Search Results (66)

Lacticaseibacillus (L.) rhamnosus CRL75 is a lactic acid bacterium (LAB) isolated from local dairy products, demonstrating significant adaptation in skimmed milk (FM75). In this context, CRL75 exhibited high microbial growth (3.63 ± 0.18 log CFU·mL⁻¹), strong acidification (9.20 ± 0.10 g·L⁻¹ lactic acid, and 2.40 ± 0.10 pH units), and increased viscosity in FM75 after 16 h of fermentation. Additionally, this LAB strain produces both capsular polysaccharides (CPS+) and extracellular polysaccharides (EPS75), contributing to a ropy phenotype (>10 cm). The purified EPS75 (70.70 ± 3.25 mg·L⁻¹) displayed low molecular weight (12.7 kDa), with galactose and glucose as its primary monomers in a 4:1 ratio. In aqueous environments, EPS75 exhibited an extended size (147 nm), a random coil structure, and macromolecular aggregation. Furthermore, vibrational spectroscopy confirmed the presence of a neutral EPS with high thermal stability. Additionally, EPS75 exhibited dose-dependent antioxidant activity, effectively reducing metal ions (Fe³⁺, Mo⁶⁺, and Mn⁷⁺) and stabilizing radicals (ABTS^•+, HO^•, O₂^•−, and HOO^•). The biopolymer also demonstrated immunostimulatory and anti-inflammatory effects in RAW 264.7 cells. In vivo assays using Balb/c mice indicated that both EPS75 and FM75 prevented constipation, suggesting their potential as natural and safe agents for constipation-related disorders. Due to its viscosifying and health-promoting attributes, CRL75 offers promising applications for functional dairy products. Full article

Fermentation is one of the oldest food processing techniques and is widely utilized in dairy product processing, during which nutrient availability and bioactive compounds are altered. However, the complete mode of action by which fermented dairy exerts beneficial effects on the host remains unknown. The present study investigated the effect of milk and yogurt ingestion alone or combined with prebiotic inulin on the transcriptome of colonic mucosa, liver, and femur in healthy rats. Young growing male rats were fed one of four experimental diets containing (1) skimmed milk, (2) skimmed milk supplemented with inulin (5% w/w), (3) yogurt, or (4) yogurt supplemented with inulin (5% w/w) for 6 weeks. Microarray results revealed that yogurt consumption resulted in 2195 upregulated differential expressed genes (DEGs) and 1474 downregulated DEGs in colonic mucosa as compared with milk consumption. According to Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, tight junction-, immune system-related pathways in the colonic mucosa and metabolic pathways in the liver were enriched with yogurt consumption. No evident differences were identified in the bone transcriptome between the diet groups. In conclusion, the study found that the intake of fermented dairy exerts more pronounced effects on gene expression in the intestinal tissue than prebiotics supplementation. Full article

Background/Objectives: Intestinal alkaline phosphatase (IAP) is an enzyme expressed in the intestinal brush border, which may exert anti-inflammatory effects by detoxifying lipopolysaccharides (LPSs), thereby preventing metabolic disorders. Various food components have been reported to influence IAP activity. However, few studies have evaluated the effects of fermented milk on IAP activity. In this study, we aimed to investigate fermented milk with high IAP-activating capacity and investigate its effect. Methods: We screened a skim milk culture (SC), a fermented milk model, using differentiated Caco-2 cells. We investigated the effect of SC on IAP activity and gene expression in the Drosophila midgut. Quantitative PCR and immunoblot assays were conducted to examine gene and protein levels. Results: Among the SC samples from different lactic acid bacteria or bifidobacteria, the SC of Lactobacillus johnsonii SBT0309 (LJ0309 SC) demonstrated a particularly strong capacity to activate IAP in Caco-2 cells, demonstrated by significantly increased IAP gene expression and protein levels in Caco-2 cells. Additionally, LJ0309 SC inhibited increased secretion of IL-8 in LPS-stimulated Caco-2 cells. Finally, in Drosophila melanogaster fed LJ0309 SC, we observed an increase in both IAP activity and gene expression in the midgut. Conclusions: LJ0309 SC increased IAP activity and gene expression in both Caco-2 cells and the Drosophila midgut, and inhibited the inflammatory response in LPS-stimulated Caco-2 cells. Although further in vivo studies are required, LJ0309 SC might help to ameliorate LPS-induced inflammation and disease via IAP activation. Full article

