Mikrobiol. Z. 2017; 79(5):23-33. Russian.
doi: https://doi.org/10.15407/microbiolj79.05.023

Lipopolysaccharide of Pantoea agglomerans 7604: Chemical Identification and Biological Activity

Bulyhina T.V., Varbanets L.D., Pasichnyk L.A.

Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

The goal of the present work was isolation, chemical characterization of the lipopolysaccharide (LPS) of P. agglomerans 7604, investigation of its biological activities and also estimation of serological relationships with other strains of this species. Methods. The amounts of the following components were determined: carbohydrates by the Dubois method; nucleic acids by the method of Spirin; protein by the Lowry method; 2-keto-3-deoxyoctulosonic acid by the Osborn method. Monosaccharide and fatty acid composition were analyzed on an Agilent 6890N/5973 inert chromato-mass-spectrometric system. The sensitivity of microbial cultures of P. agglomerans to polymyxin B was determined by disco-diffusion method. The antigenic activity of LPS was studied by the method of double immunodiffusion in agar according to Ouchterlony. Adhesive activity was determined by express-method for Briles. Results and conclusions. Lipopolysaccharide (LPS) of Pantoea agglomerans 7604 was purified and characterized chemically. The predominant monosaccharides were mannose (69.9 %) and arabinose (17.0 %). The LPS included fatty acids with chain lengths from 12 to 16 carbon atoms. In addition to saturated acids, monounsaturated acids have also been detected: palmitoleic, oleic and its cis- and trans-isomers. Since the studied strain of P. agglomerans was sensitive to polymixin B, it can be concluded that the LPS didn`t contain in the structure of lipid A such a substitute as 4-amino-4-deoxy-L-arabinose. The results of thermometry showed that the investigated LPS displays a pyrogenic effect. The antisera to the test strain reacted with LPS strains of 8606 and 8674, indicating the presence the common antigenic determinants. This may indicate that this strains belong to one and the same serogroup. The studied LPS reduces the index of adhesiveness, suggesting a possible competition between molecules of P. agglomerans 7604 LPS and adhesions of E. coli F-50, involved on the erythrocytes of the rabbit.

Key words: Pantoea agglomerans, lipopolysaccharide, monosaccharide and fatty acid composition, 4-amino-4-deoxy-L-arabinose, adhesion.

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