A Lactobacillus plantarum strain producing exopolysaccharides (EPSs) was isolated from corn silage. When this strain, named L. plantarum EP56, was grown on a chemically defined medium, two EPS fractions were isolated. The cell-bound EPS fraction (EPS-b) was composed of a single high-molecular-mass polymer of 8.5x10(5) Da containing glucose, galactose and N-acetylgalactosamine in a molar ratio of approximately 3:1:1 and traces of glycerol and phosphoglycerol. The released EPS fraction (EPS-r) was composed of the high-molecular-mass bound polysaccharide and a second polymer of 4x10(4) Da containing glucose, galactose and rhamnose in a molar ratio of 3:1:1 and traces of glycerol and phosphoglycerol. EPS-b and EPS-r contained phosphate which contributes to their negative net charge. Studies on polysaccharide production and location showed that both polymers were synthesized during the exponential growth phase and that the EPS-b polymer was progressively released into the culture medium during the stationary growth phase. Carbon source and temperature influenced EPS synthesis when L. plantarum EP56 was grown in a chemically defined medium. Lactose was the most efficient carbon source among the five tested (glucose, galactose, fructose, lactose and sucrose). EPS production was also increased when the incubation temperature is lowered.