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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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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