#METABOLOMICS WORKBENCH katherine_heal_20251107_131821 DATATRACK_ID:6637 STUDY_ID:ST004353 ANALYSIS_ID:AN007268 PROJECT_ID:PR002738
VERSION             	1
CREATED_ON             	November 15, 2025, 1:23 pm
#PROJECT
PR:PROJECT_TITLE                 	Conserved pathway for homarine catabolism in environmental bacteria
PR:PROJECT_SUMMARY               	Homarine (N-methylpicolinic acid) is a ubiquitous marine metabolite produced by
PR:PROJECT_SUMMARY               	phytoplankton and noted for its bioactivity in marine animals, yet its microbial
PR:PROJECT_SUMMARY               	degradation pathways are uncharacterized. Here, we identify a conserved operon
PR:PROJECT_SUMMARY               	(homABCDER) that mediates homarine catabolism in bacteria using comparative
PR:PROJECT_SUMMARY               	transcriptomics, mutagenesis, and targeted knockouts. Phylogenetic and genomic
PR:PROJECT_SUMMARY               	analyses show this operon distributed across abundant bacterial clades,
PR:PROJECT_SUMMARY               	including coastal copiotrophs (e.g., Rhodobacterales) and open-ocean oligotrophs
PR:PROJECT_SUMMARY               	(e.g., SAR11, SAR116). High-resolution mass spectrometry revealed
PR:PROJECT_SUMMARY               	N-methylglutamic acid and glutamic acid as key metabolic products of homarine in
PR:PROJECT_SUMMARY               	both model and natural systems, with N-methylglutamate dehydrogenase catalyzing
PR:PROJECT_SUMMARY               	their conversion. Metatranscriptomics showed responsive and in situ expression
PR:PROJECT_SUMMARY               	of hom genes aligned with homarine availability. These findings uncover the
PR:PROJECT_SUMMARY               	genetic and metabolic basis of homarine degradation, establish its ecological
PR:PROJECT_SUMMARY               	relevance, and highlight homarine as a versatile growth substrate that feeds
PR:PROJECT_SUMMARY               	into central metabolism via glutamic acid in diverse marine bacteria.
PR:INSTITUTE                     	University of Washington, School of Oceanography
PR:LAST_NAME                     	Heal
PR:FIRST_NAME                    	Katherine
PR:ADDRESS                       	1501 NE Boat Street, Marine Science Building, Room G, Seattle
PR:EMAIL                         	katherine.heal@pnnl.gov
PR:PHONE                         	612-616-4840
PR:PUBLICATIONS                  	DOI 10.21203/rs.3.rs-7359689/v1
#STUDY
ST:STUDY_TITLE                   	Homarine catabolism: Marine microbial isotope-tracing metabolomics experiments
ST:STUDY_TITLE                   	for cruise TN412 in the Winter of 2023 at two different stations in the North
ST:STUDY_TITLE                   	Pacific
ST:STUDY_SUMMARY                 	Stable-isotope probing was performed to track homarine degradation products in
ST:STUDY_SUMMARY                 	natural marine microbial communities from three locations (Figure 2D of DOI
ST:STUDY_SUMMARY                 	10.21203/rs.3.rs-7359689/v1). Seawater incubated with isotopically-labeled
ST:STUDY_SUMMARY                 	13C715N-homarine was analyzed for the compounds enriched in our model organisms
ST:STUDY_SUMMARY                 	(see other studies within this project), with the isotopic labels.
