RESUMEN
Dextransucrases play a crucial role in the production of dextran from economical sucrose; therefore, there is a pressing demand to explore novel dextransucrases with better performance. This study characterized a dextransucrase enzyme, LmDexA, which was identified from the Leuconostoc mesenteroides NN710. This bacterium was isolated from the soil of growing dragon fruit in Guangxi province, China. We successfully constructed six different N-terminal truncated variants through sequential analysis. Additionally, a truncated variant, ΔN190LmDexA, was constructed by removing the 190 amino acids fragment from the N-terminal. This truncated variant was then successfully expressed heterologously in Escherichia coli and purified. The purified ΔN190LmDexA demonstrated optimal hydrolysis activity at a pH of 5.6 and a temperature of 30 °C. Its maximum specific activity was measured to be 126.13 U/mg, with a Km of 13.7 mM. Results demonstrated a significant improvement in the heterologous expression level and total enzyme activity of ΔN190LmDexA. ΔN190LmDexA exhibited both hydrolytic and transsaccharolytic enzymatic activities. When sucrose was used as the substrate, it primarily produced high-molecular-weight dextran (>400 kDa). However, upon the addition of maltose as a receptor, it resulted in the production of a significant amount of oligosaccharides. Our results can provide valuable information for enhancing the characteristics of recombinant dextransucrase and potentially converting sucrose into high-value-added dextran and oligosaccharides.
Asunto(s)
Clonación Molecular , Glucosiltransferasas , Leuconostoc mesenteroides , Glucosiltransferasas/genética , Glucosiltransferasas/metabolismo , Glucosiltransferasas/química , Leuconostoc mesenteroides/enzimología , Leuconostoc mesenteroides/genética , Dextranos/química , Dextranos/biosíntesis , Dextranos/metabolismo , Hidrólisis , Concentración de Iones de Hidrógeno , Escherichia coli/genética , Mutación , Especificidad por Sustrato , Sacarosa/metabolismo , Cinética , TemperaturaRESUMEN
Globally colorectal cancer ranks as the third most widespread disease and the third leading cause of cancer-associated mortality. Immunotherapy treatments like PD-L1 blockade have been used to inhibit the PD-L1 legend, which boosts the activity of cytotoxic T lymphocytes. Recently, studies suggest that some probiotics could potentially enhance the effectiveness of immunotherapy treatments for cancer patients. We found that in Caco-2 and HT-29 cells, the live Leuconostoc mesenteroides treatment resulted an increase in the PD-L1 expression and this treatment stimulated interferon-gamma (IFN-γ) production in Jurkat T-cells. Due to the well-established ability of IFN-γ to enhance PD-L1 expression, the combination of IFN-γ and L. mesenteroides was used in colon cancer cell lines and a resulting remarkable increase of over tenfold in PD-L1 expression was obtained. Interestingly, when L. mesenteroides and IFN-γ are present, the blockage of PD-L1 using PD-L1 antibodies not only improved the viability of Jurkat T-cells but also significantly boosted the levels of IFN-γ and IL-2, the T-cells activation marker cytokines. In addition to upregulating PD-L1, L. mesenteroides also activated Toll-like receptors (TLRs) and NOD-like receptors (NODs) pathways, specifically through TLR2 and NOD2, while also exerting a suppressive effect on autophagy in colon cancer cell lines. In conclusion, our findings demonstrate a significant upregulation of PD-L1 expression in colon cancer cells upon co-culturing with L. mesenteroides. Moreover, the presence of PD-L1 antibodies during co-culturing activates Jurkat T cells. The observed enhancement in PD-L1 expression may be attributed to the inhibition of the Autophagy pathway or activation of the hippo pathway. KEY POINTS: Co-culturing L. mesenteroides increases PD-L1 gene and protein transaction in colon cancer. L. mesenteroides existing enhances T cells viability and activity. GPCR41/42 is a possible link between L. mesenteroides, YAP-1 and PD-L1.
