Mikrobiol. Z. 2019; 81(2):3-13. Russian.
Influence of Isolation Methods on Composition and Biological Properties
of Escherichia coli 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 study the influence of isolation methods on composition and biological activity of Escherichia coli lipopolysaccharides (LPS). Methods. Microbiological, biochemical and serological methods were used. LPS were obtained from the cells by the classic Westphal method (LPSw), Boivin (LPSb), and an alkaline lipopolysaccharide (LPSa). Results. LPSw, LPSb and LPSa were characterized by a different yield of 0.33, 0.25 and 0.05%, respectively, from the mass of microbial cells. All LPS preparations contained the main components of LPS: carbohydrates, KDO and fatty acids of lipid A. It was also shown that the method of Boivin and Westphal makes it possible to isolate higher molecular weight, and alkaline extraction method - lower molecular weight fractions of LPS. The main components of the hydrophobic part of the LPS in lipid A were 3-OH-C14:0, C12:0, C14:0, C16:0 acids, and the polysaccharide part of the LPS was represented by glucose, galactose and heptose, as well as by one unidentified monosaccharide. LPSw exhibited the largest serological and toxic activity, and LPSb - less toxic, but more pyrogenic. LPSa characterized the smallest pyrogenicity and was slightly toxic. Conclusions. The method of extraction of LPS influences its composition and biological properties.
Keywords: Escherichia coli 58, lipopolysaccharide, monosaccharide and fatty acid composition, biological activity.
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