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MB Sample ID: SA300839
| Local Sample ID: | HPLT_S1 |
| Subject ID: | SU002911 |
| Subject Type: | Bacteria |
| Subject Species: | Microbacterium sediminis YLB-01 |
| Species Group: | Bacteria |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002911 |
| Subject Type: | Bacteria |
| Subject Species: | Microbacterium sediminis YLB-01 |
| Species Group: | Bacteria |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| HPLT_S1 | SA300839 | FL036541 | HPLT | Treatment |
Collection:
| Collection ID: | CO002904 |
| Collection Summary: | In this study, a single colony of the YLB-01 strain was inoculated from an agar plate into individual test tubes containing 5 mL of TSB medium. The test tubes were then incubated in a shaker at 28°C for 12 h. Subsequently, the seed cultures were transferred to 150-mL Erlenmeyer flasks at a ratio of 1:20, with each flask containing 100 mL of TSB medium. To ensure an adequate cell count, YLB-01 cultures were initially cultivated under optimal conditions (28°C, 0.1 MPa) for 24 h. The optical density at 600 nm (OD600 nm) of the cell cultures was measured using an automatic growth curve analyzer (FP-1100-C, Growth Curves Ab Ltd., Finland). Once the cultures reached the stationary phase of growth, the cells were transferred into a 100-mL sterile saline bag and placed in a high-pressure culture kettle. This kettle, obtained from Nantong Feiyu Company, China, was specifically designed to simulate the high-pressure conditions of the deep sea (Zhang et al. 2015). The cells were then exposed to either 30 MPa (the HPLT group, N=8) or 0.1 MPa (the NPLT group, N=7) at 4°C for 7 days. Hydrostatic pressure was generated by injecting pure water into the vessel. |
| Sample Type: | Bacterial cells |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR002920 |
| Treatment Summary: | To ensure an adequate cell count, YLB-01 cultures were initially cultivated under optimal conditions (28°C, 0.1 MPa) for 24 h. The optical density at 600 nm (OD600 nm) of the cell cultures was measured using an automatic growth curve analyzer (FP-1100-C, Growth Curves Ab Ltd., Finland). Once the cultures reached the stationary phase of growth, the cells were transferred into a 100-mL sterile saline bag and placed in a high-pressure culture kettle. The cells were then exposed to either 30 MPa (the HPLT group, N=8) or 0.1 MPa (the NPLT group, N=7) at 4°C for 7 days. |
Sample Preparation:
| Sampleprep ID: | SP002917 |
| Sampleprep Summary: | After culturing the YLB-01 cells, each 100 mL of the culture was transferred into a 250-mL centrifuge bottle and centrifuged at 4°C (6000 g, 5 min). The supernatant was carefully decanted, and the cell pellets were rapidly cooled to -40°C using 100 mL of a buffer composed of a 3:2 methanol/water mixture containing 0.85% (wt/vol) NaCl. The mixture was centrifuged again at 4°C (6000 g, 5 min). The cell pellets were washed three times with 5 mL of cold phosphate-buffered saline (PBS) and centrifuged at 4°C (6000 g, 5 min) after each wash. Finally, the cell pellets were stored at -80°C until further use. Initially, 600 μL of a cold extraction buffer consisting of a 1:1 mixture of distilled water and acetonitrile was added to homogenize the samples. The mixtures were then sonicated on wet ice for 180 cycles, with each cycle consisting of 2 seconds of ultrasound followed by a 3-second pause. After centrifugation at 4°C (12000 g, 10 min), the supernatants were collected and lyophilized, resulting in an extract powder that was stored at -80°C for further analysis. |
| Processing Storage Conditions: | Described in summary |
| Extract Storage: | Described in summary |
Analysis:
| MB Sample ID: | SA300839 |
| Analysis ID: | AN004560 |
| Analysis Type: | NMR |
| Results File: | ST002804_AN004560_Results.txt |
| Units: | ppm |
NMR:
| NMR ID: | NM000266 |
| Analysis ID: | AN004560 |
| Instrument Name: | Bruker Avance III 600 MHz |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| Standard Concentration: | 1 mM TSP |
| Spectrometer Frequency: | 600 MHz |
| NMR Solvent: | H2O+D2O |
| NMR Tube Size: | 5 mm |
| Pulse Sequence: | noesygppr1d [(RD)-90°-t1-90°-τm-90°-ACQ] |
| Receiver Gain: | 71.8 |
| Offset Frequency: | 15.1 ppm |
| Temperature: | 25 |
| Number Of Scans: | 32 |
| Dummy Scans: | 4 |
| Acquisition Time: | 2 s |
| Spectral Width: | 20 ppm |
| Num Data Points Acquired: | 64 K |
| Line Broadening: | 0.3 Hz |
| Baseline Correction Method: | Auto-baseline correction of integral by abs |
| Chemical Shift Ref Std: | TSP (0.000 ppm) |
| Binned Data Excluded Range: | δ 4.700-5.100 ppm |
| NMR Results File: | microHP.txt UNITS:ppm |
