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1.
J Agric Food Chem ; 72(27): 15198-15212, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38941263

RESUMO

Numerous studies have highlighted the potential of Lactic acid bacteria (LAB) fermentation of whey proteins for alleviating allergies. Nonetheless, the impact of LAB-derived metabolites on whey proteins antigenicity during fermentation remains uncertain. Our objective was to elucidate the impact of small molecular metabolites on the antigenicity of α-lactalbumin (α-LA) and ß-lactoglobulin (ß-LG). Through metabolomic analysis, we picked 13 bioactive small molecule metabolites from Lactobacillus delbrueckii subsp. bulgaricus DLPU F-36 for coincubation with α-LA and ß-LG, respectively. The outcomes revealed that valine, arginine, benzoic acid, 2-keto butyric acid, and glutaric acid significantly diminished the sensitization potential of α-LA and ß-LG, respectively. Moreover, chromatographic analyses unveiled the varying influence of small molecular metabolites on the structure of α-LA and ß-LG, respectively. Notably, molecular docking underscored that the primary active sites of α-LA and ß-LG involved in protein binding to IgE antibodies aligned with the interaction sites of small molecular metabolites. In essence, LAB-produced metabolites wield a substantial influence on the antigenic properties of whey proteins.


Assuntos
Lactobacillus delbrueckii , Simulação de Acoplamento Molecular , Proteínas do Soro do Leite , Lactobacillus delbrueckii/metabolismo , Lactobacillus delbrueckii/química , Lactobacillus delbrueckii/imunologia , Proteínas do Soro do Leite/química , Proteínas do Soro do Leite/metabolismo , Fermentação , Lactoglobulinas/química , Lactoglobulinas/imunologia , Lactoglobulinas/metabolismo , Lactalbumina/química , Lactalbumina/imunologia , Lactalbumina/metabolismo , Animais , Bovinos , Antígenos/imunologia , Antígenos/química
2.
Biotechnol Appl Biochem ; 68(6): 1421-1431, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33125785

RESUMO

Probiotics are nonpathogenic bacterial strains that exert beneficial effects on the host. Previous studies have shown that topical use of some strains of probiotic bacteria have good effects on the healing of cutaneous wounds. In the current study, the wound healing potentials of bacterial probiotics on diabetic cutaneous wounds were evaluated. The effects of probiotics on migration, the viability of fibroblasts, and macrophage proliferation were measured through using wound healing assay, methylthiazol tetrazolium assay, and bromodeoxyuridine, respectively. In this regard, in vivo diabetic wound healing experiments in Wistar rats following treatment with nontoxic concentrations of Lactobacillus bulgaricus and Lactobacillus plantarum were conducted. The histopathological and gene expression analyses were performed following removal of wound sites 3, 7, and 14 days postwounding. Results showed that treatment with probiotics accelerated the healing process of diabetic wounds and modulated the inflammatory cells in wound sites during a 14-day period postwounding. The altered mRNA levels of inflammatory cytokines were observed in wound sites following treatment with probiotics. The findings of the current study reveal that L. bulgaricus and L. plantarum could improve the healing of diabetic wounds via regulation of inflammation.


Assuntos
Diabetes Mellitus/tratamento farmacológico , Inflamação/tratamento farmacológico , Lactobacillus delbrueckii/química , Lactobacillus plantarum/química , Probióticos/farmacologia , Cicatrização/efeitos dos fármacos , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Diabetes Mellitus/metabolismo , Humanos , Inflamação/metabolismo , Interleucina-10 , Interleucina-1beta/metabolismo , Camundongos , Ratos , Ratos Wistar , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
3.
Molecules ; 25(22)2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33207534

RESUMO

Lactobacillus bulgaricus is a LAB strain which is capable of producing bacteriocin substances to inhibit Staphylococcus aureus. The aim of this study was to purify a bacteriocin-like inhibitory substance (BLIS) produced by L. bulgaricus FTDC 1211 using an aqueous impregnated resins system consisting of polyethylene-glycol (PEG) impregnated on Amberlite XAD4. Important parameters influencing on purification of BLIS, such as the molecular weight and concentration of PEG, the concentration and pH of sodium citrate and the concentration of sodium chloride, were optimized using a response surface methodology. Under optimum conditions of 11% (w/w) of PEG 4000 impregnated Amberlite XAD4 resins and 2% (w/w) of sodium citrate at pH 6, the maximum purification factor (3.26) and recovery yield (82.69% ± 0.06) were obtained. These results demonstrate that AIRS could be used as an alternate purification system in the primary recovery step.