ST:INSTITUTE                     	University of Washington, School of Oceanography
ST:LAST_NAME                     	Heal
ST:FIRST_NAME                    	Katherine
ST:ADDRESS                       	1501 NE Boat Street, Marine Science Building, Room G, Seattle
ST:EMAIL                         	katherine.heal@pnnl.gov
ST:PHONE                         	612-616-4840
ST:STUDY_COMMENTS                	Part of project 6620
ST:PUBLICATIONS                  	DOI 10.21203/rs.3.rs-7359689/v1
#SUBJECT
SU:SUBJECT_TYPE                  	Water sample
#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           	-	230717_Smp_G5_Hom1_C_A	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_C_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_C_B	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_C_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_C_C	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_C_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T0_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T0_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T0_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T0_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T0_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T0_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T6_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T6_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T6_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T6_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T6_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T6_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T12_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T12_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T12_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T12_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T12_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T12_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T24_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T24_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T24_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T24_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T24_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T24_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T48_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T48_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T48_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T48_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T48_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T48_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T96_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T96_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T96_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T96_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom1_T96_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	cruise_id=TN412; experiment_id=G51; date=20230128; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom1_T96_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_C_A	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_C_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_C_B	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_C_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_C_C	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_C_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T0_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T0_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T0_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T0_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T0_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T0_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T6_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T6_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T6_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T6_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T6_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T6_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T12_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T12_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T12_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T12_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T12_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T12_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T24_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T24_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T24_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T24_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T24_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T24_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T48_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T48_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T48_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T48_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T48_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T48_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T96_A	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T96_A.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T96_B	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T96_B.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Smp_G5_Hom2_T96_C	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140	cruise_id=TN412; experiment_id=G52; date=20230204; RAW_FILE_NAME(raw_data_files)=230717_Smp_G5_Hom2_T96_C.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full1	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full1.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full2	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full2.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full3	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full3.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full4	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full4.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full5	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full5.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full6	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full6.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate1_Full7	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate1_Full7.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full1	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full1.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full2	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full2.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full3	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full3.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full4	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full4.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full5	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full5.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full6	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full6.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Poo_G5-HomFate2_Full7	Sample source:QC | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Poo_G5-HomFate2_Full7.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_1	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_2	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_3	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_4	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_5	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix1InH2O_6	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix1InH2O_6.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_1	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_2	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_3	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_4	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_5	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	230717_Std_4uMStdsMix2InH2O_6	Sample source:Standards | treatment:N/A | timepoint:N/A | lat:N/A | lon:N/A	cruise_id=N/A; experiment_id=N/A; date=N/A; RAW_FILE_NAME(raw_data_files)=230717_Std_4uMStdsMix2InH2O_6.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Experiments using 13C7,15N-homarine-homarine were performed on research cruise
CO:COLLECTION_SUMMARY            	TN412 in the Winter of 2023 at two different stations in the North Pacific
CO:COLLECTION_SUMMARY            	(described in Table S8 and displayed in Figure 2 of
CO:COLLECTION_SUMMARY            	10.21203/rs.3.rs-7359689/v1). The preparation of the 13C7-15N-labeled homarine
CO:COLLECTION_SUMMARY            	is described in detail in 10.21203/rs.3.rs-7359689/v1. Seawater was collected
CO:COLLECTION_SUMMARY            	through a trace metal clean stayfish system suspended at a depth of 8 m
CO:COLLECTION_SUMMARY            	prefiltered through 100 µm nylon mesh. Triplicate samples were collected into
CO:COLLECTION_SUMMARY            	acid washed 2 L polycarbonate bottles, spiked with 90 nM of 13C7-15N-labeled
CO:COLLECTION_SUMMARY            	homarine, and incubated in temperature and light-controlled incubators for 5
CO:COLLECTION_SUMMARY            	different timepoints (6, 12, 24, 48 and 96 hours). Triplicates of spiked and
CO:COLLECTION_SUMMARY            	unspiked samples were filtered as quickly as possible, no more than 30 minutes
CO:COLLECTION_SUMMARY            	(T0, and unamended control samples, respectively). All particulate samples (4 L)
CO:COLLECTION_SUMMARY            	were collected using peristaltic pumps onto Durapore® 0.22 μm, 47 mm,
CO:COLLECTION_SUMMARY            	hydrophilic PVDF membrane filters, flash frozen in liquid nitrogen, and stored
CO:COLLECTION_SUMMARY            	at -80°C.
