New bacteriocin from Bacillus clausii strainGM17: purification, characterization, and biological activity

A bacteriocin-producing strain (9,000 AU/ml) was isolated from the rhizosphere of Algerian healthy plants Ononis angustissima Lam. and identified as Bacillus clausii strain GM17. The bacteriocin, called Bac-GM17, was purified from the culture supernatant after heat treatment, ammonium sulfate precipitation, Sephadex G-50 chromatography and Mono Q fast-performance liquid chromatography (FPLC). Based on matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis, the purified Bac-GM17 is a monomer protein with a molecular mass of 5,158.11 Da. The N-terminal sequencing allowed for the straightforward identification of its first 20 residues, which were of pure bacteriocin. It also revealed that this bacteriocin contained a unique sequence, namely DWTCSKWSCLVCDDCSVELT, which suggests the identification of a novel compound. Bac-GM17 was extremely heat stable (20 min at 120 °C) and was stable within the pH range (3-9). It was found to be resistant to the proteolytic action of trypsin, pepsin, papain, pronase E, and proteinase K. It was also noted to display a bactericidal mode of action against Agrobacterium tumefaciens C58 and a fungistatic mode of action against Candida tropicalis R2 CIP203.