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Pediococcus acidilactici

Pediococcus acidilactici is a of Gram-positive, facultatively anaerobic bacterium belonging to the family within the order Lactobacillales. It is characterized by its spherical (cocci) morphology, typically arranged in pairs or tetrads, and is catalase-negative and mesophilic, with optimal growth temperatures between 25°C and 50°C. As a homofermentative organism, it primarily produces L(+)- from , enabling it to thrive in acidic environments and tolerate conditions such as 4.0–8.0 and up to 6.5% NaCl. This bacterium plays a pivotal role in as a starter culture for processes, contributing to the preservation, , and of various products. It is commonly employed in the production of fermented meats like and sausages, where it facilitates rapid acidification and inhibits spoilage organisms. Additionally, P. acidilactici is used in vegetable fermentations such as and , as well as in dairy items like cheese and bio-yogurts, enhancing sensory attributes through compounds like and . Its production of , notably pediocin PA-1, provides biopreservation by targeting Gram-positive pathogens, including , thereby extending shelf life and improving food safety. Beyond food applications, certain strains of P. acidilactici demonstrate potential, surviving gastrointestinal transit and offering health benefits such as inhibition, reduction, and modulation. Studies have shown its efficacy in improving growth and immunity in species like and calves, as well as alleviating disorders in animal models. Furthermore, it exhibits antimicrobial activity against pathogens like and supports applications in additives to enhance disease resistance.

Taxonomy and characteristics

Classification

Pediococcus acidilactici is classified within the domain Bacteria, phylum Bacillota, class Bacilli, order Lactobacillales, family Lactobacillaceae, genus Pediococcus, and species acidilactici. This taxonomic placement reflects its position among Gram-positive, lactic acid-producing bacteria, with the phylum Bacillota encompassing low-GC-content Firmicutes. The genus Pediococcus was proposed by N.H. Claussen in 1903 based on isolates from beer fermentation processes, where these bacteria were observed causing spoilage through acid production. The species P. acidilactici was formally described earlier by P. Lindner in 1887, drawing from strains identified in brewing contexts for their ability to produce lactic acid from glucose, and it was later validated under the International Code of Nomenclature of Bacteria in 1980. The name "Pediococcus" derives from the Greek "pedion" (plane surface) and "kokkos" (berry or grain), alluding to the characteristic flat, tetrad arrangement of the coccal cells that divide in two perpendicular planes. Similarly, "acidilactici" is a New Latin term indicating production of lactic acid (from "acidum lacticum"). Phylogenetically, P. acidilactici clusters closely with other in the family, particularly within the broader group, based on 16S rRNA and whole-genome analyses that highlight shared evolutionary adaptations to nutrient-poor, acidic environments. It is distinguished from genera like , which typically form rods or chains, by its unique tetrad-forming and homofermentative metabolism. This relation underscores the ecological overlap among these bacteria in fermented substrates.

Morphology and physiology

Pediococcus acidilactici is a , that typically occurs in pairs or tetrads, with cells measuring approximately 1.5–2.0 μm in . The bacterium is non-motile, non-spore-forming, and catalase-negative, characteristics that distinguish it within the group. These morphological traits enable its identification through standard microbiological techniques, such as and , where tetrad formation is particularly evident in solid media. The species exhibits facultative anaerobic metabolism and thrives under a range of suited to fermented and intestinal niches. Optimal occurs at temperatures between 37°C and 40°C, with tolerance extending up to 45°C in many strains, though some can briefly withstand higher heat during processing. It prefers a slightly with an optimal of 5.5–6.5 but demonstrates acid tolerance down to pH 4.0, facilitating survival in gastric-like conditions. Additionally, P. acidilactici is -tolerant, with viable cells persisting in 0.3% bile salts for several hours, a trait essential for potential transit through the gut. Physiologically, P. acidilactici is homofermentative, primarily producing L(+)- from via the Embden-Meyerhof pathway, resulting in nearly homolactic yields under optimal conditions. This metabolic profile contributes to its aciduric nature, allowing persistence in low-pH environments better than many other , with survival rates exceeding 70% at pH 2.5 for select strains after short exposures. The bacterium also exhibits strong to intestinal mucosa, mediated by surface proteins that promote to epithelial cells like monolayers, enhancing colonization potential. Furthermore, it maintains viability at longer than typical lactobacilli, supporting stability in non-refrigerated formulations. Notable strains include GR-1, which has been evaluated in human clinical studies for modulating and reducing bioavailability, demonstrating robust physiological resilience . For food applications, strain NCIMB 30005 is utilized in processes due to its heat and stability, contributing to consistent production in and products.

