Lactic Acid Bacterial Strains Research Articles

Lactococcus lactis subsp. cremoris YRC3780 modifies function of mesenteric lymph node dendritic cells to modulate the balance of T cell differentiation inducing regulatory T cells

IntroductionThe intestinal immune system plays a pivotal role in the induction of immune responses against food. In the case of T cell response, dendritic cells (DCs) are especially important. However, the regulation of immune responses to food by intestinal DCs has been poorly described. In this study, we analyzed the effect of Lactococcus lactis subsp. cremoris YRC3780, a lactic acid bacterial strain isolated from kefir, a traditional fermented milk product, on the immune responses induced by antigen presentation by intestinal DCs to T cells as well as the mechanism of action of these immunomodulatory effects. It has been shown that L. cremoris YRC3780 ameliorates the symptoms of pollinosis in both animal and human studies.MethodsCD11c+ cells from mesenteric lymph nodes (MLNs) of BALB/c mice were cultured as MLN DCs with L. cremoris YRC3780 and expression of genes inducing regulatory T cells (Tregs) was examined by qPCR. In addition, MLN DCs were cocultured with CD4+ T cells from DO11.10 transgenic mice expressing an ovalbumin (OVA)-specific TCR and the OVA antigen peptide and L. cremoris YRC3780. Induction of Tregs was examined by flow cytometry, gene expression was analyzed by DNA microarray and qPCR, and the production of cytokines was measured by ELISA. MLN DCs from TLR2-deficient mice and components of L. cremoris YRC3780 were used to examine the recognition of YRC3780 by MLN DCs.ResultsL. cremoris YRC3780 enhanced the expression of genes involved in Treg induction in MLN DCs and induced Foxp3+CD4+T cells in an MLN DC and CD4+ T-cell co-culture system. The effect on MLN DCs was likely mediated by receptors other than TLR2. Together with microarray analyses of CD4+ T cell gene expression and cytokine ELISA, it was demonstrated that L. cremoris YRC3780 promoted the induction of Th1 and Tregs, and regulated the balance of Th1/Th2 and Treg/Th17 cells involving multiple genes via the antigen-presentation of MLN DCs.DiscussionOur findings provide insights into the modulation of intestinal immune responses mediated by DCs and the antiallergic effects of lactic acid bacteria.

Lactococcus lactis subsp. cremoris YRC3780 modifies function of mesenteric lymph node dendritic cells to modulate the balance of T cell differentiation inducing regulatory T cells.

The intestinal immune system plays a pivotal role in the induction of immune responses against food. In the case of T cell response, dendritic cells (DCs) are especially important. However, the regulation of immune responses to food by intestinal DCs has been poorly described. In this study, we analyzed the effect of Lactococcus lactis subsp. cremoris YRC3780, a lactic acid bacterial strain isolated from kefir, a traditional fermented milk product, on the immune responses induced by antigen presentation by intestinal DCs to T cells as well as the mechanism of action of these immunomodulatory effects. It has been shown that L. cremoris YRC3780 ameliorates the symptoms of pollinosis in both animal and human studies. CD11c+ cells from mesenteric lymph nodes (MLNs) of BALB/c mice were cultured as MLN DCs with L. cremoris YRC3780 and expression of genes inducing regulatory T cells (Tregs) was examined by qPCR. In addition, MLN DCs were cocultured with CD4+ T cells from DO11.10 transgenic mice expressing an ovalbumin (OVA)-specific TCR and the OVA antigen peptide and L. cremoris YRC3780. Induction of Tregs was examined by flow cytometry, gene expression was analyzed by DNA microarray and qPCR, and the production of cytokines was measured by ELISA. MLN DCs from TLR2-deficient mice and components of L. cremoris YRC3780 were used to examine the recognition of YRC3780 by MLN DCs. L. cremoris YRC3780 enhanced the expression of genes involved in Treg induction in MLN DCs and induced Foxp3+CD4+T cells in an MLN DC and CD4+ T-cell co-culture system. The effect on MLN DCs was likely mediated by receptors other than TLR2. Together with microarray analyses of CD4+ T cell gene expression and cytokine ELISA, it was demonstrated that L. cremoris YRC3780 promoted the induction of Th1 and Tregs, and regulated the balance of Th1/Th2 and Treg/Th17 cells involving multiple genes via the antigen-presentation of MLN DCs. Our findings provide insights into the modulation of intestinal immune responses mediated by DCs and the antiallergic effects of lactic acid bacteria.