CO:SAMPLE_TYPE                   	Marine particulate matter
#TREATMENT
TR:TREATMENT_SUMMARY             	Triplicate samples were collected into acid washed 2 L polycarbonate bottles,
TR:TREATMENT_SUMMARY             	spiked with 90 nM of 13C7-15N-labeled homarine, and incubated in temperature and
TR:TREATMENT_SUMMARY             	light-controlled incubators for 5 different timepoints (6, 12, 24, 48 and 96
TR:TREATMENT_SUMMARY             	hours). Triplicates of spiked and unspiked samples were filtered as quickly as
TR:TREATMENT_SUMMARY             	possible, no more than 30 minutes (T0, and unamended control samples,
TR:TREATMENT_SUMMARY             	respectively).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	For particulate metabolomics, cell pellets were extracted using a combination of
SP:SAMPLEPREP_SUMMARY            	mechanical and chemical disruption techniques as described in previous work
SP:SAMPLEPREP_SUMMARY            	(Boysen et al 2018). Metabolites from the supernatant were extracted using a
SP:SAMPLEPREP_SUMMARY            	cation-exchange-based solid phase extraction technique as described previously
SP:SAMPLEPREP_SUMMARY            	(Sacks et al 2022), with 1 mL of supernatant diluted into 10 mL of HPLC grade
SP:SAMPLEPREP_SUMMARY            	water. To prevent confusion using the isotope labels, we used a subset of
SP:SAMPLEPREP_SUMMARY            	isotopically-labeled internal standards, as reported in Table S17 of
SP:SAMPLEPREP_SUMMARY            	https://doi.org/10.21203/rs.3.rs-7359689/v1.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	For HILIC chromatography, a SeQuant ZIC-pHILIC column (5 um particle size, 2.1
CH:CHROMATOGRAPHY_SUMMARY        	mm x 150 mm, from Millipore) was used with 10 mM ammonium carbonate in 85:15
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile to water (Solvent A) and 10 mM ammonium carbonate in 85:15 water to
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile (Solvent B) at a flow rate of 0.15 mL/min. This column was compared
CH:CHROMATOGRAPHY_SUMMARY        	with a Waters UPLC BEH amide and a Millipore cHILIC column; the pHILIC showed
CH:CHROMATOGRAPHY_SUMMARY        	superior reproducibility and peak shapes. The column was held at 100% A for 2
CH:CHROMATOGRAPHY_SUMMARY        	minutes, ramped to 64% B over 18 minutes, ramped to 100% B over 1 minute, held
CH:CHROMATOGRAPHY_SUMMARY        	at 100% B for 5 minutes, and equilibrated at 100% A for 25 minutes (50 minutes
CH:CHROMATOGRAPHY_SUMMARY        	total). The column was maintained at 30 C. The injection volume was 2 µL for
CH:CHROMATOGRAPHY_SUMMARY        	samples and standard mixes. When starting a batch, the column was equilibrated
CH:CHROMATOGRAPHY_SUMMARY        	at the starting conditions for at least 30 minutes. To improve the performance
CH:CHROMATOGRAPHY_SUMMARY        	of the HILIC column, we maintained the same injection volume, kept the
CH:CHROMATOGRAPHY_SUMMARY        	instrument running water blanks between samples as necessary, and injected
CH:CHROMATOGRAPHY_SUMMARY        	standards in a representative matrix in addition to standards in water. After
CH:CHROMATOGRAPHY_SUMMARY        	each batch, the column was flushed with 10 mM ammonium carbonate in 85:15 water
CH:CHROMATOGRAPHY_SUMMARY        	to acetonitrile for 20 to 30 minutes.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Waters Acquity I-Class
CH:COLUMN_NAME                   	SeQuant ZIC- pHILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	85% acetonitrile/15% water; 10 mM ammonium carbonate