The biotechnological production of carotenoids offers a promising alternative to their chemical synthesis or extraction from plants. Mycolicibacterium species have shown potential as pigment-producing microorganisms. However, bacterial strains typically exhibit lower productivity compared to fungal and yeast strains. Earlier, we enhanced the β-carotene biosynthesis in M. neoaurum strain VKM Ac-3067D by modifying the cultivation medium. Key changes included replacing glucose with glycerol and soybean meal with skimmed milk powder (SMP) and increasing the urea content from 0.5 to 1.0 g/L. To further optimize β-carotene yield, a steepest ascent method was applied combining factorial design with a gradient-based optimization (Wilson–Box method). The resulting regression model showed that the most influential factors were the glycerol concentration and SPM use. The in-flask fermentation of the Ac-3067D strain in a medium containing 25.5 g/L of glycerol (carbon source) and 12.80 g/L of SMP (nitrogen source) increased β-carotene yield to 318.4 ± 8.3 mg/kg. In a 15 L bioreactor, β-carotene yield increased to 432.3 ± 10.4 mg/kg, while the biomass concentration reached 23.2 ± 1.2 g/L. The further scaling up to a 100 L bioreactor increased both β-carotene yield (450.4 ± 8.2 mg/kg) and biomass concentration (25.2 ± 1.1 g/L). Thus, β-carotene production technology using the M. neoaurum strain AC-3067D was successfully scaled up from 750 mL flasks to a 100 L bioreactor, confirming its potential for industrial-scale application. Full article

The quality of Sichuan paocai in natural fermentation is often inconsistent due to the complexity of its microbial community and environmental influences. To address this, dominant microbial strains were selectively inoculated to improve the product’s quality and safety. However, vacuum freeze-drying, commonly used to prepare direct vat set (DVS) starters, can significantly damage strains due to freezing stress. This study aimed to optimize a freeze-drying protection system for Lactiplantibacillus plantarum and Bacillus subtilis to enhance their survival. Using response surface methodology, combinations of cryoprotectants were evaluated. A formulation comprising skim milk powder, glycerol, sucrose, and L-proline significantly improved strain viability after lyophilization, outperforming single cryoprotectants. Further investigation into storage conditions revealed that low temperatures (−20 °C) provided the best preservation for DVS starters. Furthermore, the optimized DVS starters demonstrated excellent fermentation performance in Sichuan paocai, enhancing its color, flavor, and sensory quality compared to natural fermentation. These findings offer a reliable freeze-drying protection strategy for survival and viability of L. plantarum and B. subtilis. Full article

A high degree of hydrolysis can reduce the allergenicity of milk, while lactic acid bacteria (LAB) fermentation can further enhance the antioxidant ability of enzymatic milk. LAB with a strong antioxidant ability was screened, and the effects of LAB on the bitterness, taste and flavor of enzymatic skim milk (ESM) with a high degree of hydrolysis were investigated in this paper, in addition to the response surface methodology optimized the conditions of the LAB fermentation of ESM. The results indicate that the skim milk hydrolyzed by Protamex has a higher degree of hydrolysis and lower bitterness. The scavenging rate of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical, the inhibition rate of hydroxyl radical (·OH) and the superoxide dismutase (SOD) activity of Lactiplantibacillus plantarum 16 and Lactococcus lactis subsp. lactis m16 are significantly higher than those of other strains (p < 0.05), while the improvement effect of L. plantarum 16 on the bitterness and flavor of ESM is better than that of L. lactis subsp. lactis m16. The fermented ESM has a strong antioxidant ability and low bitterness when the inoculum quantity of L. plantarum 16 is 5%, fermentation at 37 °C for 18 h and the pH of the ESM is 6.5, for which the DPPH free radical scavenging rate is 61.32%, the ·OH inhibition rate is 83.35%, the SOD activity rate is 14.58 and the sensory evaluation is 4.25. The contents of amino acids related to bitterness and antioxidants were reduced and increased, respectively. The ESM fermented by L. plantarum 16 has a good flavor, antioxidant ability and low bitterness. Full article

