Postbiotics as natural antimicrobials: a novel biocontrol strategy for food safety

Postbiotics are preparations of inanimate microorganisms and/or their components, with or without associated metabolites or cell structures, that confer health benefits to the host. Compared to probiotics, postbiotics offer enhanced safety, stability, and processing benefits for food applications. Their antimicrobial activity is mainly attributed to compounds like bacteriocins, organic acids, enzymes, and peptides, which can inhibit spoilage and pathogenic microorganisms and suppress biofilms. Lactic acid bacteria–derived cell-free supernatants and similar metabolite-rich preparations have been increasingly applied to foods to control foodborne pathogens and spoilage. This review highlights the emerging role of postbiotics as biocontrol agents in food safety, emphasizing their antimicrobial properties, while also outlining their definition, development, legal status, and current evidence on functionality.

1 Introduction

Foods are sources of macro and micronutrients that support body functions and health. They are mainly consumed as cellular substrates for energy, cell differentiation and proliferation, as well as serving as the foundation for chemical barriers against cellular oxidation (Munteanu and Schwartz, 2022). Non-nutritional factors such as fiber, phytochemicals, antioxidants, vitamins, minerals, probiotics, prebiotics etc. benefit the host’s physiology. The investigation of these non-nutrient factors is of great importance for various reasons, including enhancing health, preventing diseases, addressing nutritional deficiencies, and increasing consumer awareness. Functional foods are one of the main categories that focus on these factors and have gained significant attention in today’s world (Granato et al., 2020; Essa et al., 2023).

Functional foods are foods that beyond their nutritional effects beneficially influence specific body functions, contributing to improved health, well-being, and reduced disease risk. They can be natural or modified through technological or biotechnological means by adding or removing ingredients, with effects evident at consumption levels typical of a regular diet (European Commission, 2010). In addition, randomized, double-blind, placebo-controlled clinical trials are necessary to establish the functional efficacy of foods before making health claims. Furthermore, it has been underlined that no fresh, unprocessed or processed food can be considered functional without appropriate clinical research and substantial experimental evidence confirming its toxicological safety and functionality (Granato et al., 2020). Dietary interventions that modulate the microbiota include various fermented foods and fiber-rich diets, along with probiotics, prebiotics, synbiotics, and postbiotics, all of which are interrelated within the same conceptual framework (Mills et al., 2019). The International Scientific Association for Probiotics and Prebiotics (ISAPP) has set out the definitions of these terms in its consensus statements (Figure 1).

Figure 1

Figure 1. Definition of the International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement. (1. Hill et al., 2014; 2. Gibson et al., 2017; 3. Swanson et al., 2020; 4. Marco et al., 2021; 5. Salminen et al., 2021).

Postbiotics may exhibit stronger stability than probiotics and prebiotics due to their non-replicable abiotic activities (Ma et al., 2023). Hence, postbiotics have emerged as promising candidates for functional food development, offering practical benefits such as improved transport and storage conditions, while also enhancing product safety and supporting targeted health benefits (Isaac-Bamgboye et al., 2024; Wei et al., 2024). Moreover, postbiotics have been associated with a variety of health-promoting activities, such as antioxidant, anti-inflammatory (Rezaie et al., 2024), antihypertensive (Bartolomaeus et al., 2019), antiobesogenic (Seo et al., 2022), immunomodulatory (Yilmaz, 2024), antiproliferative (Nowak et al., 2022) and hypocholesterolemic (Bhat and Bajaj, 2019) activities. These properties suggest that postbiotics may contribute to host health by supporting specific physiological functions; however, the underlying mechanisms by which they exert these effects remain to be fully elucidated. While current findings are promising, they should be interpreted with caution and not generalized to all postbiotic compounds, given the diversity in their composition and sources (Aguilar-Toalá et al., 2018).