CH:SOLVENT_B                     	15% acetonitrile/85% water; 10 mM ammonium carbonate
CH:FLOW_GRADIENT                 	The column was held at 100% A for 2 minutes, ramped to 64% B over 18 minutes,
CH:FLOW_GRADIENT                 	ramped to 100% B over 1 minute, held at 100% B for 5 minutes, and equilibrated
CH:FLOW_GRADIENT                 	at 100% A for 25 minutes
CH:FLOW_RATE                     	0.15 mL/min
CH:COLUMN_TEMPERATURE            	30
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF-X Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	MS acquisition Comments: Polarity switching was used with a scan range of 60 to
MS:MS_COMMENTS                   	900 m/z and a resolution of 60,000. MS parameters were as follows: capillary
MS:MS_COMMENTS                   	temperature was 320 ∞C, the H-ESI spray voltage was 3.3 kV, and the auxiliary
MS:MS_COMMENTS                   	gas heater temperature was 100 ∞C. The S-lens RF level was 65. Sheath gas,
MS:MS_COMMENTS                   	auxiliary gas, and sweep gas flow rates were maintained at 16, 3, and 1,
MS:MS_COMMENTS                   	respectively.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	normalized peak area
MS_METABOLITE_DATA_START
Samples	230717_Smp_G5_Hom1_C_A	230717_Smp_G5_Hom1_C_B	230717_Smp_G5_Hom1_C_C	230717_Smp_G5_Hom1_T0_A	230717_Smp_G5_Hom1_T0_B	230717_Smp_G5_Hom1_T0_C	230717_Smp_G5_Hom1_T6_A	230717_Smp_G5_Hom1_T6_B	230717_Smp_G5_Hom1_T6_C	230717_Smp_G5_Hom1_T12_A	230717_Smp_G5_Hom1_T12_B	230717_Smp_G5_Hom1_T12_C	230717_Smp_G5_Hom1_T24_A	230717_Smp_G5_Hom1_T24_B	230717_Smp_G5_Hom1_T24_C	230717_Smp_G5_Hom1_T48_A	230717_Smp_G5_Hom1_T48_B	230717_Smp_G5_Hom1_T48_C	230717_Smp_G5_Hom1_T96_A	230717_Smp_G5_Hom1_T96_B	230717_Smp_G5_Hom1_T96_C	230717_Smp_G5_Hom2_C_A	230717_Smp_G5_Hom2_C_B	230717_Smp_G5_Hom2_C_C	230717_Smp_G5_Hom2_T0_A	230717_Smp_G5_Hom2_T0_B	230717_Smp_G5_Hom2_T0_C	230717_Smp_G5_Hom2_T6_A	230717_Smp_G5_Hom2_T6_B	230717_Smp_G5_Hom2_T6_C	230717_Smp_G5_Hom2_T12_A	230717_Smp_G5_Hom2_T12_B	230717_Smp_G5_Hom2_T12_C	230717_Smp_G5_Hom2_T24_A	230717_Smp_G5_Hom2_T24_B	230717_Smp_G5_Hom2_T24_C	230717_Smp_G5_Hom2_T48_A	230717_Smp_G5_Hom2_T48_B	230717_Smp_G5_Hom2_T48_C	230717_Smp_G5_Hom2_T96_A	230717_Smp_G5_Hom2_T96_B	230717_Smp_G5_Hom2_T96_C
Factors	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Control | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:19.3316 | lon:-138.71	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Control | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:0 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:6 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:12 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:24 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:48 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140	Sample source:Particulate | treatment:Homarine (13C7,15N-labelled) | timepoint:96 | lat:2.6222 | lon:-140
L-Cysteic acid	92008		73568	96096	158694			41170		158853		64062				115626	113242	254467	1114534	294432	464419	86403	71547	121373	170108	180906	196854	67225	121341	40126			63127		150828	105234	276628		174649	1347466	1463470	2134720
UDP-N-acetylglucosamine	76500		56746		47112			147195		53507								63589									94854	138807	96917				42052				53823		83192	206801	260061	276037
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	molecule_id	stable_isotope_version	retention_time	mz
L-Cysteic acid	Cysteic acid		12.15	167.9967
UDP-N-acetylglucosamine	UDP-N-acetylglucosamine		12.75	606.0738
METABOLITES_END
#END