The PsdRSAB and ApsRSAB detoxification modules, together with the antimicrobial peptides (AMPs)-resistance determinants Dlt system and MprF protein, play major roles in the response to AMPs in ...Lacticaseibacillus paracasei BL23. Sensitivity assays with a collection of mutants showed that the PsdAB ABC transporter and the Dlt system are the main subtilin resistance determinants. Quantification of the transcriptional response to subtilin indicate that this response is exclusively regulated by the two paralogous systems PsdRSAB and ApsRSAB. Remarkably, a cross-regulation of the derAB, mprF and dlt-operon genes-usually under control of ApsR-by PsdR in response to subtilin was unveiled. The high similarity of the predicted structures of both response regulators (RR), and of the RR-binding sites support this possibility, which we experimentally verified by protein-DNA binding studies. ApsR-P shows a preferential binding in the order P
> P
> P
> P
. However, PsdR-P bound with similar apparent affinity constants to the four promoters. This supports the cross-regulation of derAB, mprF and the dlt-operon by PsdR. The possibility of cross-regulation at the level of RR-promoter interaction allows some regulatory overlap with two RRs controlling the expression of systems involved in maintenance of critical cell membrane functions in response to lantibiotics.
The use of probiotic lactobacilli has been proposed as a strategy to mitigate damage associated with exposure to toxic metals. Their protective effect against cationic metal ions, such as those of ...mercury or lead, is believed to stem from their chelating and accumulating potential. However, their retention of anionic toxic metalloids, such as inorganic arsenic, is generally low. Through the construction of mutants in phosphate transporter genes (
) in
and
strains, coupled with arsenate As(V) uptake and toxicity assays, we determined that the incorporation of As(V), which structurally resembles phosphate, is likely facilitated by phosphate transporters. Surprisingly, inactivation in
of PhoP, the transcriptional regulator of the two-component system PhoPR, a signal transducer involved in phosphate sensing, led to an increased resistance to arsenite As(III). In comparison to the wild type, the
strain exhibited no differences in the ability to retain As(III), and there were no observed changes in the oxidation of As(III) to the less toxic As(V). These results reinforce the idea that specific transport, and not unspecific cell retention, plays a role in As(V) biosorption by lactobacilli, while they reveal an unexpected phenotype for the lack of the pleiotropic regulator PhoP.
Preterm microbial colonization is affected by gestational age, antibiotic treatment, type of birth, but also by type of feeding. Breast milk has been acknowledged as the gold standard for human ...nutrition. In preterm infants breast milk has been associated with improved growth and cognitive development and a reduced risk of necrotizing enterocolitis and late onset sepsis. In the absence of their mother's own milk (MOM), pasteurized donor human milk (DHM) could be the best available alternative due to its similarity to the former. However, little is known about the effect of DHM upon preterm microbiota and potential biological implications. Our objective was to determine the impact of DHM upon preterm gut microbiota admitted in a referral neonatal intensive care unit (NICU). A prospective observational cohort study in NICU of 69 neonates <32 weeks of gestation and with a birth weight ≤1,500 g was conducted. Neonates were classified in three groups according to feeding practices consisting in their MOM, DHM, or formula. Fecal samples were collected when full enteral feeding (defined as ≥150 cc/kg/day) was achieved. Gut microbiota composition was analyzed by 16S rRNA gene sequencing. Despite the higher variability, no differences in microbial diversity and richness were found, although feeding type significantly influenced the preterm microbiota composition and predictive functional profiles. Preterm infants fed MOM showed a significant greater presence of Bifidobacteriaceae and lower of Staphylococcaceae, Clostridiaceae, and Pasteurellaceae compared to preterm fed DHM. Formula fed microbial profile was different to those observed in preterm fed MOM. Remarkably, preterm infants fed DHM showed closer microbial profiles to preterm fed their MOM. Inferred metagenomic analyses showed higher presence of
genus in mother's milk group was related to enrichment in the Glycan biosynthesis and metabolism pathway that was not identified in the DHM or in the formula fed groups. In conclusion, DHM favors an intestinal microbiome more similar to MOM than formula despite the differences between MOM and DHM. This may have potential beneficial long-term effects on intestinal functionality, immune system, and metabolic activities.
Human milk oligosaccharides (HMOs) are a mixture of structurally diverse carbohydrates that contribute to shape a healthy gut microbiota composition. The great diversity of the HMOs structures does ...not allow the attribution of specific prebiotic characteristics to single milk oligosaccharides. We analyze here the utilization of four disaccharides, lacto-N-biose (LNB), galacto-N-biose (GNB), fucosyl-α1,3-GlcNAc (3FN) and fucosyl-α1,6-GlcNAc (6FN), that form part of HMOs and glycoprotein structures, by the infant fecal microbiota. LNB significantly increased the total levels of bifidobacteria and the species Bifidobacterium breve and Bifidobacterium bifidum. The Lactobacillus genus levels were increased by 3FN fermentation and B. breve by GNB and 3FN. There was a significant reduction of Blautia coccoides group with LNB and 3FN. In addition, 6FN significantly reduced the levels of Enterobacteriaceae family members. Significantly higher concentrations of lactate, formate and acetate were produced in cultures containing either LNB or GNB in comparison with control cultures. Additionally, after fermentation of the oligosaccharides by the fecal microbiota, several Bifidobacterium strains were isolated and identified. The results presented here indicated that each, LNB, GNB and 3FN disaccharide, might have a specific beneficial effect in the infant gut microbiota and they are potential prebiotics for application in infant foods.