Despite advanced technology comprehensive quality control and improved sterilization methods, pathogens continue to survive in the environment by developing resistance, forming biofilms, and using other survival mechanisms, even with multiple preventive measures in place (Newell et al., 2010; Bridier et al., 2015). Microbial intervention refers to the general, non-specific inhibition or elimination of a microorganism by other members within the same environment. Biocontrol, on the other hand, is based on the ability of certain agents to carry out microbial intervention. It involves using one or more organisms to suppress or manage others. In contrast to many control methods, biocontrol techniques originate from natural and environmentally friendly sources (Bale et al., 2008). Lactic acid bacteria (LAB) (Linares-Morales et al., 2018; Tarifa et al., 2023), bacteriocins (McManamon et al., 2019; Chen et al., 2022), endolysins (Cho et al., 2021; Li et al., 2023) and bacteriophages (Ayaz et al., 2018; Cufaoglu and Ayaz, 2019; Goncuoglu et al., 2021) have been commonly used in food biocontrol studies due to their effectiveness against foodborne pathogens. In this context, postbiotics, defined as a preparation of inanimate microorganisms and/or their components, stand out as promising contributors to biocontrol strategies. This is due to their complex composition, which includes bioactive compounds and cellular structures capable of exerting antimicrobial effects without the need for viable cells (Salminen et al., 2021). In recent years, research on the definition, production, and potential applications of postbiotics has been steadily increasing, showing that these agents may serve as innovative biocontrol agents (Aggarwal et al., 2022). However, as postbiotics represent a chemically diverse group of compounds synthesized by various microorganisms, it cannot be generalized that every postbiotic exhibits antimicrobial or preservative properties (Isaac-Bamgboye et al., 2024). This review explores postbiotics and postbiotic compounds with documented or potential antimicrobial activity, with a particular focus on their roles as biocontrol agents in foods and their implications for food safety.

2 From where to where?

The consumption of live bacterial cultures by human’s dates back to the times when ancient Egyptian and Middle Eastern civilizations, where fermentation was used to extend the shelf life of food. The modern history of probiotics began in the early 1900s with the pioneering work of Elie Metchnikoff, who introduced the concept of “pro-bios” (later known as probiotics) describing lactobacilli as beneficial bacteria that support the host’s health, unlike antibiotics (Gasbarrini et al., 2016; Krawczyk and Banaszkiewicz, 2021). In the early 20th century, French scientists conducted a study that initiated the development and commercialization of postbiotics. The research team, led by Dr. Pierre Boucard, demonstrated the therapeutic potential of heat-treated Lactobacillus-containing feces in alleviating symptoms of digestive diseases. Their findings showed that non-living microbial preparations could still retain bioactive properties (Liévin-Le Moal, 2016). In the late 20th century, advances in microbiology and immunology have led scientists to focus on bioactive compounds produced by probiotics during fermentation, such as short-chain fatty acids (SCFA), enzymes, peptides, and cell wall components. The term “postbiotic” has gained more importance in the 21st century, parallel to scientists’ efforts to distinguish inanimate microbial products from probiotics and prebiotics. In 2013, a consensus report was published on the proper definition and use of term “probiotic” was published, which indirectly increased interest in postbiotics (Hill et al., 2014). By 2021, the definition of postbiotics had become clearer, highlighting their bioactive role in health and disease management, as they are derived from well-defined microorganisms (Salminen et al., 2021) (Figure 1).

Probiotics are widely used in food products; however, their application is limited in some cases due to challenges such as maintaining cell viability, interactions with matrix compounds, and potential adverse effects in high-risk individuals (Gurunathan et al., 2023). Postbiotic is a term derived from the words “post” (after) and “bios” (life). In addition, the family of “biotic” terms, such as probiotics, prebiotics, synbiotics and postbiotics, is structured around microorganisms or their substrates.

In this context, the term postbiotic refers to substances derived from microorganisms that are “inanimate” which means lifeless and refers to killed microorganisms without implying a loss of function (Salminen et al., 2021). Moreover, ISAPP (Salminen et al., 2021) excludes purified microbial metabolites from the definition of postbiotics. Compounds such as butyric acid, lactic acid, or bacteriocins (once purified and separated from cellular biomass) are defined as chemically characterized substances with independent biological effects. While these compounds may accompany postbiotic preparations and contribute to their functionality, they cannot be considered postbiotics on their own.

