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1.
World J Microbiol Biotechnol ; 40(4): 114, 2024 Feb 29.
Artigo em Inglês | MEDLINE | ID: mdl-38418710

RESUMO

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.


Assuntos
Leuconostoc mesenteroides , Animais , Ovinos , Dextranos/metabolismo , Dextranase/química , Dextranase/metabolismo , Manitol/metabolismo , Mali , Glucosiltransferases/metabolismo , Oligossacarídeos/química , Sacarose/metabolismo , Vitaminas/metabolismo , Leuconostoc/metabolismo
2.
Food Microbiol ; 115: 104337, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37567639

RESUMO

Leuconostoc spp. is often regarded as the flavor producer, responsible for the production of acetoin and diacetyl in dairy cheese. In this study, we investigate seven plant-derived Leuconostoc strains, covering four species, in their potential as a lyophilized starter culture for flavor production in fermented soy-based cheese alternatives. We show that the process of lyophilization of Leuconostoc can be feasible using a soy-based lyoprotectant, with survivability up to 63% during long term storage. Furthermore, the storage in this media improves the subsequent growth in a soy-based substrate in a strain specific manner. The utilization of individual raffinose family oligosaccharides was strain dependent, with Leuconostoc pseudomesenteroides NFICC99 being the best consumer. Furthermore, we show that all investigated strains were able to produce a range of volatile flavor compounds found in dairy cheese products, as well as remove certain dairy off-flavors from the soy-based substrate like hexanal and 2-pentylfuran. Also here, NFICC99 was strain producing most cheese-related volatile flavor compounds, followed by Leuconostoc mesenteroides NFICC319. These findings provide initial insights into the development of Leuconostoc as a potential starter culture for plant-based dairy alternatives, as well as a promising approach for generation of stable, lyophilized cultures.


Assuntos
Laticínios , Leuconostoc , Fermentação , Leuconostoc/metabolismo , Concentração de Íons de Hidrogênio , Açúcares/metabolismo
3.
Int J Biol Macromol ; 246: 125599, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37385309

RESUMO

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.


Assuntos
Leuconostoc mesenteroides , Ressonância de Plasmônio de Superfície , Leuconostoc mesenteroides/metabolismo , Soroalbumina Bovina/química , Espectroscopia de Infravermelho com Transformada de Fourier , Termodinâmica , Leuconostoc/metabolismo
4.
Bioresour Technol ; 385: 129399, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37380039

RESUMO

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.


Assuntos
Leuconostoc mesenteroides , Leuconostoc mesenteroides/metabolismo , Glicerol , Sacarose/metabolismo , Biotransformação , Leuconostoc/genética , Leuconostoc/metabolismo
5.
Int J Mol Sci ; 24(8)2023 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-37108530

RESUMO

Probiotics provide a range of health benefits. Several studies have shown that using probiotics in obesity treatment can reduce bodyweight. However, such treatments are still restricted. Leuconostoc citreum, an epiphytic bacterium, is widely used in a variety of biological applications. However, few studies have investigated the role of Leuconostoc spp. in adipocyte differentiation and its molecular mechanisms. Therefore, the objective of this study was to determine the effects of cell-free metabolites of L. citreum (LSC) on adipogenesis, lipogenesis, and lipolysis in 3T3-L1 adipocytes. The results showed that LSC treatment reduced the accumulation of lipid droplets and expression levels of CCAAT/ enhancer-binding protein-α & ß (C/EBP-α & ß), peroxisome proliferator-activated receptor-γ (PPAR-γ), serum regulatory binding protein-1c (SREBP-1c), adipocyte fatty acid binding protein (aP2), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), resistin, pp38MAPK, and pErk 44/42. However, compared to control cells, adiponectin, an insulin sensitizer, was elevated in adipocytes treated with LSC. In addition, LSC treatment increased lipolysis by increasing pAMPK-α and suppressing FAS, ACC, and PPAR-γ expression, similarly to the effects of AICAR, an AMPK agonist. In conclusion, L. citreum is a novel probiotic strain that can be used to treat obesity and its associated metabolic disorders.


