Nisin Purification from a Cell-Free Supernatant by Electrodialysis in a Circular Economy Framework.

Nisin, an antimicrobial peptide produced by Lactococcus lactis strains, is a promising natural preservative for the food industry and an alternative to antibiotics for the pharmaceutical industry against Gram-positive bacteria. Nisin purification is commonly performed using salting out and chromatographic techniques, which are characterized by their low yields, the use of solvents and the production of large volumes of effluents. In the present work, the purification of nisin from a cell-free supernatant (CFS), after the production of nisin by fermentation on a whey permeate medium, was studied using ammonium sulfate precipitation and electrodialysis (ED) as a promising eco-friendly process for nisin purification. Results showed an increase in nisin precipitation using a 40% ammonium sulfate saturation (ASS) level with a purification fold of 73.8 compared with 34.5 and no purification fold for a 60% and 20% ASS level, respectively. The results regarding nisin purification using ED showed an increase in nisin purification and concentration fold, respectively, of 21.8 and 156 when comparing the final product to the initial CFS. Nisin-specific activity increased from 75.9 ± 4.4 to 1652.7 ± 236.8 AU/mg of protein. These results demonstrated the effectiveness of ED coupled with salting out for nisin purification compared with common techniques. Furthermore, the process was noteworthy for its relevance in a circular economy scheme, as it does not require any solvents and avoids generating polluting effluents. It can be employed for the purification of nisin and the recovery of salts from salting out, facilitating their reuse in a circular economy.

In vitro assessment of skin sensitization, irritability and toxicity of bacteriocins and reuterin for possible topical applications.

Bacteriocins and reuterin are promising antimicrobials for application in food, veterinary, and medical sectors. In the light of their high potential for application in hand sanitizer, we investigated the skin toxicity of reuterin, microcin J25, pediocin PA-1, bactofencin A, and nisin Z in vitro using neutral red and LDH release assays on NHEK cells. We determined their skin sensitization potential using the human cell line activation test (h-CLAT). Their skin irritation potential was measured on human epidermal model EpiDerm™. We showed that the viability and membrane integrity of NHEK cells remained unaltered after exposure to bacteriocins and reuterin at concentrations up to 400 µg/mL and 80 mg/mL, respectively. Furthermore, microcin J25 and reuterin showed no skin sensitization at concentrations up to 100 µg/mL and 40 mg/mL, respectively, while pediocin PA-1, bactofencin A, and nisin Z caused sensitization at concentrations higher than 100 µg/mL. Tissue viability was unaffected in presence of bacteriocins and reuterin at concentrations up to 200 µg/mL and 40 mg/mL, respectively, which was confirmed by measuring cytokine IL-1α and IL-8 levels and by histological analysis. In conclusion, the current study provides scientific evidence that some bacteriocins and reuterin, could be safely applied topically as sanitizers at recommended concentrations.

A Syst-OMICS Approach to Ensuring Food Safety and Reducing the Economic Burden of Salmonellosis.

The Salmonella Syst-OMICS consortium is sequencing 4,500 Salmonella genomes and building an analysis pipeline for the study of Salmonella genome evolution, antibiotic resistance and virulence genes. Metadata, including phenotypic as well as genomic data, for isolates of the collection are provided through the Salmonella Foodborne Syst-OMICS database (SalFoS), at https://salfos.ibis.ulaval.ca/. Here, we present our strategy and the analysis of the first 3,377 genomes. Our data will be used to draw potential links between strains found in fresh produce, humans, animals and the environment. The ultimate goals are to understand how Salmonella evolves over time, improve the accuracy of diagnostic methods, develop control methods in the field, and identify prognostic markers for evidence-based decisions in epidemiology and surveillance.

A new structure-based classification of gram-positive bacteriocins.

Bacteriocins are ribosomally-synthesized peptides or proteins produced by a wide range of bacteria. The antimicrobial activity of this group of natural substances against foodborne pathogenic and spoilage bacteria has raised considerable interest for their application in food preservation. Classifying these bacteriocins in well defined classes according to their biochemical properties is a major step towards characterizing these anti-infective peptides and understanding their mode of action. Actually, the chosen criteria for bacteriocins' classification lack consistency and coherence. So, various classification schemes of bacteriocins resulted various levels of contradiction and sorting inefficiencies leading to bacteriocins belonging to more than one class at the same time and to a general lack of classification of many bacteriocins. Establishing a coherent and adequate classification scheme for these bacteriocins is sought after by several researchers in the field. It is not straightforward to formulate an efficient classification scheme that encompasses all of the existing bacteriocins. In the light of the structural data, here we revisit the previously proposed contradictory classification and we define new structure-based sequence fingerprints that support a subdivision of the bacteriocins into 12 groups. The paper lays down a resourceful and consistent classification approach that resulted in classifying more than 70% of bacteriocins known to date and with potential to identify distinct classes for the remaining unclassified bacteriocins. Identified groups are characterized by the presence of highly conserved short amino acid motifs. Furthermore, unclassified bacteriocins are expected to form an identified group when there will be sufficient sequences.