Search Results (26)

This study employed shotgun metagenomics to investigate microbial dynamics, phage-bacteria interactions, and functional genes throughout a three-month apple vinegar fermentation process. A total of 5621 microbial species were identified, revealing three distinct phases: (i) Enterobacteria and non-Saccharomyces species dominated the initial substrate; (ii) S. cerevisiae and Leuconostoc pseudomesenteroides prevailed in the intermediate phase; and (iii) acetic acid bacteria (Acetobacter ghanesis and Gluconobacter spp.), alongside non-Saccharomyces species (Pichia kudriavzevii and Malassezia restricta), dominated the final stages. Bacteriophage analysis revealed the presence of phages targeting spoilage bacteria, such as Pseudomonas and Erwinia, suggesting a role in regulating microbial stability and enhancing fermentation control. Functional metagenomic analysis highlighted key pathways associated with microbial growth and metabolite production, including carbohydrate and amino acid metabolism, energy production, and glycan biosynthesis. Enzymes involved in stress adaptation and secondary metabolism, including oxidative phosphorylation and phenolic compound synthesis, demonstrated microbial resilience and their potential role in shaping the product’s sensory and functional properties. Moreover, Enterobacteriaceae species were associated with pectin degradation during the early stages, aiding substrate breakdown. These findings are crucial for microbial and phage management in fermentation technology, offering valuable insights for innovation in the vinegar industry. Full article

Baking leavening agents and fermentation conditions may influence the gastrointestinal fate of nutrients in baked goods, thereby affecting their bioavailability. This study aimed to evaluate the digestibility of sourdough pizza fermented with lactic acid bacteria species (Levilactobacillus brevis, Fructilactobacillus sanfranciscensis, Leuconostoc pseudomesenteroides) and yeast, compared to traditional pizza fermented with baker′s yeast. The effects of leavening time (up to 48 h) and microbial leavening agents on the nutritional profile and digestibility of baked pizzas were investigated by examining the microbiological and physico-chemical changes in the doughs, with a particular focus on the sugar content. Additionally, the degree of protein hydrolysis and the levels of FODMAPs (fermentable oligosaccharides, disaccharides, monosaccharides, and polyols) were quantified on the cooked pizzas both before and after in vitro gastrointestinal digestion. In vitro protein digestibility was not significantly influenced by the type of microbial leavening agent used or fermentation time. However, extended fermentation, particularly with lactic acid bacteria sourdough, resulted in a notable decrease in FODMAPs, thereby enhancing the digestibility and overall health profile of the pizza for individuals sensitive to these compounds. Future research should further explore the mechanisms behind these changes and their implications for dietary recommendations. Full article

Integrating heterogeneous genes is widely used in metabolic engineering to produce D-lactic acid (D-LA), an essential compound in bioplastics and pharmaceuticals. However, research on the effects of integrating various loci on gene expression, especially regarding flocculation behavior, remains limited. This study constructed Saccharomyces cerevisiae strains by incorporating a codon-optimized D-LDH gene from Leuconostoc pseudomesenteroides (LpDLDH) into the specific genomic loci of the CYB2, PDC1, MPC1, PDC6, ADH1, and PDC5 genes to redirect pyruvate toward lactic acid. Strains with the LpDLDH gene integrated at the PDC1 locus achieved the highest D-LA titers (51 g/L) with minimal ethanol byproduct, followed by strains with integrations into the CYB2 locus at 31.92 g/L, the MPC1 locus at 10 g/L, and the PDC6 locus at 0.026 g/L. In contrast, strains with LpDLDH integrated at the ADH1 and PDC5 loci failed to produce detectable levels of D-LA and exhibited a complete loss of flocculation. Gene expression analysis revealed a significant expression of genes related to flocculation (FLO5), stress adaptation (HSP150), and cell wall integrity (YGP1, SED1, and SCW11). The CYB2-integrating strain showed strong flocculant properties, contributing to its robustness. These findings highlight the influence of genomic locus selection on metabolic flux and stress adaptation, offering insights into optimizing D-LA production in flocculant S. cerevisiae yeast. Full article