Assuntos
Bacteriocinas/farmacologia , Lactobacillus delbrueckii/química , Polietilenoglicóis/química , Poliestirenos/química , Polivinil/química , Resinas Sintéticas/química , Análise de Variância , Anti-Infecciosos/farmacologia , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Peso Molecular , Citrato de Sódio/análise
4.
Int J Biol Macromol ; 163: 1361-1368, 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-32745547

RESUMO

To promote the application of probiotics that are beneficial to hosts, calcium-alginate (Ca-Alg) coated whey protein isolate microcapsules were prepared for protection and delivery of L. bulgaricus and L. paracasei. The internal layer was formed with transglutaminase-induced gelation of whey protein isolate (WPI). Sodium alginate (SA) was applied to form outer layer with external Ca2+ gelation method. The microcapsules loaded with probiotics were characterized by scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FT-IR), and differential scanning calorimetry (DSC). The results showed that the co-encapsulation efficiency was 90.54% and 84.46% of WPI micro-beads and Ca-Alg-coated microcapsules, respectively. The trehalose was added as cryoprotectant to improve the survival rate of probiotics in freeze-dried Ca-Alg-coated microcapsules from 3% to 41.26%. Ca-Alg-coated microcapsules have regular morphology and intensified structure. The protection of Ca-Alg-coated microcapsule for probiotics was improved under simulated gastrointestinal and thermal conditions. Ca-Alg-WPI microcapsules showed a useful way for protection and delivery of L. bulgaricus and L. paracasei.


Assuntos
Alginatos/química , Cálcio/química , Cápsulas/química , Lacticaseibacillus paracasei/química , Lactobacillus delbrueckii/química , Proteínas do Soro do Leite/química , Composição de Medicamentos/métodos , Liofilização/métodos , Probióticos/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos
5.
J Dairy Sci ; 103(7): 5816-5829, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32418689

RESUMO

Fermented milk is an effective carrier for probiotics, the consumption of which improves host health. The beneficial effects of probiotics, prebiotics, and synbiotics on gut dysbiosis have been reported previously. However, the way in which specific probiotics, prebiotics, and synbiotics regulate intestinal microbes remains unclear. Therefore, the probiotics Lactobacillus rhamnosus AS 1.2466 and Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 and the prebiotics xylooligosaccharide and red ginseng extracts were fed to mice to determine their effects on the intestinal microbiota. Then, mice were administered xylooligosaccharide and L. rhamnosus (synthesis) by gavage, and the number of L. rhamnosus was determined in the intestine at different times. The results show that probiotics and prebiotics can quickly reduce the Firmicutes/Bacteroidetes ratio, inhibit harmful bacteria (such as Klebsiella and Escherichia coli), and accelerate the recovery of beneficial intestinal microorganisms (such as Lactobacillus). In a complex intestinal microecology, different probiotics and prebiotics have different effects on specific intestinal microorganisms that cannot be recovered in the short term. In addition, after 20 d of intragastric xylooligosaccharide addition at 0.12 g/kg of body weight, L. rhamnosus colonization in the mouse ileum was 7.48 log cfu/mL, which was higher than in the low-dose group, prolonging colonization time and increasing the number of probiotics in the intestine. Therefore, this study demonstrated that probiotics and prebiotics can promote the balance of intestinal microbiota by regulating specific microbes in the intestine, and the effects of a suitable combination of synbiotics are beneficial, laying the foundation for the development of new dairy products rich in synbiotics.