Asunto(s)
Antígeno B7-H1 , Neoplasias del Colon , Interferón gamma , Leuconostoc mesenteroides , Linfocitos T , Humanos , Antígeno B7-H1/metabolismo , Antígeno B7-H1/genética , Células CACO-2/microbiología , Línea Celular Tumoral , Neoplasias del Colon/inmunología , Neoplasias del Colon/microbiología , Células HT29/microbiología , Interferón gamma/metabolismo , Interleucina-2/metabolismo , Células Jurkat , Leuconostoc mesenteroides/metabolismo , Leuconostoc mesenteroides/genética , Activación de Linfocitos , Proteína Adaptadora de Señalización NOD2/metabolismo , Proteína Adaptadora de Señalización NOD2/genética , Probióticos/farmacología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Linfocitos T/microbiología , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 2/genética , Regulación hacia ArribaRESUMEN
Spicy cabbage is a popular fermented vegetable food. The study aimed to determine the physicochemical properties, volatile flavor components, sensory evaluation, and microbial diversity of spicy cabbage prepared using different methods. Three methods were used: single-bacteria fermentation with Lactiplantibacillus plantarum YB-106 and Leuconostoc mesenteroides YB-23, mixed fermentation (LMP) using both strains, and natural fermentation as the blank control (CON). The LMP group has the best quality of spicy cabbage and the highest sensory score. Esters and alkenes were the main volatile flavor components of the spicy cabbage by GC-MS. The fermentation time of LMP group was shorter, and the nitrite degradation rate was >60 %, which was significantly higher than that of other groups (p < 0.05). From the perspective of microbial diversity, the dominant bacteria genera in each group were Lactobacillus, Pantoea, Enterococcus and Pseudomonas. However, mixed fermentation decreased the abundance of pathogenic bacteria, of which the abundance of Serratia was <0.1 %. In conclusion, mixed fermentation can significantly improve the quality of spicy cabbage and shorten the fermentation time. These findings laid the theoretical foundation for the industrial production of high-quality spicy cabbage.
Asunto(s)
Brassica , Fermentación , Alimentos Fermentados , Microbiología de Alimentos , Leuconostoc mesenteroides , Brassica/microbiología , Leuconostoc mesenteroides/metabolismo , Alimentos Fermentados/microbiología , Lactobacillus plantarum/metabolismo , Lactobacillus plantarum/clasificación , Gusto , BiodiversidadRESUMEN
Biocontrol of phytopathogens involving the use of bioactive compounds produced by lactic acid bacteria (LAB), is a promising approach to manage many diseases in agriculture. In this study, a lactic acid bacterium designated YB1 was isolated from fermented olives and selected for its antagonistic activity against Verticillium dahliae (V. dahliae) and Agrobacterium tumefaciens (A. tumefaciens). Based on the 16S rRNA gene nucleotide sequence analysis (1565 pb, accession number: OR714267), the new isolate YB1 bacterium was assigned as Leuconostoc mesenteroides YB1 (OR714267) strain. This bacterium produces an active peptide "bacteriocin" called BacYB1, which was purified in four steps. Matrix-assisted lasers desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) based approach was performed to identify and characterize BacYB1. The exact mass was 5470.75 Da, and the analysis of the N-terminal sequence (VTRASGASTPPGTASPFKTL) of BacYB1 revealed no significant similarity to currently available antimicrobial peptides. The BacYB1 displayed a bactericidal mode of action against A. tumefaciens. The potentiel role of BacYB1 to supress the growth of A. tumefaciens was confirmed by live-dead cells viability assay. In pot experiments, the biocontrol efficacy of BacYB1 against V. dahliae wilt on young olive trees was studied. The percentage of dead plants (PDP) and the final mean symptomes severity (FMS) of plants articifialy infected by V. dahliae and treated with the pre-purified peptide BacYB1 (preventive and curative treatments) were significantly inferior to untreated plants. Biochemical analysis of leaves of the plants has shown that polyophenols contents were highly detected in plants infected by V. dahliae and the highest contents of chlorophyl a, b and total chlorophyll were recorded in plants treated with the combination of BacYB1 with the biofertilisant Humivital. BacYB1 presents a promising alternative for the control of Verticillium wilt and crown gall diseases.
