.01E-09 3.12E-09 1.12E-09 1.50E-09 1.19E-09 1.31E-09 3.17E-10 6.40E-10 9.87E-10 8.27E-10 1.48E-09 1.27E-09 1.36E-09 1.21E-09 glycine 0 0 0 0 0 0 0 0 0 0 4.78E-13 0 9.41E-12 1.46E-11 8.61E-12 8.48E-12 1.55E-11 1.49E-11 1.54E-11 1.57E-11 8.26E-12 8.94E-12 1.05E-11 6.61E-12 glutamate 1.85E-10 1.70E-10 1.85E-10 2.02E-10 2.45E-10 2.31E-10 2.17E-10 1.94E-10 2.55E-10 2.27E-10 2.17E-10 1.81E-10 1.37E-10 2.01E-10 1.29E-10 1.31E-10 2.40E-10 2.19E-10 1.98E-10 1.92E-10 2.68E-10 2.29E-10 2.46E-10 1.76E-10 glutamine 8.38E-11 9.95E-11 9.60E-11 1.45E-10 1.09E-10 9.65E-11 1.04E-10 8.81E-11 1.49E-10 1.15E-10 8.44E-11 9.74E-11 3.85E-11 4.73E-11 4.41E-11 4.53E-11 9.43E-11 1.11E-10 1.01E-10 1.24E-10 7.04E-11 6.38E-11 9.71E-11 7.92E-11 alanine 5.97E-12 5.68E-12 4.70E-12 5.53E-12 7.51E-12 6.78E-12 5.43E-12 5.05E-12 6.67E-12 4.54E-12 5.42E-12 3.96E-12 8.03E-12 9.17E-12 6.53E-12 6.75E-12 9.97E-12 1.21E-11 1.14E-11 9.70E-12 1.06E-11 1.21E-11 1.85E-11 7.33E-12 aspartate 3.40E-11 2.76E-11 2.38E-11 2.90E-11 4.70E-11 4.09E-11 3.64E-11 3.38E-11 6.47E-11 6.57E-11 7.27E-11 5.43E-11 3.06E-11 4.33E-11 3.29E-11 3.37E-11 6.79E-11 6.85E-11 6.36E-11 7.10E-11 5.62E-11 4.11E-11 4.68E-11 6.74E-11 leucine 8.46E-12 1.37E-11 1.04E-11 1.45E-11 2.08E-11 2.30E-11 2.52E-11 1.36E-11 3.17E-11 2.70E-11 2.90E-11 2.60E-11 3.88E-11 5.40E-11 3.44E-11 3.30E-11 3.85E-11 4.06E-11 4.76E-11 4.11E-11 4.64E-11 3.80E-11 4.42E-11 2.90E-11 lysine 1.92E-10 2.03E-10 2.30E-10 2.75E-10 1.28E-10 1.21E-10 1.34E-10 1.15E-10 2.34E-10 2.10E-10 1.06E-10 1.38E-10 4.51E-11 5.76E-11 3.89E-11 3.78E-11 3.90E-11 4.42E-11 6.00E-11 5.09E-11 5.68E-11 4.76E-11 5.72E-11 4.09E-11 choline 2.75E-11 3.34E-11 3.24E-11 4.01E-11 6.76E-11 6.61E-11 6.24E-11 4.09E-11 7.32E-11 6.69E-11 6.63E-11 5.77E-11 7.26E-11 1.04E-10 6.78E-11 6.25E-11 8.87E-11 8.69E-11 8.35E-11 7.33E-11 6.97E-11 5.50E-11 4.24E-11 5.22E-11 G3P 1.79E-11 1.64E-11 2.00E-11 2.25E-11 5.11E-11 5.75E-11 5.18E-11 6.41E-11 4.74E-11 4.40E-11 6.79E-11 3.30E-11 2.79E-11 4.48E-11 2.72E-11 2.81E-11 5.07E-11 3.88E-11 3.79E-11 3.42E-11 2.75E-11 2.49E-11 2.71E-11 2.19E-11 iseleucine 3.40E-12 6.68E-12 5.55E-12 8.15E-12 1.06E-11 1.05E-11 1.24E-11 9.69E-12 2.04E-11 1.83E-11 1.75E-11 1.78E-11 1.50E-11 1.96E-11 1.21E-11 1.45E-11 1.67E-11 1.84E-11 2.09E-11 1.76E-11 1.75E-11 1.57E-11 2.24E-11 1.86E-11 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 1H_ppm 13C_ppm PubChem CID Glycine-betaine 3.25 55.90 247 Proline 3.32 49.00 145742 DHPS 3.11 56.50 n.a. DMSP 2.91 27.80 n.a. Homarine 4.37 49.30 3620 beta-1,3-glucan 3.78 86.90 n.a. glycine 3.55 44.30 750 glutamate 2.34 36.40 33032 glutamine 2.44 33.90 5961 alanine 3.78 53.60 5950 aspartate 3.91 55.10 5960 leucine 0.94 23.60 6106 lysine 3.02 42.10 5962 choline 4.05 58.50 305 G3P 3.82 67.60 439162 iseleucine 1.00 17.40 6306 METABOLITES_END #END