Habitat and ecology

Natural environments

Pediococcus acidilactici is commonly found in environments, where it has been isolated from various samples demonstrating its ability to persist in terrestrial habitats. It also inhabits plant surfaces, such as grains, contributing to the natural microbial communities on . In animal gastrointestinal tracts, including the of , cecum of chickens, pigs, and rabbits, P. acidilactici forms part of the native in healthy, free-ranging animals. Although it can transiently colonize the gut during short-term interventions, it is not a dominant member of the intestinal . This bacterium thrives in acidic and low-oxygen conditions, exhibiting tolerance to low pH levels and facultative metabolism, which enable its survival in oxygen-limited niches. It has been isolated from naturally fermented materials, such as . In microbiomes, it contributes to modulation. P. acidilactici exhibits a worldwide distribution, with frequent isolations from temperate regions associated with agriculture-prone areas, reflecting its adaptation to and -associated ecosystems.

Role in fermentation processes

Pediococcus acidilactici plays a pivotal role in spontaneous and traditional fermentation processes, primarily as a lactic acid bacterium that drives acidification through the homolactic fermentation of carbohydrates, producing DL-lactic acid and lowering pH to below 4.5, which inhibits spoilage organisms and enhances food preservation. In vegetable fermentations, such as pickles and sauerkraut, it contributes to rapid pH reduction (e.g., to 3.6) by metabolizing glucose and fructose, fostering an anaerobic environment suitable for long-term storage. Similarly, in kimchi production, it accelerates the fermentation of cabbage, balancing acidity with flavor development. In meat fermentations, particularly dry sausages, P. acidilactici accelerates acidification to levels around 4.5–5.0, improving , color , and by curbing pathogenic growth, as demonstrated in studies on traditional European salami production. For dairy products like artisanal cheeses, it acts as a non-starter during , with isolates from cheeses enhancing sensory qualities. These processes rely on its to low and , allowing dominance in mixed microbial consortia. P. acidilactici often co-ferments with and species, where it complements heterofermentative pathways by providing consistent production, thus stabilizing the dynamics in sourdoughs and vegetable mixes. It also produces exopolysaccharides () during growth, which improve and viscosity in fermented foods; for instance, strain BCB1H yields up to 271.5 mg/L EPS under optimized conditions, acting as a natural thickener in and vegetable products. Strains are commonly isolated from artisanal cheeses, , sourdoughs, and traditional Asian products like Thai fermented (e.g., Plasom), where P. acidilactici F3 was recovered from local fish ferments. Historically, it has been integral to pre-industrial in (e.g., fish and soy ferments) and (e.g., cheese and sausages) since ancient times, predating formalized .

