Characterization and antibacterial activity of a novel exopolysaccharide produced by Lactobacillus kefiranofaciens DN1 isolated from kefir

- A new exopolysaccharide (EPS), EPS_DN1, was isolated from kefir-derived bacteria.
- Lactobacillus kefiranofaciens DN1 was the best EPS producer among 22 kefir isolates.
- Lb. kefiranofaciens DN1 produced EPS_DN1 in glucose concentration-dependent manner.
- EPS_DN1 exerted bactericidal effects against S. Enteritidis and L. monocytogenes.
- EPS_DN1 is a heteropolysaccharide composed of five kinds of monosaccharide.

Kefir is fermented milk produced by co-cultures of lactic and acetic acid bacteria and yeast that are encapsulated by an exopolysaccharide (EPS) called kefiran. Probiotic kefir provides many benefits, including antimicrobial effects against many pathogens. The mechanisms underlying these effects, however, are not yet completely understood. In this study, a total of 22 strains of lactic acid bacteria (LAB), including eight strains of Lactobacillus kefiranofaciens, two strains of Lactobacillus kefiri, seven strains of Lactococcus lactis, and five strains of Leuconostoc mesenteroides, were isolated from kefir and classified by 16S ribosomal RNA gene sequencing. The ability of each strain to produce EPS was determined, and Lactobacillus kefiranofaciens DN1 was found to have the highest EPS yield among all isolates. Lactobacillus kefiranofaciens DN1 produced EPS, using glucose and lactose, and EPS yield rose to 2.2 g/L in modified MRS broth (60 g/L glucose). The antimicrobial activities of the EPS produced by Lactobacillus kefiranofaciens DN1 (EPS_DN1) against Listeria monocytogenes and Salmonella Enteritidis were assessed by growth curve analysis. We found that EPS_DN1 at a concentration of at least 1% was able to exert bactericidal effects against both pathogens. Notably, results of high-performance size-exclusion chromatography (HPSEC) analysis indicated that EPS_DN1 was not kefiran, the major EPS of kefir grains, suggesting that EPS_DN1 represents a novel bioactive compound in kefir.