Rapid purification, partial characterization, and antimicrobial spectrum of the bacteriocin, Pediocin AcM, from...

The bacteriocin from Pediococcus acidilactici M, designated as Pediocin AcM, was rapidly purified to homogeneity by the pH mediated cell adsorption-desorption method and semi-preparative reversed-phase HPLC. The purification yield was 40.4% and the specific activity was increased by 2450-fold. It gave a single band and a single peak on SDS-PAGE and HPLC analysis, respectively. When subjected to electrospray LC-MS analysis, the protein was found to be highly pure and the molecular weight was determined as 4,618 Da. High concentration of the bacteriocin (> 50 micrograms/ml) showed good resistance to extremes of pH (1-12) and temperature (121 degrees C). Pediocin AcM was deduced to be a monomer with an intra-peptide disulfide bond from the results of reversed-phase analytical HPLC analyses after reduction, oxidation and trypsin digestion. P. acidilactici M inhibited a large number of bacteria, including Staphylococcus aureus, Listeria monocytogenes, Clostridium perfringens, Bacillus coagulans, B. cereus, and Aeromonas hydrophila.