Structural studies on the lipopolysaccharide from a rough strain of Ochrobactrum anthropi containing a 2,3-diamino-2,3-dideoxy-D-glucose disaccharide lipid A backbone.
A degradation protocol using de-O-acylation and subsequent alkaline de-N-acylation was applied to the lipopolysaccharide of Ochrobactrum anthropi rough strain LMG 3301. Three main oligosaccharide bisphosphates containing core-lipid A backbone structures were obtained after fractionation by anion-exchange HPLC. Using 1H and 13C NMR spectroscopy, including two-dimensional COSY, TOCSY, and NOE spectroscopy (ROESY and NOESY), the following structures were established: [formula: see text] where Kdo is 3-deoxy-D-manno-octulosonic acid, D-GlcN3N is 2,3-diamino-2,3-dideoxy-D- glucose and R is H or alpha-D-GalpA or 4-deoxy-beta-L-threo-hex-4-enopyranuronic acid, the latter sugar being derived from alpha-D-GalpA by beta-elimination of a substituent attached to 0-4. This is the first report on the isolation from a lipopolysaccharide of an oligosaccharide containing GlcN3N in the lipid A backbone [beta-D-GlcpN3N4P-(1-->6)-alpha-D-GlcpN3N1 P]. Sugar and methylation analysis confirmed the presence of the GalA-->Kdo disaccharide and non-stoichiometric substitution of GalA. It is suggested that Glc is the substituent at 0-4 in GalA and that in the non-degraded lipopolysaccharide the amino group of GlcN is not acylated.
Forschungszentrum Borstel, Zentrum für Medizin und Biowissenschaften, Germany.
J
J
Velasco
H
H
Moll
Y A
YA
Knirel
V
V
Sinnwell
I
I
Moriyón
U
U
Zähringer
D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000737:chemistry; D016946:Gram-Negative Aerobic Rods and Cocci / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008050:Lipid A / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification
D008050:Lipid A; D008070:Lipopolysaccharides; D009844:Oligosaccharides; C040138:2,3-diamino-2,3-dideoxyglucose; D005944:Glucosamine