Characterization of lipopolysaccharides of Pectinatus cerevisiophilus

Helander, Ilkka M.

doi:10.1016/0378-1097(83)90331-2

Cited by 8 publications

(17 citation statements)

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“…The absence of heptose in Pectinatus LPS, already noted in a previous report (15), was verified in the present study. 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.…”

Section: Resultssupporting

confidence: 93%

“…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.…”

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confidence: 99%

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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: Resultssupporting

confidence: 93%

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confidence: 99%

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

et al. 1992

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“…Among this family, the inner core regions of lipopolysaccharides of members of the genus Bacteroides have been reported to contain phosphorylated Kdo [9, 161, which renders Kdo undetectable with the thiobarbituric acid method. In this respect the genus Pectinatus clearly differs from Bacteroides, as it also does in the lipopolysaccharide fatty-acid composition [17].…”

Section: Discussionmentioning

confidence: 97%

4‐O‐(2‐Amino‐2‐deoxy‐α‐d‐glucopyranosyl)‐3‐deoxy‐d‐manno‐2‐octulosonic acid, a constituent of lipopolysaccharides of the genus Pectinatus

Helander

Moll²,

Zähringer³

1993

European Journal of Biochemistry

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A disaccharide containing GlcN and 3-deoxy-D-manno-2-octu~osonic acid (Kdo) was detected after acidic methanolysis and peracetylation of hydrofluoric-acid-treated smooth-type and roughtype lipopolysaccharide of Pectinatus cerevisiiphilus and Pectinatus frisingensis, which are strictly anaerobic bacteria capable of growing in packaged beer. The disaccharide was also found in alkalitreated lipopolysaccharide, but was not directly detectable from intact lipopolysaccharide. This suggested that the disaccharide carried a phosphate residue. The position of this phosphate was shown, by GLCMS of appropriately degraded and derivatized samples, to be 0 6 of the GlcN. Methylation analysis of the purified disaccharide revealed that GlcN was linked to position 4 of Kdo. The acetylated derivative of the disaccharide was isolated in pure form, and, by 'H-NMR and I3C-NMR spectroscopy, it was confirmed to possess the structure a-~-GlcpN-(1'+4)-Kdo. In the lipopolysaccharide the amino group of GlcN is free.The Gram-negative bacterial genus Pectinatus contains two species, I? cerevisiiphilus and I? frisingensis. Both species are strict anaerobes, and they occur as contaminants and spoilage organisms in packaged beer [ 1, 21. The cell-surface lipopolysaccharides of the type strains of Pectinatus were recently reported to possess exceptional heterogeneity, and in fact two distinct lipopolysaccharide populations could be extracted from both species [3]. One of these lipopolysaccharides was of low-molecular-mass (rough, R) type, whereas the other contained additional sugar chains resembling smooth ( S ) lipopolysaccharide. Chemical characterization of the lipopolysaccharides indicated possible differences in the core oligosaccharide regions of the R and S type, particularly in the substitution of the 3-deoxyoctulosonic-acid region.These findings prompted us to structurally study the core region of Pectinatus lipopolysaccharides. In the present communication we describe a novel saccharide structure in the lipopolysaccharides of Pectinatus. This saccharide consists of a 3-deoxy-D-manno-2-octulosonic acid (Kdo) carrying at its position 4 an a-pyranosidic GlcN, which is substituted at position 6' by phosphate. MATERIALS AND METHODS Reference compoundsMethyl 2-acetarnido-3,4,6-triri-O-acetyl-2-deoxy-a-~-glucopyranoside was prepared from methyl 2-acetamido-2-de- oxy-a-D-glucopyranoside (a-D-GlcpNAc-OMe, Sigma) by standard derivatization (peracetylation) and purification (silica gel HPLC) procedures. Methyl (methyl 4,5,7,8-tetra-O-acetyl-3-deoxy-a-~-manno-2-octulopyranos~d)onate (aKdop-OMe methyl ester) was kindly provided by Prof. Paul Kosma, Universitat fur Bodenkultur, Vienna, Austria. Lipopoly saccharidesLipopolysaccharides were extracted from P. cerevisiiphilus ATCC 29359= and I? frisingensis ATCC 33332T by the phenol/chloroform/petroleum-ether method (5 : 5 :8, by vol. modification [4]) as described previously [3]. By sequential precipitations with water and acetone, R-type and S-type lipopolysaccharides, respectively, were obtained fr...

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“…The unrelatedness is also evident on the basis of the lipopolysaccharide composition of representatives of the genus Pectinatus (Helander et al, 1983;Helander et al, 1992).…”

Section: Pectinatusmentioning

confidence: 93%