NoteO-chain structure from the lipopolysaccharide of the human pathogen Halomonas stevensii strain S18214
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Section snippets
Growth bacteria, isolation and purification of the LPS
Halomonas stevensii strain S18214 was grown for 4 days at 35 °C in brain heart infusion broth (3% NaCl added) adjusted to pH 8.0; cells were checked for purity, harvested by centrifugation and lyophilized. Dried bacteria cells (6 g) were extracted by PCP method,20 obtaining 90 mg of LPS (yield 1.5% of dried cells). The pellet was then extracted by phenol–water method.21
SDS–PAGE electrophoresis
PAGE was performed by using the system of Laemmli and Favre24 with sodium dodecyl sulfate as the detergent. The separating gel
Acknowledgments
NMR experiments were carried out on instruments at Centro Interdipartimentale Metodologie Chimico Fisiche Università di Napoli. Authors are also grateful to BioTekNet for the use of 600 MHz NMR instrument.
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2016, Carbohydrate PolymersCitation Excerpt :The three EPS investigated herein have structural elements in common, viz., first of all the disaccharide β-lactose, second the trisaccharide α-d-Glcp-(1 → 4)-β-d-Galp-(1 → 4)-β-d-Glcp making up the backbone of the K16 EPS and branching regions in the THS and ST1 polysaccharides and thirdly the sequence β-d-Galp-(1 → 4)-β-d-Glcp-(1 → 6)-α-d-Glcp-(1 → 4)-β-d-Galp-(1 → 4)-β-d-Glcp, which is terminated by β-lactose, occurs in both THS and K16, where in the latter structure an additional β-(1 → 2)-linked glucose residue is present forming a branched side-chain. These structural elements are also part of other polysaccharides, e.g. in the O-antigens from Halomonas magadii strain 21 MI (de Castro, Molinaro, Wallace, Grant, & Parrilli, 2003) and Halomonas stevensii strain S18214 (Pieretti et al., 2011) both of which have a trisaccharide structure of the repeating unit corresponding to the EPS from THS devoid of the terminal lactose entity in the side-chain, i.e., →3)-[α-d-Glcp-(1 → 4)]β-d-Galp-(1 → 4)-β-d-Glcp-(1→, as well as in the CPS from Klebsiella type 37 which has a tetrasaccharide RU containing this trisaccharide structural element (Lindberg, Lindqvist, Lönngren, & Nimmich, 1977). The EPS from Lactobacillus delbrueckii subsp. bulgaricus 291 (Faber, Kamerling, & Vliegenthart, 2001) has a pentasaccharide RU that corresponds to that of the EPS from K16 devoid of the β-(1 → 2)-linked glucose residue in the side-chain and this is also the case for the EPS from Lactobacillus lactis subsp. cremoris B891 (van Casteren, de Waard, Dijkema, Schols, & Voragen, 2000) which in addition carries an O-acetyl group on the side-chain glucose residue.