Asunto(s)
Agrobacterium tumefaciens , Bacteriocinas , Leuconostoc mesenteroides , Olea , Enfermedades de las Plantas , ARN Ribosómico 16S , Agrobacterium tumefaciens/metabolismo , Bacteriocinas/farmacología , Bacteriocinas/metabolismo , Olea/microbiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , ARN Ribosómico 16S/genética , Leuconostoc mesenteroides/metabolismo , Leuconostoc mesenteroides/genética , Agentes de Control Biológico/metabolismo , Agentes de Control Biológico/farmacología , Verticillium/efectos de los fármacos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Antibiosis , Filogenia , Antibacterianos/farmacología , Antibacterianos/metabolismoRESUMEN
This study investigated the effects of two distinct probiotics, Leuconostoc mesenteroides B4 (B4) and Bacillus pumilus D5 (D5), along with their combination, on the diet of white shrimp (Litopenaeus vannamei) during an eight-week feeding trial. The diets tested included B4 + dextran at 107 CFU/g feed (the B4 group), D5 alone at 107 CFU/g feed (the D5 group), and a combination of B4 + dextran and D5 at 5 × 106 CFU/g feed each (the B4+dextran + D5 group). Relative to the control group, those administered probiotics exhibited moderate enhancements in growth. By the eighth week, the weight gain for the B4, D5, and B4+D5 groups was 696.50 ± 78.15%, 718.53 ± 130.73%, and 693.05 ± 93.79%, respectively, outperforming the control group's 691.66 ± 31.10% gain. The feed conversion ratio was most efficient in the B4 group (2.16 ± 0.06), closely followed by B4+D5 (2.21 ± 0.03) and D5 (2.22 ± 0.06), with the control group having the highest ratio (2.27 ± 0.03). While phenoloxidase activity was somewhat elevated in the B4 and D5 groups, no significant differences were noted in respiratory burst activity or total hemocyte count across all groups. Challenge tests at weeks 4 and 8 showed that the B4 + D5 combination offered superior protection against AHPND-causing Vibrio parahaemolyticus. The 4-week cumulative survival rate was highest in shrimp treated with B4 + dextran + D5 (56.25%), followed by B4 + dextran (31.25%), control (18.75%), and lowest in D5 (12.5%). By week 8, the B4 + dextran + D5 (43.75%) and B4 + dextran (37.5%) groups significantly outperformed the control group (6.25%, p < 0.05), with no significant difference observed between the D5 group (37.5%) and the control group at day 56. Analysis of the shrimp's foregut microbiota revealed an increase in unique OTUs in the B4 and B4 + D5 groups. Compared to the control, Proteobacteria abundance was reduced in all probiotic groups. Potential pathogens like Vibrio, Bacteroides, Neisseria, Botrytis, Clostridioides, and Deltaentomopoxvirus were detected in the control but were reduced or absent in probiotic groups. Beneficial microbes such as Methanobrevibacter and Dictyostelium in the B4+D5 group, and Sugiyamaella in the B4 group, showed significant increases. Probiotics also led to higher transcript levels of nitric oxide synthase in the hemocytes, and lysozyme and transglutaminase in the midgut, along with lysozyme and α2-macroglobulin in the foregut. Notably, the combined B4 + D5 probiotics synergistically enhanced the expression of superoxide dismutase and prophenoloxidase in the foregut, indicating an improved immune response. In summary, this study demonstrates that the probiotics evaluated, especially when used in combination, significantly boost the expression of specific immune-related genes, enhance the bacterial diversity and richness of the intestine, and thus prevent the colonization and proliferation of Vibrio spp. in L. vannamei.
Asunto(s)
Bacillus , Dictyostelium , Leuconostoc mesenteroides , Penaeidae , Probióticos , Vibrio parahaemolyticus , Animales , Resistencia a la Enfermedad , Muramidasa/metabolismo , Leuconostoc , Dextranos/metabolismo , Vibrio parahaemolyticus/fisiología , Dieta , Inmunidad InnataRESUMEN
Six lactic acid bacteria (LAB) isolated from Algerian sheep's milk, traditional butter, date palm sap and barley, which produce dextran, mannitol, oligosaccharides and vitamin B2 have been characterized. They were identified as Leuconostoc mesenteroides (A4X, Z36P, B12 and O9) and Liquorilactobacillus mali (BR201 and FR123). Their exopolysaccharides synthesized from sucrose by dextransucrase (Dsr) were characterized as dextrans with (1,6)-D-glucopyranose units in the main backbone and branched at positions O-4, O-2 and/or O-3, with D-glucopyranose units in the side chain. A4X was the best dextran producer (4.5 g/L), while the other strains synthesized 2.1-2.7 g/L. Zymograms revealed that L. mali strains have a single Dsr with a molecular weight (Mw) of ~ 145 kDa, while the Lc. mesenteroides possess one or two enzymes with 170-211 kDa Mw. As far as we know, this is the first detection of L. mali Dsr. Analysis of metabolic fluxes from sucrose revealed that the six LAB produced mannitol (~ 12 g/L). The co-addition of maltose-sucrose resulted in the production of panose (up to 37.53 mM), an oligosaccharide known for its prebiotic effect. A4X, Z36P and B12 showed dextranase hydrolytic enzymatic activity and were able to produce another trisaccharide, maltotriose, which is the first instance of a dextranase activity encoded by Lc. mesenteroides strains. Furthermore, B12 and O9 grew in the absence of riboflavin (vitamin B2) and synthesized this vitamin, in a defined medium at the level of ~ 220 µg/L. Therefore, these LAB, especially Lc. mesenteroides B12, are good candidates for the development of new fermented food biofortified with functional compounds.