3 Why Postbiotics?

Postbiotics are defined as “a preparation of inanimate microorganisms and/or their components that confers a health benefit on the host” (Salminen et al., 2021). In this context, the term postbiotic refers to a preparation derived from well-characterized microorganisms in which the cells have been deliberately inactivated, but retain structural integrity and/or cellular components, together with any associated metabolites produced during growth or fermentation. This “inanimate” state indicates that the microorganisms are no longer viable, without implying a loss of biological activity or functionality. According to the ISAPP consensus, an effective postbiotic must therefore contain non-viable microbial cells and/or cell components (cell wall structures, pili or other surface molecules etc.), with or without co-existing metabolites, and the health effect must be demonstrated for this specific preparation (Salminen et al., 2021). By contrast, ISAPP explicitly excludes substantially purified microbial metabolites from the definition of postbiotics. Preparations consisting only of chemically well-defined molecules such as isolated short-chain fatty acids, purified lactic or butyric acid, bacteriocins, exopolysaccharides or other individual metabolites, in the absence of accompanying cellular biomass, are not considered postbiotics but should be referred to by their specific chemical names. These compounds may be present within a complex postbiotic preparation and contribute to its overall activity, yet when they are purified and separated from microbial cells or cell fragments, they fall outside the postbiotic concept. In line with this view, cell-free supernatants or filtrates that contain metabolites but no detectable cell structures are regarded as metabolite-based preparations rather than postbiotics in the strict ISAPP sense, unless they demonstrably include defined inanimate cells or cell components from a characterized microbial source. Components of a postbiotic may contain intact inanimate microbial cells and/or microbail cell fragments/structures with or without metabolites. On the other hand, formulations consisting solely of highly purified microbial metabolites, without any accompanying cellular material, would fall outside the scope of the postbiotic definition. Unlike live microorganisms, postbiotics offer an inherently safer profile due to the absence of viable cells, thereby minimizing risks such as translocation, uncontrolled proliferation, or horizontal gene transfer (Moradi et al., 2020). Their long shelf life, stability across broad pH and temperature ranges, and resistance to degradation during processing provide distinct advantages for use in both the food and pharmaceutical sectors (Liu et al., 2023). From an industrial perspective, postbiotics eliminate the need for cold-chain logistics and support cost-effective transportation, while also allowing incorporation into thermally processed products without loss of functionality (Wegh et al., 2019). Although challenges remain regarding large-scale production and standardization, the combination of technological resilience and commercial feasibility makes postbiotics a promising class of compounds for functional food and health-related applications (Thorakkattu et al., 2022).

Understanding the interplay between prebiotics, probiotics, and postbiotics is essential for contextualizing the role of postbiotics in health and food systems. Prebiotics are nutrients that support the growth of probiotic bacteria and positively change the composition of the intestinal microbiota. In simple terms, prebiotics contribute to the formation of probiotics, and probiotics contribute to the formation of postbiotics (Li and Luo, 2025). Unlike microorganisms, postbiotics do not contain live microorganisms, which makes them a safer and more stable alternative in both food and pharmaceutical applications. The absence of live bacteria reduces the risks associated with probiotic viability, antibiotic resistance gene transfer, and the production of undesirable metabolic by-products (Binda et al., 2020; Salminen et al., 2021). Additionally, postbiotics are not affected by host variability, meaning their effects are more predictable compared to probiotics, whose survival and efficacy can depend on an individual’s gut microbiota composition and overall health (Zhang et al., 2022; Fang et al., 2025). Compared to live probiotics, the use of postbiotics can be more easily controlled and standardized (Żółkiewicz et al., 2020).

