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MB Sample ID: SA300833

Local Sample ID:HPLT_S3
Subject ID:SU002911
Subject Type:Bacteria
Subject Species:Microbacterium sediminis YLB-01

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Subject:

Subject ID:SU002911
Subject Type:Bacteria
Subject Species:Microbacterium sediminis YLB-01

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
HPLT_S3SA300833FL036541HPLTTreatment

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:SA300833
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
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