Leuconostoc pseudomesenteroides is a rare pathogen that can cause bacteremia in immunocompromised patients, particularly those with hematological conditions like acute myeloid leukemia. In this case, a 56-year-old woman developed Leuconostoc bacteremia following chemotherapy and multiple infections, including invasive aspergillosis. Despite broad-spectrum antibiotic treatments, her fever persisted until the blood cultures identified Leuconostoc pseudomesenteroides. Switching to intravenous ampicillin led to the resolution of symptoms. This case highlights the challenges in diagnosing and treating rare, glycopeptide-resistant bacteria in immunosuppressed patients and underscores the importance of repeated blood cultures and automated diagnostic systems. It also highlights the need for more rapid ways of diagnosing and treating such rare infections. Full article

Lactic acid bacteria exopolysaccharide (EPS) is a large molecular polymer produced during the growth and metabolism of lactic acid bacteria. EPS has multiple biological functions and is widely used in fields such as food and medicine. However, the low yield and high production cost of EPS derived from lactic acid bacteria limit its widespread application. In this study, we used beet waste residue as a substrate to produce EPS by fermentation with Leuconostoc pseudomesenteroides to improve the utilization rate of agricultural waste and reduce the production cost of lactic acid bacterial EPS. After purification, the molecular weight (Mw) of EPS was determined to be 417 kDa using high-performance size exclusion chromatography (HPSEC). High-performance liquid chromatography (HPLC), Fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy revealed that the EPS was composed of glucose subunits with α-1,6 glycosidic linkages. The thermal analysis and heavy metal adsorption capacity revealed a relatively high degradation temperature of 315.54 °C and that the material could effectively adsorb Cu²⁺. Additionally, the findings indicated that the EPS exhibited a significant ability to neutralize free radicals, a property that was found to be concentration dependent. Furthermore, the results of the intracellular study showed the protective effect of freshly isolated EPS on tBHP-induced cellular oxidative stress at a concentration of 50 µg/mL. These results suggest that the EPS from L. pseudomesenteroides may be developed as antioxidant agents for functional food products and pharmaceutical applications due to its capacity to scavenge free radicals. Full article

Context and Objectives: There is an urgent need for a dietary shift towards an increased consumption of plant protein foods. However, some nutritional and sensory challenges are associated with whole-meal grains and pulses. Fermentation is a promising technology for reducing anti-nutrients, increasing protein digestibility, and reducing starch digestibility in plant-based matrices. This study aimed to evaluate the impact of different fermentation conditions using different bacteria on the nutritional composition of grains and pulses. Methods: Milled wheat, buckwheat, and chickpeas were fermented using different bacteria or combinations of bacteria, i.e., Lactobacillus delbrueckii + Streptococcus thermophilus (Vega), Lacticaseibacillus rhamnosus (Lrham), Leuconostoc pseudomesenteroides (Lpseu), Weissella confusa alone (Wcon) or combined with Lactococcus lactis (Wcon + Lac), or Pediococcus pentosaceus (Wcon + Pen), for 24 or 48 h. Protein hydrolysis and protein and starch digestibility were measured using the O-phtaldialdehyde and Infogest methods, respectively. Starch digestibility was evaluated using the Englyst method. Dietary fibers (DF) were quantified. Results: In buckwheat, all fermentation conditions increased protein hydrolysis, especially Vega. In chickpeas, Lrham and Wcon, alone and in combination, increased protein hydrolysis. In wheat, fermentation did not increase protein hydrolysis. Protein digestibility increased only with Wcon + Pen in buckwheat. For chickpeas and wheat, fermentation did not increase protein hydrolysis, and it was lowest with Lpseu in both cases. Lpseu, Wcon + Lac, and Wcon + Pen led to increased DF in buckwheat, especially soluble high-molecular-weight (HMW) buckwheat (dextran formation). In chickpeas, no increase in DF content was observed due to the hydrolysis of oligosaccharides, but soluble HMW DF increased with Lpseu, Wcon + Lac, and Wcon + Pen (dextran formation). No significant change was observed for wheat DF. The starch hydrolysis index (HI) increased with Lrham over 48 h and Lpseu over 24 h but decreased with Wcon + Lac over 24 h in buckwheat. In chickpeas, HI was reduced in all conditions, the lowest values being with Wcon + Pen and Wcon + Lac. In wheat, HI increased with Lrham over 48 h and decreased with Wcon + Pen. Conclusions: The impact of the fermentation conditions tested differed depending on the grains. Some conditions improved the nutritional characteristics of these grains. These results show promising effects concerning the nutritional quality of grains, which need to be confirmed in finished products. Full article

