Search Results (270)

Chickpea flour represents a valuable plant-based ingredient due to its high protein and fiber content; however, its application is limited by antinutritional factors and off-flavor compounds. Fermentation with LAB and yeasts, applied individually or in consortia, resulted in significant microbiological, nutritional, and aromatic changes. The fastest acidification (pH 3.9) and the most effective control of Enterobacteriaceae (<4 log CFU/g after 48 h) were observed in samples containing Lactiplantibacillus plantarum LP23, both as a monoculture and in combination with Debaryomyces hansenii Y15A. Peptide content significantly increased in all fermented samples compared to the control, with a synergistic effect in the co-culture Yarrowia lipolytica Y3 + Lacticaseibacillus paracasei L (around 230%). A pronounced reduction in raffinose-family oligosaccharides was observed, especially in the consortia Y. lipolytica Y3 + Lcb. paracasei L and D. hansenii Y15A + Lacp. plantarum LP23 (0.11–0.16 mmol/100 g). Samples with lower total volatile levels showed higher olfactory acceptability due to a marked reduction in aldehydes (up to 70–95% vs. control), and a balanced accumulation of alcohols, esters, ketones, and organic acids. Overall, LAB–yeast consortia effectively enhanced the nutritional quality, safety, and sensory properties of chickpea flour, supporting its use as a functional ingredient in plant-based foods. Full article

Lactic acid bacteria (LAB) are pivotal microorganisms in the food industry. Current approaches for functional gene validation and trait improvement in LAB primarily rely on traditional gene editing and homologous recombination techniques. These methods are often cumbersome, inefficient, and time-consuming, hindering the rapid and precise customization of strains. This limitation has, to some extent, constrained the rapid selection and industrial application of functional LAB strains. The engineering of LAB through gene editing technologies has significantly advanced both fundamental and applied research. Among these, CRISPR/Cas gene editing has successfully achieved precise modification of multiple genes in various LAB species. Compared to conventional methods, it offers superior editing efficiency and lower operational costs, opening new avenues for functional gene identification and genetic improvement in LAB. However, the application of exogenous CRISPR/Cas systems in LAB faces technical challenges such as high off-target rates, chromosomal abnormalities, and cytotoxicity. The development of endogenous CRISPR/Cas-based editing tools for LAB provides novel pathways for precise regulation, rational design, and flexible application. This paper first outlines the structural components and mechanistic principles of CRISPR/Cas gene editing tools. It then explores the research progress and applications of both endogenous and exogenous CRISPR/Cas systems in LAB. Finally, it provides an outlook on the future application of CRISPR/Cas gene editing technology in LAB, offering a reference for its implementation in this field. The advent of gene editing technologies has significantly propelled functional gene validation and trait improvement in lactic acid bacteria (LAB), thereby advancing both fundamental research and industrial applications. Notably, the CRISPR/Cas system has emerged as a transformative tool enabling precise genetic modification in diverse LAB species, offering marked improvements in editing efficiency and cost reduction relative to conventional approaches. CRISPR/Cas-based editing strategies in LAB are broadly classified into exogenous and endogenous systems. Exogenous systems operate independently of the host’s native immune repertoire, conferring the advantages of broad strain applicability and high editing efficiency. These systems have been successfully deployed for functional gene characterization, metabolic pathway engineering, such as augmenting antimicrobial production, and probiotic safety enhancement via virulence gene deletion. Conversely, endogenous systems leverage the intrinsic CRISPR/Cas machinery of LAB, offering superior biocompatibility and minimized off-target risks. Notable applications include precise gene knockout and integration using the native Type I-E system in Lacticaseibacillus paracasei. This review provides a concise overview of CRISPR/Cas system architecture and mechanisms, followed by a systematic synthesis of research progress and applications for both exogenous and endogenous systems in LAB. Finally, future directions are outlined to guide the continued development and application of CRISPR/Cas technologies in this field. Full article