Effect of Air Classification and Enzymatic and Microbial Bioprocessing on Defatted Durum Wheat Germ: Characterization and Use as Bread Ingredient.

Its high dietary fiber and protein contents and nutritional quality make defatted wheat germ (DWG) a valuable cereal by-product, yet its negative impact on food structure limits its use as a food ingredient. In this research, DWG underwent air classification, which identified two fractions with high fiber (HF) and low fiber/high protein (LF) contents, and a bioprocessing protocol, involving treatment with xylanase and fermentation with selected lactic acid bacterial strains. The degree of proteolysis was evaluated through electrophoretic and chromatographic techniques, revealing differences among fractions and bioprocessing options. Fermentation led to a significant increase in free amino acids (up to 6 g/kg), further enhanced by the combination with xylanase. When HF was used as an ingredient in bread making, the fiber content of the resulting bread exceeded 3.6 g/100 g, thus reaching the threshold required to make a "source of fiber" claim according to Regulation EC No.1924/2006. Meanwhile, all breads could be labeled a "source of protein" since up to 13% of the energy was provided by proteins. Overall, bioprocessed ingredients lowered the glycemic index (84 vs. 89) and increased protein digestibility (80 vs. 63%) compared to control breads. Technological and sensory analysis showed that the enzymatic treatment combined with fermentation also conferred a darker and more pleasant color to the bread crust, as well as better crumb porosity and elasticity.

Amylolytic Capability and Performance of Probiotic Strains in a Controlled Sorghum Fermentation System

This study aimed to explore the fermentative performance of nine lactic acid bacterial strains with probiotic potential during sorghum fermentation. The strain’s attributes including proliferation counts, pH levels, production of organic acid antibacterial activity, and their ability to break down starch were evaluated during the fermentation period in the presence and absence of glucose as a carbon source. In addition, the inhibitory activity of these potential probiotic strains against pathogenic bacteria (Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus) was examined through a co-culturing technique. The results demonstrated that all 4 Lactobacillus strains exhibited robust growth in both glucose and glucose-free fermentation experiments. Glucose supplementation significantly enhanced lactic acid yield which ranged from 0.19 to 0.44% compared to fermentation without glucose which ranged from 0.04 to 0.29%. The selected Lactobacillus strains effectively lowered the media pH below 4.0 after 24 h, producing substantial lactic acid. Notably, in the absence of glucose, only Lb. helveticus D7 and Lb. amylolyticus D12 achieved pH levels below 4 after 8 h, producing the highest lactic acid amounts of 0.27 and 0.29% after 24 h, respectively. Amylase activity was detected on two strains, D7 and D12. Furthermore, most of the tested Lactobacillus strains demonstrated complete inhibition (6 log to 0 Log CFU/mL) of pathogen growth after 24 h of co-culturing, suggesting their potential for enhancing the safety quality of sorghum-based fermented products.

Lactobacillus paracasei subsp. paracasei 2004 improves health and lifespan in Caenorhabditis elegans

Recent research has highlighted the importance of the gut microbiome in regulating aging, and probiotics are interventions that can promote gut health. In this study, we surveyed several novel lactic acid bacteria to examine their beneficial effect on organismal health and lifespan in C. elegans. We found that animals fed some lactic acid bacteria, including L. acidophilus 1244 and L. paracasei subsp. paracasei 2004, grew healthy. Supplementation with the lactic acid bacterial strains L. acidophilus 1244 or L. paracasei subsp. paracasei 2004 significantly improved health, including food consumption, motility, and resistance to oxidative stressor, hydrogen peroxide. Our RNA-seq analysis showed that supplementation with L. paracasei subsp. paracasei 2004 significantly increased the expression of daf-16, a C. elegans FoxO homolog, as well as genes related to the stress response. Furthermore, daf-16 deletion inhibited the longevity effect of L. paracasei subsp. paracasei 2004 supplementation. Our results suggest that L. paracasei subsp. paracasei 2004 improves health and lifespan in a DAF-16-dependent manner.

The novel probiotic preparation based on Lactobacillus spp. mixture on the intestinal bacterial community structure of Cherry Valley duck.