Assuntos
Bactérias/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Prebióticos , Probióticos/farmacologia , Simbióticos , Ampicilina/farmacologia , Animais , Antibacterianos/farmacologia , Microbioma Gastrointestinal/fisiologia , Glucuronatos/administração & dosagem , Glucuronatos/farmacologia , Lactobacillus delbrueckii/química , Lacticaseibacillus rhamnosus/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oligossacarídeos/administração & dosagem , Oligossacarídeos/farmacologia , Panax/química , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Prebióticos/administração & dosagem , Probióticos/administração & dosagem , Organismos Livres de Patógenos Específicos , Simbióticos/administração & dosagem
6.
Pak J Pharm Sci ; 33(1): 191-197, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32122848

RESUMO

Whey plays an important role in the sports nutrition because of high quality proteins and essential amino acid profile. Nine formulations of sportsman drinks were made using Cheddar, Mozzarella and Paneer whey with normal as well as additional fermentation. The developed sportsman drinks were evaluated for physico-chemical analyses, amino acid profile, viscosity and total plate count along with sensory response during two month storage. Drink having Cheddar whey (T4) with additional fermentation was better in terms of quality and nutrition. Furthermore, amino acid profile considered it a complete and balanced source of essential and non-essential amino acids. Amongst essential amino acids, highest values was recorded for branched chain amino acids like leucine (73.16±3.09) followed by lysine (61.56±0.61) and valine (44.13±1.86)mg/g protein. The dietary significance of sportsman drink can be enhanced through additional fermentation using Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophillus.


Assuntos
Bebidas , Composição de Medicamentos/métodos , Probióticos/química , Esportes , Proteínas do Soro do Leite/química , Aminoácidos/análise , Fenômenos Químicos , Armazenamento de Medicamentos , Fermentação , Humanos , Lactobacillus delbrueckii/química , Sensação/efeitos dos fármacos , Streptococcus thermophilus/química
7.
Carbohydr Polym ; 230: 115593, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887873

RESUMO

The aim of this study was to investigate the in vitro digestion and fermentation prebiotic properties of three released-exopolysaccharide fractions (r-EPS1, r-EPS2 and r-EPS3) from Lactobacillus delbrueckii ssp. bulgaricus SRFM-1. There were no free oligosaccharides and/or monosaccharides for r-EPS1 before and after simulated buccal, gastric and small intestinal (GSI) digestion in vitro. In contrast, r-EPS2 (13.4 %) and r-EPS3 (10.6 %) generated a few monosaccharides after digestion. Additionally, r-EPS1 and r-EPS2 seemed to present a strong bifidogenic effect comparing to inulin, as they exhibited high values of selectivity index (13.17 and 12.84, respectively). Furthermore, the fermentation with r-EPS1 produced the highest contents of acetic acid and lactic acid (56.3 mM and 44.29 mM, respectively), which resulted in the highest amounts of total short chain fatty acid (145.51 mM) followed by r-EPS2 (135.57 mM) and inulin (99.28 mM). These results indicated that r-EPS from L. delbrueckii ssp. bulgaricus SRFM-1 could be a good potential candidate for new functional food prebiotic.


Assuntos
Digestão/efeitos dos fármacos , Lactobacillus delbrueckii/química , Polissacarídeos Bacterianos/química , Polissacarídeos/química , Meios de Cultura/química , Fermentação , Humanos , Ácido Láctico/química , Lactobacillus delbrueckii/metabolismo , Monossacarídeos/química , Polissacarídeos/metabolismo , Polissacarídeos Bacterianos/metabolismo
8.
Int J Biol Macromol ; 148: 238-247, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31935402

RESUMO

In the study, Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) and Lactobacillus paracasei subsp. paracasei (L. paracasei) were isolated from food products and identified. The probiotics were co-encapsulated by emulsification technique with transglutaminase-induced gelation of whey protein isolate (WPI). A double layer was formed by xanthan gum (XG). The survival of both free and microencapsulated cells in each step was enumerated. The results showed that high entrapment yield (86.36 ± 1.07%) was achieved, WPI-XG provided significant protection for cells in simulated gastric juice and bile tolerance tests compared with free cells. Besides, the survivability under thermal condition also strengthened. During freeze-drying process, the survival was improved and naked cells cannot be seen on microcapsules surface when trehalose was added into WPI solution as a lyoprotectant. Additionally, the structure changes of WPI/XG in the encapsulation process were investigated by Fourier transform infrared spectrophotometer (FT-IR) and the thermal properties were studied by differential scanning calorimetry (DSC).


