The complete structure of the trifoliin A lectin-binding capsular polysaccharide of Rhizobium trifolii 843.
The complete structure of the acidic, extracellular, capsular polysaccharide of Rhizobium trifolii 843 has been elucidated by a combination of chemical, enzymic, and spectroscopic methods, confirming an earlier proposed sugar sequence and assigning the locations of the acyl substituents. The polysaccharide was depolymerized by a lyase into octasaccharide units which were uniform in carbohydrate composition and linkage. These units also contained a uniform distribution of acetyl and pyruvic acetal [O-(1-carboxyethylidene)] groups, and half of them were further acylated with D-3-hydroxybutanoyl groups. A much smaller proportion (less than 5%) of the oligomers was further acylated by a second D-3-hydroxybutanoyl group. The locations of the subtituents were determined chemically and by J-correlated, 1H-n.m.r. spectroscopy, proton nuclear Overhauser effect (n.O.e.) measurements, double-rd structure of the carbohydrate chain were determined by methylation analysis using g.l.c.-m.s. fast-atom-bombardment mass spectrometry, and n.m.r. studies on the reduced, deacylated oligomer. Structural studies were supplemented by n.m.r. analyses on the original polymer. The oligosaccharides were found to be branched octasaccharides with four sugar residues in each branch, and the carbohydrate sequence agreed well with that expected from earlier work. In the abbreviated sequence and structure (1a), the sugar residues are labelled "a" through "h". The main chain (a-d) is composed of a 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid group (a) that is linked to O-4 of a 3-O-acetyl-D-glucosyluronic acid residue (b) which is beta-linked to O-4 of a D-glucosyl residue (c). Residue c is beta-linked to O-4 of the branching D-glucose residue (d). The side chain consists of a substituted D-galactosyl group (h) which is beta-linked to O-3 of residue 9 of a beta-(1----4)-linked D-glucose trisaccharide (fragment e-f-g). The reducing end of the resulting tetrasaccharide (e-f-g-h) is beta-linked to O-6 of the branching D-glucose residue (d). In the native polymer, this branching residue is alpha-linked to O-4 of the modified D-glucuronic acid residue (a) which is the unsaturated sugar in the oligomer. A small proportion of the O-2 atoms of the acetylated D-glucosyluronic acid residues is acetylated because of ester migration. The two terminal sugars (g and h) of the branch chain bear 4,6-O-(1-carboxyethylidene) groups. The D-galactosyl groups of half of the oligomers are acylated by D-3-hydroxybutanoyl groups at O-3.(ABSTRACT TRUNCATED AT 400 WORDS)
Hollingsworth
R I
RI
Department of Microbiology and Public Health, Michigan State University, East Lansing 48824.
Dazzo
F B
FB
Hallenga
K
K
Musselman
B
B
D002240:Carbohydrate Sequence; D005419:Flavonoids / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011135:Polysaccharides, Bacterial; D011794:Quercetin / Q000031:analogs & derivatives / Q000378:metabolism*; D012231:Rhizobium / Q000032:analysis*
D005419:Flavonoids; D037102:Lectins; D011135:Polysaccharides, Bacterial; C016527:isoquercitrin; D011794:Quercetin