Structure of the sidechain of lipopolysaccharide from <i>Erwinia amylovora</i> T

Ray, T. C.; Smith, Andrew; Wait, Robin; Hignett, R. C.

doi:10.1111/j.1432-1033.1987.tb13707.x

Cited by 22 publications

(8 citation statements)

References 16 publications

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“…The major neutral sugar component of the smooth-type LPS in both Pectinatus species was fucose, and it was present exclusively in the D configuration. In nature, D-fucose is a relatively rare sugar (8), and it has been previously identified in the 0 antigen of a few bacteria, such as Pseudomonas cepacia (21), Pseudomonas syringae (22,34), and Erwinia amylovora T (36). These bacteria are plant pathogens, and the presence of D-6-deoxyhexoses in their LPS has actually been associated with plant pathogenicity (43).…”

Section: Resultsmentioning

confidence: 99%

Separation and characterization of two chemically distinct lipopolysaccharides in two Pectinatus species

et al. 1992

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Lipopolysaccharides (LPS) from the type strains of the anaerobic beer spoilage bacteria Pectinatus cerevisiiphilus and P. frisingensis were extracted with the 5:5:8 volume ratio modification of the phenolchloroform-petroleum ether method (H. Brade and C. Galanos, Eur. J. Biochem. 122:233-237, 1982). Sequential precipitations of LPS with water and acetone from the phenol phase yielded LPS which differed in that water-precipitable material (LPS-H20; 0.1 to 0.4% of the dry weight of the cells) was rough-type LPS, whereas acetone-precipitable material (LPS-Ac; 4.6 to 5.8% of the dry weight) contained both rough-type LPS and high-molecular-weight material resembling smooth LPS. The LPS were chemically characterized, and they contained D-glucosamine, 4-amino-4-deoxy-L-arabinose, 3-deoxy-D-manno-2-octulosonic acid, D-fucose, D-galactose, D-glucose, D-mannose, and phosphate. D-Fucose was present mostly in LPS-Ac, suggesting that it is a constituent of the 0 antigen. The major fatty acids were ester-and amide-linked (R)-3-hydroxytridecanoic and ester-linked undecanoic acids, with minor amounts of ester-linked tridecanoic and (R)-3-hydroxyundecanoic acids. The chemical compositions of LPS-H20 and LPS-Ac suggested that they differ not only in their smooth or rough nature but also in the structure of their core regions. This may explain their different precipitabilities from the extraction mixture. The extraction method was also shown to be applicable to the isolation of smooth-type LPS from Salmonella enterica serovar Typhimurium. Extraction of two Typhimurium strains carrying chemically different 0 antigens resulted in high yields (8% of the dry weight) of LPS. Strain SH2183, which contains the relatively hydrophobic 0-4,5,12 antigen yielded almost exclusively LPS-Ac, whereas the LPS of strain SH5770, which has a hydrophilic 0-6,7 antigen, was exclusively LPS-H20. No fractionation to smooth and rough LPS occurred with the Typhimurium strains.The gram-negative genus Pectinatus consists of obligately anaerobic rod-shaped bacteria which have been identified as spoilage-causing organisms in packaged beer (1,12,25,45). Since their initial description and establishment of the genus and the species Pectinatus cerevisiiphilus (25), another species, P. frisingensis, has been described (41).Lipopolysaccharides (LPS), also termed endotoxins, are structural components of the outer membrane of all gramnegative bacteria (for reviews, see references 27, 33, and 38). Since the chemical composition and structure of LPS show large variation between bacterial genera (27), the composition of LPS can serve as a taxonomic criterion. For this purpose, we previously performed a preliminary compositional analysis of the LPS of four isolates of Pectinatus, then classified as the species P. cerevisiiphilus (15). That study revealed exceptional properties in the LPS, such as the predominance of fatty acids with odd-numbered chain lengths of 11 and 13 carbons and the lack of heptose, a sugar which is commonly present in LPS. Because of the general in...

