The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron

Lipopolysaccharide (LPS), a cell-associated glycolipid that makes up the outer leaflet of the outer membrane of Gram-negative bacteria, is a canonical mediator of microbe-host interactions. The most prevalent Gram-negative gut bacterial taxon, Bacteroides, makes up around 50% of the cells in a typic...

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Main Authors: Amy N. Jacobson, Biswa P. Choudhury, Michael A. Fischbach, David A. Relman
Format: Article
Language:English
Published: American Society for Microbiology 2018-03-01
Series:mBio
Online Access:http://mbio.asm.org/cgi/content/full/9/2/e02289-17
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spelling doaj-9199a9a1d0c1480b9bfdb40616db38102021-07-02T05:21:32ZengAmerican Society for MicrobiologymBio2150-75112018-03-0192e02289-1710.1128/mBio.02289-17The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicronAmy N. JacobsonBiswa P. ChoudhuryMichael A. FischbachDavid A. RelmanLipopolysaccharide (LPS), a cell-associated glycolipid that makes up the outer leaflet of the outer membrane of Gram-negative bacteria, is a canonical mediator of microbe-host interactions. The most prevalent Gram-negative gut bacterial taxon, Bacteroides, makes up around 50% of the cells in a typical Western gut; these cells harbor ~300 mg of LPS, making it one of the highest-abundance molecules in the intestine. As a starting point for understanding the biological function of Bacteroides LPS, we have identified genes in Bacteroides thetaiotaomicron VPI 5482 involved in the biosynthesis of its lipid A core and glycan, generated mutants that elaborate altered forms of LPS, and used matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry to interrogate the molecular features of these variants. We demonstrate, inter alia, that the glycan does not appear to have a repeating unit, and so this strain produces lipooligosaccharide (LOS) rather than LPS. This result contrasts with Bacteroides vulgatus ATCC 8482, which by SDS-PAGE analysis appears to produce LPS with a repeating unit. Additionally, our identification of the B. thetaiotaomicron LOS oligosaccharide gene cluster allowed us to identify similar clusters in other Bacteroides species. Our work lays the foundation for developing a structure-function relationship for Bacteroides LPS/LOS in the context of host colonization.http://mbio.asm.org/cgi/content/full/9/2/e02289-17
collection DOAJ
language English
format Article
sources DOAJ
author Amy N. Jacobson
Biswa P. Choudhury
Michael A. Fischbach
David A. Relman
spellingShingle Amy N. Jacobson
Biswa P. Choudhury
Michael A. Fischbach
David A. Relman
The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
mBio
author_facet Amy N. Jacobson
Biswa P. Choudhury
Michael A. Fischbach
David A. Relman
author_sort Amy N. Jacobson
title The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
title_short The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
title_full The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
title_fullStr The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
title_full_unstemmed The Biosynthesis of Lipooligosaccharide from Bacteroides thetaiotaomicron
title_sort biosynthesis of lipooligosaccharide from bacteroides thetaiotaomicron
publisher American Society for Microbiology
series mBio
issn 2150-7511
publishDate 2018-03-01
description Lipopolysaccharide (LPS), a cell-associated glycolipid that makes up the outer leaflet of the outer membrane of Gram-negative bacteria, is a canonical mediator of microbe-host interactions. The most prevalent Gram-negative gut bacterial taxon, Bacteroides, makes up around 50% of the cells in a typical Western gut; these cells harbor ~300 mg of LPS, making it one of the highest-abundance molecules in the intestine. As a starting point for understanding the biological function of Bacteroides LPS, we have identified genes in Bacteroides thetaiotaomicron VPI 5482 involved in the biosynthesis of its lipid A core and glycan, generated mutants that elaborate altered forms of LPS, and used matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry to interrogate the molecular features of these variants. We demonstrate, inter alia, that the glycan does not appear to have a repeating unit, and so this strain produces lipooligosaccharide (LOS) rather than LPS. This result contrasts with Bacteroides vulgatus ATCC 8482, which by SDS-PAGE analysis appears to produce LPS with a repeating unit. Additionally, our identification of the B. thetaiotaomicron LOS oligosaccharide gene cluster allowed us to identify similar clusters in other Bacteroides species. Our work lays the foundation for developing a structure-function relationship for Bacteroides LPS/LOS in the context of host colonization.
url http://mbio.asm.org/cgi/content/full/9/2/e02289-17
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