Investigation of the chemical structure and biological activity of oligosaccharides isolated from rough-type Xanthomonas campestris pv. campestris B100 lipopolysaccharide

Abstract

The rough-type lipopolysaccharide (LPS) of the phytopathogenic bacterium Xanthomonas campestris pv. campestris B 100 was isolated utilizing the hot phenol-water method and successively de-acylated by treatment with hydrazine and hot potassium hydroxide. Four compounds were separated by preparative high-performance anion-exchange chromatography and studied by sugar analysis and by 1D and 2D homonuclear and heteronuclear ¹H-, ¹³C- and ³¹P-NMR spectroscopy as well as ESI FT-MS. The two main products were a heptasaccharide and a pentasaccharide of the structures α-D-Manp-(1→3)-α-D-Man p-(1→4)-β-D-Glcp-(1→4)-α-D-Manp-3P -(1→5)-α-Kdo-(2→6)-β-D-GlcpN-4P-(1→6)-α-D-Glc pN-1P (1) and β-D-Glcp-(1→4)-α-D-Man p-3P-(1→5)-α-Kdo-(2→6)-β-D-GlcpN-4 P-(1→6)-α-D-GlcpN-1P (2), respectively. The products in smaller amounts were a heptasaccharide and pentasaccharide possessing the above structures plus a phosphate group at C-4 of the Kdo residue (compounds 3 and 4). Both, heptasaccharide 1 and pentasaccharide 2 were able to induce an oxidative burst in cell cultures of the non-host plant tobacco.

Keywords

Lipopolysaccharides Xanthomonas campestris pv. campestris structural analysis NMR spectroscopy ESI FT-MS oxidative burst

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