Mikrobiol. Z. 2018; 80(2):56-66. Ukrainian.
doi: https://doi.org/10.15407/microbiolj80.02.056

Lipopolysaccharide of Pantoea agglomerans 9649: 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 Pantoea agglomerans 9649, investigation its biological activities and also to establish 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 Osbourne method. Monosaccharide and fatty acid composition was 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 of P. agglomerans 9649 was purifed and characterized chemically. The predominant monosaccharides were rhamnose (36.47 %), mannose (20.17 %), heptoses (14.87 %) and glucose (13.57 %). In the lipid part of the LPS studied were identified saturated, monounsaturated and hydroxy acids with chain lengths from C12 to C16. Since the studiet strain of P. agglomerans were resistant to polymixin B, it can be assume that the LPS 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 display a pyrogenic effect. The antisera against to the tested strain reacted with LPS strain of П1а, indicating the presence of the common antigenic determinants belonging to one of the same serogroup. The studied LPS reduces the index of adhesiveness, suggesting a possible competition between molecules of P. agglomerans 9649 LPS and E. coli F-50 adhesin, involved in the erythrocytes of the rabbit.

Keywords: Pantoea agglomerans, lipopolysaccharide, monosaccharide and fatty acid composition, 4-amino-4-deoxy-L-arabinose, serological and adhesive activity.

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