From a technological standpoint, the application of live probiotic or starter cultures in food matrices poses several challenges due to their sensitivity to environmental stresses and potential incompatibilities with other ingredients (Liu et al., 2023). As an alternative, the use of postbiotics can help prevent unfavorable interactions between primary and secondary starters and food components. Their high thermal stability, compatibility with processing conditions, and broad-spectrum antimicrobial activity mediated by bioactive compounds enable their effective integration into food systems (Moradi et al., 2020; Żółkiewicz et al., 2020). The synergistic effects between these compounds further enhance their functional properties, making postbiotics a natural preservative in food technology (Tong et al., 2025). Moreover, due to the increasing threat of antibiotic resistance, postbiotics have the potential to emerge as alternative agents in the fight against drug-resistant pathogens. The WHO underlines the urgent need for novel antimicrobial strategies, while its 2023 report emphasizes that despite limited progress, the current global antibiotic pipeline remains insufficient to address the escalating threat of antimicrobial resistance (WHO, 2023). The potential for postbiotics to be used in clinical settings, particularly in the management of inflammatory and metabolic diseases, is an exciting area of ongoing exploration (Zhang et al., 2022; Ali et al., 2023). Advances in bioprocessing technology and precision fermentation are expected to improve the scalability and cost-effectiveness of postbiotic production, making them more accessible for widespread use (Thorakkattu et al., 2022). The advantages of postbiotics are summarized in Figure 2.

Figure 2

Figure 2. Advantages of postbiotics.

Despite their advantages, postbiotics are not entirely free from safety concerns. The mere absence of viability does not inherently guarantee safety, particularly in preparations derived from Gram-negative bacteria, where lipopolysaccharides (LPS, endotoxins) released from dead cell walls may trigger adverse effects such as sepsis or toxic shock (Periti and Mazzei, 1998). Therefore, specific safety assessments are essential for each postbiotic formulation prior to use. Moreover, clinical and preclinical data on postbiotics remain limited; randomized controlled trials evaluating their efficacy in both children and adults are scarce and often show considerable heterogeneity in quality (Salminen et al., 2021). Notably, while some clinical studies have evaluated the effects of heat-inactivated strains such as Lactobacillus rhamnosus GG and Lactobacillus acidophilus in children, the evidence remains inconclusive (Szajewska et al., 2014). A 2015 systematic review found no clear additional benefits of fermented formula over standard infant formula; however, it did not document any negative health effects, and some potential improvements in gastrointestinal symptoms could not be excluded (Szajewska et al., 2015). These findings underscore the importance of cautious interpretation of postbiotic efficacy data and highlight the need for well-designed clinical trials to better understand their safety and functional outcomes. On the other hand, the commercialization of postbiotics for food applications remains limited, primarily due to regulatory uncertainties and the need for further research on their long-term safety and efficacy. While numerous in vitro studies have demonstrated their benefits, more in vivo and clinical trials are required to confirm their effects in human populations (Moradi et al., 2020; Vinderola et al., 2022). Additionally, the scalability of postbiotic production presents challenges, as optimizing fermentation conditions and ensuring consistent bioactive compound yield remain areas for further technological development (Aguilar-Toalá et al., 2018). A further complicating factor is the structural diversity and compositional complexity of postbiotic preparations, which makes it difficult to establish a single “gold standard” analytical method for their characterization (Salminen et al., 2021). Therefore, the development of analytical standards in this field should remain flexible and adaptable, taking into account the inherent variability in postbiotic structure and composition. Considering all these factors, while postbiotics stand out in terms of stability and safety, probiotics are still regarded as a practical and familiar option in the production of functional foods aimed at supporting human health.