The development and utilization of probiotics have many environmental benefits when they are used to replace antibiotics in animal production. In this study, intestinal lactic acid bacteria were isolated from the intestines of Cherry Valley ducks. Probiotic lactic acid bacterial strains were screened for antibacterial activity and tolerance to produce a Lactobacillus spp. mixture. The effects of the compound on the growth performance and intestinal flora of Cherry Valley ducks were studied. Based on the results of the antibacterial activity and tolerance tests, the highly active strains Lactobacillus casei 1.2435, L. salivarius L621, and L. salivarius L4 from the intestines of Cherry Valley ducks were selected. The optimum ratio of L. casei 1.2435, L. salivarius L621, and L. salivarius L4 was 1:1:2, the amount of inoculum used was 1%, and the fermentation time was 14h. In vivo experiments showed that compared with the control group, the relative abundances of intestinal Lactobacillus and Blautia were significantly increased in the experimental group fed the lactobacilli compound (P < 0.05); the relative abundances of Parabacteroides, [Ruminococcus]_torques_group, and Enterococcus were significantly reduced (P < 0.05), and the growth and development of the dominant intestinal flora were promoted in the Cherry Valley ducks. This study will provide more opportunities for Cherry Valley ducks to choose microecological agents for green and healthy breeding.

Health Benefits and Antibacterial Properties of Probiotics Isolated From Fermented Food Products

As a result of the continuously expanding scientific evidence probiotic bacteria have become increasingly popular due to their beneficial effects on human health. The main objective of this study was to isolate, identify, and characterize some lactic acid bacterial strains from fermented food products as potential probiotics with antimicrobial activity against some test strains. Total four samples of fermented food products were taken (Sauerkraut, Fermented Cucumber, Idli batter and Dosa batter). The Lactic acid bacteria were isolated from these samples and the probiotics were further tested for their ability to tolerate pH, bile salt at different concentrations. The probiotic strains were found to grow at different pH and also at different conc. of bile and NaCl. These isolates exhibited high antibacterial activity when tested against the test organism (Pseudomonas, Streptococcus, Escherichia coli and Klebsiella) in well diffusion method. The present study concludes that the isolated probiotic strain from the fermented food qualifies pH, bile and NaCl tolerance tests that make an isolate to be an ideal probiotic. Probiotic strains were found to carry antimicrobial property and can be used to control gut microbiota. The result of the present study showed that the fermented food products can be used as a good source of probiotic culture.

Isolation of 60 strains from fermented milk of mares and donkeys in Algeria and identification by 16S rRNA sequencing of lactobacilli: Assessment of probiotic skills of important strains and aromatic productivity power.

Donkey and mare milk have high nutritional and functional values, but their lactic acid bacteria (LAB) content remains poorly studied and undervalued in the Algerian dairy industry. This study aimed to isolate and select LAB strains that produce antimicrobial substances during fermentation and to characterize the probiotic profiles of each extracted strain to indicate their potential for antioxidant and proteolytic activity. This study focuses on isolating and identifying lactic acid bacterial strains from 10 Equid-fermented milk samples collected in two regions of El Bayed Wilaya (Algeria). Identification of LAB strains was obtained by 16S rRNA sequencing. The probiotic properties of important strains and their aromatic productivity power are assessed. To evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, Chryseobacterium joostei, Pseudomonas aeruginosa, and Escherichia coli, we selected 21 strains. Different induction methods have been used to amplify the antibacterial effects against these pathogenic strains. Among a total of 60 identified strains, 31 had a probiotic profile, and most were catalase-negative. Aromatic productivity power was observed in eight strains: Lactiplantibacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Weissella confusa, Weissella cibaria, Leuconostoc mesenteroides, Leuconostoc lactis, and Lactobacillus sp1. Our results provide insight into the considerable diversity of LAB present in fermented donkey and mare milk. To meet the expectations of the Algerian dairy industry, it is important that the probiotic skills of the nine selected strains are met. In addition, a significant number of these strains may have important probiotic activity and biotechnological potential.

