Open AccessIsolation and structural characterization of an R‐form lipopolysaccharide from Yersinia enterocolitica serotype O:8
Author(s) -
Oertelt Clemens,
Lindner Buko,
Skurnik Mikael,
Holst Otto
Publication year - 2001
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.2001.01891.x
Subject(s) - lipid a , chemistry , amide , yersinia enterocolitica , heptose , stereochemistry , chloroform , nuclear magnetic resonance spectroscopy , lipopolysaccharide , bacteria , mutant , biochemistry , organic chemistry , biology , genetics , gene , endocrinology
The lipopolysaccharide (LPS) of strain 8081‐c‐R2, a spontaneous R‐mutant of Yersinia enterocolitica serotype O:8, was isolated using extraction with phenol/chloroform/light petroleum. Its compositional analysis indicated the presence of d ‐GlcN, d ‐Glc, l glycero d manno ‐ and d glycero d manno ‐heptose, 3‐deoxy‐ d manno ‐oct‐2‐ulosonic acid (Kdo) and phosphate. From deacylated LPS obtained after successive treatment with hydrazine and potassium hydroxide, three oligosaccharides (1–3) were isolated using high‐performance anion‐exchange chromatography, the structures of which were determined by compositional analysis and one‐ and two‐dimensional NMR spectroscopy as in which all sugars are pyranoses, and R and R′ represent β‐ d ‐Glc (in 1 and 2) and β‐ d ‐GlcN (in 1 only), respectively. d ‐α‐ d ‐Hep is d glycero ‐α‐ d manno ‐heptose, l ‐α‐ d ‐Hep is l glycero ‐α‐ d manno‐ heptose, Kdo is 3‐deoxy‐ d manno ‐oct‐2‐ulosonic acid, and P is phosphate. The liberated lipid A was analyzed by compositional analyses and MALDI‐TOF MS. Its β‐ d ‐GlcN4P‐(1→6)‐α‐ d ‐GlcN‐1→P backbone is mainly tetra‐acylated with two amide‐ and one ester‐linked (at O3 of the reducing GlcN) ( R )‐3‐hydroxytetradecanoic acid residues, and one tetradecanoic acid that is attached to the 3‐OH group of the amide‐linked ( R )‐3‐hydroxytetradecanoic acid of the nonreducing GlcN. Additionally, small amounts of tri‐ and hexa‐acylated lipid A species occur.