Metabolic and genetic properties

Fermentation pathways

Pediococcus acidilactici is an obligate homofermentative bacterium that metabolizes carbohydrates primarily through the Embden-Meyerhof-Parnas (EMP) glycolysis pathway, converting glucose to with high efficiency. In this process, one molecule of glucose is catabolized to two molecules of pyruvate, which are then reduced to DL-, regenerating NAD⁺ and yielding a net gain of 2 ATP molecules per glucose consumed. The overall balanced equation for this homolactic fermentation is: \ce{C6H12O6 -> 2 CH3CH(OH)COOH} This pathway ensures that over 90% of the fermented substrate is directed toward lactic acid production under optimal conditions, with minimal byproducts such as CO₂. The bacterium exhibits efficient utilization of hexose sugars, including glucose and fructose, as primary carbon sources via the phosphoenolpyruvate:phosphotransferase system for uptake. Pentose sugars, such as ribose, arabinose, and xylose, are fermented to a limited extent, often resulting in additional products like ethanol and acetate alongside lactate, reflecting a partial shift toward heterolactic metabolism for these substrates. Overall, CO₂ production remains minimal during hexose fermentation due to the dominance of the homofermentative route. Acid production by P. acidilactici occurs rapidly during the phase, typically acidifying culture media to a of 3.5–4.0 within 24 hours at optimal temperatures of 30–40°C. This kinetic profile supports its role in quick environmental acidification, with strains achieving up to 0.4% from 0.5% initial glucose in nutrient-rich broths. While primarily homofermentative, P. acidilactici can exhibit adaptive shifts toward heterolactic under environmental stresses such as nutrient limitation or high acidity, producing small amounts of CO₂ and other metabolites to maintain balance. This flexibility, though secondary to its core metabolism, enhances survival in fluctuating conditions like those in fermented environments.

Bacteriocin production and genetics

_Pediococcus acidilactici produces several , primarily pediocins belonging to the class IIa group of unmodified, heat-stable with molecular weights under 10 kDa. The most studied is pediocin PA-1, a 44-amino-acid that exhibits potent antilisterial activity by disrupting the cytoplasmic of target cells, particularly against Gram-positive pathogens such as , , and other foodborne spoilage organisms like . These share a conserved N-terminal YGNGV essential for specificity and a flexible C-terminal domain for receptor binding. The genetic basis of pediocin PA-1 production is encoded by the ped operon, comprising four genes: pedA (structural gene for the pre-pediocin), pedB (immunity protein), pedC (accessory protein aiding disulfide bond formation), and pedD (ATPase for precursor processing and secretion). This operon is typically plasmid-borne in producer strains, such as pSMB74 in P. acidilactici PAC1.0, facilitating horizontal transfer and strain variability. Transcription is regulated by quorum sensing mechanisms involving two-component systems, where the mature pediocin acts as an autoinducer to activate expression at high cell densities, ensuring coordinated production during late exponential growth. Bacteriocin production is optimized under specific environmental conditions, with maximal yields achieved at 30–37°C and an initial of 6.0–6.2 in MRS broth, where activity can reach up to 320 AU/mL after 18–24 hours of incubation. Glucose serves as a primary carbon source, supporting both growth and synthesis alongside production. The complete of P. acidilactici typically spans 1.9–2.1 with a G+C content of approximately 42%, featuring CRISPR-Cas systems (often Type II-A) that confer phage resistance by targeting invading viral DNA. Recent sequencing of the probiotic strain BCB1H in 2025 revealed a 1.92 harboring clusters for and other beneficial traits, including exopolysaccharide production genes.

