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The fruit-origin strain Fructobacillus tropaeoli CRL 2034 can biotransform selenium into seleno-nanoparticles and selenocysteine. The proteomic analysis of F. tropaeoli CRL 2034 exposed to 5 and 100 ppm of Se showed a dose-dependent response since 19 and 77 proteins were deregulated, respectively. In the presence of 5 ppm of Se, the deregulated proteins mainly belonged to the categories of energy production and conversion or had unknown functions, while when cells were grown with 100 ppm of Se, most of the proteins were grouped into amino acid transport and metabolism, nucleotide transport and metabolism, or into unknown functions. However, under both Se conditions, glutathione reductases were overexpressed (1.8–3.1-fold), while mannitol 2-dehydrogenase was downregulated (0.54–0.19-fold), both enzymes related to oxidative stress functions. Mannitol 2-dehydrogenase was the only enzyme found that contained SeCys, and its activity was 1.27-fold increased after 5 ppm of Se exposure. Our results suggest that F. tropaeoli CRL 2034 counteracts Se stress by overexpressing proteins related to oxidative stress resistance and changing the membrane hydrophobicity, which may improve its survival under (food) storage and positively influence its adhesion to intestinal cells. Selenized cells of F. tropaeoli CRL 2034 could be used for producing Se-enriched fermented foods. Key points: • Selenized cells of F. tropaeoli showed enhanced resistance to oxidative stress. • SeCys was found in the Fructobacillus mannitol 2-dehydrogenase polypeptide chain. • F. tropaeoli mannitol 2-dehydrogenase activity was highest when exposed to selenium. [ABSTRACT FROM AUTHOR]

The enzymes D- and L-lactate dehydrogenase are involved in the reduction of pyruvate to D(+)- and L(−)-lactate, respectively. The fig-origin strain Fructobacillus tropaeoli CRL 2034 produces D- and L-lactic acids in a 9:1 ratio. In this work, two D-ldh (ldh1 and ldh2) and one L-ldh (ldh3) genes were found in the CRL 2034 genome. ldh1 and ldh2 are homologous (79% identity) and organized as contiguous operons, each gene containing 996 base pair (bp) and encoding for a 331-amino acid (aa) protein (74% identity). In contrast, ldh3 is a 927-bp gene coding for a 308-aa protein. The identity between ldh1/ldh2 and ldh3 was lower than 48%. To elucidate the role of these genes in the synthesis of lactic acid by the Fructobacillus strain, plasmid insertion mutants in each gene were generated and characterized. The growth kinetic parameters were affected only in CRL2034 ldh1::pRV300 cells, this mutant showing the lowest total lactic acid production (4.50 ± 0.15 versus 6.36 ± 0.67 g/L of wild-type strain), with a D/L ratio of 7.1:2.9. These results showed that the ldh1 gene is primarily responsible for lactic acid production by the studied strain. A comparative analysis among strains of the five Fructobacillus species revealed that the identity of D-LDH proteins was higher than 70%, while the identity of L-LDH was over 60%. Finally, phylogenetic analysis of D- and L-LDHs revealed that only D-LDH phylogeny was consistent to the phylogenetic evolution among Fructobacillus and evolutionarily related genera. Key Points •F. tropaeoli CRL 2034 harbors three ldh genes in its genome. •ldh1 and ldh2 encode D-lactate dehydrogenase; ldh3 encodes L-lactate dehydrogenase. •Gene ldh1 plays the major role in lactic acid production by strain CRL 2034. •Fructobacillus D-LDH phylogeny was consistent to phylogenetic evolution. [ABSTRACT FROM AUTHOR]

Adequate fruit consumption helps to prevent several chronic age-related diseases. Selenium (Se) is an elemental micronutrient with antioxidant capacity. In general, fruits and Se ingest by humans are below the recommended daily intake value. Fresh fruits are highly susceptible to deterioration during storage. Fermentation can improve the storage period, sensory profile, and bioactive compound content of foods; moreover, some lactic acid bacteria can accumulate organic Se intracellularly. In this work, microbial growth and Se accumulation by Levilactobacillus brevis CRL2051 and Fructobacillus tropaeoli CRL2034 in tropical fruit juices were evaluated. The strains could grow 1–2 log cfu/mL, alone or combined, in mango, passion fruit, and mango/passion fruit juices, although they could not completely eliminate native microorganisms in unpasteurised juices. In pasteurised trials, both strains consumed fruit carbohydrates producing lactic acid, in addition to acetic acid, and mannitol by F. tropaeoli. Both strains accumulated Se intracellularly in the fruit juices, especially (123.0 μg/L) in the passion fruit juice fermented by the mixed culture. Finally, the Fructobacillus strain increased 3.42 times the phenolic compound concentration in the mango/passion fruit juice with added Se after 24 h. The assayed LAB strains could be used for preparing functional fermented fruit beverages bio-enriched in Se. • The cultures grew in the juices but could not eliminate fruit native microorganisms • F. tropaeoli produced the natural sweetener mannitol during fruit fermentation • Selenium concentration was higher in mixed and passion fruit fermented juices • Mixed fruit juice phenolics increased when F. tropaeoli was used as starter culture • A fermented beverages bio-enriched in selenium was formulated using tropical fruits [ABSTRACT FROM AUTHOR]