Isolation and Molecular Characterization of Exo-Polysaccharide Producing Weissella Confusa from Buffalo Ruminal Gut
Lactobacilli are the most important colonizers of the ruminant’s intestinal tract and are an important source of probiotics. Among Lactic acid bacteria, Exopolysaccharide producing Weissella confusa has gained considerable interest due to its importance in the food industry and several health attributes. The present analysis was carried out to isolate and characterize exopolysaccharide producing W. confusa from buffalo ruminal gut. The initial characterization of the isolate was done morphologically and biochemically. Molecular detection of the isolate was done by PCR based amplification of 16s rRNA gene. Temperature, pH, incubation-time and media conditions were optimized for the enzyme assay. A total of 35 LAB isolates were cultured for exopolysaccharide production on MRS agar having ruthenium red dye. The pure isolates were also screened for proteolytic, amylase, lipolytic, and antibiotic susceptibility testing. Two Catalase and Oxidase negative strains were selected which shows their probiotic potential.. The two strains identified as W.confusa (by GenBank accession number MK128505, Mk212944), which was based on 99% nucleotide homology and phylogenetic analysis. Isolates produced amylase, protease, lipase and also displayed a high resistance range against selected antibiotics. These isolates also showed antimicrobial activity against pathogens Listeria monocytogenes as well as against Staphylococcus aureus. The isolates also produce EPS when grown on ruthenium red MRS agar. The temperature 30°C-37°C and pH 4 were found optimum for enzyme assay. This study unravelled the capability and safety of EPS producing W. confusa for industrial use, and other potential applications.