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Antimicrobial resistance presents a substantial threat to global public health, demanding urgent attention and action. This study focuses on lanthipeptides, ribosomally encoded peptides that display significant structural diversity and hold promising potential as antibiotics. Genome mining was employed to locate biosynthetic gene clusters (BGCs) containing class II lanthipeptide synthetases encoded by lanM genes. A phylogenetic study analyzing homologous sequences of functional LanM sequences revealed a unique evolutionary clade of 17 LanM proteins associated with 12 Clostridium bacterial genomes. In silico exploration identified nine complete BGCs, including one super-cluster containing two co-localized operons from Clostridium cellulovorans 743B, that encode for two new peptides named clostrisin and cellulosin. Each operon was heterologously expressed in Escherichia coli. Molecular weights associated with the expected post-translational modifications of the purified lanthipeptide were confirmed by MS-MS/MS analysis for cellulosin, while clostrisin was not post-translationally modified. Both peptides demonstrated antimicrobial activity against multidrug-resistant bacteria, such as a clinical strain of Staphylococcus epidermidis MIQ43 and Pseudomonas aeruginosa PA14. This is the first report of lanthipeptides from the Clostridium genus produced with its native biosynthetic machinery, as well as chemically and biologically characterized. This study showcases the immense potential of genome mining in identifying new RiPP synthetases and associated bioactive peptides.

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Bacteriocins is the name given to products of the secondary metabolism of many bacterial genera that must display antimicrobial activity. Although there are several bacteriocins described today, it has not been possible to reach a consensus on the method of classification for these biomolecules. In addition, many of them are not yet authorized for therapeutic use against multi-drug-resistant microorganisms due to possible toxic effects. However, recent research has achieved considerable progress in the understanding, classification, and elucidation of their mechanisms of action against microorganisms, which are of medical and biotechnological interest. Therefore, in more current times, protocols are already being conducted for their optimal use, in the hopes of solving multiple health and food conservation problems. This review aims to synthetize the information available nowadays regarding bacteriocins, and their classification, while also providing an insight into the future possibilities of their usage for both the pharmaceutical, food, and biotechnological industry.

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In this work, a comparative analysis of the peripheral cell component (PCC) proteins of Listeria monocytogenes was carried out. The study was conducted on two set of samples consisting of bacteria treated with sub-lethal concentration of nisin and untreated bacteria as control. PCC proteins were extracted by Tris-Urea-EDTA treatment and then subjected to trypsin digestion and mass spectrometry analysis. The whole cell proteome was analyzed through label-free quantitative proteomics approach. Proteomic analysis was carried out using OrbiTrap Mass Spectrometer coupled to nanoflow liquid chromatography. The treatment with sub-lethal nisin concentration resulted in 62 up regulated and 97 down regulated proteins compared to untreated samples. Using PSORTb 3.0, 19 and 18 surface proteins were detected among the up regulated and down regulated proteins, respectively. Proteins related with increased biofilm formation by L.monocytogenes, such as moonlight proteins of the pyruvate dehydrogenase complex and flagellin-related proteins, were identified as up regulated surface proteins. Proteins associated with virulence of L.monocytogenes, including listeriolysin O, internalin B and actin assembly-inducing protein, were detected among the down regulated proteins. To confirm proteomics data, increased production of biofilm was experimentally confirmed in nisin-treated cells through crystal violet method. BIOLOGICAL SIGNIFICANCE: Proteosurfaceomics can be defined as the "omics" science applied to the proteins of the peripheral cell component (PCC). The surface proteins of Listeria monocytogenes, an important foodborne pathogen were investigated after treatment with nisin, a bacteriocin approved as a natural food preservative by regulatory agencies. Recent cases of nisin tolerance by Listeria spp. were documented, and deeper studies on the molecular process behind the bacterial survival may help in both understanding the development of tolerance process and comparing nisin effect with other antimicrobial compounds.

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One of the biggest challenges faced presently by clinicians is the emergence of multidrug -resistant pathogens that can infect humans and animals. To control the infections caused by such pathogens the development of new drugs is required. Bacteria are a rich source of ribosomally -synthesized antimicrobial peptides known as bacteriocins, which are characterized by the presence of a self-defense immunity system. Labionin-containing lantibiotics and sactibiotics are posttranslationally modified bacteriocins with peculiar features. Labionin-containing peptides belong to subclass Ic lantibiotics in which the carbacyclic triamino triacid labionin, a structural variant of lanthionine, and a methyl-substitute labionin derivative are found, giving the molecule a labyrinthine structure. Sactibiotics are circular or linear peptides belonging to a distinct bacteriocin class (class V) which is characterized by the presence of cross-linkages formed by the thiol group of cysteine residues and the α-carbon of acceptor amino acids. A few examples of these bacteriocins have been described in the literature to date, although putative gene clusters with the potential to encode such peptides can be found in the genome of many bacterial species. Some peptides already under study exhibit potential biotechnological applications because of their remarkable antibacterial or antiviral activities, as well as their analgesic activity. Therefore, in this review, the main findings concerning these peptides will be addressed and discussed, with an emphasis on their potential use in clinical settings.

