Manganese privation induced transcriptional upregulation of the class IIa bacteriocin plantaricin 423 in Lactobacillus plantarum 423

Plantaricin 423 is produced by Lactobacillus plantarum 423 using the pla biosynthetic operon located on the 8188 bp plasmid, pPLA4. As with many class IIa bacteriocin operons, the pla operon encodes biosynthetic genes ( plaA : precursor peptide, plaB : immunity, plaC : accessory and plaD : ABC transporter) but does not encode local regulatory genes. Little is known about the regulatory mechanisms involved in the expression of the apparently regulationless class IIa bacteriocins such as plantaricin 423. In this study, phylogenetic analysis of class IIa immunity proteins indicated that at least three distinct clades exist, which were then used to subgroup the class IIa operons. It became evident that the absence of classical quorum sensing genes on mobile bacteriocin encoding elements is a predisposition of the subgroup which includes plantaricin 423, pediocin AcH/PA-1, divercin V41, enterocin A, leucocin-A and -B, mesentericin Y105 and sakacin G. Further analysis of the subgroup suggested that the regulation of these class IIa operons may be linked to transition metal homeostasis in the host. By using a fluorescent promoter-reporter system in Lactobacillus plantarum 423, transcriptional regulation of plantaricin 423 was shown to be upregulated in response to manganese privation. IMPORTANCE Lactic acid bacteria hold huge industrial application and economic value, especially bacteriocinogenic strains which further aids in the exclusion of specific foodborne pathogens. Since bacteriocinogenic strains are sought after it is equally important to understand the mechanism of bacteriocin regulation. This is currently an understudied aspect of class IIa operons. Our research suggests the existence of a previously undescribed mode of class IIa bacteriocin regulation, whereby bacteriocin expression is linked to management of the producer’s transition metal homeostasis. This delocalized metalloregulatory model may fundamentally affect the selection of culture conditions for bacteriocin expression and change our understanding of class IIa bacteriocin gene transfer dynamics in a given microbiome.

Leuconostoc citreumMB1 as biocontrol agent ofListeria monocytogenesin milk

Cell-free supernatant fromLeuconostoc citreumMB1 revealed specific antilisterial activity. Preliminary studies demonstrated the proteinaceous, heat-stable, bacteriocin-like trait of the antimicrobial components present in the supernatant. Determination of the genes encoding bacteriocins by PCR and DNA sequencing led to amplification products highly homologous with leucocin A (found in diverseLeuconostocspecies) and UviB (found inLeuc. citreumKM20) sequences. Additionally, antimicrobial activity of cell-free supernatant fromLeuc. citreumMB1 was revealed by an inhibition halo of the SDS-PAGE gel subjected to a direct detection usingListeria monocytogenesas indicator strain. Different assays were carried out to assess the capacity ofLeuc.citreumMB1 to controlList. monocytogenesgrowth: (i) inactivation kinetics of the pathogen by antilisterial compounds present in concentrated cell-free supernatant fromLeuc. citreumMB1, (ii) evaluation of optimalLeuc. citreumMB1 initial concentration to obtain maximumList. monocytogenesATCC 15313 inhibition, and (iii) biocontrol ofList. monocytogenesATCC 15313 withLeuc. citreumMB1 during growth in milk at refrigeration temperature. According to our results, it is unquestionable that at least one bacteriocin is active inLeuc. citreumMB1, since important antilisterial activity was verified either in its cell-free supernatant or in co-culture experiments. Co-culture tests showed that ∼107 CFU/mlLeuc. citreumMB1 was the optimal initial concentration to obtain maximum pathogen inhibition. Moreover,Leuc. citreumMB1 was able to delayList. monocytogenesgrowth at refrigerated temperature.

Approaches to the discovery of new antibacterial agents based on bacteriocinsThis paper is one of a selection of papers published in this Special Issue, entitled CSBMCB — Systems and Chemical Biology, and has undergone the Journal's usual peer review process.

The development of antibiotic resistance in pathogenic bacteria has led to a search for novel classes of antimicrobial drugs. Bacteriocins are peptides that are naturally produced by bacteria and have considerable potential to fulfill the need for more effective bacteriocidal agents. In this mini-review, we describe research aimed at generating analogues of bacteriocins from lactic acid bacteria, with the goal of gaining a better understanding of structure–activity relationships in these peptides. In particular, we report recent findings on synthetic analogues of leucocin A, pediocin PA1, and lacticin 3147 A2, as well as on the significance of these results for the design and production of new antibiotics.