Assuntos
Cápsulas/química , Suco Gástrico/química , Lacticaseibacillus paracasei/química , Lactobacillus delbrueckii/química , Polissacarídeos Bacterianos/química , Proteínas do Soro do Leite/química , Composição de Medicamentos/métodos , Liofilização/métodos , Trato Gastrointestinal/microbiologia , Probióticos/química
9.
Nutr Cancer ; 72(8): 1390-1399, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31707847

RESUMO

Many beneficial effects of probiotic Lactobacilli on cancer prevention and therapy were previously presented. So finding probiotics with proapoptotic activities is a promising approach for cancer drug discovery. Here, the antiproliferative and antioxidant activities of cell-free extracts of Lactobacillus acidophilus and Lactobacillus delbrueckii on HT-29 cell line were evaluated employing MTT and DPPH assays. The induction of apoptosis was assessed by Hoechst staining and flow cytometry analysis which was further confirmed by expression analysis of BCL-2, BAX, caspase-3, caspase-8, and caspase-9 genes using real-time quantitative PCR. Caspase-3 activity was also analyzed. Results showed that cell viability was significantly reduced to 42.2 ± 0.01% and 19.40 ± 0.01% by 5 and 8 mg ml-1 of L. acidophilus and L. delbrueckii extracts, respectively. Apoptosis induction was shown with both bacterial extracts. Caspase-9 and caspase-3 overexpression as well as Bax/Bcl-2 ratio increase revealed the ability of both probiotics to induce intrinsic pathway-dependent apoptosis. The extrinsic pathway was also activated by L. acidophilus. At the concentration of 198 µg ml-1, L. acidophilus and L. delbrueckii had a DPPH scavenging activity of 59.37 ± 3.97% and 71.19 ± 3.64%, respectively. Taken together, these findings provide evidence for antiproliferative, proapoptotic, and antioxidant effects driven by these probiotic lactic acid bacteria (LAB) strains.


Assuntos
Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Neoplasias do Colo/tratamento farmacológico , Lactobacillus acidophilus/química , Lactobacillus delbrueckii/química , Probióticos/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Células HT29 , Humanos
10.
J Dairy Sci ; 102(7): 5913-5921, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31030932

RESUMO

Angiotensin-converting enzyme inhibitory peptides were isolated and identified from milk fermented using Lactobacillus delbrueckii QS306. The peptide with the highest angiotensin-converting enzyme inhibitory activity (C5) was purified using ultrafiltration with 10 and 3 kDa molecular mass cut-off membranes, Sephadex G-15 (Sigma-Aldrich, St. Louis, MO) gel filtration chromatography, reversed-phase HPLC, and Orbitrap Elite (Thermo Fisher Scientific Inc., Waltham, MA) liquid chromatography-tandem mass spectrometry. We obtained peptide LPYPY by microbial fermentation, which was derived from κ-casein f (AA 77-81). We synthesized LPYPY using an Fmoc solid-phase synthesis method and explored the secondary structure of the pentapeptide. The half maximal inhibitory concentration for the angiotensin-converting enzyme inhibitory activity of LPYPY was 12.87 µg/mL. The results provide additional information for ongoing research and the development of functional foods having antihypertensive effects.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/análise , Proteínas de Bactérias/análise , Lactobacillus delbrueckii/química , Leite/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Anti-Hipertensivos/análise , Anti-Hipertensivos/farmacologia , Fermentação , Leite/microbiologia , Peptídeos/análise
11.
Elife ; 72018 05 29.
Artigo em Inglês | MEDLINE | ID: mdl-29809140

RESUMO

Vitamin B12 (cobalamin) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Although its biosynthesis and role as co-factor are well understood, knowledge about uptake of cobalamin by prokaryotic auxotrophs is scarce. Here, we characterize a cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, and demonstrate that it mediates the specific, ATP-dependent uptake of cobalamin. We solved the crystal structure of ECF-CbrT in an apo conformation to 3.4 Å resolution. Comparison with the ECF transporter for folate (ECF-FolT2) from the same organism, reveals how the identical ECF module adjusts to interact with the different substrate binding proteins FolT2 and CbrT. ECF-CbrT is unrelated to the well-characterized B12 transporter BtuCDF, but their biochemical features indicate functional convergence.