Mechanistic Insights into Nitrite Degradation by Metabolites of L. plantarum A50: An LC-MS-Based Untargeted Metabolomics Analysis

Nitrites are universally acknowledged natural toxic substances that frequently lead to poisoning in humans and animals. During fermentation, certain microorganisms utilize a portion of the nitrogen element and reduce nitrates to nitrites through specific metabolic pathways. In this study, a highly effective lactic acid bacterial strain, Lactiplantibacillus plantarum A50, was isolated and screened from alfalfa silage for its remarkable ability to degrade nitrites. L. plantarum A50 exhibits exceptional nitrite removal capacity, with a degradation rate of 99.06% within 24 h. Furthermore, L. plantarum A50 demonstrates normal growth under pH values ranging from 4 to 9 and salt concentrations of 5%, displaying excellent tolerance to acidity, alkalinity, and salinity. Additionally, it undergoes fermentation using various carbon sources. Within the first 6–12 h of culture, L. plantarum A50 primarily achieves nitrite degradation through non-acidic processes, resulting in a degradation rate of 82.67% by the 12th hour. Moreover, the metabolites produced by L. plantarum A50 exhibit a synergistic interaction with acidity, leading to a nitrite degradation rate of 98.48% within 24 h. Notably, both L. plantarum A50 and MRS broth were found to degrade nitrites. Consequently, a non-targeted metabolomic analysis using LC-MS was conducted to identify 342 significantly different metabolites between L. plantarum A50 and MRS broth. Among these, lipids and lipid-like molecules, organic acids and derivatives, organic oxygen compounds, and organoheterocyclic compounds emerged as the main constituents. Lipids and lipid-like molecules, derivatives of glucose and galactose, amino acids and their derivatives, as well as organoheterocyclic compounds, are likely to play a role in nitrite elimination. Through the enrichment analysis of differential metabolic pathways using KEGG, nine distinct pathways were identified. These pathways provide essential nutrients, maintain cellular structure and function, participate in substance transport, regulate metabolic activities, and enhance resistance against pathogenic microorganisms in L. plantarum A50.

Bifidobacterium species viability in dairy-based probiotic foods: challenges and innovative approaches for accurate viability determination and monitoring of probiotic functionality.

Bifidobacterium species are essential members of a healthy human gut microbiota. Their presence in the gut is associated with numerous health outcomes such as protection against gastrointestinal tract infections, inflammation, and metabolic diseases. Regular intake of Bifidobacterium in foods is a sustainable way of maintaining the health benefits associated with its use as a probiotic. Owing to their global acceptance, fermented dairy products (particularly yogurt) are considered the ideal probiotic carrier foods. As envisioned in the definition of probiotics as "live organisms," the therapeutic functionalities of Bifidobacterium spp. depend on maintaining their viability in the foods up to the point of consumption. However, sustaining Bifidobacterium spp. viability during the manufacture and shelf-life of fermented dairy products remains challenging. Hence, this paper discusses the significance of viability as a prerequisite for Bifidobacterium spp. probiotic functionality. The paper focuses on the stress factors that influence Bifidobacterium spp. viability during the manufacture and shelf life of yogurt as an archetypical fermented dairy product that is widely accepted as a delivery vehicle for probiotics. It further expounds the Bifidobacterium spp. physiological and genetic stress response mechanisms as well as the methods for viability retention in yogurt, such as microencapsulation, use of oxygen scavenging lactic acid bacterial strains, and stress-protective agents. The report also explores the topic of viability determination as a critical factor in probiotic quality assurance, wherein, the limitations of culture-based enumeration methods, the challenges of species and strain resolution in the presence of lactic acid bacterial starter and probiotic species are discussed. Finally, new developments and potential applications of next-generation viability determination methods such as flow cytometry, propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR), next-generation sequencing, and single-cell Raman spectroscopy (SCRS) methods are examined.

Novel Placenta-Derived Liquid Product Suitable for Cosmetic Application Produced by Fermentation and Digestion of Porcine or Equine Placenta Using Lactic Acid Bacterium Enterococcus faecalis PR31

Since ancient times, the placenta has been used to produce cosmetic and health food products, whereas fermentation is a technology that has been used to produce foods and cosmetics. For application in cosmetics, traditional placental extracts produced solely by proteolysis have not had enough moisturizing properties or the ability to stimulate the proliferation of epidermal keratinocytes. We combined these two traditional approaches to produce raw materials without such drawbacks that are suitable for cosmetic applications. Using a unique lactic acid bacterial strain, Enterococcus faecalis PR31, to directly ferment and digest both porcine and equine placentas, we produced the following liquid products: placenta ferment filtrates. The ferment filtrates stimulated the proliferation of not only normal human dermal fibroblasts but also epidermal keratinocytes. The ferments had higher equilibrium water content properties than traditional placental extracts, and the ferment derived from the porcine placenta maintained high stratum corneum water content levels for up to 6 h after its application on the skin. Metabolome analysis revealed various molecules that were increased by fermentation, among which lactic acid was assumed to play a central role in the high moisturizing properties. To conclude, the placenta ferment filtrates developed in this study are beneficial for cosmetic applications.

