Summary of Study ST001938

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001227. The data can be accessed directly via it's Project DOI: 10.21228/M83Q5Z This work is supported by NIH grant, U2C- DK119886.

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Study IDST001938
Study TitleMetabolomics characterized concentration-dependent metabolic influence of magnesium on biofilm formation in Escherichia coli (Part1)
Study SummaryBiofilms are broadly formed by a diversity of microorganisms that enable them to adapt stressful environments. Biofilms often impose harmful influences in many niches, as they can cause food contamination, antibiotics resistance, and environmental issues. However, eradicating biofilms remains difficult since the formation mechanism of biofilms are still incompletely clarified. In this study, we aimed at exploring the regulatory role of magnesium (Mg2+) on biofilm formation in Escherichia coli (E. coli) using phenotype visualization combined with targeted metabolomics method. We found that Mg2+ could exert significant influence on biofilm formation in a concentration-dependent manner by regulating phenotypic morphology and triggering metabolic modifications of biofilm. Phenotypic imaging revealed that increasing concentration of Mg2+ gradually inhibited biofilm formation, Mg2+ was observed to restore the microstructure of E. coli strain in biofilms to that in the relevant planktonic cells. In addition, our metabolomics analysis characterized 20 differential metabolites and associated 2 metabolic pathways including nucleotide metabolism and amino acid metabolism that were notably modified during biofilm formation under the treatments of different concentrations of Mg2+. Altogether, our work provides a novel insight into the influence of Mg2+ on biofilm formation at a metabolic level, which are implicated in the novel solution to disturb biofilm formation through the regulation of Mg2+ and functional metabolite interaction, then biofilms associated harmful impacts in different niches could be well tangled accordingly.
Institute
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DepartmentShanghai Center for Systems Biomedicine
LaboratoryLu Group
Last NameLu
First NameHaitao
Address800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Emailhaitao_lu@sjtu.edu.cn
Phone15221478139
Submit Date2021-09-07
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2021-11-04
Release Version1
Haitao Lu Haitao Lu
https://dx.doi.org/10.21228/M83Q5Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001227
Project DOI:doi: 10.21228/M83Q5Z
Project Title:Metabolomics characterized concentration-dependent metabolic influence of magnesium on biofilm formation in Escherichia coli
Project Type:Targeted MS quantitative analysis
Project Summary:Metabolomics characterized concentration-dependent metabolic influence of magnesium on biofilm formation in Escherichia coli
Institute:Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
Department:Shanghai Center for Systems Biomedicine
Laboratory:Lu Group
Last Name:Lu
First Name:Haitao
Address:800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Email:haitao_lu@sjtu.edu.cn
Phone:15221478139

Subject:

Subject ID:SU002016
Subject Type:Bacteria
Subject Species:Escherichia coli
Taxonomy ID:562

Factors:

Subject type: Bacteria; Subject species: Escherichia coli (Factor headings shown in green)

mb_sample_id local_sample_id Concentration of MgSO4(w/v)
SA18312920210306-4-C-10.01%
SA18313020210306-4-C-60.01%
SA18313120210306-4-C-50.01%
SA18313220210306-4-C-20.01%
SA18313320210306-4-C-40.01%
SA18313420210306-4-30-60.15%
SA18313520210306-4-30-50.15%
SA18313620210306-4-30-20.15%
SA18313720210306-4-30-40.15%
SA18313820210306-4-30-10.15%
SA18313920210306-4-50-40.25%
SA18314020210306-4-50-60.25%
SA18314120210306-4-50-50.25%
SA18314220210306-4-50-30.25%
SA18314320210306-4-50-10.25%
SA18314420210306-4-50-20.25%
SA18314520210306-4-80-50.40%
SA18314620210306-4-80-40.40%
SA18314720210306-4-80-60.40%
SA18314820210306-4-80-30.40%
SA18314920210306-4-80-10.40%
SA18315020210306-4-80-20.40%
SA18315120210306-4-100-60.50%
SA18315220210306-4-100-50.50%
SA18315320210306-4-100-30.50%
SA18315420210306-4-100-10.50%
SA18315520210306-4-100-20.50%
SA18315620210306-4-100-40.50%
Showing results 1 to 28 of 28

Collection:

Collection ID:CO002009
Collection Summary:After 72 hours of culture, the planktonic cells were isolated, and the biofilms were washed with PBS solvent for three times. Then, planktonic cells and biofilm solutions were centrifuged at 2000 rpm and 4°C for 15 minutes, to collect the strain pellets.
Sample Type:Biofilm

Treatment:

Treatment ID:TR002028
Treatment Summary:Colony-forming antigen (CFA) medium (1% casamino acids, 0.15% yeast extract, 0.005% magnesium sulfate, and 0.0005% manganese chloride) was used to form biofilms. The E. coli strain was incubated in LB-agar plate for 12 hours, one colony was isolated to LB broth for further 4 hours incubation, then diluted the solution into CFA medium at a ratio of 1:1000 and the cultures were incubated for another 72 h at 30°C to form mature biofilms. To test the influence of MgSO4 on biofilm formation, the purified compound of MgSO4 was added to the CFA medium at different concentrations for the preparation of treated culture medium; The left cultural procedures were kept as the same as the protocol listed above.

Sample Preparation:

Sampleprep ID:SP002022
Sampleprep Summary:Each pellet sample was mixed with 2 mL of 80% ice-cold methanol, and the mixed solution was homogenized on ice for 2 minutes and repeated the same procedure for three times. Next, the collected samples were centrifuged at 12000 rpm and 4°C for 15 minutes, to collect the supernatants. The supernatants were further mixed with 800 μL of ice-cold acetonitrile for 20 minutes before further centrifuge at 12000 rpm and 4°C for 15 minutes. At last, the isolated supernatants were gone through a 0.22 μm filter membrane (nylon) before the lyophilization. The lyophilized samples were stored at -80°C for metabolomics analysis.

Combined analysis:

Analysis ID AN003151 AN003152
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name Agilent 6495 QQQ Agilent 6495 QQQ
Ion Mode NEGATIVE POSITIVE
Units - -

Chromatography:

Chromatography ID:CH002331
Chromatography Summary:The chromatographic separation of biological samples was implemented on a Waters ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 μm) with an optimized gradient-elution program (mobile phase A and B were 0.1% formic acid in water and acetonitrile (v/v)): 0-2 minutes, 98% A; 2-10 minutes, 98%-65% A; 10-12 minutes, 65%-20% A; 12-14 minutes, 20%-2% A; 14-30 minutes, 2% A. The column temperature was at 40°C and the flow rate was set at 0.3 mL/ minutes.
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um)
Chromatography Type:Reversed phase

MS:

MS ID:MS002931
Analysis ID:AN003151
Instrument Name:Agilent 6495 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ)
Ion Mode:NEGATIVE
  
MS ID:MS002932
Analysis ID:AN003152
Instrument Name:Agilent 6495 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ)
Ion Mode:POSITIVE
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