Listeria monocytogenes is a food-borne pathogen. Pediocin is a group IIα bacteriocin with anti-listeria activity that is naturally produced by Pediococcus acidilactic and Lactobacillus plantarum. The pedA/papA gene encodes pediocin/plantaricin. In native hosts, the expression and secretion of active PedA/PapA protein rely on the accessory protein PedC/PapC and ABC transporter PedD/PapD on the same operon. The excretion machines were also necessary for pediocin protein expression in heterologous hosts of E. coli, Lactobacillus lactis, and Corynebacterium glutamicum. In this study, two vectors carrying the codon sequence of the mature PapA peptide were constructed, one with and one without a His tag. Both fragments were inserted into the plasmid pHT43 and transformed into Bacillus subtilis WB800N. The strains were induced with IPTG to secrete the fused proteins PA1 and PA2. Supernatants from both recombinant strains can inhibit Listeria monocytogenes ATCC54003 directly. The fused protein possesses inhibition activity as a whole dispense with removal of the leading peptide. This is the first report of active pediocin/PapA expression without the assistance of PedCD/PapCD in heterogeneous hosts. In addition, the PA1 protein can be purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography.

Listeria moniocytogenes is food-borne pathogen. Pediocin is group II α bacteriocin with anti-listeria activity, naturally produced by Pediococuus acidilactic and Lactobacillus plantarum. Gene pepA/papA encode for pediocin. Expression and secretion of active papA was relayed on transporter papC and accessary protein papD on the same operon in native host. The excretion machines were also necessary for pediocin protein expression in heterologous host of E. coli, Lactobacillus lactis, and Corynebacterium glutamicum. In this study, two vector carrying codon sequence of papA mature peptide was constructed, with or without His tag. Both fragments were inserted into plasmid pHT43 and transformed Bacillus subtilis WB800N. The strains were induced with IPTG to secrete recombination protein PA1 and PA2 respectively. Supernatant from both recombination strains can inhibit Listeria monocytogenes ATCC54003 directly. The fused protein possesses inhibition activity as a whole, exempting from cleavage of leading peptide. Protein PA1 can be purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography. This is the first report for active pediocin expression without assistance of papCD in heterogenous host.