Bacterial exopolysaccharides (EPSs) share their compositional and structural features with plant dietary fibers. Therefore, analysis of bacterial EPSs produced during fermentation of fruit or vegetables interferes with that of plant fibers. To get rid of this effect, bacteria were grown on a solid medium containing fruit juice or purée and EPSs were recovered in a quantitative dependent manner and were quantified with the phenol-sulfuric acid colorimetric method. The protocol was assayed both on MRS medium and fruit-based media, with three bacterial strains from two species, Leuconostoc pseudomesenteroides 56 and Weissella cibaria 21 and 64. With that method, differences in EPS production levels were shown according to the strain and cultivation conditions, such as sucrose content and pH. Complementary analysis with NMR indicated that glucose and sucrose were partly recovered with EPSs, pointing out the requirement for a further purification step. It also showed that EPSs’ ramified structure differed according to the strain and the fruit used in the medium. This method for EPS recovery is helpful to select strains and to pilot EPS production during lactic fermentation of fruit or vegetable foods. Full article

Texture and flavour are the key attributes determining sensory quality and are highly affected by starter cultures. A selection of phenotypic strains is needed to create diverse texture and flavour to meet consumers’ preferences. In this study, the use of five lactic acid bacteria strains in the production of fermented milk, along with the metabolite profiles, microstructure, and rheological properties of the fermented milk samples, was investigated. Our results showed that Lactobacillus helveticus (LH) and Streptococcus thermophilus (ST) had a stronger acidification during fermentation but resulted in products with a coarser protein network compared to Lactococcus lactis (BL1) and Leuconostoc mesenteroides (CL3). Milk fermented by LH had the highest viscosity and exopolysaccharide concentration, while milk fermented by ST had the highest concentration of diacetyl. Although Leuconostoc pseudomesenteroides (CL3ST) had a minimal acidification capability, it produced high levels of ethyl-derived compounds associated with sweet, fruity, and floral fragrances. The results demonstrated that LH and ST could be used as starter cultures targeting fermented milks with different viscosities, while BL1, CL3, and CL3ST are suitable as adjunct cultures to impact different acidic sharpness and flavour notes. Full article

d-lactic acid, a chiral organic acid, can enhance the thermal stability of polylactic acid plastics. Microorganisms such as the yeast Pichia pastoris, which lack the natural ability to produce or accumulate high amounts of d-lactic acid, have been metabolically engineered to produce it in high titers. However, tolerance to d-lactic acid remains a challenge. In this study, we demonstrate that cell flocculation improves tolerance to d-lactic acid and increases d-lactic acid production in Pichia pastoris. By incorporating a flocculation gene from Saccharomyces cerevisiae (ScFLO1) into P. pastoris KM71, we created a strain (KM71-ScFlo1) that demonstrated up to a 1.6-fold improvement in specific growth rate at high d-lactic acid concentrations. Furthermore, integrating a d-lactate dehydrogenase gene from Leuconostoc pseudomesenteroides (LpDLDH) into KM71-ScFlo1 resulted in an engineered strain (KM71-ScFlo1-LpDLDH) that could produce d-lactic acid at a titer of 5.12 ± 0.35 g/L in 48 h, a 2.6-fold improvement over the control strain lacking ScFLO1 expression. Transcriptomics analysis of this strain provided insights into the mechanism of increased tolerance to d-lactic acid, including the upregulations of genes involved in lactate transport and iron metabolism. Overall, our work represents an advancement in the efficient microbial production of d-lactic acid by manipulating yeast flocculation. Full article

Glucansucrase was purified from Leuconostoc pseudomesenteroides. The glucansucrase exhibited maximum activity at pH 5.5 and 30 °C. Ca²⁺ significantly promoted enzyme activity. An exopolysaccharide (EPS) was synthesized by this glucansucrase in vitro and purified. The molecular weight of the EPS was 3.083 × 10⁶ Da. Fourier transform infrared (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy showed that the main structure of glucan was 97.3% α-(1→6)-linked D-glucopyranose units, and α-(1→3) branched chain accounted for 2.7%. Scanning electron microscopy (SEM) observation of dextran showed that its surface was smooth and flaky. Atomic force microscopy (AFM) of dextran revealed a chain-like microstructure with many irregular protuberances in aqueous solution. The results showed that dextran had good thermal stability, water holding capacity, water solubility and emulsifying ability (EA), as well as good antioxidant activity; thus it has broad prospects for development in the fields of food, biomedicine, and medicine. Full article