During the last few decades, an urgent need for sustainable and health-promoting food products has been witnessed. In this vein, the development of functional foods enriched with probiotics has gained considerable interest from both the food industry and consumers. However, the maintenance of high cell viability until the time of consumption remains a significant challenge. In this study, freeze-dried immobilized Lacticaseibaciilus paracasei FBM_1327 cells on sunflower seeds were evaluated as a functional food ingredient, and their ability to survive during simulated digestion and storage at ambient and refrigerated temperatures in comparison to free cells was assessed. Cell immobilization resulted in higher survival rates (>70%) after in vitro digestion compared to free cells (<40%), while the freeze-dried immobilized cells maintained in cell levels >7.5 log cfu/g during storage for 6 months at 4 °C. In the next step, freeze-dried free or immobilized cells were incorporated in cream cheese (CCF and CCI samples, respectively) at a concentration of >8 log cfu/g. Cell viability of the immobilized cells remained stable (>8.1 log cfu/g) during storage, while live cell counts of free cells dropped to 7.51 ± 0.11 log cfu/g after 28 days. The fortification of cream cheese with immobilized L. paracasei FBM_1327 cells on sunflower seeds improved the volatile compounds profile, while all samples were accepted by the panel during the sensory evaluation. Full article

This study presents a genome-resolved shotgun metagenomic analysis of artisanal raw-milk goat cheese from the Masurian region of Poland, addressing the limited understanding of strain-level diversification and functional restructuring during traditional cheese ripening. While microbial succession in cheese has been widely described, comprehensive genome-resolved analyses integrating strain-level genomic heterogeneity, pathway reweighting, and mobile genetic elements in artisanal goat cheese remain scarce. By combining taxonomic profiling with metagenome-assembled genome (MAG) reconstruction and pathway-level functional analysis, we characterised microbial succession and genome plasticity across ripening stages. Genome reconstruction yielded 37 MAGs during early ripening and 141 MAGs in mature cheese, revealing increased genome recoverability and pronounced strain-level heterogeneity within dominant taxa, including Lactiplantibacillus plantarum, Lacticaseibacillus paracasei, and Lactococcus lactis. Alpha diversity increased in mature samples, consistent with progressive community restructuring. Functional profiling demonstrated coordinated metabolic reweighting, particularly within carbohydrate metabolism, while amino acid and lipid metabolism remained proportionally stable. Genome-resolved analyses further identified tetracycline- and sulfonamide-associated resistance determinants and diverse bacteriophages targeting lactic acid bacteria, highlighting the role of mobile genetic elements in horizontal gene transfer and microevolutionary adaptation during ripening. Full article

This study evaluated the probiotic, technological, and safety properties of 124 lactic acid bacteria isolated from traditional Beyaz cheese, and the in situ survival of selected strains in fermented milk. Eighteen isolates showing over 80% tolerance in simulated gastric juice (pH 3.0) were subjected to further characterization. On the basis of 16S rRNA gene sequencing, most isolates belong to Lacticaseibacillus paracasei and Lactiplantibacillus plantarum, while Lactobacillus helveticus, Lentilactobacillus kefiri, and Limosilactobacillus fermentum were also identified. L. plantarum EH140 showed the greatest resistance to the simulated gastric environment (pH 2.0), whereas L. paracasei EH131 exhibited the highest bile salt tolerance. L. plantarum EH106 demonstrated strong auto-aggregation, and L. fermentum EH132 displayed notable hydrophobicity. Nine isolates exhibited bile salt hydrolase activity, but none showed γ-hemolysis, gelatinase, or DNase activity. All the isolates were susceptible to ampicillin, erythromycin, clindamycin, and chloramphenicol. Organic acid analysis revealed lactic acid as the major metabolite, followed by acetic acid. Virulence gene screening identified the efaAfs gene only in L. paracasei strains, and no biogenic amine genes were detected. The selected isolates maintained viability above 6 log CFU/mL in milk during storage. Overall, L. plantarum EH106, EH109, EH140, and EH141 were identified as the most promising candidates because of their safety and superior probiotic potential. Full article