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Section: Resultsmentioning

confidence: 99%

Separation and characterization of two chemically distinct lipopolysaccharides in two Pectinatus species

et al. 1992

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“…Nevertheless, furanosides in general and those of glucose are found in nature. [27][28][29][30] Studies carried out with disaccharides in the negativeion mode [10][11][12][13][14][15][16] prompted four questions concerning their dissociation and the potential of garnering additional information about their structures: (1) Do any of the product ions of disaccharide dissociation, upon isolation and further fragmentation, permit differentiation of anomeric configuration and/or ring form? Of the above studies in the negative-ion mode, [10][11][12][13][14][15] only one further dissociated the ions derived by cross-ring cleavage of the disaccharides.…”

Section: Introductionmentioning

confidence: 99%

Differentiation of the anomeric configuration and ring form of glucosyl-glycolaldehyde anions in the gas phase by mass spectrometry: isomeric discrimination between m/z 221 anions derived from disaccharides and chemical synthesis of m/z 221 standards

Fang¹,

Zirrolli

Bendiak³

2007

Carbohydrate Research

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“…1-5 The structures of O polysaccharides were es tablished only for two strains, viz., E. carotovora ssp. atroseptica GSPB 436 1 and Erwinia amylovara T. 4 The classification scheme of bacteria Erwinia is lacking. With the aim of classifying strains of phytopathogenic bacteria, we perform a series of structural studies of their O poly saccharides.…”

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Structure of the O-polysaccharide of Erwinia carotovora ssp. atroseptica GSPB 9205 containing a new higher branched monosaccharide

et al. 2005

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An O polysaccharide was prepared by mild acid hydrolysis of a lipopolysaccharide of Erwinia carotovora ssp. atroseptica GSPB 9205 and was found to contain a new higher branched monosaccharide, viz., 3,6,8 trideoxy 4 C (R 1 hydroxyethyl) D gulo octose, which we called erwiniose. The structure of the pentasaccharide repeating unit of the O polysaccharide was established by monosaccharide analysis, including the determination of absolute configura tions, methylation analysis, O deacetylation, Smith degradation, and one and two dimen sional NMR spectroscopy. The configuration of erwiniose was determined based on the cou pling constants of vicinal protons combined with the nuclear Overhauser effect data and the results of periodate oxidation of the polysaccharide followed by reduction of the resultant 4 keto sugar (the C(1)-C(4)-C(1´)-C(2´) fragment) to give 3,6 dideoxy D ribo hexose (paratose) and oxidation of 3 hydroxybutyraldehyde (the C(5)-C(8) fragment) to give R 3 hydroxybutyric acid.Lipopolysaccharide (LPS) forms the outer layer of the outer membrane of the Gram negative bacteria cell walls. The fine structure of the O polysaccharide chain of the LPS is responsible for intraspecies diversity of strains and specificity of interactions of bacterial cells with other bio logical systems. Classification schemes of many Gram negative bacteria necessary for epidemiologic monitoring are based on O polysaccharide structures.Lipopolysaccharides of phytopathogenic bacteria are involved in plant pathogenesis, for example, in soft rot caused by Erwinia carotovora strains. The structure of LPS of bacteria belonging to the Erwinia genus was poorly studied. 1-5 The structures of O polysaccharides were es tablished only for two strains, viz., E. carotovora ssp. atroseptica GSPB 436 1 and Erwinia amylovara T. 4 The classification scheme of bacteria Erwinia is lacking. With the aim of classifying strains of phytopathogenic bacteria, we perform a series of structural studies of their O poly saccharides. In the present study, we established the structure of the O polysaccharide of E. carotovora ssp. atroseptica GSPB 9205. We found and identified a new higher branched monosaccharide in this O polysaccharide and called it erwiniose. Results and DiscussionMild acid degradation of LPS, which was isolated from E. carotovora ssp. atroseptica GSPB 9205 cells by aque ous phenol extraction, afforded an O polysaccharide (1). Monosaccharide analysis by GLC of alditol acetates ob tained upon total acid hydrolysis of polysaccharide 1 re vealed the presence of galactose, mannose, and rhamnose in a ratio of 2 : 1 : 1. The GLC analysis of acetylated glycosides with an optically active alcohol, viz., S octan 2 ol, revealed D configuration for galactose and mannose and L configuration for rhamnose. Methylation analysis of the polysaccharide revealed 3 substituted galactose resi dues, a 4 substituted rhamnose residue, and a 2,3 disub stituted mannose residue. The presence of the latter monosaccharide is indicative of a branched structure of the polys...

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Structure of the sidechain of lipopolysaccharide from Erwinia amylovora T

Cited by 22 publications

References 16 publications

Separation and characterization of two chemically distinct lipopolysaccharides in two Pectinatus species

Separation and characterization of two chemically distinct lipopolysaccharides in two Pectinatus species

Differentiation of the anomeric configuration and ring form of glucosyl-glycolaldehyde anions in the gas phase by mass spectrometry: isomeric discrimination between m/z 221 anions derived from disaccharides and chemical synthesis of m/z 221 standards

Structure of the O-polysaccharide of Erwinia carotovora ssp. atroseptica GSPB 9205 containing a new higher branched monosaccharide

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