Assuntos
Transportadores de Cassetes de Ligação de ATP/química , Apoproteínas/química , Proteínas de Bactérias/química , Ácido Fólico/química , Lactobacillus delbrueckii/química , Vitamina B 12/química , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Trifosfato de Adenosina/química , Trifosfato de Adenosina/metabolismo , Apoproteínas/genética , Apoproteínas/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Transporte Biológico , Clonagem Molecular , Cristalografia por Raios X , Escherichia coli/genética , Escherichia coli/metabolismo , Ácido Fólico/metabolismo , Expressão Gênica , Teste de Complementação Genética , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Cinética , Lactobacillus delbrueckii/metabolismo , Modelos Moleculares , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia Estrutural de Proteína , Especificidade por Substrato , Vitamina B 12/metabolismo
12.
Mol Immunol ; 93: 253-265, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28800975

RESUMO

Previous studies demonstrated that the extracellular polysaccharides (EPSs) produced by Lactobacillus delbrueckii OLL1073R-1 (LDR-1) improve antiviral immunity, especially in the systemic and respiratory compartments. However, it was not studied before whether those EPSs are able to beneficially modulate intestinal antiviral immunity. In addition, LDR-1-host interaction has been evaluated mainly with immune cells while its interaction with intestinal epithelial cells (IECs) was not addressed before. In this work, we investigated the capacity of EPSs from LDR-1 to modulate the response of porcine IECs (PIE cells) to the stimulation with the Toll-like receptor (TLR)-3 agonist poly(I:C) and the role of TLR2, TLR4, and TLR negative regulators in the immunoregulatory effect. We showed that innate immune response triggered by TLR3 activation in porcine IECs was differentially modulated by EPS from LDR-1. EPSs treatment induced an increment in the expression of interferon (IFN)-α and IFN-ß in PIE cells after the stimulation with poly(I:C) as well as the expression of the antiviral factors MxA and RNase L. Those effects were related to the reduced expression of A20 in EPS-treated PIE cells. EPS from LDR-1 was also able to reduce the expression of IL-6 and proinflammatory chemokines. Although further in vivo studies are needed, our results suggest that these EPSs or a yogurt fermented with LDR-1 have potential to improve intestinal innate antiviral response and protect against intestinal viruses.


Assuntos
Células Epiteliais/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Interferon beta/biossíntese , Mucosa Intestinal/citologia , Lactobacillus delbrueckii/imunologia , Polissacarídeos Bacterianos/farmacologia , Sus scrofa/imunologia , Animais , Células Cultivadas , Citocinas/biossíntese , Citocinas/genética , Células Epiteliais/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Interferon beta/genética , Lactobacillus delbrueckii/química , Poli I-C/farmacologia , Polissacarídeos Bacterianos/isolamento & purificação , Transdução de Sinais , Suínos , Doenças dos Suínos/imunologia , Receptor 2 Toll-Like/imunologia , Receptor 3 Toll-Like/imunologia , Receptor 4 Toll-Like/imunologia , Viroses/imunologia , Viroses/veterinária
13.
Carbohydr Polym ; 173: 654-664, 2017 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-28732910

