Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.

Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic...

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Autores principales:	Clothilde Bertin, Corinne Pau-Roblot, Josiane Courtois, Lucía Manso-Silván, François Thiaucourt, Florence Tardy, Dominique Le Grand, François Poumarat, Patrice Gaurivaud
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3a5df021e69e42da8441003488efe3d7
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spelling	oai:doaj.org-article:3a5df021e69e42da8441003488efe3d72021-11-18T07:37:40ZCharacterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.1932-620310.1371/journal.pone.0068373https://doaj.org/article/3a5df021e69e42da8441003488efe3d72013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23869216/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS), also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical β(1->6)-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan) and associated with an antigenic switch constitutes a finely tuned mechanism that may be involved in virulence.Clothilde BertinCorinne Pau-RoblotJosiane CourtoisLucía Manso-SilvánFrançois ThiaucourtFlorence TardyDominique Le GrandFrançois PoumaratPatrice GaurivaudPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 7, p e68373 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Clothilde Bertin Corinne Pau-Roblot Josiane Courtois Lucía Manso-Silván François Thiaucourt Florence Tardy Dominique Le Grand François Poumarat Patrice Gaurivaud Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
description	Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS), also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical β(1->6)-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan) and associated with an antigenic switch constitutes a finely tuned mechanism that may be involved in virulence.
format	article
author	Clothilde Bertin Corinne Pau-Roblot Josiane Courtois Lucía Manso-Silván François Thiaucourt Florence Tardy Dominique Le Grand François Poumarat Patrice Gaurivaud
author_facet	Clothilde Bertin Corinne Pau-Roblot Josiane Courtois Lucía Manso-Silván François Thiaucourt Florence Tardy Dominique Le Grand François Poumarat Patrice Gaurivaud
author_sort	Clothilde Bertin
title	Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
title_short	Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
title_full	Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
title_fullStr	Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
title_full_unstemmed	Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.
title_sort	characterization of free exopolysaccharides secreted by mycoplasma mycoides subsp. mycoides.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/3a5df021e69e42da8441003488efe3d7
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Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.

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