Bacteriocin from Lacticaseibacillus rhamnosus sp. A5: Isolation, Purification, Characterization, and Antibacterial Evaluation for Sustainable Food Processing

A new Lacticaseibacillus rhamnosus strain A5 was isolated from pickle soup and characterized for its probiotic suitability. Strain A5 was Gram-positive, catalase-negative, acid-producing, and exhibited potential antibacterial activity against Escherichia coli (inhibition zone 17.3 mm), Bacillus subtilis (inhibition zone 14.5 mm), Salmonella enterica (zone of inhibition 16.1 mm) and Staphylococcus aureus (zone of inhibition 14.2 mm) by performing investigations on the disc diffusion. The cell-free supernatant of newly isolated strain A5 retained its inhibition ability of the growth of test bacteria at pH 2.0 to 5.0, temperature 121 degrees C for 30 min and UV irradiation for 8 h. However, the inhibitory effects of cell-free supernatant disappeared when subjected to papain, trypsin, and pepsin enzymatic treatments. By eliminating the interferences of organic acid and hydrogen peroxide, the cell-free supernatant possessed antibacterial activity against two indicator bacteria (E. coli and B. subtilis) and showed high thermal tolerance. These results indicated that the antibacterial substances produced by strain A5 were proteinaceous in nature, namely bacteriocin. The antibacterial bacteriocins in the supernatant of the strain A5 culture were further purified by ammonium sulfate fractionation and gel filtration chromatography. The purified bacteriocins also showed a pronounced inhibitory effect against E. coli and B. subtilis. The approximated molecular weight of bacteriocins was less than 14 kDa after determining by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In conclusion, the newly isolated strain A5 and its bacteriocins could be potentially applied in food preservation to prevent the risk of foodborne illness.