Asunto(s)
Leuconostoc mesenteroides , Animales , Ovinos , Dextranos/metabolismo , Dextranasa/química , Dextranasa/metabolismo , Manitol/metabolismo , Malí , Glucosiltransferasas/metabolismo , Oligosacáridos/química , Sacarosa/metabolismo , Vitaminas/metabolismo , Leuconostoc/metabolismoRESUMEN
Polyethylene microplastics (PE-MPs) has become a global concern due to their widespread distribution and hazardous properties in aquatic habitats. In this study, the accumulation effect of PE-MPs in the intestine of large-scale loach (Paramisgurnus dabryanus) was explored by adding different concentrations of PE-MPs to the water, the destination of PE-MPs after breaking the intestinal barrier and the effects caused. The collected data showed that PE-MPs accumulation for 21d altered the histomorphology and antioxidant enzyme activity of the intestine, induced dysbiosis of the intestinal flora. 10 mg/L of PE-MPs induced a significant increase in the transcript levels of intestinal immunity factors in loach after 21d of exposure. Moreover, the levels of diamine oxidase (DAO) and d-lactic acid (D-Lac) in the gut and serum of loach were significantly increased after exposure to PE-MPs at all concentrations (1, 5, 10 mg/L). Subsequently, the presence of PE-MPs was detected in the blood, suggesting that the disruption of the intestinal multilayer barrier allowed PE-MPs to spill into the circulation. The accumulation of PE-MPs (1,5,10 mg/L) in the blood led to massive apoptosis and necrosis of blood cells and activated phagocytosis in response to PE-MPs invasion. To alleviate the damage, this study further exposure the effect of probiotics on PE-MPs treated loach by adding Leuconostoc mesenteroides DH (109 CFU/g) to the feed. The results showed that DH significantly increased the intestinal index and reduced the levels of DAO and D-Lac. To investigate the reason, we followed the PE-MPs in the intestine and blood of the loach and found that the number of PE-MPs particles was significantly reduced in the probiotic group, while the PE-MPs content in the feces was elevated. Thus, we concluded that DH reducing the accumulation of PE-MPs in the intestinal by increases fecal PE-MPs, which in turn mitigates the damage to the intestinal barrier caused by PE-MPs, and reduces the amount of PE-MPs in the blood. This work offers a robust analysis to understand the mechanisms of damage to the intestinal barrier by MPs and the fate of MPs after escaping the intestinal barrier and provide a new perspective on the application of probiotics in mitigating PE-MPs toxicity.
Asunto(s)
Cipriniformes , Leuconostoc mesenteroides , Animales , Polietileno , Microplásticos , Plásticos , Antioxidantes , Intestinos , Células Sanguíneas , InmunidadRESUMEN
Yoghurt fermented by Leuconostoc mesenteroides XR1 from Kefir grains was found to produce a unique silk drawing phenomenon. This property was found to be associated with the exopolysaccharides (EPS), X-EPS, produced by strain XR1. In order to better understand the mechanism that produced this phenomenon, the X-EPS was extracted, purified and characterized. The molecular weight and monosaccharide composition were determined by size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) and ion chromatography (IC) analysis, respectively. The results showed that its molecular weight was 4.183 × 106 g/mol and its monosaccharide composition was glucose, and glucuronic acid, with the contents of 567.6148 and 0.2096 µg/mg, respectively. FT-IR and NMR analyses showed that X-EPS was an α-pyranose polysaccharide and was composed of 92.22 % α-(1 â 6) linked d-glucopyranose units and 7.77 % α-(1 â 3) branching. Furthermore, it showed a chain-like microstructure with branches in atomic force microscopy (AFM) and scanning electron microscopy (SEM) experiments. These results suggested that the unique structure of X-EPS, gave the yoghurt a strong viscosity and cohesiveness, which resulted in the silk drawing phenomenon. This work suggested that X-EPS holds the potential for food and industrial applications.
