Mikrobiol. Z. 2018; 80(4):28-40. Russian.
doi: https://doi.org/10.15407/microbiolj80.04.028

The Influence of Рseudomonas chlororaphis subsp. chlororaphis UCМ В-106 Cultivation Temperature
on Composition and Biological Properties of Lipopolysaccharides

Brovarskaya O.S., Varbanets L.D.

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

The purpose of this work was to conduct a comparative study of the chemical composition of Pseudomonas chlororaphis subsp. chlororaphis UCM B-106 lipopolysaccharides (LPS), isolated from bacteria grown at different temperature regimes (10, 28 and 37 °C). Methods. Phenol-water extraction of LPS, electrophoresis in polyacrylamide gels, chromato-mass spectrometric determination of monosaccharide and fatty acid compositions, study of the antigenic structure by immunodiffusion in agar by Ouchterlony. Results and conclusions. It is shown that an increase in the temperature of cultivation causes changes in the biochemical, immunochemical and biological properties of LPS obtained from cells growing at different temperatures (LPS-10, LPS-28 and LPS-37). The yield of LPS from the “cold” variant was almost 5 times higher than that of cells cultivated at 37 °С. One of the reasons for the low yield of LPS-37 (2.3 %), as compared to 11.3 % of LPS-10 yield, may be the presence of the “thermal” variant of LPS with short O-polysaccharide chains (as we showed by PAGE) and which because of their hydrophobicity poorly extracted by phenol-water procedure. Analysis of monosaccharide composition of LPS showed that the preparations are similar in quality composition. At the same time, with increasing cell growth temperature, the percentage of heptose, a characteristic component of core oligosaccharide, increased (47.5, 52.5, 52.9 % for LPS-10, LPS-28, LPS-37, respectively). The results of the determination of the fatty acid composition of the studied LPS showed that the qualitative composition of fatty acids is very close, and their quantitative content varies significantly. The dominant were 3-OH-C14 : 0 (35.5, 47.2, 32.4 %), and also C14 : 0 (31.6, 34.4, 42.1 %, respectively for LPS-10, LPS-28, LPS-37) the content of which increases with an increase in the growing temperature. Of unsaturated acids, C16: 1 (12.0%) was detected only in LPS-10. The toxicity study showed that the LD50 values decreased in the range of LPS-10, LPS-20 and LPS-37 (2.0, 20.0 and 35.56 mg/kg animals, respectively). That is, the greatest toxicity was shown by LPS “cold variant”, and the least toxic - “thermal” (LPS-37). When studying the antigenic structure by immunodiffusion in Ouchterloni agar, the presence of common antigenic determinants in the composition of the LPS studied was shown. Summary. The increasing of cultivation temprerature of cells growing causes the changes in biochemical, immunochemical and biological properties of LPS.

Keywords: Рseudomonas chlororaphis subsp. chlororaphis УКМ В-106, different temperatures of cultivation, lipopolysaccharide, monosaccharide, fatty acid composition, biological activity.

Full text (PDF, in Russian)

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