4 Source

Postbiotics are generally derived from some known microorganisms and their components. As indicated by ISAAP (Salminen et al., 2021), an inactivated microorganism does not need to originate from a previously classified probiotic strain in order to be considered a postbiotic. For instance, fermented foods produced with undefined and mixed microbial communities cannot serve as suitable sources for postbiotic preparations. In contrast, postbiotics may be obtained from fermented products formulated with well-characterized, defined microbial strains. Lactobacillaceae family members and certain species of the Bifidobacterium genus are common sources of inanimate strains used in postbiotic formulations, as they belong to well-established probiotic taxa frequently studied for their health-promoting properties (Cicenia et al., 2014; Andresen et al., 2020). A large part of their probiotic effect is due to bacterial components or metabolites. In addition to Lactobacillus and Bifidobacterium, other genera such as Streptococcus, Leuconostoc, Eubacterium, Pediococcus, Enterococcus, and Saccharomyces are also prominent in the production postbiotic-associated components (Ma et al., 2023; Prajapati et al., 2023). Beyond these genera, researchers are also exploring microorganisms that are not classified as probiotics or whose live use is limited due to safety and health concerns as potential sources of postbiotics. These include bacterial species such as Akkermansia muciniphila, Faecalibacterium prausnitzii, Eubacterium hallii, and heat-killed strains of Mycobacterium manresensis, as well as the yeast Saccharomyces boulardii (Liu et al., 2020).

In the food industry, postbiotics are generally obtained through fermentation. Microbial strains such as Lactobacillus, Bifidobacterium, Streptococcus, Akkermansia, Eubacterium, and Saccharomyces contribute not only to the fermentation of traditional foods (such as kefir, kombucha, sourdough bread, yogurt, and pickled vegetables) but also to the formation of inactivated microbial cells and their associated components, which may qualify as postbiotics when properly characterized (Collado et al., 2019; Cufaoglu and Erdinc, 2023; Asqardokht-Aliabadi et al., 2024). While these microorganisms also enhance the sensory and physicochemical qualities of fermented products, natural fermentation lacks control over the consistency and quantity of bioactive postbiotic compounds. This limitation has driven interest in standardized, industrial-scale fermentation as a means to reliably produce preparations that contain both inanimate microorganisms and their beneficial cellular or metabolic components, in alignment with the accepted postbiotic definition (Thorakkattu et al., 2022). Industrial fermentation thus provides a sustainable and reproducible platform for developing postbiotic-based applications in the food, pharmaceutical, and nutraceutical sectors. During fermentation, for instance, Bifidobacterium and Lactobacillus produce bioactive substances such organic acids, peptides, and EPSs. These components can be utilized in functional foods and dietary supplements to promote health and well-being (Wegh et al., 2019; Rafique et al., 2023). To maximize their potential, various methods are applied to extract these bioactive compounds and enhance their stability. Thermal (e.g. pasteurization, sterilization, ohmic heating) and non-thermal (e.g., pulsed electric fields, ultrasound, irradiation, supercritical CO₂) methods are widely employed for microbial inactivation in postbiotic production. These technologies help preserve the integrity and functionality of bioactive compounds, thereby enhancing the overall efficiency and effectiveness of postbiotics. In particular, innovative processes such as high hydrostatic pressure, ultraviolet rays, and pulsed light are emerging as advanced and sustainable technologies that hold great promise for reliable, large-scale postbiotic production (Pimentel et al., 2023).

5 Classification of postbiotics

Postbiotics can be categorized according to their structural characteristics, elemental composition, microbiota-derived metabolites, and physiological benefits (Figure 3). This categorization is critical for comprehending the health impacts of postbiotics and assessing their diverse uses. According to their structural features, they may contain components such as peptides, teichoic acids and plasmalogens. They can also be classified based on the chemical nature of the metabolites generated from the microbiota; SCFA, EPS, cell wall components, enzymes/proteins and other metabolites. They can also be grouped according to the elements they contain: proteins (lactocepin, p40, p75 molecules), organic acids (propionic acid, acetic acid, lactic acid), lipids (butyrate, dimethyl acetyl derivative plasmalogens), carbohydrates (teichoic acids, galactose-rich polysaccharides), vitamins/cofactors (B group vitamins), other complex molecules (lipoteichoic acids, peptidoglycan-derived muropeptides). Furthermore, postbiotics can be classified according to physiological benefits, such as anti-obesity, antioxidant, anti-inflammatory, hypocholesterolemic, anti-allergenic, anti-microbial, anti-mold, anti-cancer, anti-hypertensive, immunomodulatory and anti-proliferative (Aguilar-Toalá et al., 2018; Thorakkattu et al., 2022; Rafique et al., 2023; Asqardokht-Aliabadi et al., 2024).

