A novel cell-bound exopolysaccharide (c-EPS) was isolated from Lactobacillus helveticus MB2-1 by ultrasonic extraction, anion exchange, and gel filtration chromatography before being structurally characterized. The c-EPS is a heteropolysaccharide with an average molecular weight of 1.83 × 10(5) Da and is composed of glucose, mannose, galactose, rhamnose, and arabinose at a molar ratio of 3.12:1.01:1.00:0.18:0.16. Methylation analysis and nuclear magnetic resonance analysis revealed that the c-EPS is a linear glucomannogalactan containing repeating units of → 6)-β-D-Manp-(1 → 3)-β-D-Glcp-(1 → 3)-β-D-Glcp-(1 → 3)-β-D-Glcp-(1 → 4)-α-D-Galp-(1 → and trace amounts of Rhap-(1 → and (1 → 4)-Arap residues. Complex formation with Congo red demonstrated a triple-strand helical conformation for the c-EPS. Scanning electron microscopy of the c-EPS revealed many regular feather-like structural units. Topographical examination of c-EPS by atomic force microscopy revealed that the c-EPS formed rounded-to-spherical lumps with different sizes and chain formations. Furthermore, preliminary in vitro tests revealed that c-EPS significantly inhibited the proliferation of HepG-2, BGC-823, and especially HT-29 cancer cells.
Keywords: Lactobacillus helveticus MB2-1; anticancer activity; cell-bound exopolysaccharide (c-EPS); structural characterization.