RESUMO

Three released exopolysaccharide fractions (r-EPS1, r-EPS2 and r-EPS3) were isolated from the fermented milk of Lactobacillus delbrueckii ssp. bulgaricus SRFM-1 and purified by anion exchange chromatography, and characterizations of the structures were conducted. The r-EPS1 and r-EPS2 were homogenous with the average molecular weights of 3.97×105Da and 3.86×105Da, respectively. Three r-EPS fractions were composed of galactose and glucose with a molar ratio of 1.23: 1.00, 1.33: 1.00 and 1.00: 1.34, respectively. Structural characterization indicated that the r-EPS1 contained a backbone of →6-ß-d-Galp-(1→4)-ß-d-Glcp-(1→4)-α-d-Galp-(1→4)-ß-d-Galp-(1→6)-ß-d-Galp-(1→4)-ß-d-Glcp-(1→4)-α-d-Galp-(1→4)-ß-d-Galp-(1→4)-α-d-Glcp-(1→, and had three branching points which existed in terminal with D-Glcp residues with α/ß-d-(1→6) linkages. The r-EPS2 was composed of →6-ß-d-Galp-(1→4)-ß-d-Glcp-(1→6)-α-d-Galp-(1→ as the backbone chain with a branching point which also existed in terminal D-Glcp residue with ß-(1→6) linkage. In addition, three r-EPS fractions exhibited strong scavenging activities on superoxide radical, hydroxyl radical, DPPH radical and chelating activity on ferrous ion, and their antioxidant activities decreased in the order of r-EPS1>r-EPS2>r-EPS3.


Assuntos
Antioxidantes/química , Lactobacillus delbrueckii/química , Polissacarídeos Bacterianos/química , Animais , Produtos Fermentados do Leite/microbiologia , Peso Molecular
14.
Fish Shellfish Immunol ; 68: 84-91, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28698125

RESUMO

The aim of the present study was to investigate effects of dietary Lactobacillus delbrueckii (L. delbrueckii) on immune response, disease resistance against Aeromonas hydrophila (A. hydrophila), antioxidant capability and growth performance of Cyprinus carpio Huanghe var. 450 fish (mean weight of 1.05 ± 0.03 g) were randomly distributed into five groups that fed diets containing different levels of L. delbrueckii (0, 1 × 105, 1 × 106, 1 × 107 and 1 × 108 CFU g-1) for 8 weeks. The results showed that intestinal immune parameters such as lysozyme, acid phosphatase, and myeloperoxidase activities, immunoglobulin M content, and the survival rate were improved in fish fed with 1 × 106 and 1 × 107 CFU g-1L. delbrueckii. In addition, 1 × 107 CFU g-1L. delbrueckii supplementation down-regulated mRNA levels of TNF-α, IL-8, IL-1ß and NF-κBp65, and up-regulated IL-10 and TGF-ß mRNA levels in the intestine. The survival rate was significantly (P < 0.05) higher (68.33%) in fish fed 1 × 106 CFU g-1L. delbrueckii than the control diet-fed group (40%) after challenge by A. hydrophila. Fish fed with diet containing 1 × 106 CFU g-1L. delbrueckii showed higher antioxidant enzyme activities such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and total antioxidant capacity (T-AOC) and lower MDA concentrations than those of the control group (P < 0.05). The relative gene expression (SOD, CAT, GPX) showed the same trend with their activities. In addition, the growth performance was significantly improved in fish fed with the diet containing 1 × 106 and 1 × 107 CFU g-1L. delbrueckii (P < 0.05). These results demonstrated that dietary optimal levels of L. delbrueckii enhanced immunity, disease resistance against A. hydrophila antioxidant capability and growth performance in Cyprinus carpio Huanghe var.


Assuntos
Carpas , Suplementos Nutricionais , Resistência à Doença , Doenças dos Peixes/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterinária , Imunidade Inata , Lactobacillus delbrueckii , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Carpas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Lactobacillus delbrueckii/química , Lactobacillus delbrueckii/imunologia , Distribuição Aleatória
15.
AAPS PharmSciTech ; 18(5): 1605-1616, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27620196

