Osmoregulated periplasmic glucans of the free-living photosynthetic bacterium Rhodobacter sphaeroides.
The osmoregulated periplasmic glucans (OPGs) produced by Rhodobacter sphaeroides, a free-living organism, were isolated by trichloracetic acid treatment and gel permeation chromatography. Compounds obtained were characterized by compositional analysis, matrix-assisted laser desorption ionization mass spectrometry and nuclear magnetic resonance. R. sphaeroides predominantly synthesizes a cyclic glucan containing 18 glucose residues that can be substituted by one to seven succinyl esters residues at the C6 position of some of the glucose residues, and by one or two acetyl residues. The glucans were subjected to a mild alkaline treatment in order to remove the succinyl and acetyl substituents, analyzed by MALDI mass spectrometry and purified by high-performance anion-exchange chromatography. Methylation analysis revealed that this glucan is linked by 17 1,2 glycosidic bonds and one 1,6 glycosidic bond. Homonuclear and (1)H/(13)C heteronuclear NMR experiments revealed the presence of a single alpha-1,6 glycosidic linkage, whereas all other glucose residues are beta-1,2 linked. The different anomeric proton signals allowed a complete sequence-specific assignment of the glucan. The structural characteristics of this glucan are very similar to the previously described OPGs of Ralstonia solanacearum and Xanthomonas campestris, except for its different size and the presence of substituents. Therefore, similar OPGs are synthesized by phytopathogenic as well as free-living bacteria, suggesting these compounds are intrinsic components of the Gram-negative bacterial envelope.
Talaga
Philippe
P
Unité de Glycobiologie Structurale et Fonctionnelle, CNRS UMR8576, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Cogez
Virginie
V
Wieruszeski
Jean-Michel
JM
Stahl
Bernd
B
Lemoine
Jérôme
J
Lippens
Guy
G
Bohin
Jean-Pierre
JP
D000595:Amino Acid Sequence; D002852:Chromatography, Ion Exchange; D005936:Glucans / Q000096:biosynthesis*; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D008969:Molecular Sequence Data; D019897:Periplasm / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D014867:Water / Q000378:metabolism
D005936:Glucans; D014867:Water