Anthracnose disease is still a threat to avocado fruit quality, and the use of fungicide (Plochloraz^®) for its control has generated safety concerns that necessitate the search for alternatives. Therefore, the efficiency of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against Colletotrichum gloeosporioides was investigated in this study. Weissella cibaria 21 (LAB 21), Leuconostoc pseudomesenteroides 56 (LAB 56), Weissella confusa 17 (LAB 17), Lactiplantibacillus plantarum 75 (LAB 75), and Lactiplantibacillus plantarum 171 (LAB 171) were evaluated in vitro as potential biocontrol agents to replace the Prochloraz^® that is currently used in susceptible avocado (Persea americana Miller) Fuerte fruit. To confirm the biocontrol activity of the selected LAB strains, the antagonistic growth, spore germination, LAB recovery, nutrient competition, acid tolerance, and biofilm formation were assessed. In fruit treated with a LAB cell suspension, curatively inoculated with C. gloeosporioides, or naturally infected avocado cv Fuerte fruit, the epicatechin content and expression of defense-related genes (PAL, LOX, AVFADl 2–3, AVFAEL, and FLS) were compared with Prochloraz^® and sterile water (control) treatments. With LAB 56, LAB 75, and LAB 21, significant inhibition of radial mycelial growth (MGI) (>90%) and spore germination (100%) was observed similar to those due to Prochloraz^®. The MGI increased with a reduction in nutrient concentration. LAB strains reduced anthracnose disease incidence and severity compared with Prochloraz^® and were the highest in LAB 21 and LAB 56. The LAB 21 and LAB 56 strains produced strong biofilms against C. gloeosporioides. In contrast to LAB 56, the control, and Prochloraz^®, and LAB 21 had the highest epicatechin content (406 mg/g) and upregulated the PAL, AVFADl 2–3, AVFAEl, and FLS genes, thereby reducing the incidence of anthracnose in avocado fruit. As a result, LAB 21 suspensions can be used as an alternative to Prochloraz^® in the control of anthracnose disease. Full article

The purpose of this study was to investigate the bioaccessibilities of total phenolic compounds, carotenoid profile, antioxidant activity, and Lactic acid bacteria (LAB) survival in fermented mango juice (MJs) obtained from three mango cultivars after exposure to an in vitro gastrointestinal digestion model. The MJs from three cultivars (‘Sabre’, ‘Peach’, and ‘Tommy Atkins’) were fermented using Lactiplantibacillus plantarum 75 (L75), Leuconostoc pseudomesenteroides 56 (L56), and their combination (L56 + 75). Fermented MJs were digested and fractions: gastric (GF), intestinal (IF), and dialysis (DF) were analyzed for total polyphenolic content (TPC), antioxidant activity (FRAP), 1-diphenyl-2-picrylhydrazyl (DPPH), and 2.2-azinobis-3-ethyl-benzothiazoline–6-sulfonic acid (ABTS). In addition, the carotenoid content and the LAB population were determined from the GF and IF. After digestion, TPC decreased while fermentation improved its bioaccessibility. L75-fermented ‘Sabre’ MJs had the highest bioaccessible TPC in the GF (75.65%), IF (50.10%), and DF (32.52%) while L56 ‘Peach’ MJs increased the β-carotene bioaccessibility by 1.32-fold at GF and IF (1.21-fold). When compared to the other two juices, ‘Sabre’ and ‘Peach’ MJs fermented with L75 showed the highest IC₅₀ values for DPPH and ABTS. Generally, L75-fermented ‘Sabre’ MJs had the highest LAB survival at both GF (7.57 Log CFU/mL) and IF (7.45 Log CFU/mL) and hold potential as probiotic juices. L56-fermented ‘Sabre’ MJs would ensure the delivery of four times the carotenoid recommended dietary allowance (RDA) to a target site in the body while L75-fermented ‘Peach’ MJs could be used to effectively counteract oxidants in the body system. Full article