Asunto(s)
Leuconostoc mesenteroides , Leuconostoc/química , Yogur , Espectroscopía Infrarroja por Transformada de Fourier , Polisacáridos Bacterianos/química , MonosacáridosRESUMEN
Excessive alcohol consumption can have serious negative consequences on health, including addiction, liver damage, and other long-term effects. The causes of hangovers include dehydration, alcohol and alcohol metabolite toxicity, and nutrient deficiency due to absorption disorders. Additionally, alcohol consumption can slow reaction times, making it more difficult to rapidly respond to situations that require quick thinking. Exposure to a large amount of ethanol can also negatively affect a person's righting reflex and balance. In this study, we evaluated the potential of lactic acid bacteria (LAB) to alleviate alcohol-induced effects and behavioral responses. Two LAB strains isolated from kimchi, Levilactobacillus brevis WiKim0168 and Leuconostoc mesenteroides WiKim0172, were selected for their ethanol tolerance and potential to alleviate hangover symptoms. Enzyme activity assays for alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) were then conducted to evaluate the role of these bacteria in alcohol metabolism. Through in vitro and in vivo studies, these strains were assessed for their ability to reduce blood alcohol concentrations and protect against alcohol-induced liver damage. The results indicated that these LAB strains possess significant ethanol tolerance and elevate ADH and ALDH activities. LAB administration remarkably reduced blood alcohol levels in rats after excessive alcohol consumption. Moreover, the LAB strains showed hepatoprotective effects and enhanced behavioral outcomes, highlighting their potential as probiotics for counteracting the adverse effects of alcohol consumption. These findings support the development of functional foods incorporating LAB strains that can mediate behavioral improvements following alcohol intake.
Asunto(s)
Alcohol Deshidrogenasa , Aldehído Oxidorreductasas , Etanol , Lactobacillales , Probióticos , Animales , Etanol/metabolismo , Alcohol Deshidrogenasa/metabolismo , Ratas , Masculino , Probióticos/administración & dosificación , Lactobacillales/metabolismo , Nivel de Alcohol en Sangre , Hígado/metabolismo , Hígado/efectos de los fármacos , Administración Oral , Leuconostoc mesenteroides , Aldehído Deshidrogenasa/metabolismo , Levilactobacillus brevis/metabolismo , Ratas Sprague-Dawley , Alimentos Fermentados/microbiologíaRESUMEN
The current study was conducted to statistically compare the SYBR® Green quantitative polymerase chain reaction (qPCR) assay and the conventional plate counting (PC) method to construct growth curves of a cocktail of Weissella viridescens in pure culture under different isothermal storage conditions (4, 8, 14, and 30 °C) and in mixed culture with Leuconostoc mesenteroides at 8 °C. The efficiency and specificity of the qPCR standard curves were confirmed, and both methods were adequate to quantify the growth kinetics of W. viridescens at all isothermal temperatures, demonstrating a good correlation and agreement. The efficiencies of the standard curves varied between 98% and 102%. The SYBR® Green qPCR assay was also able to differentiate the growth curves of W. viridescens and L. mesenteroides in the mixed culture at 8 °C. Additionally, the SYBR® Green qPCR method was considered a faster and more sensitive alternative to construct growth curves under different isothermal conditions and differentiate morphologically similar lactic acid bacteria. Overall, the results suggest that the SYBR® Green qPCR method is a reliable and efficient tool to study microbial growth kinetics in pure and mixed cultures.
Asunto(s)
Lactobacillales , Leuconostoc mesenteroides , Weissella , Lactobacillus , Weissella/genética , Leuconostoc/genéticaRESUMEN
There has been an increasing interest in biocatalysts over the past few decades in order to obtain high efficiency, high yield, and environmentally benign procedures aiming at the manufacture of pharmacologically relevant chemicals. Lactic Acid Bacteria (LAB), a microbial group, can be employed as biocatalysts while performing asymmetric reduction of prochiral ketones. In this study, Leuconostoc mesenteroides N6 was used for the asymmetric bioreduction 1-indanone. And then, a novel and innovative face-centered design-based multi-objective optimization model was used to optimize experimental conditions. Also, the experimental design factors were defined as agitation speed, incubation period, pH, and temperature for optimization to acquire the maximum enantiomeric excess (ee) and conversion rate (cr) values. When using the face-centered design-based multi-objective optimization model, the optimum culture conditions corresponded to 96.34 and 99.42%, ee and cr responses, respectively, were pH = 5.87, incubation temperature = 35 °C, incubation period = 50.88 h, and agitation speed = 152.60 rpm. Notably, the validation experiment under the optimum model conditions confirmed the model results. This study demonstrated the importance of the optimization and the efficiency of the face-centered design-based multi-objective model.