Sodium carboxymethyl celluloses as a cryoprotectant for survival improvement of lactic acid bacterial strains subjected to freeze-drying

This study investigated the possibility of sodium carboxymethyl celluloses (Na-CMC) in protecting the viability of lactic acid bacteria (LAB) against freeze-drying stress. 1 % concentration of Na-CMC with a 0.7 substitution degree and viscosity of 1500 to 3100 (MPa.s) was found to protect Lactobacillus delbrueckii subsp. bulgaricus CICC 6098 best, giving a high survival rate of 23.19 ± 0.88 %, high key enzymatic activities, and 28-day storage stability. Additionally, Na-CMC as cryoprotectant provided good protection for other 7 lactic acid bacterial strains subjected to freeze-drying. The highest survival rate was 48.79 ± 0.20 U/mg for β-GAL, 2.75 ± 0.15 U/mg for Na+-K+-ATPase, and 2.73 ± 0.41 U/mg for Ca2+-Mg2+-ATPase as 48.48 ± 0.46 % for freeze-dried Pediococcus pentosaceus CICC 22228. It was Interesting to note that the presence of Na-CMC reduced the freezable water content of the lyophilized powders containing the tested strains through its hydroxyl group, and supplied micro-holes and fibers for protecting the integrated structure of LAB cell membrane and wall against the freezing damage. It is clear that addition of Na-CMC should be promising as a new cryoprotective agent available for processing the lyophilized stater cultures of LAB strains.

Gut production of GABA by a probiotic formula: an in vitro study.

The objective of this study was to evaluate the influence of a probiotic formula on the production of neuroactive compounds in different parts of the colon in batch culture and in vitro gut simulator experiments. Thirteen lactic acid bacterial strains, belonging to the species Levilactobacillus brevis, Lactiplantibacillus plantarum, Lacticaseibacillus paracasei, Ligilactobacillus salivarius, Streptococcus thermophilus, were characterised for their in vitro ability to produce neurotransmitters. L. brevis P30021 and L. plantarum P30025 were selected based on their capability to produce γ-aminobutyric acid (GABA) and acetylcholine in vitro. A probiotic formulation with potential psychobiotic activity was prepared and tested in a batch culture of human microbiota monitoring the formation of GABA and acetylcholine. Samples of the three colonic tracts were taken from the Simulator of the Human Intestinal Microbiota (SHIME®) evaluating the production of GABA and other neurotransmitters by LC-MS. Short chain fatty acids (SCFA) by GC and microbiota composition by 16S rRNA gene sequencing were also determined. Probiotic supplementation led to the formation of GABA and acetylcholine with a decrease in glutamate concentrations in the in vitro batch fermentation. Production of GABA after the treatment with probiotics was confirmed in the SHIME® short-term experiment. No differences in short-chain fatty acids were observed up to 72h of fermentation. Different microbiota composition was found in the three different parts of the colon, with a higher abundance of Veillonellaceae in the ascending colon vessels. The probiotic-exposed microbiota showed higher levels of Bacteroides, a gut microbe associated with anti-inflammatory activities and a potential GABA producer. Results demonstrate the impact of the tested probiotic formula on gut microbiota structure and GABA production. In conclusion, the probiotic treatment changed the microbiota composition and increased neuroactive metabolites production, indicating promising potential as psychobiotics, even if further clinical evidence is needed to confirm the effectiveness of these probiotics in improving mental health.

Analysis of the key genes of Lactobacillus reuteri strains involved in the protection against alcohol-induced intestinal barrier damage.

Gastrointestinal inflammation and intestinal barrier function have important effects on human health. Alcohol, an important foodborne hazard factor, damages the intestinal barrier, increasing the risk of disease. Lactobacillus reuteri strains have been reported to reduce gastrointestinal inflammation and strengthen the intestinal barrier. In this study, we selected three anti-inflammatory L. reuteri strains to evaluate their role in the protection of the intestinal barrier and their immunomodulatory activity in a mouse model of gradient alcohol intake. Among the three strains tested (FSCDJY33M3, FGSZY33L6, and FCQHCL8L6), L. reuteri FSCDJY33M3 was found to protect the intestinal barrier most effectively, possibly due to its ability to reduce the expression of interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) and increase the expression of tight junction proteins (occludin, claudin-3). Genomic analysis suggested that the protective effects of L. reuteri FSCDJY33M3 may be related to functional genes and glycoside hydrolases associated with energy production and conversion, amino acid transport and metabolism, carbohydrate transport and metabolism, and DNA replication, recombination, and repair. These genes include COG2856, COG1804, COG2071, and COG1061, which encode adenine deaminase, acyl-CoA transferases, glutamine amidotransferase, RNA helicase, and glycoside hydrolases, including GH13_20, GH53, and GH70. Our results identified functional genes that may be related to protection against alcohol-induced intestinal barrier damage, which might be useful for screening lactic acid bacterial strains that can protect the intestinal barrier.

