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Lipid-bound Saccharides Containing Glucose and Galactose in Agrobacterium tumefaciens

Instituto de Investigations Bioquimicas ‘Fundacion Campomar’ and Facultad de Ciencias Exactas y Naturales, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina

ABSTRACT

Summary: The incubation of an enzyme preparation of Agrobacterium tumefaciens with UDP-[¹⁴C]Glc led to the formation of radioactive substances soluble in chloroform/methanol/water (1 : 1 : 0·3, by vol.). These substances had properties similar to the polyprenol diphosphate saccharides. The lipid moiety appears to be an unsaturated polyprenol. Mild acid hydrolysis of the substances liberated five water-soluble products which were separated by TLC and characterized by borohydride reduction-acid hydrolysis, partial acid hydrolysis, methylation analysis, TLC and paper chromatography and paper electrophoresis. The products behaved like: galactose; the disaccharide Glc(β1-3)Gal; the tetrasaccharide Glc(β1-4)Glc(β1-4)Glc(β1-3)Gal; an octasaccharide and a pyruvylated octasaccharide. The last two substances were compared by paper electrophoresis, TLC and methylation analysis with an octasaccharide obtained by the action of a hydrolytic enzyme on the exopolysaccharide of Agrobacterium tumefaciens. The octasaccharide and the pyruvylated octasaccharide from the lipid behaved like those obtained by enzyme action on the A. tumefaciens exopolysaccharide. These results suggest that the lipid-bound saccharides are involved in the biosynthesis of the extracellular polysaccharides.