Purification, characterization and mode of action of enterocin, a novel bacteriocin produced by Enterococcus faecium TJUQ1.

In this study, enterocin TJUQ1 was purified by ammonium sulfate precipitation, reversed-phase chromatography (Sep-Pak C8) and cation-exchange chromatography. The activity of the purified bacteriocin was 44,566.41 ± 874.69 AU/mg, which corresponds to a purification fold of 35.89 ± 2.34. The molecular mass was 5520 Da by MALDI-TOF MS and Tris-Tricine SDS-PAGE. The result of LC-MS/MS showed that the bacteriocin shared 59.15% identity with enterocin produced by E. faecium GN (accession no. O34071). PCR amplification revealed that E. faecium TJUQ1 possesses a gene encoding enterocin B with 60% identity to enterocin B. Circular dichroism (CD) spectroscopy showed that the molecular conformation was 32.6% helix, 19.5% beta, 12.9% turn and 35.0% random. The stability of enterocin TJUQ1 was measured. After exposure at 121 °C for 15 min, the residual antimicrobial activity of enterocin TJUQ1 was 85.95 ± 1.32%. The antimicrobial activity of enterocin TJUQ1 was still active over a pH range of 3-11. Enterocin TJUQ1 was inactivated after exposure to proteolytic enzymes but was not inactivated by lipase or amylase. These results showed that enterocin TJUQ1 was a novel class II bacteriocin. Enterocin TJUQ1 showed wide antibacterial activity against food-borne gram-negative and gram-positive pathogens, such as Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Salmonella enterica. The MIC was 5.26 ± 0.24 μg/mL against L. monocytogenes CMCC 1595....
Source: International Journal of Biological Macromolecules - Category: Biochemistry Authors: Tags: Int J Biol Macromol Source Type: research