This experiment was designed to determine the effect of the Lacticaseibacillus paracasei (paracasei) strain NSMJ27, isolated from Korean fermented vegetable food (home-made kimchi), on laying performance, egg quality, intestinal histology, cecal short-chain fatty acids, and ileal antioxidant/immunity indicators of laying hens. Ninety-six 55-week-old Hy-Line Brown hens were randomly assigned to two dietary treatments with each treatment comprising eight replicates of six hens each. Experimental diets were prepared by mixing corn and soybean meal basal diets without or with L. paracasei NSMJ27 at 2.5 × 10⁹ CFU/kg. The experiment lasted 4 weeks. Laying hens fed with the NSMJ27-supplemented diet were not affected (p > 0.05) in their laying performance or egg quality. With respect to ileal morphology, villus height: crypt depth ratio tended to be higher (p = 0.067) in laying hens fed with L. paracasei NSMJ27 vs. control diets. Dietary L. paracasei did not affect (p > 0.05) the activities of glutathione peroxidase and catalase, malondialdehyde contents, or secretory immunoglobulin A in ileal mucosa, but increased (p = 0.048) the activity of superoxide dismutase compared with the control diet-fed laying hens. Dietary L. paracasei elevated (p = 0.016) the relative percentage of butyrate but lowered (p = 0.057) that of isovalerate in cecal digesta. Dietary L. paracasei did not affect the percentages of cells expressing macrophages, B cells, CD4⁺ T cells, CD8⁺ T cells, or TCRγδ T cell surface markers (p > 0.05). Overall, these results suggest that dietary L. paracasei NSMJ27 could enhance gut health via increasing gut antioxidant capacity and butyrate production in the cecal digesta of laying hens. Full article

The growing demand for high-quality gluten-free bread requires innovative technological and functional approaches. This study investigates, for the first time, the use of Lacticaseibacillus paracasei SP5 in free and immobilized form on traditional Greek trahanas for producing GF sourdough bread based on rice and buckwheat flour. Sourdough breads produced with free and immobilized cells displayed greater performance than the control sourdough bread in microbial stability and physicochemical properties, with the immobilized form showing the best results. In particular, the application of immobilized L. paracasei SP5 led to gluten-free sourdough bread with enhanced acidification (pH 4.55; total titratable acidity 12.9 mL) and remarkable lactic (2.20 g/kg) and acetic acid (0.76 g/kg) levels, extending the shelf life to 7.0 days against mold and 7.5 days against rope spoilage, statistically significantly higher than the other two gluten-free sourdough samples. It also showed the strongest antifungal activity and improved technological characteristics, including higher loaf volume (2.84 mL/g), greater height (5.50 cm), and lower baking loss (17.42%). Total phenolic content (87.3 mg GAE/100 g) and antioxidant activity were also statistically significantly increased. Overall, immobilized L. paracasei SP5 on trahanas appears to be a promising clean-label strategy to improve the quality, shelf life, and functional value of GF bread. Full article

