Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin.

This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing Enterococcus faecium BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against L. monocytogenes. The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in Escherichia coli ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against L. monocytogenes in a milk model.

Insight into the Postbiotic Potential of the Autochthonous Bacteriocin-Producing Enterococcus faecium BGZLM1-5 in the Reduction in the Abundance of Listeria monocytogenes ATCC19111 in a Milk Model.

This study aimed to explore the probiogenomic characteristics of artisanal bacteriocin-producing Enterococcus faecium BGZLM1-5 and its potential application in reducing Listeria monocytogenes in a milk model. The BGZLM1-5 strain was isolated from raw cow's milk from households in the Zlatar Mountain region. The whole genome sequencing approach and bioinformatics analyses reveal that the strain BGZLM1-5 is non-pathogenic to humans. Bacteriocin-containing supernatant was thermally stable and antimicrobial activity retained 75% of the initial activity compared with that of the control after treatment at 90 °C for 30 min. Antimicrobial activity maintained relative stability at pH 3-11 and retained 62.5% of the initial activity compared with that of the control after treatment at pH 1, 2, and 12. The highest activity of the partially purified bacteriocin was obtained after precipitation at 40% saturation with ammonium sulfate and further purification by mixing with chloroform. Applying 3% and 5% (v/v) of the bacteriocin-containing supernatant and 0.5% (v/v) of the partially purified bacteriocin decreased the viable number of L. monocytogenes ATCC19111 after three days of milk storage by 23.5%, 63.5%, and 58.9%, respectively.

Ancient Wheat Varieties and Sourdough Fermentation as a Tool to Increase Bioaccessibility of Phenolics and Antioxidant Capacity of Bread.

This study aimed to determine the impact of ancient wheat varieties (emmer, spelt and khorasan) and spontaneous sourdough fermentation on the bioaccessibility of total phenolic content (TPC) and the DPPH antioxidant capacity evolution during breadmaking and in vitro digestion. Sourdough and yeast-fermented modern wheat breads were used as controls. After 6 h of fermentation, the total titrable acidity of the sourdough increased from 139 to 167%. The wheat variety, type of fermentation and processing affected TPC, antioxidant activity and bioaccessibility. Antioxidant activity and TPC were reduced by dough mixing, increased after sourdough fermentation and slightly decreased or remained the same after baking. Although wheat flour had the highest TPC, the modeling of TPC kinetic revealed that emmer and spelt sourdough exhibited a higher bound phenolics release rate due to the higher acidity, which contributed to increased phenolics solubility. Although wheat bread, both before and after digestion, had the lowest TPC, especially the one prepared with yeast, high TPC bioaccessibilities and antioxidant activities after the digestion suggested that, except phenolics, digestion process improved the release of additional compounds with different bioaccessibility and biological activity. The results of this study proved that the application of sourdough fermentation can increase the potential of ancient wheats in the developing of functional bakery products.