Degradation of native and 13C-labelled linolenic acid and linoleic acid in Nitrosomonas europaea Nm 50 cultures
Ontology highlight
ABSTRACT:
- concentrated ~600 ml culture to 280 ml mother culture
- let acclimate back to 26C for ~1h without substrate
- add 1 mM NH4Cl (final concentration) to mother culture bottle:
- T0 M1 - 15:30,
T1 M1 - 16:30 (working, already nitrite visible),
T2 - 18:00 (working, more nitrite clearly),
T3 - 19:20 (still increasing, time to split)
Split mother culture to 15 subcultures for treatment:
LNH - linolenic 8 ul 98% (triplicates, 17 ml culture)
LNL - linolenic 2 ul 98% (triplicates, 17 ml culture)
LEH - linoleic 8 ul 98% (triplicates, 17 ml culture)
LEL - linoleic 2 ul 98% (triplicates, 17 ml culture)
C - control, nothing added (just 17 ml, culture)
shake up really well after addition:
T1 19:40 (500 ul samples for MS)
T2 21:30 (500 ul samples for MS)
T3 23:00 (500 ul samples for MS)
T4 1:30 - 1:50 (500 ul samples for MS)
Harvest at T4 by filtration 15 ml from each, freeze @ -80C
LC-HRMS measurments according to 10.1007/s00216-025-05818-y
INSTRUMENT(S): Q Exactive HF
ORGANISM(S): Nitrosomonas Europaea (ncbitaxon:915)
SUBMITTER: 
christoph bueschl
PROVIDER: MSV000100368 | MassIVE | Mon Jan 05 07:08:00 GMT 2026
REPOSITORIES: MassIVE
Similar Datasets
Project description:Nitrosomonas europaea Nm50 BNI degradation experiment:<br>
- concentrate ~ 600 ml culture<br>
- resuspend to make motherculture - 270 ml (need 255 for experiments)<br>
- let acclimate back to 26C for ~20min- 1h without substrate<br>
- add 1 mM NH4Cl (final concentration) to mother culture bottle:<br>
- take T0 mother culture; take T1 mother culture (cca 20 min); T2 mother culture (40min); <br>
split to treatments after 60 min<br>
<br>
Treatments:<br>
prepare 150 ul Linolenic acid (LNL) mix (50% unlabeled; 50% 13C LNL) on ice; after using<br>
120 for inoculation of treatments; freeze the 30 ul mix @ -80<br>
prepare just medium with 0.5 mM NH4<br>
<br>
Biotic:<br>
LNL; 8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml culture each (5)<br>
PHA-LNL - 1 mM PHA+8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml<br>
culture each (5)<br>
control - nothing added - 5 replicates; 17 ml culture each (5)<br>
<br>
Abiotic:<br>
LNL - 8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml medium with 0.5 mM<br>
NH4+ (5)<br>
<br>
Sampling every 30-45 min; 5 timepoints<br>
<br>
A: LE + culture<br>
B: PHA-LE + culture<br>
C: culture only<br>
D: LE in media only<br>
<br>
LC-HRMS according to 10.1007/s00216-025-05818-y<br>
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