Background/Objectives: This study investigated the systemic metabolic effects of two probiotic strains, Lacticaseibacillus paracasei MG5012 and Bifidobacterium animalis subsp. lactis MG741, on metabolic dysfunction-associated steatotic liver disease (MASLD) and obesity-related muscle dysfunction in high-fat-diet (HFD)-induced obese mice. Methods: Obesity was induced in C57BL/6 mice via high-fat diet (HFD) feeding for 6 weeks. Subsequently, the mice were orally administered MG5012 or MG741 for 8 weeks. We assessed systemic metabolic parameters, including body weight, adiposity, and serum biomarkers. Additionally, histological and molecular analyses were performed to evaluate hepatic steatosis, intestinal barrier integrity, and muscle oxidative status. Results: Both strains significantly attenuated body weight gain and adiposity, reduced serum liver injury markers (γ-GTP, ALT, AST), and improved systemic metabolic parameters by restoring serum GLP-1 levels and reducing hyperinsulinemia. Crucially, MG5012 and MG741 strengthened intestinal barrier integrity by upregulating the tight junction proteins Occludin and Claudin-1. In the liver, histological analyses revealed reductions in hepatic steatosis and triglyceride content, accompanied by the downregulation of lipogenic genes (SREBP-1c, FAS). Furthermore, the probiotics preserved skeletal muscle integrity; while muscle weight remained unchanged, the strains increased muscle fiber cross-sectional area (CSA) and reduced serum markers of muscle damage (CPK, LDH). This protective effect was associated with significantly enhanced expression of antioxidant enzymes (SOD, CAT, GPx) in muscle tissue. Conclusions: These findings suggest that MG5012 and MG741 confer systemic metabolic benefits through the modulation of the gut–liver–muscle axis and may serve as promising functional food ingredients for the management of MASLD and obesity-associated muscle atrophy. Full article

Evidence suggests that gut dysbiosis may contribute to acute myocardial infarction (AMI) and its complications, including reduced physical performance and muscle weakness. We hypothesized that probiotic supplementation could improve muscle strength during post-AMI recovery. In a randomized, controlled, triple-blind clinical trial, adults and older adults undergoing myocardial revascularization received either a multistrain probiotic formulation (Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, Lacticaseibacillus acidophilus, and Bifidobacterium lactis) or placebo for 90 days. The primary outcome was handgrip strength (HGS). Forty-five participants completed the study. No significant between-group differences were observed in the main analysis. However, in an exploratory subgroup of men aged 50 years and older with low baseline HGS (n = 30), probiotic supplementation led to a greater improvement in non-dominant HGS after 90 days compared with placebo (mean difference: +4.6 kg/f; p = 0.04). A baseline-adjusted ANCOVA confirmed a significant baseline-by-treatment interaction for the non-dominant hand (β = +0.33; 95% CI: +0.02 to +0.62; p = 0.038), indicating greater improvements among participants with lower initial strength. Although the primary analysis yielded null results, these exploratory findings indicate a potential benefit of probiotic supplementation in a clinically vulnerable subgroup of revascularized men with low baseline strength. Larger and prospectively powered trials are warranted to confirm these observations. Trial registration: RBR-6ztyb7. Full article

Identifying probiotics that modulate the gut–brain axis is vital for non-pharmacological Alzheimer’s disease (AD) therapy. Through a staged screening from transgenic Drosophila to a D-galactose/AlCl₃-induced murine model, Lactobacillus acidophilus LA4 and Lacticaseibacillus paracasei F5 were prioritized for their ability to improve climbing indices and reduce Aβ deposition and AChE activity. In AD mice, LA4 and F5 significantly ameliorated cognitive deficits and anxiety-like behaviors. Mechanistically, both strains reduced hippocampal Aβ_1–42 and p-Tau levels, inhibited AChE, suppressed pro-inflammatory cytokines (TNF-α, IL-6, IL-1β), and enhanced antioxidant enzymes (SOD, GSH-Px). 16S rRNA analysis revealed restored Firmicutes/Bacteroidetes ratios and enrichment of SCFA-producers (Muribaculaceae, Dubosiella). Metabolomics highlighted remodeled purine and arginine pathways, with strain-specific effects on primary bile acid biosynthesis/sphingolipid metabolism (LA4) and butanoate metabolism/nicotinate and nicotinamide metabolism (F5). Consequently, LA4 and F5 alleviate AD pathology by restructuring microbial and metabolic profiles, thereby mitigating neuroinflammation and oxidative stress. These findings confirm the potential of specific probiotics as functional food ingredients for the prevention and adjuvant treatment of neurodegenerative diseases. Full article

