Characterization and partial purification of a bacteriocin-like substance produced by thermophilic Bacillus licheniformis H1 isolated from cow manure compost

The production and partial characterization of bacteriocin-like substances (BLSs) produced by bacteria isolated from cow manure compost were investigated. Eight BLS producers, which exhibited inhibitory activity against pathogenic bacteria, were isolated from cow manure compost at different stages of the composting process. The pile temperature ranged from 9.1°C to 73.2°C. The BLSs showed thermostability, but the BLS producers were not thermostable except for the H1 producer. Thermophilic Bacillus licheniformis H1 was further characterized. The culture supernatant of B. licheniformis H1 exhibited antagonistic activity against various species of Gram-positive bacteria such as Listeria monocytogenes ATCC19111 but not against Gram-negative bacteria except Pseudomonas fluorescens ATCC11251. Inactivation of bacteriocin-like activity by α-chymotrypsin, trypsin, and papain was highly significant (P<0.001). The BLS was found to be stable under a pH range from 3 to 9 and at temperatures up to 75°C for 60min, but it lost activity after being autoclaved at 121°C for 15min. The optimum production of BLS by B. licheniformis H1 was obtained at a temperature of 55°C. Sodium dodecyl sulfate - polyacrylamide electrophoresis analysis of concentrated partially purified supernatants collected after resting the bacterial cells at 55°C revealed a bacteriocin-like protein with a molecular mass of approximately 3.5kDa. This study is the first report of a BLS from thermophilic B. licheniformis with an animal compost origin.