#METABOLOMICS WORKBENCH chichen@umn.edu_20210125_140613_mwtab.txt DATATRACK_ID:2422 STUDY_ID:ST001664 ANALYSIS_ID:AN002716 PROJECT_ID:000000 VERSION 1 CREATED_ON January 28, 2021, 11:02 am #PROJECT PR:PROJECT_TITLE Effect of IPL on E.coli Metabolome amino acids PR:PROJECT_TYPE Untargeted LC-MS metabolomic study PR:PROJECT_SUMMARY Intense pulsed light (IPL) is becoming a new technical platform for disinfecting PR:PROJECT_SUMMARY food against pathogenic bacteria. Metabolic changes are deemed to occur in PR:PROJECT_SUMMARY bacteria as either the causes or the consequences of IPL-elicited bactericidal PR:PROJECT_SUMMARY and bacteriostatic effects. However, little is known about the influences of IPL PR:PROJECT_SUMMARY on bacterial metabolome. In this study, the IPL treatment was applied to E. coli PR:PROJECT_SUMMARY K-12 for 0-20s, leading to time- and dose-dependent reductions in colony-forming PR:PROJECT_SUMMARY units (CFU) and morphological changes. Cytoplasmic metabolites of the control PR:PROJECT_SUMMARY and IPL-treated E. coli were examined by the liquid chromatography-mass PR:PROJECT_SUMMARY spectrom-etry (LC-MS)-based metabolomic fingerprinting. The results from PR:PROJECT_SUMMARY multivariate modeling and marker identification indicated that the metabolites PR:PROJECT_SUMMARY in redox response, glycolysis, amino acid and nucleotide metabolism were PR:PROJECT_SUMMARY selectively affected by the IPL treatments. PR:INSTITUTE University of Minnesota PR:LAST_NAME Chen PR:FIRST_NAME Chi PR:ADDRESS 1334 Eckles Ave, St Paul, Minnesota, 55108, USA PR:EMAIL chichen@umn.edu PR:PHONE 6126247704 #STUDY ST:STUDY_TITLE E.coli K-12 treated by IPL - analysis of polar phase (part-II) ST:STUDY_SUMMARY E.coli K-12 cells were treated by IPL, extracted and separated into ST:STUDY_SUMMARY organic/lipid phase and polar phase. Chemical derivatization with dansyl ST:STUDY_SUMMARY chloride was applied for analysis of amino acids in the polar phase extraction. ST:INSTITUTE University of Minnesota ST:LAST_NAME Chen ST:FIRST_NAME Chi ST:ADDRESS 1334 Eckles Ave, St Paul, Minnesota, 55108, USA ST:EMAIL chichen@umn.edu ST:PHONE 6126247704 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Escherichia coli SU:TAXONOMY_ID 316407 SU:SUBJECT_COMMENTS E. coli strain K-12 W3110 #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 - ctl_1 Treatment:control RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_ctl_1 SUBJECT_SAMPLE_FACTORS - ctl_2 Treatment:control RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_ctl_2 SUBJECT_SAMPLE_FACTORS - ctl_3 Treatment:control RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_ctl_3 SUBJECT_SAMPLE_FACTORS - ctl_4 Treatment:control RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_ctl_4 SUBJECT_SAMPLE_FACTORS - 5s_1 Treatment:IPL 5 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_5s_1 SUBJECT_SAMPLE_FACTORS - 5s_2 Treatment:IPL 5 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_5s_2 SUBJECT_SAMPLE_FACTORS - 5s_3 Treatment:IPL 5 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_5s_3 SUBJECT_SAMPLE_FACTORS - 5s_4 Treatment:IPL 5 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_5s_4 SUBJECT_SAMPLE_FACTORS - 10s_1 Treatment:IPL 10 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_10s_1 SUBJECT_SAMPLE_FACTORS - 10s_2 Treatment:IPL 10 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_10s_2 SUBJECT_SAMPLE_FACTORS - 10s_3 Treatment:IPL 10 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_10s_3 SUBJECT_SAMPLE_FACTORS - 10s_4 Treatment:IPL 10 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_10s_4 SUBJECT_SAMPLE_FACTORS - 15s_1 Treatment:IPL 15 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_15s_1 SUBJECT_SAMPLE_FACTORS - 15s_2 Treatment:IPL 15 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_15s_2 SUBJECT_SAMPLE_FACTORS - 15s_3 Treatment:IPL 15 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_15s_3 SUBJECT_SAMPLE_FACTORS - 15s_4 Treatment:IPL 15 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_15s_4 SUBJECT_SAMPLE_FACTORS - 20s_1 Treatment:IPL 20 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_20s_1 SUBJECT_SAMPLE_FACTORS - 20s_2 Treatment:IPL 20 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_20s_2 SUBJECT_SAMPLE_FACTORS - 20s_3 Treatment:IPL 20 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_20s_3 SUBJECT_SAMPLE_FACTORS - 20s_4 Treatment:IPL 20 s RAW_FILE_NAME=180312_QM_ecoli_extraction_polar phase_20s_4 #COLLECTION CO:COLLECTION_SUMMARY E.coli strain K-12 was cultured in LB broth and harvested at OD=1. After CO:COLLECTION_SUMMARY centrifuge, cell pellets were harvested and washed with PBS twice. Then the CO:COLLECTION_SUMMARY pellet was re-suspended in PBS for further treatment. CO:COLLECTION_PROTOCOL_FILENAME Cell_culture_IPL treatment_Sample preparation_method.docx CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY For each IPL treatment, a 30 mL K-12 suspension in PBS was loaded in a petri TR:TREATMENT_SUMMARY dish (15 cm diameter) and then placed under the broad-spectrum light (wavelength TR:TREATMENT_SUMMARY 190-1100 nm) in a Z-1000 SteriPulse-XL system (Xenon Corporation, Woburn, MA). TR:TREATMENT_SUMMARY The samples were treated with the IPL for 0, 5, 10, 15, and 20 s. TR:TREATMENT_PROTOCOL_FILENAME Cell_culture_IPL treatment_Sample preparation_method.docx #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The control and IPL-treated E. coli samples were centrifuged for obtaining the SP:SAMPLEPREP_SUMMARY pellets, which were then washed with PBS twice. After extraction with SP:SAMPLEPREP_SUMMARY methanol-water-chloroform, the polar phase was separated and derivatized with SP:SAMPLEPREP_SUMMARY dansyl chloride (DC) for amino acid analysis. SP:SAMPLEPREP_PROTOCOL_FILENAME Cell_culture_IPL treatment_Sample preparation_method.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (50 x 2.1mm, 1.7 um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Xevo-G2-S MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Features were captured by MarkerLynx (Waters) and processed. The relative MS:MS_COMMENTS intensity of each ion was calculated by normalizing the single ion counts (SIC) MS:MS_COMMENTS versus the total ion counts (TIC) in the whole chromatogram, while setting the MS:MS_COMMENTS TIC arbitrarily to 10,000. MS:MS_RESULTS_FILE ST001664_AN002716_Results.txt UNITS:no applicable unites Has m/z:Yes Has RT:Yes RT units:Minutes #END