Assuntos
Adipogenia , Lipogênese , Camundongos , Animais , Proteínas Quinases Ativadas por AMP/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Diferenciação Celular , Transdução de Sinais , Obesidade , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Leuconostoc/metabolismo , Células 3T3-L1 , PPAR gama/metabolismo
6.
J Sci Food Agric ; 103(7): 3521-3530, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36799142

RESUMO

BACKGROUND: In industrial production of suancai, baijiu is commonly used to inhibit the spoilage bacteria and enhance the flavor. However, the effects of baijiu on the microbial diversity and metabolic pathways of suancai are rarely reported in the literature. This study aimed to explore the microbial community, its predicted functional roles, and the metabolites formed during fermentation of Chinese Dongbei suancai fermented using a mixed starter with Chinese baijiu as supplementary material. RESULTS: Results showed that Lactobacillus, Enterobacter, and Leuconostoc were the major bacterial genera in the Dongbei suancai fermented by adding baijiu. Linear discriminant analysis effect size indicated that Leuconostoc was the major biomarker in the early stage of fermentation, whereas Lactococcus, Weissella, and Lactobacillus plantarum were biomarkers in the middle and later stages of fermentation. A total of 638 metabolites were detected in suancai fermented by adding baijiu. However, the principal component analysis showed that baijiu significantly affected the metabolites of suancai in the early and later stages of fermentation. Furthermore, 58, 22, and 26 significantly differential metabolites (P < 0.01) were found on day 0, day 2, and day 30 of fermentation respectively. Moreover, Lactobacillus, Lactococcus, and Enterobacter had positive correlations with amino acids, nucleotides, organic acids, alcohols, and esters. Functional analysis implied that carbohydrate, amino acid, energy, and nucleotide metabolism were the major determinants of the characteristics of suancai fermented with baijiu as supplementary material. CONCLUSION: Baijiu changed the metabolites of inoculated fermented Dongbei suancai. © 2023 Society of Chemical Industry.


Assuntos
Lactobacillus plantarum , Microbiota , Fermentação , Bactérias , Lactobacillus/metabolismo , Leuconostoc/metabolismo
7.
Probiotics Antimicrob Proteins ; 15(5): 1447-1463, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-36227534

RESUMO

This study aimed to select and characterize lactic acid bacteria (LAB) with potential antifungal activities against the filamentous fungi Alternaria alternata ATCC MYA-4642, Aspergillus flavus KACC 45470, Aspergillus niger KACC 42589, Cladosporium sphaerospermum ATCC MYA-4645, Penicillium chrysogenum ATCC MYA-4644, and Penicillium expansum KACC 40815. Initial screening of the antifungal activity has identified six LAB strains belonging to the genera Enterococcus and Leuconostoc, selected by their antagonistic activities against at least three of the filamentous fungi in the test panel. Preliminary prediction of bioactive compounds was carried out to narrow down the possible identity of the antagonistic metabolites produced by the studied LAB. Furthermore, metabolic profiles were assessed and used as a basis for the identification of key metabolites based on VIP scores and PCA plot scores. Key metabolites were identified to be ß-phenyllactic acid, ⍺-hydroxyisobutyric acid, 1,3-butanediol, phenethylamine, and benzoic acid. Individual assessment of each metabolic compound against the test panel showed specificity inhibitory patterns; yet, combinations between them only showed additive, but not synergetic effects. The pH neutralization significantly reduced the antifungal activity of the cell-free supernatant (CFS), but no bioactive compounds were found to be stable in high temperatures and pressure. This study will be beneficial as an additional building block on the existing knowledge and future antifungal application of LAB produced metabolites. Furthermore, this study also provides a new bio-preservative perspective on unexplored antifungal metabolites produced by LAB as biocontrol agents.


Assuntos
Antifúngicos , Lactobacillales , Antifúngicos/química , Lactobacillales/metabolismo , Aspergillus niger/metabolismo , Leuconostoc/metabolismo , Aspergillus flavus/metabolismo
8.
Food Res Int ; 160: 111622, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36076372

