The use of multifunctional yeast-lactobacilli starter cultures improves fermentation performance of Spanish-style green table olives.

In this work, Lactobacillus pentosus LPG1, Lactobacillus pentosus Lp13, Lactobacillus plantarum Lpl15, and Wickerhanomyces anomalous Y12, all of them previously isolated from fermented table olive biofilms, were used (alone or in combination) as multifunctional starters for Manzanilla Spanish-style green table olive fermentations. Their performances were evaluated through the changes in the key physico-chemical and microbiological parameters, correlation between AI-2 production and biofilm formation, inoculum imposition, metataxonomic analysis and sensory characteristics of the finished products. Inoculation only with lactic acid bacteria (LAB) strains led to higher titratable acidities and lower pH values than the spontaneous fermentation (non-inoculated control), mainly during the first steps of processing. However, the sequential inoculation of the yeast and then the combination of the 3 LAB strains showed the most favourable evolution. LPG1 strain and, particularly Lp13, were excellent biofilms former and showed the major imposition on the fruit epidermis, as corroborated by rep-PCR analysis. Production of AI-2 was lower in the treatment inoculated exclusively with yeast Y12 but had the highest presence in the sequential yeast-LAB inoculum, with its maximum concentration and maximum LAB population on fruits (19th days) strongly related. Metataxonomic analysis of the biofilms at the end of the fermentation revealed, in addition to Lactobacillus, high proportions of sequences from genera Marinilactobacillus, Alkalibacterium, Halolactobacillus, and low levels of Halomonas and Aerococcus. Compositional data analysis of the omics data revealed that Lpl15 was scarcely efficient for controlling the spontaneous microbiota since its treatment presented the highest proportions of Aerococcus genus. Finally, the sensory analysis showed similar characteristics for the treatment inoculated with LPG1 and the spontaneous process, with olives inoculated with the yeast (alone or in combination with Lactobacillus strains) showing attractive scores. Then, inoculation of Spanish-style table olive fermentations with a sequential yeast and LAB combination could be an advisable practice.

The use of starter cultures in the table olive fermentation can modulate the antiadhesive properties of brine exopolysaccharides against enterotoxigenic Escherichia coli.

The present study aimed to evaluate different mates of Candida boidinii and Lactobacillus pentosus strains as starters in green table olive fermentation. Changes in fermentation characteristics as well as changes in the functional properties of the microbial exopolysaccharides (EPS) produced during the process were registered. The in vitro adhesion test demonstrated that most EPS samples could specifically attach ETEC K88. In vitro studies with porcine intestinal cells showed the improved blocking activity of the fimbria (blocking test) when the mutant strain L. pentosus 119-14MT was used alone as a starter. All EPS samples showed the ability to block receptors in the cells (exclusion test) although without differences between starter treatments. In the displacement test, EPS samples failed to remove the pathogen once attached. According to these results, L. pentosus 119-14MT, a high EPS variant, seemed to be the most effective starter improving the anti-adhesive properties of brine EPS and increasing its ability to block the ETEC K88 fimbria. These results illustrate that the anti-adhesive properties of the EPSs produced during the traditional fermentation of olives could be modulated by the use of defined starters. This opens the door to new fermentation processes aimed to produce green table olives as functional food to prevent ETEC diarrhea.

Biodiversity and Multifunctional Features of Lactic Acid Bacteria Isolated From Table Olive Biofilms.

In the present study, a total of 554 lactic acid bacteria (LAB) isolates were obtained from the olive surface of Manzanilla, Gordal, and Aloreña cultivars processed as green Spanish-style or directly brined (natural) olives. The isolates obtained from industrial processes were genotyped by rep-PCR with primer GTG5, collecting a total of 79 different genotypes. The α-biodiversity indexes showed that the LAB diversity was higher in the biofilms on the fruits which followed the Spanish-style process than in those just brined. Sixteen genotypes had a frequency higher >1% and were identified, by multiplex PCR recA gene and 16S gene sequencing, as belonging to Lactobacillus pentosus (n = 13) and Lactobacillus plantarum (n = 3) species. A multivariate analysis based on a dataset with 89,744 cells, including technological (resistance to salt and pH, production of lactic acid, auto and co-aggregation with yeast species, ß-glucosidase and esterase activities), and potential probiotic characteristics (survival to gastric and pancreatic digestions, resistance to antibiotics, inhibition of pathogens, presence of bsh genes, cholesterol removal, hemolytic, α-glucosidase, ß-galactosidase, and phytase activities) showed that the 16 genotypes could be grouped into 3 great phenotypes. Thus, the genotype biodiversity in table olive biofilms was limited but, at phenotype level, it was even lower since L. pentosus predominated clearly (80.15% isolates). L. pentosus Lp13 was the genotype with the most promising characteristics for its use as a multifunctional starter, with this strain being and ubiquitous microorganism present in both natural and lye-treated olive fermentations.

Multiple Genome Sequences of Lactobacillus pentosus Strains Isolated from Biofilms on the Skin of Fermented Green Table Olives.

The draft genome sequences of five Lactobacillus pentosus strains isolated from biofilms on the skin of green table olives are presented here. These genome sequences will assist in revealing the potential probiotic properties of these strains, as the intake of fermented olives implicates the passage of millions of Lactobacillus spp. throughout a consumer's gastrointestinal tract.

Draft Genome Sequences of Six Lactobacillus pentosus Strains Isolated from Brines of Traditionally Fermented Spanish-Style Green Table Olives.

Here, we report the genome sequences of six Lactobacillus pentosus strains isolated from traditional noninoculated Spanish-style green table olive brines. The total genome sizes varied between 3.77 and 4.039 Mbp. These genome sequences will assist in revealing the genes responsible for both technological and probiotic properties of these strains.

Genome overview of eight Candida boidinii strains isolated from human activities and wild environments.

Candida boidinii is an Ascomycota yeast with important biotechnological applications. In this paper we present the genome sequencing and annotation of eight strains of this species isolated from human activities and wild environments. The produced assemblies revealed several strain specific features in terms of genomic GC content (ranging from 30.9 to 32.7%), genome size (comprised between 18,791,129 and 19,169,086 bp) and total number of protein coding genes (ranging from 5819 to 5998), with putative assignation to their general KOG functional categories. The obtained data underlined the presence of two different groups for this species. The results reported herein provide new insights into the plasticity of the genome of this yeast species and represent a starting point for further studies in view of its biotechnological applications.