Structures of the O-specific Polysaccharide Chains of Pectinatus cerevisiiphilus and Pectinatus frisingensis Lipopolysaccharides

Abstract

Mild acid hydrolysis of the smooth-type lipopolysaccharide (LPS) of Pectinatus ,frisingensis afforded no polysaccharide but monomeric 6-deoxy-~-altrose (L-6dAlt) which was identified by anion-exchange chromatography in borate buffer, GLC/MS, 'H-NMR spectroscopy, and optical rotation. LPS was degraded with alkali under reductive conditions to give a completely 0-deacylated polysaccharide, which was studied by methylation analysis, 'H-NMR and IT-NMR spectroscopy, including sequential, selective spin-decoupling, two-dimensional correlation spectroscopy (COSY), COSY with relayed coherence transfer, two-dimensional heteronuclear "C,'H-COSY, one-dimensional NOE and two-dimensional rotating-frame NOE spectroscopy. It was found that the 0-specific polysaccharide chain of l? frisingensis LPS is a homopolymer of 6-deoxy-~-altrofuranose built up of tetrasaccharide-repeating units having the following structure:Similarly, mild acid degradation of smooth-type LPS of Pectinatus cerevisiiphilus resulted in depolymerisation of the polysaccharide chain to give a disaccharide consisting of D-glUCoSe and D-fucose. Study of the disaccharide by methylation analysis and alkali-degraded LPS by one-dimensional and twodimensional 'H-NMR and "C-NMR spectroscopy showed that the 0-specific polysaccharide of I? cerevisiiphilus has the following structure : --.2)-~-~-Fucf-(l-+2)-a-~-Glcp-(l--,Keywords: Pectinatus ; lipopolysaccharide; 0-specific polysaccharide; structure; methylation.Recently, outer-membrane lipopolysaccharides (LPS) of two species of the strictly anaerobic beer spoilage bacteria of the genus Pectinatus have been isolated and chemically characterised [l]. Both species (l? cerevisiiphilus and 19 frisingensis) elaborated mainly smooth-type LPS, but small amounts of chemically distinct rough-type LPS was also extracted from the cells. Sugar analysis showed that the 0-specific polysaccharide chains of the smooth-type Pectinatus LPSs are rich in 6-deoxyhexoses [I]. We now report the structures of the 0-specific polysaccharides of P. cerevisiiphilus and 19 frisingensis. The former contains, amongst others, D-fucofuranose, whereas the latter is a homopolymer of 6-deoxy-~-altrofuranose. EXPERIMENTAL PROCEDURESChromatography. Gel-permeation chromatography was carried out on a column (40 cmX2.S cm) of Sephadex G-50 or on a column (40 cmX1.5 cm) of TSK HW-40 using 0.5 M pyridine acetate, pH 4.5, or water, respectively as eluent; monitoring was performed using a Knauer differential refractometer. Anionexchange chromatography was performed with a column (15 cmX0.6 cm) of Durrum DAX4 resin using 0.S M sodium borate, pH 9, at 55 "C and monitored using a Biotronik LC-2000 sugar analyser. GLC was performed using a Hewlett-Packard 5890 instrument equipped with a glass capillary column (25 mX0.2 mm) coated with Ultra 1 stationary phase.NMR spectroscopy. NMR spectra of D,O solutions were run with a Bruker WM-250 instrument at 60°C. Acetone was used as internal standard (~3~2 . 2 3 ppm, 6,31.45 ppm). Selective spin decoupling [2], one-dimensiona...

Antibacterial activity of a disaccharide isolated fromStreptomycessp. strain JJ45 againstXanthomonassp.

“…l ‐Sorbose is the precursor of 2‐keto‐ l ‐gluconic acid, which is the precursor of vitamin C (Sugisawa et al , 2005). Although l ‐altrose is present in many bacterial cell wall components or precursors (Senchenkova et al , 1995; Liu & Tanner, 2006), d ‐altrose is found rarely. Judging by its narrow spectrum of action, α‐ l ‐sorbofuranose (3→2)‐β‐ d ‐altrofuranose may disturb a pathway involving sorbose or altrose, perhaps a pathway unique to Xanthomonas sp.…”

Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E‐82164

“…This unusual construction would be better named lipo-oligosaccharide or LOS, although usually the term LOS is used to denote LPS from natural rough strains with core-lipid A parts only [18]. This conclusion is supported by the discovery of a similar core part in the polysaccharide O-chain-containing strain E-79100 T (O-chain structure described in [5]).…”

“…The lipopolysaccharides (LPS) of type strains of P. cerevisiiphilus and P . frisingensis possess a number of remarkable properties, including the predominance of odd‐numbered fatty acids in lipid A [4] and the presence of furanosidic 6‐deoxysugars in the O‐specific chains [5]. The lipid A was shown to be quantitatively substituted at the 4′‐phosphate and partially at the glycosidic phosphate by 4‐amino‐4‐deoxy‐β‐ l ‐arabinose [6].…”

Lipopolysaccharides of anaerobic beer spoilage bacteria of the genusPectinatus– lipopolysaccharides of a Gram-positive genus

“…The structures of the repeating units of these strains have been analyzed [53]. In P. cerevisiiphilus ATCC 29359 T the structure of the repeating unit is the disaccharide !…”

“…Structure of the O-specific repeating unit of P. frisingensis ATCC 33332 T was shown [53] to consist of a single monosaccharide unit, the rare 6-deoxy-L L -altrofuranose in a branched tetrasaccharide structure…”