Repeating unit structure of Enterobacter sakazakii ZORB A 741 O-polysaccharide

Janusz Szafranek , Małgorzata Czerwicka , Jolanta Kumirska , Monika Paszkiewicz , Ewa Łojkowska

Abstract

The O-specific polysaccharide from Enterobacter sakazakii cell was isolated and structurally characterized. Lipopolysaccharide (LPS) was obtained from cell mass by hot phenol-water extraction procedure. Mild acid hydrolysis followed by gel filtration provided pure O-antigen (OPS). Two-stage sugar analysis detected tyvelose, rhamnose and galactose in the molar ratio of 1:1:2, and their linkages were established by means of methylation analysis. Sugar configurations, D or L, were determined by gas-liquid chromatography on an achiral liquid phase for (S)-(+)-2-butyl glycosides. D configuration was determined for galactose and 3,6-dideoxy-mannose (tyvelose), but L for rhamnose. Repeating unit structure was deduced by analysis of 1H and 13C NMR spectra. 1H and 13C NMR resonances have been assigned by homonuclear (COSY, TOCSY) and heteronuclear (HSQC, HMBC) correlations spectra. Anomeric configurations were determined from anomeric proton chemical shifts and 3JH1-H2 and JC-H coupling constants. Sugar sequences were established from comparisons of specific carbon chemical shifts with those in literature, two-dimensional nuclear Overhauser effect spectroscopy (NOESY), and heteronuclear multiple-bond correlation experiments (HMBC). The repeating unit structure of Enterobacter sakazakii was found to be as: (Equation Presented).
Author Janusz Szafranek (FCh)
Janusz Szafranek,,
- Faculty of Chemistry
, Małgorzata Czerwicka (FCh / DBCh / LSB)
Małgorzata Czerwicka,,
- Laboratory of Structural Biochemistry
, Jolanta Kumirska (FCh / DEA / LChAD)
Jolanta Kumirska,,
- Laboratory of Chemical Analytics and Diagnostics
, Monika Paszkiewicz (FCh / DEA / PAiMŚ)
Monika Paszkiewicz,,
- Pracownia Analityki i Monitoringu Środowiska
, Ewa Łojkowska (IFB / IB / DBiotech)
Ewa Łojkowska,,
- Department of Biotechnology
Journal seriesPolish Journal of Chemistry, [Roczniki Chemii], ISSN 0137-5083, (A 15 pkt, Not active)
Issue year2005
Vol79
No2
Pages287-295
Publication size in sheets0.5
ASJC Classification1600 General Chemistry
Languageen angielski
Score (nominal)15
Score sourcejournalList
Publication indicators Scopus Citations = 12; Scopus SNIP (Source Normalised Impact per Paper) [Not active]: 2005 = 0.368; WoS Impact Factor: 2006 = 0.491 (2) - 2007=0.536 (5)
Citation count*15 (2020-02-11)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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