Exploring peptidomes of by-products generated during chhurpi production using Lactobacillus delbrueckii WS4 for identification of novel bioactive peptides.

The considerable value of whey is evident from its significant potential applications and contributions to the functional food and nutraceutical market. The by-products were individually obtained during functional chhurpi and novel soy chhurpi cheese production using defined lactic acid bacterial strains of Sikkim Himalaya's traditional chhurpi. Hydrolysis of substrate proteins by starter proteinases resulted in a comparable peptide content in whey and soy whey which was associated with antioxidant and ACE inhibition potential. Peptidome analysis of Lactobacillus delbrueckii WS4 whey and soy whey revealed the presence of several bioactive peptides including the multifunctional peptides PVVVPPFLQPE and YQEPVLGPVRGPFPIIV. In silico analyses predicted the antihypertensive potential of whey and soy whey peptides with strong binding affinity for ACE active sites. QSAR models predicted the highest ACE inhibition potential (IC50) for the β-casein-derived decapeptide PVRGPFPIIV (0.95 μM) and the Kunitz trypsin inhibitor protein-derived nonapeptide KNKPLVVQF (16.64 μM). Chhurpi whey and soy whey can be explored as a valuable source of diverse and novel bioactive peptides for applications in designer functional foods development.

ANTIFUNGAL ACTIVITY OF Pediococcus pentosaceus ISOLATED FROM BAMBARA GROUNDNUT (Vigna subterranea (L.) Verdc.) SEEDS AGAINST Aspergillus flavus

In this study, 22 lactic acid bacterial (LAB) strains were isolated from Bambara groundnut seeds and assessed for their ability to suppress the development of two strains of aflatoxigenic Aspergillus flavus. The antifungal activity of LAB strains was assessed via the overlay assay. Also, HPLC methodology was used to quantify the production of lactic and acetic acid by the LAB strains with high antifungal activity. After incubation at 30°C for 72 hours, five of these LAB strains (52B1, 52C1, 74B3, 67B1, 67A9) showed high inhibitory activity against Aspergillus flavus strains. These LAB strains exhibiting antifungal potential were identified as Pediococcus pentosaceus through 16S rDNA sequencing. Furthermore, the lactic acid bacteria (LAB) strains reported lactic acid production ranging from 12.08 mg/ml to 15.38 mg/ml and acetic acid production ranging from 0.06 mg/ml to 1.68 mg/ml. Notably, antifungal experiments are conducted in a controlled setting. Further, it would be worthwhile to explore the direct application of Pediococcus pentosaceus strains or their antimicrobial agents to food products to prevent fungal growth during storage. By doing so, the use of fungicides can be avoided and lead to safer food products.

Purine nucleosidase (PNase) activity, probiotics potential, and food applicability of a newly-isolated Levilactobacillus brevis LAB42.

Hyperuricemia, a condition characterized by elevated levels of uric acid in the blood, is known as a risk factor for gout disease. In this study, we isolated a total of 72 MRS-grown colonies and evaluated their purine nucleosidase (PNase) activity. Among the isolated bacteria, Levilactobacillus (L.) brevis LAB42 displayed the highest PNase activity. Our findings also indicate that PNase activity can vary among lactic acid bacterial strains and during different growth phases. Based on the kinetics study, LAB42 consistently exhibits the highest PNase activity. Due to its ability to attach to Caco-2 cells and its resistance to acidic environments and bile exposure, L. brevis LAB42 was chosen for further studies and showed that with the right combination of additives, it has the potential to be an appropriate starter for milk fermentation.