Fermented products have recently garnered substantial interest in both research and commercial contexts. Although probiotic fermentation is predominantly practiced with dairy, fruits, vegetables, and grains, its application to dual-purpose food-medicine materials like Ganoderma lucidum has been comparatively underexplored. In this study, Ganoderma lucidum fermented juice (GFJ) served as the substrate and was fermented with five probiotic strains. The optimal inoculation ratios—determined by employing a uniform design experiment—were as follows: Bifidobacterium animalis 6.05%, Lacticaseibacillus paracasei 9.52%, Lacticaseibacillus rhamnosus 6.63%, Pediococcus pentosaceus 21.38%, and Pediococcus acidilactici 56.42%. Optimal fermentation parameters established by response surface methodology included 24 h of fermentation at 37 °C, a final cell density of 5 × 10⁶ CFU/mL, and a sugar content of 4.5 °Brix. Experiments with RAW264.7 macrophages revealed that GFJ significantly promoted both phagocytic activity and nitric oxide (NO) secretion, indicating enhanced immune characteristics as a result of fermentation. Untargeted metabolomics profiling of GFJ across different fermentation stages showed upregulation of functional metabolites, including polyphenols, prebiotics, functional oligosaccharides, and Ganoderma triterpenoids (GTs)—notably myricetin-3-O-rhamnoside, luteolin-7-O-glucuronide, raffinose, sesamose, and Ganoderma acids. These increments in metabolic compounds strongly correlate with improved functional properties in GFJ, specifically heightened superoxide dismutase activity and immunomodulatory capacity. These results highlight an effective approach for developing functionally enriched fermented products from medicinal fungi, with promising applications in functional food and nutraceutical industries. Full article

Fortifying sourdough bread with functional ingredients is an effective strategy to enhance nutritional value, bioactive content, and sensory quality. The novel strain Lactiplantibacillus paracasei SP5 was incorporated into mother sponges together with cereal-based derivatives, such as trahanas or delignified wheat bran (2% and 5% w/w), and their effects on sourdough bread functionality were compared with a control bread without supplementation. Sourdough bread containing 5% w/w trahanas exhibited the highest lactic acid bacteria (10.4 log cfu/g), approximately 22% higher than the control, and the strongest fermentation activity, leading to acetic acid levels 27% higher than the control. This sourdough bread sample also had 73% higher total phenolic content (112.1 mg GAE/100 g) and approximately 27% higher antioxidant activity compared to control bread. In addition, phytic acid degradation exceeded 91%, representing a 16% higher reduction than the control, thereby improving mineral bioavailability. Shelf-life was also markedly extended, in terms of mould and rope spoilage compared with the control. Wheat bran-supplemented breads showed moderate improvements, with LAB counts, antioxidant activity, and phytic acid reduction about 10–25% higher than the control. Overall, these findings demonstrate that L. paracasei SP5 in combination with nutrient-rich, low-cost cereal-based derivatives can significantly enhance the technological, nutritional, and bioactive properties of sourdough breads. This approach provides a simple, industrially feasible strategy for producing functional breads with improved health-promoting and preservative attributes. Full article

With growing emphasis on antibiotic-free poultry production, functional probiotics represent a promising strategy to improve gut health and reduce pathogen transmission. This study characterized three lactic acid bacteria (LAB) strains Lactobacillus delbrueckii subsp. bulgaricus NRRL-B-548 (LD), Lacticaseibacillus paracasei DUP-13076 (LP), and Lacticaseibacillus rhamnosus NRRL-B-442 (LR) for their probiotic potential and evaluated their efficacy against Salmonella enterica in poultry. The LAB strains were assessed for acid and bile tolerance, lysozyme resistance, cholesterol assimilation, antimicrobial activity, surface hydrophobicity, epithelial adherence, hemolysis, and antibiotic susceptibility. Genomic analysis was performed to identify genes associated with probiotic functionality. The protective potential of LR and LP was further validated in hatchlings using a hatchery spray model challenged with Salmonella Enteritidis. All strains survived simulated gastric and intestinal conditions, exhibited strong adhesion to epithelial cells, and demonstrated high hydrophobicity, indicating robust colonization capacity. The LAB significantly inhibited Salmonella Enteritidis, S. Typhimurium, and S. Heidelberg growth in vitro and remained sensitive to clinically relevant antibiotics. In vivo application of LR and LP to hatching eggs markedly reduced S. Enteritidis colonization in the liver, spleen, and ceca of hatchlings. Further, genomic profiling of the LAB strains revealed genes for bacteriocin production, exopolysaccharide synthesis, and carbohydrate metabolism supporting probiotic function. In summary, the evaluated LAB strains exhibit multiple probiotic attributes and strong anti-Salmonella activity, confirming their potential as safe, hatchery-applied probiotics for improving gut health and biosecurity in poultry production systems. Full article