RESUMO

Material barrier properties to microbes are an important issue in many pharmaceutical applications like wound dressings. A wide range of biomaterials has been used to manage the chronic inflamed wounds. Eight hydrogel membranes of poly vinyl alcohol (PVA) with κ-carrageenan (KC) and Lactobacillus bulgaricus extract (LAB) have been prepared by using freeze-thawing technique. To evaluate the membranes efficiency as wound dressing agents, various tests have been done like gel fraction, swelling behavior, mechanical properties, etc. The antibacterial activities of the prepared membranes were tested against the antibiotic-resistant bacterial isolates. In addition, the safety usage of the prepared hydrogel was checked on human dermal fibroblast cells. The anti-inflammatory properties of the prepared hydrogel on LPS-PBMC cell inflammatory model were quantified using enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-qPCR). The analysis data of TGA, SEM, gel fraction, and swelling behavior showed changes in properties of prepared PVA\KC\LAB hydrogel membrane than pure PVA hydrogel membrane. The antibacterial activities of the prepared membranes augmented in LAB extract-prepared membranes. Out of the eight used hydrogel membranes, the PVAKC4 hydrogel membrane is the safest one on fibroblast cellular proliferation with a maximum proliferation percentage 97.3%. Also, all the used hydrogel membrane showed abilities to reduce the concentration of IL-2 and IL-8 compared with both negative and positive control. In addition, almost all the prepared hydrogel membrane showed variable abilities to downregulate the expression of TNF-α gene with superior effect of hydrogel membrane KC1. PVA/KC/LAB extract hydrogel membrane may be a promising material for wound dressing application and could accelerate the healing process of the chronic wound because of its antimicrobial and anti-inflammatory properties.


Assuntos
Curativos Hidrocoloides , Carragenina , Lactobacillus delbrueckii/química , Álcool de Polivinil , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Carragenina/química , Carragenina/farmacologia , Humanos , Teste de Materiais/métodos , Membranas Artificiais , Permeabilidade , Álcool de Polivinil/química , Álcool de Polivinil/farmacologia
16.
J Agric Food Chem ; 64(49): 9421-9427, 2016 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-27960296

RESUMO

Many bacteria in nature use quorum sensing (QS) to regulate gene expression. The quorum sensing system plays critical roles in the adaptation of bacteria to the surrounding environment. Previous studies have shown that during high-density fermentation, the autolysis of lactic acid bacteria was regulated by the QS system, and the two-component system (TCS, LBUL_RS00115/LBUL_RS00110) is involved in the autolysis of Lactobacillus delbrueckii subsp. bulgaricus. However, the QS signal molecule, which regulates this pathway, has not been identified. In this study, we compared the genome of Lactobacillus bulgaricus ATCC BAA-365 with the locus of seven lactobacillus QS systems; the position of the QS signal molecule of Lactobacillus bulgaricus ATCC BAA-365 was predicted by bioinformatics tool. Its function was identified by in vitro experiments. Construction of TCS mutant by gene knockout of LBUL_RS00115 confirmed that the signal molecule regulates the density of the flora by the TCS (LBUL_RS00115/LBUL_RS00110). This study indicated that quorum quenching and inhibition based on the signal molecule might serve as an approach to reduce the rate of autolysis of LAB and increase the number of live bacteria in fermentation.


Assuntos
Proteínas de Bactérias/genética , Lactobacillus delbrueckii/fisiologia , Percepção de Quorum , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Bacteriólise , Fermentação , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/metabolismo , Lactobacillus delbrueckii/química , Lactobacillus delbrueckii/genética , Dados de Sequência Molecular , Filogenia
17.
J Microbiol Methods ; 126: 72-5, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27184086

RESUMO

Whole cell extracts of two Lactobacillus strains were tested with primary antibodies from two pooled sera from allergic patients. Fluorescently labelled anti-human IgG and anti-human IgE secondary antibodies applied in Western blotting, together with an appropriate image acquisition protocol facilitated imagining bacterial proteins that reacted with human IgG and IgE.