Industrial and probiotic applications

Food industry uses

Pediococcus acidilactici serves as a key starter culture in the production of dry fermented sausages, such as , where it facilitates to develop characteristic flavors and aromas through the production of organic acids and volatile compounds. Strains like P. acidilactici PE1 enhance sensory attributes, including and overall acceptability, while reducing indicators of spoilage such as volatile basic and lipid oxidation during ripening. By rapidly lowering pH and inhibiting pathogens like , it improves and allows for reduced levels in formulations, minimizing reliance on chemical preservatives. In dairy processing, P. acidilactici acts as an adjunct culture in , particularly for varieties like cheddar, where it promotes the breakdown of proteins and fats to generate free (e.g., , ) and (e.g., acetic and butyric acids) that contribute to mature flavor profiles. Strains such as AS185 support uniform acidification and increase total free to levels exceeding 4,000 mg/kg after 90 days of without adversely affecting cheese or sensory qualities. Additionally, it metabolizes like serine to produce compounds such as and α-aminobutyrate, further enriching the biochemical diversity during extended maturation. For vegetable processing, P. acidilactici is employed as a starter in brine fermentations for products like and , ensuring consistent acidification and preventing spoilage by off-flavors from undesirable microbes. Its tolerance to high salt (up to 6.5%) and concentrations enables rapid conversion of sugars to , achieving stable levels in and cabbage fermentations. In biopreservation, commercial strains such as P. acidilactici 1.0 produce pediocin PA-1, a that extends by targeting Gram-positive spoilers and pathogens in various foods, including meats and ready-to-eat products. This antimicrobial peptide inhibits Listeria monocytogenes growth by up to 3 log cycles in vacuum-packaged sausages during refrigerated storage, often in combination with other hurdles like low temperature. The strain's freeze-drying stability facilitates its incorporation into formulations for enhanced preservation without synthetic additives. P. acidilactici is also utilized in production, where inoculants like strain G24 accelerate lactic in high-carbohydrate forages, reducing by 12-14% and preserving crude protein content compared to uninoculated controls. These applications contribute to the global fermented food market, valued at over $788 billion in 2025 and projected to reach $1,122 billion by 2032, by enabling efficient, safe production of traditional and commercial products.

Probiotic formulations

Pediococcus acidilactici strains selected for use, such as GR-1 and MA18/5M, are evaluated for their ability to survive gastrointestinal , with a minimum viability threshold of greater than 10^6 CFU/g required to ensure efficacy in formulations. To enhance survival in acidic environments, these strains are often encapsulated using materials like whey protein isolate and , which protect against low pH and improve delivery to the gut. Probiotic formulations of P. acidilactici are delivered in various forms, including capsules, fermented dairy products like , and animal feeds, allowing for targeted human or veterinary applications. Typical dosages for human consumption range from 10^8 to 10^9 CFU per day, while veterinary uses employ higher levels, such as 10^9 to 10^10 CFU per kg of feed, to support animal health. Commercially, P. acidilactici is incorporated into synbiotic products combining the bacterium with prebiotics like galactooligosaccharides or to enhance microbial activity, and it has been a key component in veterinary for poultry gut health since the 1980s, as seen in products like Bactocell. Maintaining viability poses challenges, with most formulations requiring at to preserve counts over extended periods; however, advancements in spray-drying encapsulation developed after 2020 have enabled room-temperature stable strains that retain over 7 log CFU/g for at least three weeks at 25°C.

Health benefits

Gastrointestinal effects

Pediococcus acidilactici strains demonstrate potential for relieving by modulating and promoting short-chain (SCFA) production. In a 2021 preclinical study on loperamide-induced constipated mice, of four P. acidilactici strains (CCFM18, CCFM28, NT17-3, 102H8) improved gastrointestinal rates (up to 55.10% for CCFM28, P < 0.05) and reduced the time to first defecation (from 6.21 h to 5.38–6.10 h, though P > 0.05 overall), with bacteriocin-producing strains showing greater regulation of harmful bacteria like and . This alleviation was linked to increased abundance of SCFA producers such as (4- to 21-fold) and (1- to 37-fold). The bacterium also aids in preventing diarrhea, particularly by inhibiting enteric pathogens in gut environments. In weaned piglets challenged with Shiga toxin-producing Escherichia coli, dietary P. acidilactici supplementation (0.1%) reversed pathogen-induced depletion of beneficial Lactobacillus, preserved microbial diversity, and reduced diarrhea incidence through enhanced beneficial genera like Prevotella and Succinivibrio. In vitro assessments confirm antimicrobial activity against pathogens including Salmonella enterica and Listeria monocytogenes via bacteriocin production from strains like Kp10, supporting inhibition of E. coli adhesion and growth in simulated gut conditions. P. acidilactici enhances gut barrier integrity, which contributes to overall digestive health. In vitro studies using fish intestinal epithelial cells showed that pre-treatment with P. acidilactici MA18/5M upregulated tight junction protein ZO-1 expression (P < 0.05) and maintained transepithelial electrical resistance against Vibrio challenge, while in vivo supplementation in Chinook salmon increased mucin 2 expression (P < 0.05). A 2024 human clinical trial in patients with diarrhea-predominant or mixed irritable bowel syndrome (IBS) reported that a probiotic formulation including P. acidilactici CECT 7483 reduced IBS severity scores by a mean of 146.6 points (P < 0.0001), with 62.9% of participants achieving ≥50% symptom improvement, alongside enhanced quality of life. Additionally, P. acidilactici supports gastrointestinal detoxification of via modulation. A 2022 randomized, double-blind trial in 152 workers exposed to occupational found that 12 weeks of containing P. acidilactici GR-1 reduced blood levels by 34.45% (from 1246 to 817 μg/L, P < 0.0001) and by 38.34% (from 4.855 to 2.994 μg/L, P < 0.0001), primarily through increased fecal excretion and elevated SCFA levels (e.g., P = 0.0322) that bolstered gut and .