Asunto(s)
Leuconostoc mesenteroides , Cetonas , Lactobacillales/químicaRESUMEN
Microcystis aeruginosa is ubiquitously found in various water bodies and can produce microcystins (MCs), which threaten the health of aquatic animals and human beings. The elimination of excessive M. aeruginosa is beneficial for the protection of the ecosystems and public health. In this regard, algae-lysing bacteria have been extensively studied as an effective measure for their eradication. However, the active substances generated by algae-lysing bacteria are limited. For this study, we reveal that the phenyllactic acid (PLA) produced by Leuconostoc mesenteroides DH exhibits high efficacy for the removal of M. aeruginosa, and explore the elimination mechanism of strain DH on M. aeruginosa. It was found that a cell-free supernatant of strain DH possessed high removal activities against M. aeruginosa. Abundant reactive oxygen species were induced in algal cells following exposure to strain DH supernatant, as well as superoxide dismutase and catalase responses. Furthermore, the integrity of algal cell membranes and photosynthesis was seriously damaged. Interestingly, added exogenous eugenol significantly inhibited the synthesis of active substance produced by strain DH, which further identified that PLA is one of the active substances that contribute to the eradication of M. aeruginosa on the basis of metabolomics analysis. Our finding demonstrated, for the first time, that PLA (as an anti-cyanobacterial compound) can be used for the removal of M. aeruginosa, which provides a theoretical basis for the control of M. aeruginosa.
Asunto(s)
Cianobacterias , Leuconostoc mesenteroides , Microcystis , Animales , Humanos , Microcystis/fisiología , Leuconostoc mesenteroides/metabolismo , Ecosistema , Cianobacterias/metabolismo , Plantas/metabolismo , Microcistinas/metabolismo , PoliésteresRESUMEN
Lactic acid bacteria (LAB) are currently being investigated for their potential use as probiotics and starter cultures. Researchers have developed powdering processes for the commercialization of LAB. Previous studies have focused on identifying innovative cryoprotective agents and freeze-drying (FD) techniques to enhance the stability of LAB. In this study, adaptive laboratory evolution (ALE) was employed to develop a strain with high FD tolerance and enhanced storage stability. Leuconostoc mesenteroids WiKim33 was subjected to heterotypic shock (heat and osmosis shock) to induce the desired phenotype and genotype. An FD-tolerant enhanced Leu. mesenteroides WiKim33 strain (ALE50) was obtained, which harbored a modified fatty acid composition and cell envelope characteristics. Specifically, ALE50 showed a lower unsaturated fatty acid (UFA)/saturated fatty acid (SFA) ratio and a higher cyclic fatty acid (CFA) composition. Moreover, the exopolysaccharide (EPS) thickness increased significantly by 331% compared to that of the wild type (WT). FD tolerance, which was evaluated using viability testing after FD, was enhanced by 33.4%. Overall, we demonstrated the feasibility of ALE to achieve desirable characteristics and provided insights into the mechanisms underlying increased FD tolerance.
Asunto(s)
Lactobacillales , Leuconostoc mesenteroides , Leuconostoc mesenteroides/genética , Leuconostoc mesenteroides/metabolismo , Liofilización/métodos , Ácidos Grasos/metabolismo , Congelación , Lactobacillales/genética , Lactobacillales/metabolismoRESUMEN
BACKGROUND: Lactobacillales including L mesenteroides have beneficial effects on human health, including improvement of psychological status and alleviation of allergic rhinitis. In mice, L mesenteroides subsp. strain NTM048 (NTM048) increased intestinal s-IgA. In humans, however, the effects of NTM048 on s-IgA secretion have been unclear. STUDY: This 16-week trial was performed using a double-blind, placebo-controlled, parallel group design. We aimed to establish whether Leuconostoc mesenteroides subsp. strain NTM048 increases the secretion of s-IgA in saliva. Forty healthy adults and forty patients with Japanese cedar pollinosis were recruited. Participants took either 2 test capsules including NTM048 (1010 CFU/day), or 2 placebo capsules per day, for 16 weeks. They were asked to collect their saliva and answered POMS2, a questionnaire about psychological status. The patients also answered questions about nasal symptoms. Blood samples were collected from the patients with Japanese Cedar pollinosis. Stool samples were collected at the start and on the last day of the trial. RESULTS: All subjects completed the trial. It was conducted during the season when Japanese cedar pollen is most scattered. Serum concentration of Japanese cedar pollen-specific IgE wasâ >â 2.0 UA/mL in patients with Japanese cedar pollinosis. The amount of s-IgA in saliva was not increased by NTM048 in overall subjects, and Japanese cedar pollen-specific IgE was not changed by NTM048 in patients with Japanese cedar pollinosis. The symptom of nasal blockage was improved by NTM048 12 weeks after the start of trial. post hoc analysis indicated a positive correlation between improving psychological status and the increase in occupation ratio of lactobacillus including NTM048. CONCLUSION: The amount of s-IgA in saliva was not increased by NTM048, but nasal blockage was improved by it. Psychological status might be improved if dosage of NTM048 is raised to the degree that NTM048 might be increased in the intestinal tract.