Assuntos
Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Western Blotting/métodos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Lactobacillus/química , Proteínas de Bactérias/química , Corantes Fluorescentes , Humanos , Hipersensibilidade , Lacticaseibacillus casei/química , Lactobacillus delbrueckii/química
18.
Carbohydr Res ; 413: 115-22, 2015 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-26117825

RESUMO

The neutral exopolysaccharide (NPS) of Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 was purified and characterized. The molecular mass was 5.0×10(6) g/mol. Sugar and absolute configuration analyses gave the following composition: d-Glc, 1; d-Gal, 1.5. The NPS was also submitted to periodate oxidation followed by borohydride reduction and Smith degradation. Sugar and methylation analyses, (1)H and (13)C nuclear magnetic resonance, and mass spectrometry of the NPS or of its specifically modified products allowed determining the repeating unit sequence: {2)Glc(α1-3)Glc(ß1-3)[Gal(ß1-4)]Gal(ß1-4)Gal(α1-}n. The structure is compared to that of exopolysaccharides produced by other Lactobacillus bulgaricus strains.


Assuntos
Lactobacillus delbrueckii/química , Polissacarídeos Bacterianos/química , Sequência de Carboidratos , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular
19.
Carbohydr Res ; 413: 93-9, 2015 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-26117227

RESUMO

Lactobacilli are valuable strains for commercial (functional) food fermentations. Their cell surface-associated polysaccharides (sPSs) possess important functional properties, such as acting as receptors for bacteriophages (bacterial viruses), influencing autolytic characteristics and providing protection against antimicrobial peptides. The current report provides an elaborate molecular description of several surface carbohydrates of Lactobacillus delbrueckii subsp. bulgaricus strain 17. The cell surface of this strain was shown to contain short chain poly(glycerophosphate) teichoic acids and at least two different sPSs, designated here as sPS1 and sPS2, whose chemical structures were examined by 2D nuclear magnetic resonance spectroscopy and methylation analysis. Neutral branched sPS1, extracted with n-butanol, was shown to be composed of hexasaccharide repeating units (-[α-d-Glcp-(1-3)-]-4-ß-l-Rhap2OAc-4-ß-d-Glcp-[α-d-Galp-(1-3)]-4-α-Rhap-3-α-d-Galp-), while the major component of the TCA-extracted sPS2 was demonstrated to be a linear d-galactan with the repeating unit structure being (-[Gro-3P-(1-6)-]-3-ß-Galf-3-α-Galp-2-ß-Galf-6-ß-Galf-3-ß-Galp-).


Assuntos
Parede Celular/química , Lactobacillus delbrueckii/citologia , Polissacarídeos Bacterianos/química , Sequência de Carboidratos , Glicerofosfatos/química , Lactobacillus delbrueckii/química , Dados de Sequência Molecular , Polissacarídeos Bacterianos/isolamento & purificação
20.
Appl Biochem Biotechnol ; 175(8): 3657-72, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25666370

RESUMO

Biomass degradation compounds significantly inhibit biochemical conversion of biomass prehydrolysates to biofuels and chemicals, such as lactic acid. To characterize the structure-activity relationship of carbonyl inhibition on lactic acid fermentation, we examined effects of eight carbonyl compounds (furfural, 5-hydroxymethyl furfural, vanillin, syringaldehyde, 4-hydroxybenzaldehyde, phthalaldehyde, benzoic acid, and pyrogallol aldehyde) and creosol on lactic acid production by Lactobacillus delbrueckii. Pyrogallol aldehyde reduced the cell growth rate by 35 % at 1.0 mM and inhibited lactic acid production completely at 2.0 mM. By correlating the molecular descriptors to the inhibition constants in lactic acid fermentation, we found a good relationship between the hydrophobicity (Log P) of aldehydes and their inhibition constants in fermentation. The inhibitory effect of carbonyl inhibitors appeared to correlate with their thiol reactivity as well. In addition, we found that H2O2 detoxified pyrogallol aldehyde and phthalaldehyde inhibitory activity. H2O2 detoxification was applied to real biomass prehydrolysates in lactic acid fermentation.


Assuntos
Biomassa , Ácido Láctico/metabolismo , Lactobacillus delbrueckii/química , Biocombustíveis , Etanol/química , Fermentação , Peróxido de Hidrogênio/química , Hidrólise , Ácido Láctico/química , Lactobacillus delbrueckii/metabolismo , Relação Estrutura-Atividade
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