The development of functional dairy products has increasingly become a focus of the dairy industry, with goat milk gaining prominence due to its nutritional properties and digestibility. This study aimed to evaluate the effects of freeze concentration processes on skimmed goat milk, observing its potential prebiotic effects and impacts on the physical, chemical, microbiological, rheological, and sensory profiles of fermented milk, using the Preferred Attributes Elicitation (PAE) methodology. Skimmed goat milk was initially concentrated using the gravitational block freeze concentration technique. A fermented milk containing probiotics (FM1) was produced from this concentrate. In addition, two other samples were developed: one with skimmed goat’s milk, 6% inulin, and probiotics (FM2) and another using whole goat’s milk with probiotics as a control (FM3). The results indicated that the freezing concentration process resulted in a concentrate with 14.70 ± 0.06 g 100 g⁻¹ of total solids. Among the three types of fermented milk, FM1 presented the highest values of total solids and titratable acidity. Regarding color, both FM1 and FM2 tended towards yellowish and greenish tones, while FM3 presented a greater luminosity. During storage, all fermented milks maintained their probiotic properties. The freeze concentration process increased the viscosity of FM1, a characteristic also evidenced in the sensory evaluations using PAE. In contrast, FM2 presented a rheological behavior similar to that of the control (FM3). Regarding sensory acceptance, FM1 had lower acceptance regarding aroma, being described as having notes of “goat flavor” and “acid” and being “salty”. The PAE methodology proved effective in characterizing the sensory qualities of the products, providing valuable information for developing new dairy products. These results offer an important theoretical basis for the industrial production of functional dairy products based on goat’s milk, helping to evaluate quality characteristics and optimize manufacturing processes. Full article

Cosmetically applicable soluble agonists for Toll-like receptor 2 (TLR2), which can strengthen skin barrier function, were produced by fermentation of asparagus (Asparagus officinalis L.) extract supplemented with skimmed milk using Lactobacillus delbrueckii subsp. lactis TL24. Their molecular size was estimated to be >100 kDa. Their TLR2-stimulating activity was stable over 1 year at 4 °C, but it decreased by more than 95% within 10 and 4 months at 25 °C and 40 °C, respectively. The possibility of stabilization of TLR2-stimulating activity by powdering was tested, and we found that lyophilization with 10% or a higher amount of dextrin could stabilize the activity even at 40 °C. The powdered fermented product dose-dependently stimulated TLR2. It augmented the formation of tight junctions in normal human keratinocytes, as detected by fluorescence staining of occludin and ZO-1, whereas their protein and gene expression levels did not increase, suggesting that a change in subcellular localization of these proteins without significant changes in their amounts might be responsible. The powder nature has some benefits over the aqueous, besides stability, e.g., it can be dissolved just before application, allowing fresh material to be used each time, and it may widen a range of cosmetic applications in non-aqueous types of cosmetics. Full article

A Leuconostoc mesenteroides strain (SJC113) isolated from cheese curd was found to produce large amounts of a mucoid exopolysaccharide (EPS). An analysis revealed the glucan nature of the EPS with 84.5% (1→6)-linked α-d-glucose units and 5.6% (1,3→6)-linked α-d-glucose units as branching points. The EPS showed 52% dextranase resistance and a yield of 7.4 ± 0.9 g/L from MRS medium supplemented with 10% sucrose within 48 h. Ln. mesenteroides SJC113 was also characterized and tested for the production of EPS as a fat substitute in fresh cheese. Strain SJC113 showed high tolerance to a wide range of NaCl concentrations (2, 5 and 10%), high β-galactosidase activity (2368 ± 24 Miller units), cholesterol-reducing ability (14.8 ± 4.1%), free radical scavenging activity (11.7 ± 0.7%) and hydroxyl scavenging activity (15.7 ± 0.4%). The strain had no virulence genes and was sensitive to clinically important antibiotics such as ampicillin, tetracycline and chloramphenicol. Ln. mesenteroides SJC113 produced highly viscous EPS during storage at 8 °C in skim milk with 5% sucrose. Therefore, these conditions were used for EPS production in skim milk before incorporation into fresh cheese. Four types of fresh cheese were produced: full-fat cheese (FF) made from pasteurized whole milk, non-fat cheese (NF) made from pasteurized skim milk, non-fat cheese made from skim milk fermented with Ln. mesenteroides without added sugar (NFLn0) and non-fat cheese made from skim milk fermented with Ln. mesenteroides with 5% sucrose (NFLn5). While the NF cheeses had the highest viscosity and hardness, the NFLn5 cheeses showed lower firmness and viscosity, higher water-holding capacity and lower weight loss during storage. Overall, the NFLn5 cheeses had similar rheological properties to full-fat cheeses with a low degree of syneresis. It was thus shown that the glucan-type EPS produced by Ln. mesenteroides SJC113 can successfully replace fat without altering the texture of fresh cheese. Full article