Asunto(s)
Leuconostoc mesenteroides , Obstrucción Nasal , Rinitis Alérgica Estacional , Adulto , Humanos , Animales , Ratones , Rinitis Alérgica Estacional/tratamiento farmacológico , Japón , Inmunoglobulina E , Inmunoglobulina A/uso terapéuticoRESUMEN
The colour of meat typically fades as it decays. However, it has been observed that certain vacuum-packaged spoiled hams can maintain a pink colour even when the packaging is bulged. A large amount of Zinc protoporphyrin IX (ZnPP) was found in these hams, compared to fresh red hams or spoiled and grey hams. Combined with high-throughput sequencing and cultural isolation, the potential cultures of Leuconostoc mesenteroides S-13 (LM), Leuconostoc citreum OCLC11 (LC), and Leuconostoc mesenteroides subsp. IMAU:80679 (LS) were selected based on their ability to produce ZnPP. Subsequently, these cultures were introduced into a fermented sausage model to assess their effect on colour conversion. The analysis of absorption and fluorescent spectra showed that Nitrite sausages contained nitrosyl heme pigment, while bacteria-inoculated sausages were predominantly composed of ZnPP. In addition, the a* value of the LS sausage was close to the Nitrite group at the end of fermentation, significantly higher than control, indicating the effect of bacterial metabolism on the redness. Meanwhile, the Ferrochelatase (FECH) activity of LM, LC and LS groups were 140 ± 13, 113 ± 16 and 201 ± 20 U/g sausage, respectively, providing a potential method on compensating for nitrite/nitrate substitution based on the presence of ZnPP in meat products.
Asunto(s)
Leuconostoc mesenteroides , Carne de Cerdo , Nitritos , Vacio , LeuconostocRESUMEN
The Leuconostoc mesenteroides subsp. IMAU:80679 (LM) was chosen for its superior capability in enhancing redness, and was incubated in a broth system containing metmyoglobin (MetMb) to investigate its mechanisms for color improvement. The a* value of LM group reached its highest level of 52.75 ± 1.04 at 24 h, significantly higher than control of 19.75 ± 0.6 (p < 0.05). The addition of LM could inhibit myoglobin oxidation to some extent. Meanwhile, higher content of nitrosylmyoglobin (NOMb) and Zn-protoporphyrin (Znpp) were observed in LM samples during the whole incubation period. Furthermore, enzymatic activity and encoded genes related to MetMb reduction and pigment formation were determined to explain its possible mechanism on color enhancement. Finally, by extracting crude enzymes and adding them to meat batters, the redness of crude enzyme group was comparable to that achieved with 20 ppm nitrite, providing a potential method on compensating for nitrite/nitrate substitution in meat products.
Asunto(s)
Leuconostoc mesenteroides , Mioglobina , Mioglobina/metabolismo , Leuconostoc mesenteroides/genética , Leuconostoc mesenteroides/metabolismo , Nitritos , Carne , Metamioglobina , Oxidación-Reducción , ColorRESUMEN
2-O-α-D-glucopyranosyl-sn-glycerol (2-αGG) is a high value product with wide applications. Here, an efficient, safe and sustainable bioprocesses for 2-αGG production was designed. A novel sucrose phosphorylase (SPase) was firstly identified from Leuconostoc mesenteroides ATCC 8293. Subsequently, SPase mutations were processed with computer-aided engineering, of which the activity of SPaseK138C was 160% higher than that of the wild-type. Structural analysis revealed that K138C was a key functional residue moderating substrate binding pocket and thus influences catalytic activity. Furthermore, Corynebacterium glutamicum was employed to construct microbial cell factories along with ribosome binding site (RBS) fine-tuning and a two-stage substrate feeding control strategy. The maximum production of 2-αGG by these combined strategies reached 351.8 g·L-1 with 98% conversion rate from 1.4 M sucrose and 3.5 M glycerol in a 5-L bioreactor. This was one of the best performance reported in single-cell biosynthesis of 2-αGG, which paved effective ways for industrial-scale preparation of 2-αGG.
