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Abstract Background Aggregation may play a main role in the adhesion of bacteria to the gastrointestinal epithelium and their colonization ability, as well as in probiotic effects through co-aggregation with intestinal pathogens and their subsequent removal. The aggregation phenomenon in lactococci is directly associated with the sex factor and lactose plasmid co-integration event or duplication of the cell wall spanning (CWS) domain of PrtP proteinase. Results Lactococcus lactis subsp. lactis BGKP1 was isolated from artisanal semi-hard homemade cheese and selected due to its strong auto-aggregation phenotype. Subsequently, non-aggregating derivative (Agg-) of BGKP1, designated as BGKP1-20, was isolated, too. Comparative analysis of cell surface proteins of BGKP1 and derivative BGKP1-20 revealed a protein of approximately 200 kDa only in the parental strain BGKP1. The gene involved in aggregation (aggL) was mapped on plasmid pKP1 (16.2 kb), cloned and expressed in homologous and heterologous lactococci and enterococci. This novel lactococcal aggregation protein was shown to be sufficient for cell aggregation in all tested hosts. In addition to the aggL gene, six more ORFs involved in replication (repB and repX), restriction and modification (hsdS), transposition (tnp) and possible interaction with mucin (mbpL) were also located on plasmid pKP1. Conclusion AggL is a new protein belonging to the collagen-binding superfamily of proteins and is sufficient for cell aggregation in lactococci.

Lactobacillus helveticus BGRA43 isolated from human intestines shows antimicrobial activity against foodborne pathogens and during fermentation in milk releases peptides with demonstrated anti-inflammatory properties. In this study, it was found that strain BGRA43 exhibits antimicrobial activity against human pathogens Yersinia enterocolitica, Shigella sonnei, S. flexneri and Streptococcus pneumoniae. Strain BGRA43 was able to survive in simulated gastric juice containing milk and retained cell number stability during the incubation in simulated intestinal conditions. In addition, LC/MS/MS analysis showed the ability of BGRA43 to hydrolyze β-lactoglobulin. Abundant growth of strain BGRA43 occurred in the presence of prebiotics inulin or concentrated oat bran β-glucan (Nutrim®), even when used as the sole carbon source. Similarly, strain BGRA43 grew satisfactorily in pure cow's or goat's milk as well as in the milk containing inulin or Nutrim®. Using the probiotic strain BGRA43 as a single starter strain, fermented milk products obtained from cow's or goat's milk with or without inulin or Nutrim® contained about 107 CFU/mL. The products were homogeneous and viscous and the best sensory scores were observed for fermented milk beverage made from reconstituted skimmed milk, whole cow's milk and whole goat's milk supplemented with 1% inulin. [ABSTRACT FROM AUTHOR]

The diversity of 87 Lactobacillus paracasei and Lactobacillus plantarum/ paraplantarum strains, previously identified from different autochthonous dairy products, was investigated by phenotypic and genotypic approaches. The increased resolution obtained using phenotypic and genotypic characterization allowed the level of strain heterogeneity detection to be widened. Phenotypic diversity was evaluated by studying biochemical characteristics of technological interest, including antimicrobial and proteinase activities, resistance to nisin, aggregation ability, production of exopolysaccharides, acetoin and diacetyl, citrate utilization, and antibiotic susceptibility. Genotypic diversity was generally evaluated by PCR amplification of repetitive bacterial DNA element fingerprinting using the (GTG) primer [(GTG)-PCR]. Moreover, in cases where strains were not discriminated by (GTG)-PCR combined with phenotypic analysis, pulsed-field gel electrophoresis (PFGE) analysis was performed. The results indicate that L. plantarum/ paraplantarum and L. paracasei natural isolates from artisanal dairy products are a gold mine in terms of diversity of strains and could be potentially interesting to dairy companies for the formulation of functional starter cultures in the production of innovative foods. [ABSTRACT FROM AUTHOR]

In this study, the plasmid content and bacteriocin production of natural isolates of lactococci were investigated. Five bacteriocin producing lactococcal strains (Lactococcus lactis subsp. lactis BGMN1-2, BGMN1-3, BGMN1-5, BGMN1-6, and BGMN2-7) were isolated as nonstarter microflora of semi-hard homemade cheese and characterized. All isolates contained a number of plasmids. It was shown that lcnB structural genes for bacteriocin lactococcin B were located on large plasmids in all isolates. In the strains BGMN1-3 and BGMN1-5 proteinase prtP genes collocated with lcnB. Furthermore, these strains produced two additional bacteriocins (LsbA and LsbB) with genes responsible for their production and immunity located on the small rolling circle-replicating plasmid pMN5. Using deletion experiments of pMN5, minimal replicon of the plasmid and involvement of a bacteriocin locus in plasmid maintenance were identified. In addition, plasmid curing experiments showed that genes for catabolism or transport of 10 carbohydrates in the strain BGMN1-5 were plasmid located.