Isolation, purification, characterization and immunostimulatory activity of an exopolysaccharide produced by Lactobacillus pentosus LZ-R-17 isolated from Tibetan kefir

Abstract

In this study, three strains of lactic acid bacteria isolated from Tibetan kefir grains, including two strains of Lactobacillus pentosus LZ-R-17 and L. helveticus LZ-R-5, and one strain of Lactococcus lactis subsp. lactis LZ-R-12. The ability of three strains to produce exopolysaccharide (EPS) was tested, and L. pentosus LZ-R-17 was found to have the highest EPS yield. One EPS (R-17-EPS) was isolated from the fermented milk by L. pentosus LZ-R-17 and purified by DEAE-52 anion exchange chromatography. Furthermore, R-17-EPS preliminary structure and macrophage immunomodulatory activity in vitro were investigated. On the basis of the analytical results of ultraviolet-visible spectrum, Fourier transform-infrared spectrum, monosaccharide composition analysis and one-dimensional and two-dimensional nuclear magnetic resonance (NMR) spectra, R-17-EPS was found to have an average molecular weight of 1.20 × 106 Da and was composed of galactose and glucose residues with a molar ratio of 1.00:3.15. NMR analysis revealed that the R-17-EPS was a linear hetero-galactoglucan containing repeating units of →2)-α-D-Galp-(1 → 4)-β-D-Glcp-(1 → 4)-β-D-Glcp-(1 → 4)-β-D-Glcp-(1→. In addition, R-17-EPS could effectively enhanced the proliferation, phagocytosis, nitric oxide and cytokines production of RAW264.7 cells, suggesting that R-17-EPS had potent immunostimulatory activity and could be explored as immunomodulator in functional food and/or medicine fields.