RESUMO

Paocai is a traditional Chinese fermented vegetable product popular in Asian countries. As an important additive, salt concentration is closely related to the quality of paocai. The aim of this study was to investigate the effect of salt concentration on the physicochemical characteristics, microbial diversity, and flavor profiles of spontaneously fermented radish, and the cross-correlation between microorganisms and flavor compounds was also revealed. Analysis of the microbial diversity of paocai showed that Firmicutes, Proteobacteria, and Ascomycota were detected as the main phyla with different salt concentrations, Weissella and Lactobacillus were the predominant bacterial genera, and Yarrowia dominated the fungal genera. Based on LEfSe analysis, Lactobacillus, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Microbacterium, Lactococcus, Staphylococcus, and Weissella were regarded as differential genera caused by differences in salinity. Analysis of the flavor compounds showed that 17 free amino acids, 5 isothiocyanates, 3 terpenes, 15 sulphur-containing compounds, 16 esters, 8 organic acids, 9 aldehydes, 8 ketones, 25 alcohols, 7 nitriles, 2 lactones, and 10 hydrocarbons were detected. Then, the correlation between the microbial community and flavor compounds was revealed, and the results indicated that several bacterial genera significantly correlated with flavors, including Lactobacillus, Kosakonia, Weissella, Leuconostoc, and Staphylococcus, while fungi had weak correlations with flavors. In addition, Metacyc pathway analysis was carried out to elucidate the effect of salt content on the metabolic pathways, showing that most flavor-related pathways were up-regulated with the increase in salt content. Results presented in this study may contribute to further understanding the role of salt in the fermentation of paocai and provide effective references for quality control of traditional fermented vegetables.


Assuntos
Microbiota , Raphanus , Weissella , Bactérias , Fermentação , Lactobacillus/metabolismo , Leuconostoc/metabolismo , Cloreto de Sódio na Dieta/metabolismo
9.
Chem Pharm Bull (Tokyo) ; 70(2): 155-161, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35110436

RESUMO

Exopolysaccharides (EPSs) occur widely in natural products made by bacteria, fungi and algae. Some EPSs have intriguing biological properties such as anticancer and immunomodulatory activities. Our group has recently found that EPSs generated from Leuconostoc mesenteroides ssp. mesenteroides strain NTM048 (NTM048 EPS) enhanced a production of mucosal immunoglobulin A (IgA) of mouse. Herein, we described the synthesis and evaluation of the tetrasaccharide fragments of NTM048 EPS to obtain information about the molecular mechanism responsible for the IgA-inducing activity.


Assuntos
Produtos Biológicos/síntese química , Produtos Biológicos/metabolismo , Leuconostoc/química , Polissacarídeos/biossíntese , Polissacarídeos/síntese química , Produtos Biológicos/química , Configuração de Carboidratos , Leuconostoc/metabolismo , Polissacarídeos/química
10.
Prep Biochem Biotechnol ; 52(3): 245-252, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34092177

RESUMO

Leuconostoc pseudomesenteroides belongs to a group of lactic acid bacteria normally isolated from fruits, which has the capacity to produce exopolysaccharides (EPS). The present study aimed to optimize the EPS production of L. pseudomesenteroides JF17, isolated from juçara fruits (palm trees threatened with extinction in the Atlantic Forest), using the response surface methodology (RSM), besides evaluating the fermentation kinetics. The maximum production of EPS 53.77 mg/mL was obtained under ideal conditions of MRS broth supplemented with sucrose at 18%, w/v, fermentation temperature of 20 °C and initial pH of 7.30. The Luedeking-Piret model suggested that the production of EPS by the JF17 strain appeared to be associated with the cell growth of the microorganism, in addition to having high efficiency in the production of the polysaccharide from the substrate (Yp/s = 17.85 ± 0.74 mg EPS/log CFU ). Thus, the ideal optimization conditions and kinetic parameters can be useful for increasing the scale up of the fermentation process in the industrial production of EPS by L. pseudomesenteroides JF17.