Polyphosphate (poly-P) biosynthesis in bacteria has been linked to many physiological processes and has been characterized as an interesting functional molecule involved in intestinal homeostasis. We ...determined the capacity for poly-P production of 18 probiotic strains mainly belonging to Bifidobacterium and former Lactobacillus genera, showing that poly-P synthesis varied widely between strains and is dependent on the availability of phosphate and the growth phase. Bifidobacteria were especially capable of poly-P synthesis and poly-P kinase (ppk) genes were identified in their genomes together with a repertoire of genes involved in phosphate transport and metabolism. In Bifidobacterium longum KABP042, the strain we found with highest poly-P production, variations in ppk expression were linked to growth conditions and presence of phosphate in the medium. Moreover, the strain produced poly-P in presence of breast milk and lacto-N-tetraose increased the amount of poly-P synthesized. Compared to KABP042 supernatants low in poly-P, exposure of Caco-2 cells to KABP042 supernatants rich in poly-P resulted in decreased epithelial permeability and increased barrier resistance, induction of epithelial protecting factors such as HSP27 and enhanced expression of tight junction protein genes. These results highlight the role of bifidobacteria-derived poly-P as a strain-dependent functional factor acting on epithelial integrity.
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•UAE limited the extraction of polyphenols from thyme and rosemary.•UAE improved the extraction of carotenoids from thyme and rosemary.•Aqueous extracts with UAE stimulated the ...activity of L. casei and Bifidobacterium.•Thyme aqueous extract with UAE inhibited Salmonella enterica growth rate.•UAE improved the aqueous extraction of antimicrobial compounds from thyme.
Mediterranean herbs, specially thyme and rosemary, are important ingredients in food preparation and more recently have been studied as natural sources of bioactive compounds. This study aimed to study the effect of matrix (thyme vs. rosemary), and extraction protocol (conventional extraction vs. ultrasound assisted extraction) solvent composition (water vs. 50:50 ethanol:water solution) on the extraction of high value compounds (phenolic compounds, flavonoids and carotenoids) and also explore the antioxidant, antimicrobial (Listeria innocua, Staphylococcus aureus, and Salmonella enterica), probiotic (Lactobacillus casei and Bifidobacterium lactis), and anti-inflammatory activities. The phenolic, flavonoid and carotenoid content of extracts was greatly influenced by extraction conditions wherein the ultrasound pre-treatment improved the extraction of carotenoids but induced the opposite effect for polyphenols and flavonoids in both herbs. Only the aqueous extract of thyme obtained from ultrasound pre-treatment was the only extract that inhibited the growth of potentially pathogenic bacteria, stimulated the probiotic bacteria and achieved high anti-inflammatory and antioxidant activity. Moreover, this extract also was rich on phenolic compounds (such as p-coumaric acid 4-O-glucoside, kaempferol 3-O-rutinoside, feruloyl glucose, and 4-vinylguaiacol) and carotenoids. Therefore, ultrasound extraction of bioactive compounds with water as solvent could be explored in food and pharmaceutical applications.
Lactobacillus casei BL23 carries 17 two-component signal transduction systems. Insertional mutations were introduced into each gene encoding the cognate response regulators, and their effects on ...growth under different conditions were assayed. Inactivation of systems TC01, TC06, and TC12 (LCABL_02080-LCABL_02090, LCABL_12050-LCABL_12060, and LCABL_19600-LCABL_19610, respectively) led to major growth defects under the conditions assayed.
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•Spelt is a traditional cereal from Garfagnana.•Spelt has high levels of proteins, carotenoids and bioactive compounds.•Spelt fermentation decreases SCCs, except for ...mannitol.•Fermented spelt stimulates Lactobacillusspp. andBifidobacteriumspp. group growth in the gut.•Fermented spelt increases SCFAs content in microbiota from normal weight and obese people.
The aim of this study has been to evaluate the impact of the fermentation process of the spelt from Garfagnana on its chemical composition and short-chain carbohydrates (SCCs) levels, andon thein vitromicrobial growth and metabolism.
The fermentation process of spelt significantly increases its protein and mannitol content, and decreases its dietary fiber content and fructose, glucose, sucrose, maltose, and raffinose concentration. Fermented spelt modulates thein vitrointestinal microbiota, promoting a stimulation ofLactobacillus andBifidobacteriumspp. growth accompanied by a high production of lactate, acetate,and propionate, both in human gut microbiota from normal weight and obese subjects. The multivariate approach (PCA) combining viable counts and metabolite concentration values has suggested that spelt fermentation could beneficially modulate the gut microbiota from normal weight and obese individuals, stimulating bacteria eliciting anti-inflammatory responses. Further, in vivo studies are recommended for the impact that fermented spelt could have in human nutrition in health and disease.
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