In three consecutive experiments, natural yogurt (NY) and fruit yogurt (FY) fortified with 5 and 10% skimmed milk powder (SMP) and 10% jam from black currant (BC), elderberry (EB), and their mixture of 1:1 (BCEB) were analyzed, and consumer acceptance was assessed. In Experiment 1, the effect of SMP (0, 5, and 10%) on selected physicochemical parameters of the NY was evaluated. With the increasing addition of SMP, a decrease in fat content (up to −19%) and, conversely, an increase in protein content (up to +82%) and viscosity were noted. Analyses of fruits and jams intended for yogurt fortification revealed a significantly higher vitamin C content in BC than in EB and higher anthocyanins in EB than in BC. In Experiment 2, NY with 5 and 10% SMP was fortified with 10% jams (BC or EB). A joint effect of SMP and the type of fruit jam was evaluated. The same trends in fat and protein contents as in Experiment 1 were detected. The sensory evaluation showed better acceptance of FY with 10% SMP and no differences between BC and EB perception. Thus, for Experiment 3, FY was prepared with only 10% SMP and 10% jam (BC, EB, BCEB). Significant differences were detected in active acidity and color measured in the CIELab system. These results were also confirmed in sensory evaluation. The overall acceptability showed that FY with different types of jam did not significantly differ. The launching of innovative fortified yogurt onto the market represents a promising way to increase the diversity of fermented dairy products with nutritionally desirable properties. Full article

Milk, on account of its abundant protein content, is recognized as a vital source of bioactive substances. In this study, the bioactive ingredients in milk were obtained by a combination of protease hydrolysis and fermentation with Lactobacillus plantarum. The compositions of protease hydrolysate (PM) and fermentation supernatant (FM) were determined, and their anti-oxidant and anti-bacterial activities were evaluated. Using LC-MS/MS, the molecular weights and sequences of the peptides were characterized, among which a total of 25 bioactive peptides were identified. The DPPH radical scavenging results demonstrated that FM exhibited an enhanced anti-oxidant capacity compared to PM. The bacterial survival rate results revealed that FM had a remarkable anti-bacterial ability compared to PM. Additionally, the anti-bacterial component and potential anti-bacterial mechanisms were determined. The results of cytoplasmic membrane depolarization, cell membrane permeability, and morphological observation indicated that FM could interact with bacterial membranes to achieve its anti-bacterial effect. These findings suggested that FM, as a bioactive substance of natural origin, holds potential applications in the functional food, pharmaceutical, and cosmetic industries. Full article

Fermented foods and ingredients, including furmenties derived from lactic acid bacteria (LAB) in dairy products, can modulate the immune system. Here, we describe the use of reconstituted skimmed milk powder to generate novel fermentates from Lactobacillus helveticus strains SC232, SC234, SC212, and SC210, and from Lacticaseibacillus casei strains SC209 and SC229, and demonstrate, using in vitro assays, that these fermentates can differentially modulate cytokine secretion via bone-marrow-derived dendritic cells (BMDCs) when activated with either the viral ligand loxoribine or an inflammatory stimulus, lipopolysaccharide. Specifically, we demonstrate that SC232 and SC234 increase cytokines IL-6, TNF-α, IL-12p40, IL-23, IL-27, and IL-10 and decrease IL-1β in primary bone-marrow-derived dendritic cells (BMDCs) stimulated with a viral ligand. In contrast, exposure of these cells to SC212 and SC210 resulted in increased IL-10, IL-1β, IL-23, and decreased IL-12p40 following activation of the cells with the inflammatory stimulus LPS. Interestingly, SC209 and SC229 had little or no effect on cytokine secretion by BMDCs. Overall, our data demonstrate that these novel fermentates have specific effects and can differentially enhance key immune mechanisms that are critical to viral immune responses, or can suppress responses involved in chronic inflammatory conditions, such as ulcerative colitis (UC), and Crohn’s disease (CD). Full article