A milk-alternative produced from lentil protein isolate was fermented with three multifunctional strains of lactic acid bacteria, Leuconostoc citreum TR116, Leuconostoc pseudomesenteroides MP070, and Lacticaseibacillus paracasei FST 6.1. As a control, a commercial starter culture containing Streptococcus thermophilus was used. The metabolic performance of these strains and the techno-functional properties of the resulting yogurt alternatives (YA) were studied. Microbial growth was evaluated by cell counts, acidification, and carbohydrate metabolization. The structure of the YA was investigated by textural and rheological analyses and confocal laser scanning microscopy (CLSM). Production of antifungal compounds, the influence of fermentation on the content of FODMAPs, and typical metabolites were analyzed, and a sensory analysis was performed. The results revealed an exponential microbial growth in the lentil base substrate supported by typical acidification, which indicates a suitable environment for the selected strains. The resulting YA showed a gel-like texture typical for non-stirred yogurts, and high water holding capacity. The tested strains produced much higher levels of antifungal phenolic compounds than the commercial control and are therefore promising candidates as adjunct cultures for shelf-life extension. The Leuconostoc strains produced mannitol from fructose and could thus be applied in sugar-reduced YA. Preliminary sensory analysis showed high acceptance for YA produced with Lacticaseibacillus paracasei FST 6.1, and a yogurt-like flavor not statistically different to that produced by the control. Overall, each tested strain possessed promising functionalities with great potential for application in fermented plant-based dairy-alternatives. Full article

The current environmental challenge is pushing food systems towards more sustainable models of production that require reorganizing of processes by re-using side products still containing nutrients. This work aimed at valorising a mix of bovine sweet whey and sunflower press cake, through targeted fermentation. After preliminary screening based on growth rate, final pH, lactose/galactose assimilation, phytase activity, six Lactic Acid Bacteria strains (Lacticaseibacillus casei, L. paracasei (2), Lactococcus lactis, Lentilactobacillus parakefiri and Leuconostoc pseudomesenteroides) and three yeasts (Kluyveromyces lactis, K. marxianus and Torulaspora delbrueckii) were co-cultivated in pairs in microcosms (1-part ground press cake: 4-parts whey). All tested microorganisms were able to grow and acidify the blend: the LAB counts increased during the incubation (26 °C for 48 h) of +2.80 log CFU/g, whereas yeasts counts were of +1.98 log CFU/g, with significant differences among the different associations (p < 0.01). Mould counts were always <3 log CFU/g. Interestingly, the bacterial contaminants count significantly varied in samples with different pairs of strains (p < 0.001). Acidification level, acetic acid and ethanol contents were the limiting factors affecting the growth of spoilage micro-organisms. Best performances were attained in microcosms inoculated with L. lactis or L. paracasei and K. lactis or K. marxianus. Full article

Due to changes in climatic conditions in Poland interest in viticulture and wine production has considerably boosted. One of the worldwide trends in winemaking is use of indigenous strains of yeast and lactic acid bacteria (LAB). The aim of the study was to analyse the microbial diversity of Polish red wines and their key properties and combine them for better understanding of the processes responsible for creating the sensory attributes. Metagenetic analysis was used to characterise the bacterial and yeast diversity of wines produced by spontaneous fermentation of grapes of the Regent variety, which came from three vineyards: “Dom Bliskowice” (DB), “Małe Dobre” (MD), and “Winnica Janowiec” (WJ). Among bacteria, Tatumella ptyseos was the most abundant species in DB and WJ wines and Leuconostoc pseudomesenteroides was the most abundant in MD wine. Among yeasts, Saccharomyces cerevisiae was found in DB and WJ wines, Saccharomyces cariocanus in MD wine, and Hanseniaspora uvarum in all samples studied. Studied wines had statistically significantly different antioxidant capacities and distinct glucose, fructose, and lactic acid concentrations. The presence of acetic and lactic acid bacteria was positively related to the concentrations of acetic and lactic acid, respectively, while the lack of malic acid was indicative of malolactic fermentation. This knowledge may be useful in the development of unique local starter cultures for the production of wines with specific characteristics. Full article