Immune modulation

Pediococcus acidilactici exerts immune modulatory effects through its production of , such as pediocin PA-1, which inhibit colonization in animal models. In a model of infection, purified pediocin PA-1 reduced fecal bacterial counts by up to 2 log units and eliminated the from the liver and within 6 days. Similarly, strains of P. acidilactici isolated from equine sources reduced serovar Typhimurium loads in the gut of infected by enhancing antibacterial activity and modulating immune responses. These contribute to broader immune enhancement by activating natural killer () cells; for instance, P. acidilactici supplementation improved cell activity in a cyclophosphamide-induced model, increasing cytotoxic potential against target cells. P. acidilactici shows promise as a by amplifying immune responses to antigens. A 2025 in chickens demonstrated that supplementation with P. acidilactici (PediGuard) alongside enhanced production and immune modulation, including elevated IgA and IgG levels, suggesting synergistic effects on . This potential is linked to (TLR) signaling pathways, as like P. acidilactici upregulate TLR expression to boost innate immune activation and antigen-specific responses in preclinical models. The bacterium exhibits properties by downregulating pro-inflammatory in inflammatory models. In a dextran sulfate sodium (DSS)-induced mouse model, a synbiotic containing P. acidilactici and significantly reduced TNF-α levels in serum (P < 0.05), alleviating histopathological damage and imbalances. A 2024 human trial evaluating a cocktail containing P. acidilactici in healthy volunteers confirmed its safety, with no significant changes observed in inflammation markers. In animal health applications, P. acidilactici mitigates stress-induced in by supporting immune . Supplementation in broilers preserved status and immune function, countering oxidative damage and suppression associated with environmental stressors. This strain has been utilized in veterinary feeds since the , with commercial products like BACTOCELL demonstrating consistent benefits in enhancing resistance to infections and stress-related immune deficits.

Other therapeutic potentials

Research has explored the potential of Pediococcus acidilactici-derived compounds, particularly like pediocin PA-1, in exhibiting anti-cancer effects against cells. In vitro studies demonstrate that pediocin PA-1 induces and in HT-29 human colon cells, leading to reduced cell viability through membrane disruption and (ROS) generation. A strain of P. acidilactici isolated from traditional cheese has also shown cytotoxic activity, inducing via regulation of Bax/ pathways in doxorubicin-resistant cells and reducing viability by approximately 75%. In contexts, P. acidilactici serves as a in veterinary applications to mitigate stress in animals. A 2024 study on dogs under transport stress found that supplementation with strain GLP06 improved composition, reduced levels, and alleviated physiological stress responses, supporting its use in companion . Additionally, produced by P. acidilactici, such as pediocin NV5, exhibit inhibitory effects against Streptococcus mutans, the primary pathogen in dental caries, with moderate antimicrobial zones indicating potential for oral health formulations to prevent formation. Preliminary research links P. acidilactici to neurological benefits through gut-brain axis modulation. In a model of chronic unpredictable mild stress, strain CCFM1344 reduced anxiety-like and depression-like behaviors by restoring balance, increasing beneficial genera like , and suppressing via lowered pro-inflammatory cytokines (e.g., TNF-α, IL-1β) in the . A 2025 clinical extension suggests similar potential in humans for mood disorders, though animal models provide foundational evidence for anxiety reduction.

