Characterization of paraplantaricin C7, a novel bacteriocin produced by, Lactobacillus paraplantarum C7 isolated from Kimchi

Abstract

A Lactobacillus paraplantarum strain producing a bacteriocin was isolated from kimichi using the spot-on-the lawn method and named L. paraplantarum C7 [15]. The bacteriocin, paraplantaricin C7, was found to inhibit certain Lactobacillus strains, including L. plantarum, L. pentosus, and L. delbrueckii subsp. lactis. It also inhibited Enterococcus faecalis, yet did not inhibit most of the other LAB (lactic acid bacteria) tested. The maximum level of paraplantaricin C7 activity was observed under the Culture conditions of 25 degrees C and a constant pH of 4.5. Paraplantaricin C7 retained 90% of its activity after 10 min of treatment at 100 degrees C and remained stable within a pI-I range of 2-8. Based on a culture supernatant, paraplantaricin C7 was purified by DEAE-Sephacel column chromatography and C-18 reverse-phase HPLC. SDS-PAGE and activity staining were then conducted using the purified paraplantaricin C7, and its molecular mass determined to be about 3,800 Da. The 28 N-terminal amino acids from the purified paraplantaricin C7 were determined, and the structural gene encoding paraplantaricin C7, ppnC7, was cloned by PCR using degenerate primers based on the N-terminal amino acid sequence. The nucleotide sequences for ppnC7 and other neighboring orfs exhibited a limited homology to the previously reported plantaricin operon genes. Paraplantaricin C7 is a novel type 11 bacteriocin containing a double glycine leader sequence.