Mikrobiol. Z. 2020; 82(1):33-42. Ukrainian.
doi: https://doi.org/10.15407/microbiolj82.01.033

Characteristic of Pseudomonas putida Lipopolysaccharide

O.S. Brovarskaya1, L.D. Varbanets1, A.F. Likhanov2

1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

2National University of Life and Environmental Sciences of Ukraine
15 Heroes of Defense Str., Kyiv, 03041, Ukraine

Pseudomonas putida, along with representatives of other species such as Pseudomonas syringae and Pseudomonas fluorescens, cause diseases of a significant number of wild and cultivated plants. One of the factors of pathogenicity of gram-negative bacteria are lipopolysaccharides. There is evidence in the literature that, depending on the medium on which the bacteria are grown, the lipopolysaccharides obtained from its may differ in their composition and biological activity. Aim. To conduct a comparative study of the composition and biological activity Pseudomonas putida lipopolysaccharide (LPS) when grown on different media. Methods. In work microbiological, biochemical and serological methods were used. LPS was obtained from cells by waterphenolic extraction, its heterogeneity was determined by SDS-PAAG electrophoresis, the monosaccharide and fatty acid composition was identified by chromatography-mass spectrometry, and serological activity was determined by immunodiffusion in agar. Results. LPS isolated from Pseudomonas putida cells grown on meat-peptone agar (LPS-MPA) or on potato agar (LPS-PA) differ both in their qualitative composition and in the quantitative content of monosaccharides. In contrast to LPS-KA, in LPSMPA identified rhamnose (20.38%) and fucose (13.33%), and also galactose, glucose, mannose and ribose. Saturated and hydroxy acids with a carbon chain length from C12 to C18 were identified as part of the studied LPS preparations. Hexadecanoic acid was the dominant acid in LPS preparations (32.3 and 40.39% respectively for LPS-MPA and LPS-PA). Also, 2-hydroxy-dodecanoic (2-OHC12:0) (8.47 and 18.79%), 3-hydroxy-dodecanoic (3-OH-C12:0) (4.29 and 9.69%), dodecanoic (C12:0) (7.97 and 19.25%) acids (respectively for LPS-MPA and LPS-PA were udentified). The LD50 of LPS-MPA was found to be 29.73 μg/mouse, while LPS-PA was less toxic: its LD50 was 84.09 μg/mouse. The studied LPS preparations was pyrogen-free compared to Pyrogenal. LPS-KA did not show antigenic activity. Conclusions. LPS obtained from P. putida cells grown on various media differ monosaccharide, fatty acid composition, toxicity and serological activity.

Keywords: Pseudomonas putida, lipopolysaccharide, monosaccharide and fatty acid composition, serological activity, toxicity, pyrogenicity.

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