Figure 3

Figure 3. Classification of postbiotics by structure, composition, metabolites, and benefits.

6 Regulatory challenges and market potential

The food industry generally uses preservatives to ensure product safety and extend shelf life. However, modern consumers’ growing skepticism toward additives has accelerated the demand for more natural and healthier alternatives. In this context, postbiotics are gaining significant interest particularly in food biopreservation, biofilm control, and pathogen inhibition (Silva et al., 2018a; Moghanjougi et al., 2020). As their applications continue to expand, regulatory authorities play an important role in assessing the safety of postbiotics. Institutions such as the European Food Safety Authority (EFSA) and the U.S. Food and Drug Administration (FDA) conduct safety assessments of the use of postbiotics. In Europe in particular, safety assessments of postbiotics are rigorously conducted. On the other hand, in countries such as South America, Japan and Brazil, regulations on postbiotics are rapidly evolving, which is accelerating the market acceptance of products (Vinderola et al., 2019; Salminen et al., 2021).

Beyond regulatory considerations, the functional properties of postbiotics further reinforce their growing importance in ensuring food quality and protection. LAB and their metabolites play a key role in enhancing microbial safety by inhibiting the growth of undesirable microbes. It has been demonstrated that particularly bacteriocins and organic acids are efficient at suppressing pathogens and preventing the production of biofilms (Gildea et al., 2022; Lima et al., 2024). For a commercial postbiotic product to maintain its bioactive properties and positive health effects, it is crucial to correctly identify the key molecules that provide these benefits. However, the commercialization of postbiotics remains a significant challenge due to ongoing research gaps in determining the target population, determining the appropriate dosage and method of administration, and identifying and isolating the safest and most effective strains (Gurunathan et al., 2023). Enhancing the yield of postbiotic production is a complex process that requires optimizing metabolic pathways, selecting appropriate probiotic strains, and carefully controlling fermentation conditions. Scaling up production from laboratory to industrial levels necessitates the development of innovative solutions while ensuring economic sustainability and maintaining product quality (Prajapati et al., 2023).

The regulatory frameworks for probiotics are based on the Generally Recognized as Safe (GRAS) and Qualified Presumption of Safety (QPS) lists established by the FDA and EFSA (Suez et al., 2019). However, new regulatory frameworks are needed to assess postbiotics in this context. The QPS status is determined at the species level, based on a history of safe use and sufficient evidence of human exposure through food (Scott et al., 2022). The QPS list, established by EFSA’s Biological Hazards Panel (BIOHAZ), is reviewed approximately every six months (EFSA, 2025). Notably, recent assessments have shown that safety evaluation of non-viable microorganisms such as potential postbiotics, may be more straightforward than for viable probiotics. For instance, Mycobacterium setense (killed bacteria + mannitol) and pasteurized Akkermansia muciniphila have undergone safety evaluation under the Novel Food Regulation (EU 2015/2283), following their QPS assessments (EFSA, 2024; EFSA, 2025). In the United States, although the FDA has not yet established a formal regulatory definition for postbiotics, several heat-inactivated microbial preparations have been assessed individually under existing regulatory pathways. Some inactivated microbial products derived from fermentation processes have been granted GRAS status, while others have been acknowledged under the New Dietary Ingredient (NDI) framework (FDA, 2024). The FDA’s 2025 Human Foods Program Guidance Agenda has indicated an intention to address novel microbial ingredients; however, a specific regulatory framework for postbiotics has yet to be finalized (FDA, 2025). On the other hand, until FDA and EFSA establish a regulatory framework specifically for postbiotics, further research is needed to determine appropriate safety and regulatory standards that can be applied to postbiotic preparations. Given these ongoing regulatory developments, it becomes increasingly important to establish clear production standards and characterization criteria to support the safe and consistent use of postbiotics across different industries.