Assuntos
Leuconostoc/metabolismo , Polissacarídeos/biossíntese , Fermentação , Florestas , Concentração de Íons de Hidrogênio , Temperatura
11.
J Sci Food Agric ; 102(5): 2023-2031, 2022 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-34558071

RESUMO

BACKGROUND: Type 1 diabetes is an autoimmune disease that results in the specific destruction of insulin-producing beta cells in the pancreas. The aim of this study was to investigate the mechanism of exopolysaccharide from Leuconostoc pseudomesenteroides XG5 (XG5 EPS) against type 1 diabetes. RESULTS: Long-term drench of XG5 EPS delayed the onset of autoimmune diabetes and had fewer islets with high-grade infiltration (an insulitis score of 3 or 4) than untreated NOD mice. Oral administration of 50 mg kg-1  d-1 XG5 EPS increased the insulin and glucagon-like peptide-1 (GLP-1) levels of serum, stimulated GLP-1 secretion and upregulated gcg mRNA expression of colon in NOD mice. Moreover, oral administration of 50 mg kg-1  d-1 XG5 EPS significantly increased total short-chain fatty acids levels in the colon contents, especially those of acetic acid and butyric acid. In NCI-H716 cells, 500 and 1000 µmol L-1 sodium butyrate promoted the secretion of GLP-1 and upregulated the mRNA expression of gcg and PC3, while XG5 EPS and sodium acetate did not stimulate the GLP-1 secretion. Therefore, long-term drench of XG5 EPS delayed the onset of autoimmune diabetes, which may be directly correlated with the increase of butyrate in the colon of NOD mice. CONCLUSION: Long-term drench of 50 mg kg-1  d-1 XG5 EPS promoted the expression and secretion of GLP-1 by increasing the production of butyric acid, thereby delaying T1D onset in NOD mice. © 2021 Society of Chemical Industry.


Assuntos
Diabetes Mellitus Tipo 1 , Peptídeo 1 Semelhante ao Glucagon , Animais , Diabetes Mellitus Tipo 1/prevenção & controle , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Insulina/metabolismo , Leuconostoc/metabolismo , Camundongos , Camundongos Endogâmicos NOD
12.
Int J Biol Macromol ; 187: 690-698, 2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34343579

RESUMO

Fermentation of Lactic Acid Bacteria (LAB) is considered to be a sustainable approach for polysaccharide production. Herein, exopolysaccharide (EPS)-producing LAB strain KM01 was isolated from Thai fermented dessert, Khao Mak, which was then identified as Leuconostoc holzapfelii. High-performance anion-exchange chromatography, nuclear magnetic resonance spectroscopy and Fourier-transform infrared spectroscopy suggested that the KM01 EPS comprises α-1,6-linked glucosides. The molecular weight of KM01 EPS was around 500 kDa, but it can form large aggregates formation (MW > 2000 kDa) in an aqueous solution, judged by transmission electron microscopy and dynamic light scattering to be around 150 nm in size. Furthermore, this KM01 EPS form highly viscous hydrogels at concentrations above 5% (w/v). The formation of hydrogels and nanoparticle of KM01 EPS was found to be reversible. Finally, the suitability of KM01 EPS for biomedical applications was demonstrated by its lack of cytotoxicity and its ability to form complexes with quercetin. Unlike the common α-1,6-linked dextran, KM01 EPS can enhance the solubility of quercetin significantly.


Assuntos
Excipientes/química , Glucanos/química , Leuconostoc/metabolismo , Nanopartículas , Polissacarídeos Bacterianos/química , Quercetina/química , Sacarose/metabolismo , Composição de Medicamentos , Excipientes/isolamento & purificação , Excipientes/toxicidade , Fermentação , Glucanos/isolamento & purificação , Glucanos/toxicidade , Hidrogéis , Peso Molecular , Polissacarídeos Bacterianos/isolamento & purificação , Polissacarídeos Bacterianos/toxicidade , Solubilidade , Viscosidade
13.
J Microbiol Biotechnol ; 31(8): 1144-1153, 2021 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-34226411

RESUMO

A released exopolysaccharide (rEPS)-producing strain (LM187) with good acid resistance, bile salt resistance, and cholesterol-lowering properties was isolated from Sichuan paocai and identified as Leuconostoc mesenteroides subsp. mesenteroides. The purified rEPS, designated as rEPS414, had a uniform molecular weight of 7.757 × 105 Da. Analysis of the monosaccharide composition revealed that the molecule was mainly composed of glucose. The Fourier transform-infrared spectrum showed that rEPS414 contained both α-type and ß-type glycosidic bonds. 1H and 13C nuclear magnetic resonance spectra analysis showed that the purified rEPS contained arabinose, galactose, and rhamnose, but less uronic acid. Scanning electron microscopy demonstrated that the exopolysaccharide displayed a large number of scattered, fluffy, porous cellular network flake structures. In addition, rEPS414 exhibited strong in vitro antioxidant activity. These results showed that strain LM187 and its rEPS are promising probiotics with broad prospects in industry.


