BaCf3: highly thermostable bacteriocin from Bacillus amyloliquefaciens BTSS3 antagonistic on food-borne pathogens.

In the present study, we characterized bacteriocin BaCf3, isolated and purified from Bacillus amyloliquefaciens BTSS3, and demonstrated its inhibitory potential on growth and biofilm formation of certain food spoilage bacteria and pathogens. Purification was by gel filtration chromatography and its molecular weight was 3028.422 Da after MALDI-TOF MS. The bacteriocin was highly thermostable withstanding even autoclaving conditions and pH tolerant (2.0-13.0). The bacteriocin was sensitive to oxidizing agent (DMSO) and reducing agent (DTT). The de novo sequence of the bacteriocin BaCf3 was identified and was found to be novel. The sequence analysis shows the presence of a disulphide linkage between C6 and C13. The microtitre plate assay proved that BaCf3 could reduce up to 80% biofilm produced by strong biofilm producers from food samples. In addition, BaCf3 did not show cytotoxicity on 3-TL3 cell line.

Characterization of Deep Sea Fish Gut Bacteria with Antagonistic Potential, from Centroscyllium fabricii (Deep Sea Shark).

The bacterial isolates from Centroscyllium fabricii (deep sea shark) gut were screened for antagonistic activity by cross-streak method and disc diffusion assay. This study focuses on strain BTSS-3, which showed antimicrobial activity against pathogenic bacteria including Salmonella Typhimurium, Proteus vulgaris, Clostridium perfringens, Staphylococcus aureus, Bacillus cereus, Bacillus circulans, Bacillus macerans and Bacillus pumilus. BTSS3 was subjected to phenotypic characterization using biochemical tests, SEM imaging, exoenzyme profiling and antibiotic susceptibility tests. Comparative 16S rDNA gene sequence analysis indicated that this strain belonged to the genus Bacillus, with high (98%) similarity to 16S rDNA sequences of Bacillus amyloliquefaciens. The chemical nature of the antibacterial substance was identified by treatment with proteolytic enzymes. The antibacterial activity was reduced by the action of these enzymes pointing out its peptide nature. It was observed from the growth and production kinetics that the bacteriocin was produced in the eighth hour of incubation, i.e., during the mid-log growth phase of the bacteria.