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Biosynthesis of osmoregulated periplasmic glucans in Escherichia coli: the phosphoethanolamine transferase is encoded by opgE.

Bontemps-Gallo S, Cogez V, Robbe-Masselot C, Quintard K, Dondeyne J, Madec E, Lacroix JM - Biomed Res Int (2013)

Bottom Line: Osmoregulated periplasmic glucans (OPGs) are oligosaccharides found in the periplasm of many Gram-negative bacteria.In E. coli, OPG are substituted by phosphoglycerol and phosphoethanolamine derived from membrane phospholipids and by succinyl residues.Both genes show structural and functional similarities without sequence similarity.

View Article: PubMed Central - PubMed

Affiliation: Unité de Glycobiologie Structurale et Fonctionnelle, UMR CNRS 8576, IFR 147, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq Cedex, France ; Université Lille Nord de France, Lille, France.

ABSTRACT
Osmoregulated periplasmic glucans (OPGs) are oligosaccharides found in the periplasm of many Gram-negative bacteria. Glucose is the sole constitutive sugar and this backbone may be substituted by various kinds of molecules depending on the species. In E. coli, OPG are substituted by phosphoglycerol and phosphoethanolamine derived from membrane phospholipids and by succinyl residues. In this study, we describe the isolation of the opgE gene encoding the phosphoethanolamine transferase by a screen previously used for the isolation of the opgB gene encoding the phosphoglycerol transferase. Both genes show structural and functional similarities without sequence similarity.

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Motility and mucoidy analysis of opg mutants of E. coli. (a) Motility: strains (107 CFU) were spotted onto swarming medium containing 0,3% agar. The plates were incubated at 37°C and motility was examined after 24 h of incubation. (b) Mucoidy: The same strains were streaked on SOBG medium. The plates were incubated at 30°C and mucoidy was examined after 30 h of incubation.
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fig5: Motility and mucoidy analysis of opg mutants of E. coli. (a) Motility: strains (107 CFU) were spotted onto swarming medium containing 0,3% agar. The plates were incubated at 37°C and motility was examined after 24 h of incubation. (b) Mucoidy: The same strains were streaked on SOBG medium. The plates were incubated at 30°C and mucoidy was examined after 30 h of incubation.

Mentions: The main phenotypes of mutant strains devoid of OPGs backbone (i.e., opgG or opgH) are loss of motility and increased exopolysaccharides synthesis (see Section 1). We decided to test these phenotypes in strains devoid of one or all kinds of substitution on OPGs backbone. For motility measurement, opgB (NFB732), opgC (NFB1887), opgE (NFB4576), and opgB opgC opgE (NFB4548) strains were spotted on swimming agar (see Section 2) for 24 h together with the wild type (JM83) and the opgH (NFB216) strains as controls. As expected for the two controls, the wild-type strain was motile while motility was severely reduced for the opgH strain. As observed for the wild-type strain, all the mutant strains tested were motile (Figure 5). For exopolysaccharides synthesis measurement, the same strains were plated onto SOB-Glycerol medium. As observed for the wild-type strain, all the mutant strains were non mucoid indicating no increase in exopolysaccharides synthesis except for the opgH mutant strain where mucoidy was increased indicating an increase in exopolysaccharides synthesis (Figure 5). Thus, no classical phenotype found in mutants devoid of OPGs backbone could be associated with mutants only devoid of substituent. Thus, the role of substituent remains to be elucidated.


Biosynthesis of osmoregulated periplasmic glucans in Escherichia coli: the phosphoethanolamine transferase is encoded by opgE.

Bontemps-Gallo S, Cogez V, Robbe-Masselot C, Quintard K, Dondeyne J, Madec E, Lacroix JM - Biomed Res Int (2013)

Motility and mucoidy analysis of opg mutants of E. coli. (a) Motility: strains (107 CFU) were spotted onto swarming medium containing 0,3% agar. The plates were incubated at 37°C and motility was examined after 24 h of incubation. (b) Mucoidy: The same strains were streaked on SOBG medium. The plates were incubated at 30°C and mucoidy was examined after 30 h of incubation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3818809&req=5

fig5: Motility and mucoidy analysis of opg mutants of E. coli. (a) Motility: strains (107 CFU) were spotted onto swarming medium containing 0,3% agar. The plates were incubated at 37°C and motility was examined after 24 h of incubation. (b) Mucoidy: The same strains were streaked on SOBG medium. The plates were incubated at 30°C and mucoidy was examined after 30 h of incubation.
Mentions: The main phenotypes of mutant strains devoid of OPGs backbone (i.e., opgG or opgH) are loss of motility and increased exopolysaccharides synthesis (see Section 1). We decided to test these phenotypes in strains devoid of one or all kinds of substitution on OPGs backbone. For motility measurement, opgB (NFB732), opgC (NFB1887), opgE (NFB4576), and opgB opgC opgE (NFB4548) strains were spotted on swimming agar (see Section 2) for 24 h together with the wild type (JM83) and the opgH (NFB216) strains as controls. As expected for the two controls, the wild-type strain was motile while motility was severely reduced for the opgH strain. As observed for the wild-type strain, all the mutant strains tested were motile (Figure 5). For exopolysaccharides synthesis measurement, the same strains were plated onto SOB-Glycerol medium. As observed for the wild-type strain, all the mutant strains were non mucoid indicating no increase in exopolysaccharides synthesis except for the opgH mutant strain where mucoidy was increased indicating an increase in exopolysaccharides synthesis (Figure 5). Thus, no classical phenotype found in mutants devoid of OPGs backbone could be associated with mutants only devoid of substituent. Thus, the role of substituent remains to be elucidated.

Bottom Line: Osmoregulated periplasmic glucans (OPGs) are oligosaccharides found in the periplasm of many Gram-negative bacteria.In E. coli, OPG are substituted by phosphoglycerol and phosphoethanolamine derived from membrane phospholipids and by succinyl residues.Both genes show structural and functional similarities without sequence similarity.

View Article: PubMed Central - PubMed

Affiliation: Unité de Glycobiologie Structurale et Fonctionnelle, UMR CNRS 8576, IFR 147, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq Cedex, France ; Université Lille Nord de France, Lille, France.

ABSTRACT
Osmoregulated periplasmic glucans (OPGs) are oligosaccharides found in the periplasm of many Gram-negative bacteria. Glucose is the sole constitutive sugar and this backbone may be substituted by various kinds of molecules depending on the species. In E. coli, OPG are substituted by phosphoglycerol and phosphoethanolamine derived from membrane phospholipids and by succinyl residues. In this study, we describe the isolation of the opgE gene encoding the phosphoethanolamine transferase by a screen previously used for the isolation of the opgB gene encoding the phosphoglycerol transferase. Both genes show structural and functional similarities without sequence similarity.

Show MeSH