Assuntos
Antioxidantes/farmacologia , Leuconostoc/metabolismo , Polissacarídeos Bacterianos/farmacologia , Probióticos , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/metabolismo , Alimentos Fermentados/microbiologia , Leuconostoc/isolamento & purificação , Peso Molecular , Monossacarídeos/química , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/isolamento & purificação , Polissacarídeos Bacterianos/metabolismo
14.
Int J Food Microbiol ; 348: 109208, 2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-33940536

RESUMO

Microbiological spoilage of meat is considered as a process which involves mainly bacterial metabolism leading to degradation of meat sensory qualities. Studying spoilage requires the collection of different types of experimental data encompassing microbiological, physicochemical and sensorial measurements. Within this framework, the objective herein was to carry out a multiblock path modelling workflow to decipher causality relationships between different types of spoilage-related responses: composition of microbiota, volatilome and off-odour profiles. Analyses were performed with the Path-ComDim approach on a large-scale dataset collected on fresh turkey sausages. This approach enabled to quantify the importance of causality relationships determined a priori between each type of responses as well as to identify important responses involved in spoilage, then to validate causality assumptions. Results were very promising: the data integration confirmed and quantified the causality between data blocks, exhibiting the dynamical nature of spoilage, mainly characterized by the evolution of off-odour profiles caused by the production of volatile organic compounds such as ethanol or ethyl acetate. This production was possibly associated with several bacterial species like Lactococcus piscium, Leuconostoc gelidum, Psychrobacter sp. or Latilactobacillus fuchuensis. Likewise, the production of acetoin and diacetyl in meat spoilage was highlighted. The Path-ComDim approach illustrated here with meat spoilage can be applied to other large-scale and heterogeneous datasets associated with pathway scenarios and represents a promising key tool for deciphering causality in complex biological phenomena.


Assuntos
Bactérias/metabolismo , Produtos da Carne/microbiologia , Carne/microbiologia , Compostos Orgânicos Voláteis/análise , Animais , Bactérias/classificação , Microbiologia de Alimentos , Embalagem de Alimentos , Lactococcus/metabolismo , Leuconostoc/metabolismo , Microbiota , Odorantes/análise , Psychrobacter/metabolismo , Perus/microbiologia
15.
Sci Rep ; 11(1): 7662, 2021 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-33828213

RESUMO

Nicotinamide mononucleotide (NMN), an intermediate in nicotinamide adenine dinucleotide biosynthesis, is recently attracting much attention for its pharmacological and anti-aging efficacies. However, current commercial products containing NMN are very high-priced because efficient and facile methods for industrial NMN production are limited. In this study, aiming for its nutraceutical application, we attempted to screen lactic acid bacteria for intracellular and/or extracellular NMN production. Using a bioassay system with an auxotrophic yeast that requires nicotinamide riboside (NR; dephosphorylated NMN), three candidates were obtained from a library of 174 strains of facultative anaerobic lactic acid bacteria. All three candidates belonged to the genus Fructobacillus and produced NR in the culture media (0.8-1.5 mg/l). Lactic acid bacteria of the genus Fructobacillus are known to use D-fructose as an electron acceptor in anaerobic lactic acid fermentation; addition of D-fructose to the medium caused intracellular accumulation of NMN and NR, but no extracellular production of these compounds was observed. Draft genome sequencing for one of the candidates suggested that nicotinamide phosphoribosyltransferase, which exists commonly in mammals but is less reported in microorganisms, is a key enzyme for NMN and NR production in the fructophilic bacteria.


