Biological activity of (Lipo)polysaccharides of the exopolysaccharide-deficient mutant Rt120 derived from <Emphasis Type="Italic">Rhizobium leguminosarum</Emphasis> bv. trifolii strain TA1 |
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Authors: | J Kutkowska A Turska-Szewczuk M Janczarek R Paduch T Kaminska T Urbanik-Sypniewska |
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Institution: | 1.Department of Genetics and Microbiology,M. Curie-Sklodowska University,Lublin,Poland;2.Department of Immunology and Virology,M. Curie-Sklodowska University,Lublin,Poland |
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Abstract: | Lipopolysaccharides (LPS) from Rhizobium leguminosarum biovar trifolii TA1 (RtTA1) and its mutant Rt120 in the pssB-pssA intergenic region as well as degraded polysaccharides (DPS) derived from the LPS were elucidated in terms of their chemical
composition and biological activities. The polysaccharide portions were examined by methylation analysis, MALDI-TOF mass spectrometry,
and 1H NMR spectroscopy. A high molecular mass carbohydrate fraction obtained from Rt120 DPS by Sephadex G-50 gel chromatography
was composed mainly of L-rhamnose, 6-deoxy-L-talose, D-galactose, and D-galacturonic acid, whereas that from RtTA1 DPS contained
L-fucose, 2-acetamido-2,6-dideoxy-D-glucose, D-galacturonic acid, 3-deoxy-3-methylaminofucose, D-glucose, D-glucuronic acid,
and heptose. Relative intensities of the major 1H NMR signals for O-acetyl and N-acetyl groups were 1: 0.8 and 1: 1.24 in DPS of Rt120 and RtTA1, respectively. The intact
mutant LPS exhibited a twice higher lethal toxicity than the wild type LPS. A higher in vivo production of TNFα and IL-6 after induction of mice with Rt120 LPS correlated with the toxicity, although the mutant LPS
induced the secretion of IL-1β and IFNγ more weakly than RtTA1 LPS. A polysaccharide obtained by gel chromatography on Bio-Gel
P-4 of the high molecular mass material from Rt120 had a toxic effect on tumor HeLa cells but was inactive against the normal
human skin fibroblast cell line. The polysaccharide from RtTA1 was inactive against either cell line. The potent inhibitory
effect of the mutant DPS on tumor HeLa cells seems to be related with the differences in sugar composition. |
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