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AIMS: To compare the action of nisin and bovicin HC5 in combination with EDTA on Salmonella Typhimurium under different environmental conditions. METHODS AND RESULTS: Salmonella Typhimurium was treated in BHI broth containing EDTA (1·5 mmol l(-1)) and nisin or bovicin HC5 (200 AU ml(-1)) under different pH and temperature conditions, and according to a central composite design with two factors (temperature and pH). Cell viability was evaluated on plate count agar for 48 h. The combination of nisin or bovicin HC5 with EDTA was able to inhibit the growth of Salmonella, but the temperature and pH conditions promoting inhibition were distinct for each bacteriocin. Nisin was bactericidal over a broad range of temperature and pH, while bovicin HC5 was bacteriostatic in most conditions and bactericidal only in specific conditions (pH >6·0 and temperature >30°C). Salmonella Typhimurium did not show tolerance to bovicin HC5 or cross-tolerance between these lantibiotics. CONCLUSIONS: Nisin and bovicin HC5 both inhibited the growth of Salmonella, but the activity of each bacteriocin was differently influenced by environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Nisin and bovicin HC5 have the potential to inhibit the growth of Salmonella, but environmental conditions should be considered to establish optimal conditions for its application.

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OBJECTIVES: To compare the in vitro activity of mutacins D-123.1 and F-59.1 against different bacteria including antibiotic-resistant strains, in order to evaluate their application potential. DESIGN AND METHODS: The antibacterial activity spectrum of purified F-59.1 and the MIC and MBC of F-59.1 and D-123.1 against target bacteria were determined. RESULTS: Most bacteria were inhibited by the purified mutacins. Mutacin F-59.1 shows a relatively wide activity spectrum. Mutacin D-123.1 has low Minimum Inhibitory Concentrations [MICs] (0.25-4 µ/ml) against human pathogens while F-59.1 has higher MICs (3.2-12.8 fig/ml) mainly against food-borne pathogens. CONCLUSION: The effectiveness of mutacins D-123.1 and F-59.1 against human and food-borne pathogens is demonstrated. Mutacin D-123.1 shows potential as a new antibiotic while F-59.1 shows promising application in food products. ABBREVIATIONS: MALDI-TOF MS, matrix assisted laser desorption ionisation-time of flight mass spectrometry; MB(I)C, minimum bactericidal (inhibitory) concentrations; OD, optical density; RP-HPLC, reverse-phase high-pressure liquid chromatography; TSBYE, trypticase soy broth yeast extract.

OBJETIVOS: Comparar la actividad in vitro de las mutacinas D-123.1 y F-59.1 frente a diferentes bacterias incluyendo las cepas resistentes a los antibióticos, a fin de evaluar el potencial de su aplicación. DISEÑO Y MÉTODOS: Se determinó el espectro de actividad antibacteriana de F-59.1 purificada y la CIM y la CBM de F-59.1 y D-123.1 frente a determinadas bacterias. RESULTADOS: La mayor parte de las bacterias eran inhibidas por las mutacinas purificadas. La mutacina F-59.1 muestra un espectro de actividad relativamente amplio. La mutacina D-123.1 posee bajas concentraciones de inhibición mínimas [CIM] (0.25-4 fig/ml) contra los patógenos humanos, mientras que el F-59.1 posee concentraciones CIM más altas (3.2-12.8 fig/ml) principalmente frente a los patógenos alimentarios. CONCLUSIÓN: Queda demostrada la efectividad de las mutacinas D-123.1 y F-59.1 frente a los patógenos humanos y alimentarios. La mutacina D-123.1 muestra poseer un potencial como nuevo antibiótico, en tanto que F-59.1 se presenta como una aplicación promisoria en relación con los productos alimentarios. ABREVIATURAS: MALDI-TOF MS, espectrometría de masas con desorción/ionización mediante láser asistida por matriz asociada a un analizador de tiempo de vuelo (del inglés: matrix assisted laser desorption ionisation-time de flight mass spectrometry). CIM: concentración inhibitoria mínima (inglés MIC). CBM: concentración bactericida mínima (inglés MBC). DO: densidad óptica (inglés OD); RP-HPLC: cromatografía líquida de alta resolución en fase revertida; TSBYE:caldo tripticasa soya- extracto de levadura.

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