Assuntos
Leuconostoc/metabolismo , Mononucleotídeo de Nicotinamida/biossíntese , Escherichia coli , Frutose/metabolismo , Lactobacillales/metabolismo , Leuconostoc/genética , Niacinamida/análogos & derivados , Niacinamida/biossíntese , Nicotinamida Fosforribosiltransferase/metabolismo , Compostos de Piridínio
16.
BMC Biotechnol ; 21(1): 14, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33541325

RESUMO

BACKGROUND: Levan is a well-known homopolymer of fructose composed predominantly of ß-(2, 6) fructofuranosyl linkages in the backbone with occasional ß-(2, 1) linkages in the branch chains with varied applications. However, high production cost due to low yield of microbial levan has become a bottleneck for its practical applications. Furthermore, factors affecting the molecular mass of the synthesized levan by Leuconostoc spp. during prolonged cultivation is not fully elucidated. METHODS: The cultivation condition for Leuconostoc citreum BD1707 to synthesize levan was optimized by single-factor experiments and subsequently with response surface methodology (RSM). The average molecular weight (Mw) of levan synthesized by the strain L.citreum BD1707 under the optimized cultivation conditions was monitored by high-performance size exclusion chromatography (HPSEC). Finally, the enzyme with levan-degrading activity was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The levan yield of BD1707 reached 34.86 g/L with a corresponding productivity of 7.47 g/L/d under the optimal cultivation conditions deduced by RSM, i.e., cultivation at 26 °C and 200 rpm for 112 h in tomato juice supplemented with 172 g/L sucrose with an initial pH value of 6.12. The Mw of levan reached a peak value of 2.320 × 107 Da at 6 h of cultivation under the optimized cultivation conditions and then gradually decreased to 8.809 × 106 Da after 120 h of cultivation. CONCLUSION: The levan yield of the strain L.citreum BD1707 could be sufficiently enhanced via cultivation condition optimization. The decrease in molecular mass of the synthesized levan was attributed predominantly to the hydrolytic activity of levansucrase secreted by L.citreum BD1707 during cultivation, with an estimated Mw of 130 KD by SDS-PAGE, while the effect of acid hydrolysis could be nearly neglected.


Assuntos
Frutanos/química , Frutanos/metabolismo , Leuconostoc/genética , Leuconostoc/metabolismo , Frutanos/genética , Frutose/metabolismo , Glucose , Hexosiltransferases/genética , Hexosiltransferases/metabolismo , Concentração de Íons de Hidrogênio , Solanum lycopersicum , Peso Molecular , Sacarose/metabolismo , Temperatura
17.
Microb Cell Fact ; 20(1): 23, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33482833

RESUMO

BACKGROUND: Lactic acid bacteria can synthesize dextran and oligosaccharides with different functionality, depending on the strain and fermentation conditions. As natural structure-forming agent, dextran has proven useful as food additive, improving the properties of several raw materials with poor technological quality, such as cereal by-products, fiber-and protein-rich matrices, enabling their use in food applications. In this study, we assessed dextran biosynthesis in situ during fermentation of brewers´ spent grain (BSG), the main by-product of beer brewing industry, with Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16. The starters performance and the primary metabolites formed during 24 h of fermentation with and without 4% sucrose (w/w) were followed. RESULTS: The starters showed similar growth and acidification kinetics, but different sugar utilization, especially in presence of sucrose. Viscosity increase in fermented BSG containing sucrose occurred first after 10 h, and it kept increasing until 24 h concomitantly with dextran formation. Dextran content after 24 h was approximately 1% on the total weight of the BSG. Oligosaccharides with different degree of polymerization were formed together with dextran from 10 to 24 h. Three dextransucrase genes were identified in L. pseudomesenteroides DSM20193, one of which was significantly upregulated and remained active throughout the fermentation time. One dextransucrase gene was identified in W. confusa A16 also showing a typical induction profile, with highest upregulation at 10 h. CONCLUSIONS: Selected lactic acid bacteria starters produced significant amount of dextran in brewers' spent grain while forming oligosaccharides with different degree of polymerization. Putative dextransucrase genes identified in the starters showed a typical induction profile. Formation of dextran and oligosaccharides in BSG during lactic acid bacteria fermentation can be tailored to achieve specific technological properties of this raw material, contributing to its reintegration into the food chain.