Background/Objectives: Impaired intestinal barrier function is a hallmark of inflammatory bowel disease (IBD). Akkermansia muciniphila and its outer membrane protein Amuc_1100 can enhance this barrier, but the clinical application of Amuc_1100 is limited by the fastidious growth of its native host. This study aimed to overcome this by utilizing the robust probiotic Lacticaseibacillus paracasei L9 for targeted Amuc_1100 delivery. Methods: We engineered Lc. paracasei L9 to express Amuc_1100 via intracellular (pA-L9), secretory (pUA-L9), and surface-display (pUPA-L9) strategies. Their efficacy was assessed in Lipopolysaccharide (LPS)-induced macrophages and a dextran sulfate sodium (DSS)-induced colitis mouse model, evaluating inflammation, barrier integrity, and mucosal repair. Results: The secretory (pUA-L9) and surface-display (pUPA-L9) strains most effectively suppressed pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) in macrophages. In mice, both strains alleviated colitis and outperformed native A. muciniphila in improving disease activity. Crucially, they exhibited distinct, specialized functions: pUA-L9 acted as a systemic immunomodulator, reducing pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), elevating anti-inflammatory mediators (IL-4 and IL-10), and promoting goblet cell differentiation; notably, the inhibitory effect of pUA-L9 on IL-6 expression was approximately 2-fold greater than that of pUPA-L9. In contrast, pUPA-L9 excelled in local barrier repair, uniquely restoring mucus layer integrity (Muc1, Muc2, and Tff3) and reinforcing tight junctions (ZO-1, Occludin, Claudin1, Claudin3, and Claudin4). In particular, pUPA-L9 increased Muc2 expression by approximately 3.6-fold compared with pUA-L9. Conclusions: We demonstrate that the subcellular localization of Amuc_1100 within an engineered probiotic dictates its therapeutic mode of action. The complementary effects of secretory and surface-displayed Amuc_1100 offer a novel, spatially targeted strategy for precision microbiome therapy in IBD. Full article

Inoculated fermentation can enhance the flavor, nutrition, and functionality of juice. The lactic acid bacteria (LAB) are commonly used as starter cultures. This study screened LAB for orange juice fermentation and optimized fermentation factors using response surface methodology (RSM) to improve GABA content in orange juice. A total of 52 LAB strains were screened, and Lacticaseibacillus paracasei ZY (Lcb. paracasei ZY) and Lacticaseibacillus rhamnosus SN12 (Lcb. rhamnosus SN12) presented higher GABA yields and adaptability to substrates. The optimized fermentation factors for GABA enhancement in orange juice were as follows: initial pH of 5.5, fermentation temperature of 37 °C, soluble solids content of 12.0 °Bx, inoculum ratio of Lcb. paracasei ZY to Lcb. rhamnosus SN12 as 1:1, inoculum size of 6 Log CFU/mL, and fermentation time of 96 h. Under these optimized conditions, the GABA content reached 0.89 g/L, representing a 39.06% increase compared to uninoculated orange juice. This indicates that RSM-based optimization is conducive to increasing GABA content in orange juice and provides a scientific basis for the development of GABA-enriched functional fermented juices. Full article