Safety and regulation

Safety assessments

Pediococcus acidilactici has been recognized as (GRAS) by the U.S. (FDA) for use in since the establishment of the GRAS notification in 1997, with specific affirmation through GRAS Notice No. 171 for its application as an antimicrobial agent in and products. Sequenced strains of P. acidilactici, including those used in and contexts, exhibit no hemolytic activity and lack virulence genes such as those encoding invasins or toxins, confirming their microbiological safety profile. Toxicity evaluations demonstrate low , with a 2025 PLOS One study reporting no in mice administered up to 10^9 CFU/kg body weight of P. acidilactici NMCC-B, alongside no sub-chronic effects at higher doses of 2 × 10^9 CFU/kg over 28 days. Genomic analyses further reveal that antibiotic resistance genes in P. acidilactici strains are either absent or located on non-transferable elements, minimizing risks of in the gut . Regarding allergenicity, P. acidilactici poses a low risk, as confirmed by a 2024 study showing no production, including , in strains isolated from fermented foods, which supports its suitability for sensitive populations. While generally safe, rare cases of opportunistic infections, including bacteremia and , have been reported, primarily in immunocompromised patients or those with underlying conditions. Clinical trials in healthy populations show no infections up to 10^10 CFU/day. Strain-specific assessments highlight safety for industrial applications; for instance, P. acidilactici strains SY21 and SY22, evaluated in 2025, showed no hemolytic activity, absence of production, and tolerability in processes without adverse microbial interactions. In human consumption, rare side effects may include mild gastrointestinal symptoms like , similar to other , typically resolving within days.

Regulatory status

Pediococcus acidilactici has been included on the European Food Safety Authority's (EFSA) list of Qualified Presumption of Safety (QPS) biological agents since the establishment of the QPS process in 2007, allowing its use in food and feed applications without case-by-case safety assessments for qualified strains. Specific strains, such as CNCM I-4622, have been authorized as a feed additive under EU Regulation (EC) No 1831/2003 for use in animal nutrition, including aquaculture, following EFSA safety evaluations. In the United States, the Food and Drug Administration (FDA) has granted Generally Recognized as Safe (GRAS) status to multiple strains of P. acidilactici for applications including antimicrobial control in meat products and as a probiotic ingredient in foods. Internationally, the Commission recognizes , including Pediococcus species, as safe starter cultures for fermented milk products under standards such as CXS 243-2003. For veterinary applications, P. acidilactici is regulated as a feed additive under (EC) No 1831/2003, with authorizations for use in animal nutrition, including , requiring demonstration of safety for target species, consumers, and the environment. Labeling requirements for probiotic products containing P. acidilactici mandate declaration of viable cell counts in colony-forming units (CFU) per serving to ensure accurate consumer information on potency. Under EU Regulation (EC) No 1924/2006, any health claims related to probiotics must be strain-specific, authorized by the European Commission following EFSA assessment, with updates post-2020 emphasizing substantiated evidence for efficacy. Restrictions apply to the use of P. acidilactici in formulas, where requires specific pre-market approval in both the and due to vulnerability of this population; the FDA has issued warnings against unapproved use in preterm s, citing risks of . In applications, ongoing monitoring for resistance is mandated under feed additive regulations to prevent environmental dissemination.

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