FolE gene expression for folic acid productivity from optimized and characterized probiotic Lactobacillus delbrueckii

BackgroundLactobacillus delbrueckii was one of the most common milk lactic acid bacterial strains (LAB) which characterized as probiotic with many health influencing properties. ResultsAmong seven isolates, KH1 isolate was the best producer of folic acid with 100 µg/ml after 48 h of incubation; FolE gene expression after 24 h of incubation was in the highest value in case of KH1 with three folds. Lactose was the best carbon source for this KH1, besides the best next isolates KH80 and KH98. The selected three LAB isolates were identified through 16S rDNA as Lactobacillus delbrueckii. These three isolates have high tolerance against acidic pH 2–3; they give 45, 10, and 22 CFUs at pH 3, besides 9, 6, and 4 CFUs at pH2, respectively. They also have resistance against elevated bile salt range 0.1–0.4%. KH1 recorded 99% scavenging against 97.3% 1000 µg/ml ascorbic acid. Docking study exhibits the binding mode of folic acid which exhibited an energy binding of − 8.65 kcal/mol against DHFR. Folic acid formed four Pi-alkyl, Pi-Pi, and Pi-sigma interactions with Ala9, Ile7, Phe34, and Ile60. Additionally, folic acid interacted with Glu30 and Asn64 by three hydrogen bonds with 1.77, 1.76, and 1.96 Å. ConclusionLAB isolates have probiotic properties, antioxidant activity, and desired organic natural source for folic acid supplementation that improve hemoglobin that indicated by docking study interaction. Graphical Abstract▪

Effect of two lyoprotectants on the survival rate and storage stability of freeze-dried probiotic lactic acid bacterial strains

Lactic acid bacteria are very important for biotechnology, pharmaceutical and food industry, and are extensively used as probiotic products and starter cultures. Freeze-drying is a widely used and effective process for preservation of lactic acid bacteria in a dried form. To ensure stability and maintain biological activity of lactic acid bacteria, suitable protective media formulations should be used during the freeze-drying process. Twelve oral strains with functional and probiotic properties were subjected to freeze-drying and assessed for viability after the process and after long-term storage. Two lyoprotective media with different composition were used and the freeze-drying process was performed, where retained stability of the studied strains in both lyoprotectors was reported. Viability after four and eight months of storage was assessed, showing that the strains maintained high viability. The two lyoprotective media ensured the obtaining of freeze-dried samples with preserved viability for all studied strains of lactic acid bacteria. Four of the tested strains, L. fermentum N 2 and TC 3-11, L. delbrueckii subsp. lactis VG 2, and W. confusa NN 1 showed well preserved stability and viability after freeze-drying and long-term storage and, prepared in freezedried formulations, are suitable to be included in the omposition of probiotic products for oral healthcare.

Optimization of culture conditions for and assessment of kimchi-originated lactic acid bacterial isolates toward their extracellular GABA-producing ability

The objectives of this study were to optimize the medium and culture conditions using a strong γ-aminobutyric acid (GABA) producer as a reference lactic acid bacterial strain, to screen and identify GABA-producing lactic acid bacterial isolates from kimchi, and to determine their extracellular GABA-producing abilities. Thin-layer chromatography was used to screen GABA-producing bacterial isolates and high-performance liquid chromatography was used to evaluate the bacterial GABA production abilities. Species-specific polymerase chain reaction analyses were used to identify GABA-producing bacterial isolates. The optimal medium and culture conditions were found to be the modified Man–Rogosa–Sharpe (MRS) broth (with an initial pH of 6.5) containing 4% sucrose, 5% glutamate, and 1% yeast extract at 37°C for 5 days. After incubation under the optimized culture conditions, 217 kimchi bacterial isolates were screened to evaluate their respective GABA-producing abilities. Screening the 217 kimchi bacterial isolates identified 24 GABA-producing lactic acid bacterial isolates (11%): Lactobacillus plantarum (17), Lactobacillus brevis (six), and Leuconostoc mesenteroides (one), indicating that only a small proportion of the strains produce GABA in the culture broth. The extracellular GABA-producing abilities of the bacterial strains identified in this study varied even within the same species, ranging from 5.8 to 101.7 mM among the 17 GABA-producing L. plantarum isolates and from 8.5 to 88.6 mM among the six GABA-producing L. brevis isolates. In summary, three species of the 24 kimchi GABA-producing bacterial isolates were identified, including one rare species (L. mesenteroides) and the two most dominant species (L. brevis and L. plantarum).&#x0D; Keywords: Lactobacillus brevis; Lactobacillus plantarum; Leuconostoc mesenteroides; Optimization; γ-aminobutyric acid (GABA)