Identification and structural determination of the capsular polysaccharides from two Acinetobacter baumannii clinical isolates, MG1 and SMAL

The structures of the capsular polysaccharides (CPSs) of the two clinical isolates Acinetobacter baumannii SMAL and MG1 were elucidated. Hot phenol/water extractions of the dry biomasses, followed by enzymatic digestions and repeated ultracentrifugations led to the isolation of polysaccharides that...

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Bibliographic Details
Published in:Carbohydrate research 2011-05, Vol.346 (7), p.973-977
Main Authors: Fregolino, Eleonora, Gargiulo, Valentina, Lanzetta, Rosa, Parrilli, Michelangelo, Holst, Otto, Castro, Cristina De
Format: Article
Language:English
Subjects:
Acinetobacter
Acinetobacter baumannii
Acinetobacter baumannii - chemistry
Acinetobacter baumannii - immunology
Acinetobacter Infections - microbiology
antibodies
antigens
Capsular polysaccharide
Carbohydrate Sequence
Gas Chromatography-Mass Spectrometry
Molecular Sequence Data
NMR spectroscopy
nuclear magnetic resonance spectroscopy
O Antigens - chemistry
phenol
polysaccharides
Polysaccharides, Bacterial - chemistry
Polysaccharides, Bacterial - isolation & purification
Structural analysis
ultracentrifugation
Western blot
western blot analysis
Western blotting
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Summary:The structures of the capsular polysaccharides (CPSs) of the two clinical isolates Acinetobacter baumannii SMAL and MG1 were elucidated. Hot phenol/water extractions of the dry biomasses, followed by enzymatic digestions and repeated ultracentrifugations led to the isolation of polysaccharides that were negative in Western blot analysis utilizing an anti-lipid A antibody, thus proving that they were not the LPS O-antigens but CPSs. Their structures were established on the basis of NMR spectroscopy and GC-MS analyses. The A. baumannii MG1 CPS consisted of a linear aminopolysaccharide with acyl substitution heterogeneity at the N-4 amino group of Qui pN4N: 4)-α- d-GlcpNAc-(1→4)-α- l-GalpNAcA-(1→3)-β- d-QuipNAc4NR-(1→ R = 3-hydroxybutyrryl or acetyl. The repeating unit of the CPS produced by strain SMAL is a pentasaccharide, already reported for the O-antigen moiety from A. baumannii strain ATCC 17961: [Display omitted]
ISSN:0008-6215
1873-426X
DOI:10.1016/j.carres.2011.03.024