The discovery and biotechnological application of new antimicrobial peptides are impeded by a lack of sensitive methods for peptide quantification. Paenibacillin is an emerging antimicrobial lantibiotic that was discovered in Paenibacillus polymyxa OSY-DF ATCC PTA-7852, isolated from the fermented vegetable Kimchee. This lantibiotic has potency against many foodborne pathogenic and spoilage bacteria. To advance the research and application of paenibacillin, a rapid, specific, and sensitive detection and quantification immunoassay was developed. After anti-paenibacillin polyclonal antibodies (pAbs) were generated and purified, a competitive enzyme-linked immunosorbent assay (cELISA) was developed and optimized for paenibacillin quantification. The dynamic range of the cELISA was determined by using a three-parameter nonlinear regression model, achieving a correlation (R²) value of 0.95. The cELISA displayed high sensitivity, with the ability to detect paenibacillin at levels as low as 15.6 ng/mL, which is significantly lower than the limit of detection of the conventional antimicrobial assay (20 µg/mL paenibacillin). The cELISA successfully differentiated paenibacillin concentrations in cell-free crude supernatants of P. polymyxa wild type and its mutant strain when grown at 30 °C and 37 °C; higher paenibacillin levels were found in the mutant (0.248–0.276 µg/mL) than in the wild type (0.122–0.212 µg/mL) culture. These findings were validated by the transcriptional analysis of 11 paenibacillin biosynthetic genes, which were significantly upregulated (≥2-fold increase) in the mutant compared with the wild strain. Additionally, the cELISA exhibited high sensitivity by recovery of paenibacillin titers spiked at 2.5 and 10 µg/mL in de Man, Rogosa, and Sharpe (MRS) broth and diluted skim milk. These results suggest that the anti-paenibacillin pAbs and the developed cELISA could be valuable in quantifying paenibacillin in complex matrices and in aiding the discovery of paenibacillin-producing natural microbiota. Full article

Lactobacillus plantarum has beneficial effects on the reduction of symptoms of poor lactose digestion and hypercholesterolemia, removal of the duration and severity of diarrheal processes, improvement of the intestinal permeability barrier, prevention of some types of cancer by adsorption or inactivation of genotoxic agents, increased resistance to intestinal and extraintestinal infections, attenuation of inflammatory bowel disease, and prevention of allergies (especially food). On the other hand, carao (Cassia grandis) has shown remarkable nutritious content with influential dietary applications. As a result, this investigation aimed to explore the effect of Cassia grandis pulp on viability of Lactobacillus plantarum under gastrointestinal conditions, immunomodulatory capacity, and probiotic potential. Adding carao to the medium under different experimental conditions, including rich and minimal culture media and a gastrointestinal digestion process of skimmed milk, did not substantially affect Lactobacillus plantarum’s growth but prolonged its viability. The administration of Lactobacillus plantarum with carao in mice did not induce a proinflammatory response at a systemic level. Still, it did cause an increase in the production of immunoregulatory cytokines. Also, the viability of TSB broth was improved by adding carao. Carao improved the growth of acid tolerance, bile tolerance, growth in TSB broth, and NaCl resistance. According to the results, carao may enhance the characteristics of L. plantarum when enriching fermented dairy products. Full article

Fermented foods have long been known to have immunomodulatory capabilities, and fermentates derived from the lactic acid bacteria of dairy products can modulate the immune system. We have used skimmed milk powder to generate novel fermentates using Lb. helveticus strains SC234 and SC232 and we demonstrate here that these fermentates can enhance key immune mechanisms that are critical to the immune response to viruses. We show that our novel fermentates, SC234 and SC232, can positively impact on cytokine and chemokine secretion, nitric oxide (NO) production, cell surface marker expression, and phagocytosis in macrophage models. We demonstrate that the fermentates SC234 and SC232 increase the secretion of cytokines IL-1β, IL-6, TNF-α, IL-27, and IL-10; promote an M1 pro-inflammatory phenotype for viral immunity via NO induction; decrease chemokine expression of Monocyte Chemoattractant Protein (MCP); increase cell surface marker expression; and enhance phagocytosis in comparison to their starting material. These data suggest that these novel fermentates have potential as novel functional food ingredients for the treatment, management, and control of viral infection. Full article