Summary of Study ST001904

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 PR001199. The data can be accessed directly via it's Project DOI: 10.21228/M8QQ5J This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST001904
Study Title	Lipidomics analysis of outer membrane vesicles and elucidation of the ceramide phosphoinositol biosynthetic pathway in Bacteroides thetaiotaomicron
Study Type	Lipidic profile in wild-type and mutant strains
Study Summary	In this work, we characterized the lipid composition of membranes and OMV from Bacteroides thetaiotaomicron VPI-5482. LC-MS analysis indicate that OMV carry sphingolipids, glycerophospholipids and serine-dipeptide lipids. Sphingolipid species represent more than 50% of the total lipid content of OMV. The most abundant sphingolipids in OMV are ceramide phosphoethanolamine (CerPE) and ceramide phosphoinositol (CerPI). Bioinformatic analysis allowed the identification of the BT1522-1526 operon putatively involved in CerPI synthesis. Mutagenesis studies revealed BT1522-1526 are essential for synthesis of PI and CerPI, confirming the role of this operon in biosynthesis of CerPI. BT1522-1526 mutant strains lacking CerPI produced OMV that were indistinguishable from the wild-type strain, indicating that CerPI sphingolipid species are not involved in OMV biogenesis. Bacteroides sphingolipids are thought to modulate host-commensal interactions, and based on our data, we propose that OMV could act as long distance delivery vehicles for these molecules.
Institute	Washington University in St. Louis
Department	Molecular Microbiology
Laboratory	Feldman lab
Last Name	Sartorio
First Name	Mariana
Address	660 S Euclid avenue, campus box 8230, 63110
Email	mgsartorio@wustl.edu
Phone	3147474477
Submit Date	2021-06-22
Analysis Type Detail	LC-MS
Release Date	2021-08-30
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001988
Sampleprep Summary:	Total lipids from OMV and TM were extracted based on the Bligh and Dyer chloroform:methanol method (1). Briefly, 2 volumes of methanol, 1 volume of chloroform, and 0.8 volumes of Milli-Q water were added to 1 volume of PBS-resuspended OMV or TM fractions into solvent-resistant glass tubes. Contents were mixed for 1 min by vortexing and 1 volume of chloroform was added to the mixture. Contents were mixed for another minute and tubes were centrifuged for 5 min at 4000 rpm. After centrifugation, bottom phase (organic) was recovered using a glass Pasteur pipette and stored in solvent-sealed vials at -80°C until lipid analysis by LC-MS. References: 1. Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959;37(8):911-7.

Sampleprep ID:

SP001988

Sampleprep Summary:

Total lipids from OMV and TM were extracted based on the Bligh and Dyer chloroform:methanol method (1). Briefly, 2 volumes of methanol, 1 volume of chloroform, and 0.8 volumes of Milli-Q water were added to 1 volume of PBS-resuspended OMV or TM fractions into solvent-resistant glass tubes. Contents were mixed for 1 min by vortexing and 1 volume of chloroform was added to the mixture. Contents were mixed for another minute and tubes were centrifuged for 5 min at 4000 rpm. After centrifugation, bottom phase (organic) was recovered using a glass Pasteur pipette and stored in solvent-sealed vials at -80°C until lipid analysis by LC-MS. References: 1. Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959;37(8):911-7.