#METABOLOMICS WORKBENCH chichen@umn.edu_20210120_131024 DATATRACK_ID:2410 STUDY_ID:ST001657 ANALYSIS_ID:AN002705 PROJECT_ID:PR001062 VERSION 1 CREATED_ON January 21, 2021, 2:59 pm #PROJECT PR:PROJECT_TITLE Effect of IPL on E.coli Metabolome 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 ap-plied to E. PR:PROJECT_SUMMARY coli K-12 for 0-20s, leading to time- and dose-dependent changes in E.coli PR:PROJECT_SUMMARY metabolome. We consider the degradation of membrane-bound quinone electron PR:PROJECT_SUMMARY carriers as the trigger of dramatic metabolis shift in IPL-treated E.coli. PR:INSTITUTE University of Minnesota PR:DEPARTMENT Food Science and Nutrition PR:LABORATORY Nutritional Metabolomics PR:LAST_NAME Chen PR:FIRST_NAME Chi PR:ADDRESS 1334 Eckles Ave W, St Paul, MN 55108 PR:EMAIL chichen@umn.edu PR:PHONE 612-624-7704 #STUDY ST:STUDY_TITLE E.coli K-12 treated by IPL_analysis of organic phase ST:STUDY_SUMMARY In this study, E.coli K-12 was treated by intense pulsed light (IPL) for 0-20 ST:STUDY_SUMMARY seconds. Then the organic/lipid phase of the cellular metabolome was extracted ST:STUDY_SUMMARY and submitted to untargeted LC-MS based metabolomic study. ST:INSTITUTE University of Minnesota ST:DEPARTMENT Food Science and Nutrition ST:LABORATORY Nutritional Metabolomics ST:LAST_NAME Chen ST:FIRST_NAME Chi ST:ADDRESS 1334 Eckles Ave ST:EMAIL chichen@umn.edu ST:PHONE 6126247704 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Escherichia coli SU:TAXONOMY_ID 562 SU:GENOTYPE_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=180309_QM_ecoli_extraction_non-polar phase_ctl_1 SUBJECT_SAMPLE_FACTORS - ctl_2 Treatment:control RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_ctl_2 SUBJECT_SAMPLE_FACTORS - ctl_3 Treatment:control RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_ctl_3 SUBJECT_SAMPLE_FACTORS - ctl_4 Treatment:control RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_ctl_4 SUBJECT_SAMPLE_FACTORS - 5s_1 Treatment:IPL 5 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_5s_1 SUBJECT_SAMPLE_FACTORS - 5s_2 Treatment:IPL 5 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_5s_2 SUBJECT_SAMPLE_FACTORS - 5s_3 Treatment:IPL 5 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_5s_3 SUBJECT_SAMPLE_FACTORS - 5s_4 Treatment:IPL 5 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_5s_4 SUBJECT_SAMPLE_FACTORS - 10s_1 Treatment:IPL 10 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_10s_1 SUBJECT_SAMPLE_FACTORS - 10s_2 Treatment:IPL 10 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_10s_2 SUBJECT_SAMPLE_FACTORS - 10s_3 Treatment:IPL 10 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_10s_3 SUBJECT_SAMPLE_FACTORS - 10s_4 Treatment:IPL 10 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_10s_4 SUBJECT_SAMPLE_FACTORS - 15s_1 Treatment:IPL 15 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_15s_1 SUBJECT_SAMPLE_FACTORS - 15s_2 Treatment:IPL 15 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_15s_2 SUBJECT_SAMPLE_FACTORS - 15s_3 Treatment:IPL 15 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_15s_3 SUBJECT_SAMPLE_FACTORS - 15s_4 Treatment:IPL 15 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_15s_4 SUBJECT_SAMPLE_FACTORS - 20s_1 Treatment:IPL 20 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_20s_1 SUBJECT_SAMPLE_FACTORS - 20s_2 Treatment:IPL 20 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_20s_2 SUBJECT_SAMPLE_FACTORS - 20s_3 Treatment:IPL 20 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_20s_3 SUBJECT_SAMPLE_FACTORS - 20s_4 Treatment:IPL 20 s RAW_FILE_NAME=180309_QM_ecoli_extraction_non-polar phase_20s_4 #COLLECTION CO:COLLECTION_SUMMARY E. coli strain K-12 W3110 (ATCC 27325) was cultured on TSA agar medium. A single CO:COLLECTION_SUMMARY bacterial colony was picked from TSA agar medium, and then used to inoculate CO:COLLECTION_SUMMARY Luria-Bertani broth (EMD Millipore, Billerica, MA). After being incubated at 37 CO:COLLECTION_SUMMARY °C for 12 h on a rotary shaker set to 200 rpm, E. coli cells were harvested at CO:COLLECTION_SUMMARY an optical density (OD600) of 1, and then centrifuged at 7,500 × g for 10 min CO:COLLECTION_SUMMARY in 50 mL centrifuge tubes. After decanting the supernatant, the pellet was CO:COLLECTION_SUMMARY washed with phosphate buffered saline (PBS), and then re-suspended to the volume CO:COLLECTION_SUMMARY of bacterial culture for further treatment and analysis. CO:COLLECTION_PROTOCOL_FILENAME Cell culture_IPL treatment_Sample preparation_method.docx CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY E.coli K-12 was treated by a customized IPL instrument for 0 - 20 seconds. TR:TREATMENT_PROTOCOL_FILENAME Cell culture_IPL treatment_Sample preparation_method.docx #SAMPLEPREP SP:SAMPLEPREP_SUMMARY After IPL treatment, the cells were centrifuged and washed with PBS twice. The SP:SAMPLEPREP_SUMMARY pellets then went though methanol-water-chloroform extraction for obtaining the SP:SAMPLEPREP_SUMMARY organic/non-polar phase. SP:SAMPLEPREP_PROTOCOL_FILENAME Cell culture_IPL treatment_Sample preparation_method.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY analysis of lipids CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C8 (100 x 2.1mm, 1.7um) CH:FLOW_RATE 0.5 mL/min CH:SOLVENT_A 40% aqueous ACN containing 0.1% formic acid and 10mM NH4OAc CH:SOLVENT_B methanol solution containing 0.1% formic acid and 10mM NH4OAc #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 Ion features were captured by MakerLynx (Waters) after eliminating noises. MS:MS_RESULTS_FILE ST001657_AN002705_Results.txt UNITS:no applicable unites Has m/z:Yes Has RT:Yes RT units:Minutes #END