Asunto(s)
Leuconostoc mesenteroides , Leuconostoc mesenteroides/metabolismo , Glicerol , Sacarosa/metabolismo , Biotransformación , Leuconostoc/genética , Leuconostoc/metabolismoRESUMEN
This paper describes the structural elucidation of Leuconostoc mesenteroides P35 exopolysaccharide (EPS-LM). Ln. mesenteroides P35 strain was isolated from a French goat cheese for its capacity to produce EPS increasing the viscosity of a whey-based fermentation medium. The chemical structure of EPS-LM analysis was elucidated by determination of optical rotation degree, macromolecular characterization, sugar units and methylation analyses, FT-IR, 1D NMR spectroscopy (1H and 13C NMR), 2D NMR spectroscopy (1H1H COSY, HSQC and HMBC). EPS-LM was a high molecular weight (ranging from 6.7 × 106 Da to 9.9 × 106 Da) dextran that is composed of only d-glucose units containing α (1 â 6) linkages and paltry α (1 â 3) branches. Since polysaccharide-protein interactions can be exploited to control and design food matrices, EPS-LM interactions with bovine serum albumin (the main constituent of bovine plasma) were investigated by surface plasmon resonance (SPR). Kinetic properties of EPS-LM binding with immobilized BSA via showed an increase of EPS-LM affinity (equilibrium constant (Kd)) for BSA from (2.50 ± 0.01) × 10-5 M-1 at 298 K to (9.21 ± 0.05) × 10-6 M-1 at to 310 K. The thermodynamic parameters revealed that van der Waals and hydrogen binding forces play a major role in the interaction of EPS-LM with BSA. However, EPS-LM-BSA interaction was non-spontaneous, entropy driven and an EPS-LM - BSA binding process was endothermic (ΔG > 0). The structural findings suggested that Ln. mesenteroides P35 α-D-glucan might find widespread technological applications in the biopolymer, medical and food industries.
Asunto(s)
Leuconostoc mesenteroides , Resonancia por Plasmón de Superficie , Leuconostoc mesenteroides/metabolismo , Albúmina Sérica Bovina/química , Espectroscopía Infrarroja por Transformada de Fourier , Termodinámica , Leuconostoc/metabolismoRESUMEN
Lactic acid bacteria (LAB) are being used for probiotic and starter cultures to prevent global damage to microbial cells. To retain the benefits of LAB in the commercially used powdered form, highly efficient cryoprotective agents are required during the manufacturing process. This study suggests a novel cryoprotective agent derived from Jerusalem artichoke (JA; Helianthus tuberous L.) and describes the mechanism of cryoprotective effect improvement by sonication treatment. The cryoprotective effect of JA extract was verified by examining the viability of Leuconostoc mesenteroides WiKim33 after freeze-drying (FD). Sonication of JA extract improved the cryoprotective effect. Sonication reduced fructose and glucose contents, which increased the induction of critical damage during FD by 15.84% and 46.81%, respectively. The cryoprotective effects of JA and sonication-treated JA extracts were determined using the viable cell count of Leu. mesenteroides WiKim33. Immediately after FD and storage for 24 weeks, the viability of Leu. mesenteroides WiKim33 with JA extract was 82.8% and 76.3%, respectively, while that of the sonication-treated JA extract was 95.2% and 88.8%, respectively. Our results show that reduction in specific monosaccharides was correlated with improved cryoprotective effect. This study adopted sonication as a novel treatment for improving the cryoprotective effect and verified its efficiency.
Asunto(s)
Helianthus , Lactobacillales , Leuconostoc mesenteroides , Crioprotectores , Helianthus/química , Monosacáridos , Extractos Vegetales/farmacologíaRESUMEN
Leuconostoc spp are lactic acid-producing bacteria closely related to the Streptococcus family. While usually associated with the fermentation of dairy products and sauerkraut, they are rarely associated with human infections. This is a case report of an immunocompetent patient found to have L. mesenteroides bacteremia associated with raw milk and poor dentition as the likely source of infection, which was treated successfully with daptomycin as this genus is intrinsically resistant to vancomycin.