Assuntos
Dextranos/biossíntese , Grão Comestível/metabolismo , Fermentação , Leuconostoc/metabolismo , Weissella/metabolismo , Cerveja , Regulação Enzimológica da Expressão Gênica , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Concentração de Íons de Hidrogênio , Lactobacillales/genética , Lactobacillales/metabolismo , Leuconostoc/genética , Leuconostoc/crescimento & desenvolvimento , Manitol/metabolismo , Oligossacarídeos/metabolismo , Polissacarídeos/metabolismo , Sacarose/metabolismo , Viscosidade , Weissella/genética , Weissella/crescimento & desenvolvimento
18.
Carbohydr Polym ; 256: 117523, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33483044

RESUMO

EPS-BMS, is to our knowledge, the first high molecular weight exopolysaccharide from potential probiotic Leuconostoc citreum-BMS strain that consists on a mixture of α-(1,6)-dextran branched at the third position and ß-(2,6)-levan. This sample exhibited interesting rheological and emulsifying properties under different conditions. Steady shear experiments proved that EPS-BMS had a pseudoplastic behavior without thixotropic properties. Interestingly, pseudoplasticity was maintained even under stress conditions of temperature, pH and salts, which could provide some sensory properties for food products such as mouth feel. Dynamic oscillatory measurements reflected a liquid-like behavior of the sample regardless of the studied EPS concentration, pH, temperature and ionic force. Results related to the emulsifying as well as interfacial properties showed that EPS-BMS had great potential to be applied as emulsifier and/or emulsion stabilizer in both neutral and acidic conditions. Based on the properties reported in this work, EPS-BMS could be potentially applied in the food industry.


Assuntos
Dextranos/química , Emulsificantes/química , Frutanos/química , Leuconostoc/química , Polissacarídeos Bacterianos/química , Probióticos/química , Culinária/métodos , Dextranos/isolamento & purificação , Emulsificantes/isolamento & purificação , Frutanos/isolamento & purificação , Humanos , Concentração de Íons de Hidrogênio , Leuconostoc/metabolismo , Polissacarídeos Bacterianos/isolamento & purificação , Probióticos/metabolismo , Reologia , Sais/química , Percepção Gustatória/fisiologia , Temperatura
19.
Carbohydr Polym ; 253: 117254, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33279004

RESUMO

High molecular weight dextrans improve the rheological properties of fermented products and have immunomodulatory and antiviral activity. We report on 5.84 × 107-2.61 × 108 Da dextrans produced by Leuconostoc lactis AV1n, Weissella cibaria AV2ou and Weissella confusa V30 and FS54 strains. Dextransucrases catalyze dextran synthesis by sucrose hydrolysis concomitant with fructose generation. The four bacteria have dextransucrases with molecular weight of about 160 kDa detected by zymograms. Each bacterium showed different interplay of dextran production and metabolic fluxes. All bacteria produced lactate, and AV2ou apart, synthesized mannitol from fructose. FS54 hydrolyzed dextran blue and the concentration of dextran produced by this bacterium decreased during the stationary phase. The AV1n binding to Caco-2 cells and polystyrene plates was higher under conditions for dextran synthesis. Thus, this is the first instance of a Weissella dextranase, associated with a dextransucrase ability, and of a positive influence of dextran on adhesion and aggregation properties of a bacterium.


Assuntos
Dextranos/química , Leuconostoc/metabolismo , Weissella/enzimologia , Weissella/metabolismo , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Células CACO-2 , Comunicação Celular/efeitos dos fármacos , Dextranase/metabolismo , Dextranos/farmacologia , Enterócitos/metabolismo , Fermentação , Glucosiltransferases/metabolismo , Humanos , Peso Molecular , Sacarose/metabolismo
20.
FEMS Microbiol Lett ; 367(20)2020 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-33107908

RESUMO

Synthesis of polysaccharides by Leuconostoc can result in improved texture of fermented products. A total of 249 Leuconostoc strains were screened for homo-polysaccharide production and for texturing capabilities in milk. A total of six Ln. mesenteroides strains with superior texturing properties had the genetic blueprint for both homo- (HoPS) and hetero-polysaccharide (HePS) synthesis. Only one strain produced texture in milk without added sucrose, suggesting HePS synthesis via the Wzy dependent pathway. In milk acidification experiments with added sucrose, all six strains depleted the sucrose and released fructose. Thus, they can be used for both texture and possibly also for sweetness enhancement.


Assuntos
Microbiologia de Alimentos/métodos , Leuconostoc/metabolismo , Polissacarídeos/biossíntese , Animais , Leite/microbiologia , Polissacarídeos/metabolismo , Sacarose/metabolismo
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