Developing lactic acid bacteria starter cultures for wholemeal rye flour bread with improved functionality, nutritional value, taste, appearance and safety.

Starter cultures composed of lactic acid bacteria (LAB) were developed based on the genotypic and phenotypic characterisation of isolates belonging to dominant groups of bacteria in spontaneous rye wholemeal sourdoughs. Combinations of strains have been evaluated on an industrial scale in the sourdough fermentation process. Wholemeal rye bread was prepared using sourdoughs obtained with 3 new starter cultures, and compared to bread made using the commercial culture (LV2). All newly developed cultures used for the preparation of wholemeal rye bread allowed to obtain better quality products as compared to the LV2 based bread. The best results were obtained when the culture containing Lactiplantibacillus plantarum 2MI8 and exopolysaccharide (EPS)-producing Weissella confusa/cibaria 6PI3 strains was applied. The addition of yeast during sourdough breads production, especially the one prepared from mentioned above starter culture, significantly improved their organoleptic properties, their volume and crumb moisture was increased, and also the crumb acidity and hardness was reduced. Fermentation of rye wholemeal dough, especially without the yeast addition, resulted in a significant reduction in the content of higher inositol phosphates as compared to the applied flour, which is associated with improved bioavailability of minerals. The results of this study prove that the investigated new starter cultures can be successfully applied in wholemeal rye bread production.

Extraction, purification and characterisation of exopolysaccharides produced by newly isolated lactic acid bacteria strains and the examination of their influence on resistant starch formation.

Exopolysaccharides (EPS) were produced by four newly isolated lactic acid bacteria strains, then further extracted, separated and characterised under standardised conditions. Using a sucrose carbon source, these LAB strains belonging to Weissella confusa/cibaria produced EPS with a dextran high molecular weight fraction. The obtained yields of EPS ranged from 3.2 g/L to 47.1 g/L and outstandingly high yields were obtained using Weissella confusa/cibaria 3MI3 isolated from spontaneous spelt sourdough. After purification the influence of EPS-dextran of molar mass 3,244,000 g/mol on resistant starch formation in wheat starch pastes and pasted samples after temperature-cycled storage was examined. Size exclusion chromatography with post-column derivatisation revealed that a 1.5% share of EPS dextran limited formation of high molar mass resistant starch in starch pastes during storage. This work provides new insight on hindering resistant starch formation by using EPS, which could be efficiently produced in sourdough, thus improving the properties of sourdough bread.

A Kayvirus Distant Homolog of Staphylococcal Virulence Determinants and VISA Biomarker Is a Phage Lytic Enzyme.

Staphylococcal bacteriophages of the Kayvirus genus are candidates for therapeutic applications. One of their proteins, Tgl, is slightly similar to two staphylococcal virulence factors, secreted autolysins of lytic transglycosylase motifs IsaA and SceD. We show that Tgl is a lytic enzyme secreted by the bacterial transport system and localizes to cell peripheries like IsaA and SceD. It causes lysis of E. coli cells expressing the cloned tgl gene, but could be overproduced when depleted of signal peptide. S. aureus cells producing Tgl lysed in the presence of nisin, which mimics the action of phage holin. In vitro, Tgl protein was able to destroy S. aureus cell walls. The production of Tgl decreased S. aureus tolerance to vancomycin, unlike the production of SceD, which is associated with decreased sensitivity to vancomycin. In the genomes of kayviruses, the tgl gene is located a few genes away from the lysK gene, encoding the major endolysin. While lysK is a late phage gene, tgl can be transcribed by a host RNA polymerase, like phage early genes. Taken together, our data indicate that tgl belongs to the kayvirus lytic module and encodes an additional endolysin that can act in concert with LysK in cell lysis.

Bacterial community dynamics in spontaneous sourdoughs made from wheat, spelt, and rye wholemeal flour.

Sourdough fermentation is a traditional process that is used to improve bread quality. A spontaneous sourdough ecosystem consists of a mixture of flour and water that is fermented by endogenous lactic acid bacteria (LAB) and yeasts. The aim of this study was to identify bacterial diversity during backslopping of spontaneous sourdoughs prepared from wheat, spelt, or rye wholemeal flour. Culture-dependent analyses showed that the number of LAB (109  CFU/ml) was higher by three orders of magnitude than the number of yeasts (106  CFU/ml), irrespective of the flour type. These results were complemented by next-generation sequencing of the 16S rDNA V3 and V4 variable regions. The dominant phylum in all sourdough samples was Firmicutes, which was represented exclusively by the Lactobacillales order. The two remaining and less abundant phyla were Proteobacteria and Bacteroidetes. The culture-independent approach allowed us to detect changes in microbial ecology during the 72-hr fermentation period. Weissella sp. was the most abundant genus after 24 hr of fermentation of the rye sourdough, but as the process progressed, its abundance decreased in favor of the Lactobacillus genus similarly as in wheat and spelt sourdoughs. The Lactobacillus genus was dominant in all sourdoughs after 72 hr, which was consistent with our results obtained using culture-dependent analyses. This work was carried out to determine the microbial biodiversity of sourdoughs that are made from wheat, spelt, and rye wholemeal flour and can be used as a source of strains for specific starter cultures to produce functional bread.