7 Antimicrobial features of postbiotics in food safety

Food safety is threatened by various factors such as physical, chemical and biological hazards. Among biological hazards, bacteria are a major cause of food spoilage and foodborne illnesses (Mafe et al., 2024). In this context, postbiotics emerge as promising alternative antimicrobial agents against foodborne pathogens (Tong et al., 2025). The antimicrobial activity of postbiotics is primarily attributed to bioactive compounds such as bacteriocins, organic acids, fatty acids, peptides and H₂O₂ (Tarique et al., 2022). Owing to these properties, postbiotics are increasingly recognized as natural and sustainable candidates in food safety strategies, offering several advantages over conventional antibiotics and synthetic preservatives in preventing both food spoilage and the proliferation of harmful bacteria (Isaac-Bamgboye et al., 2024; Singh et al., 2025).

The antibacterial efficacy of postbiotics varies depending on the producing probiotic strain, the qualitative and quantitative composition of the postbiotic preparation, the target bacterial species (and even strain), the postbiotic concentration, and environmental conditions such as pH and growth medium (Ooi et al., 2021; Jahedi and Pashangeh, 2025). Beyond these factors, interactions between active postbiotic metabolites, resident microbiota, enzymes, and food components may also inhibit their functionality (Rad et al., 2020). Food-derived proteolytic enzymes, such as trypsin, chymotrypsin, pepsin, papain, and proteinase K can alter postbiotic activity. These enzymes are typically produced by proteolytic bacteria present in the diet (Abdulhussain Kareem and Razavi, 2020; Peluzio et al., 2021). In addition, the antimicrobial mechanisms of postbiotics involve cytoplasmic acidification, disruption of energy production and regulation, and inhibition of pathogenic microorganisms by creating pores in cell membranes. They also induce morphological and functional alterations in sensitive cellular components, such as proteins and peptides, by increasing acidity within the bacterial cell membrane (Rad et al., 2021). A schematic representation of these production stages and mechanisms of action is given in Figure 4.

Figure 4

Figure 4. Schematic overview of postbiotic production and mechanisms of action. (1) Postbiotics are derived from well-characterized microbial strains, including those used in fermented foods or pure culture fermentations, rather than from undefined mixed cultures. (2) Deliberate microbial inactivation occurs through heat, pressure, or other physical and chemical stressors, leading to loss of viability and/or cell lysis. (3) The inactivation step yields non-viable biomass composed of intact cells, cell wall fragments (e.g., peptidoglycan, teichoic acids), and cell-associated or released metabolites such as organic acids, bacteriocins, and enzymes. (4) In the host, these non-viable components act together to inhibit pathogens, strengthen epithelial barrier integrity, and modulate immune responses, as illustrated in the antimicrobial, barrier, and immune panels of the figure.

Certain fermented foods, such as fermented dairy products (e.g. yogurt, kefir), fermented vegetables (e.g., pickles, kimchi), fermented soy products (e.g. miso, tempeh), and fermented cereals (e.g., sourdough bread), provide an indirectly favorable environment for postbiotic production due to their enzymatic activity (Nataraj et al., 2020). Postbiotic components may also indirectly inhibit pathogen growth by competing for essential nutrients, thereby exerting cross-species antagonistic effects through competitive exclusion (Ansari et al., 2024). Yet, only a limited number of postbiotic types have been clearly identified to date, and systematic studies on their antibacterial mechanisms and active components on foods remain insufficient. In the literature, there are mostly original experimental studies and general reviews in this area. However, comprehensive, methodologically rigorous systematic reviews and meta-analyses are still lacking.

8 Postbiotic components against foodborne pathogens

Since ancient times, fermented foods have played a significant role in human diets. They contribute to food safety and help prolong shelf life by preventing food deterioration and the growth of harmful microbes. These protective effects are mainly mediated by bioactive compounds and subsequent inactivated preparations derived from LAB and other beneficial microorganisms. LAB exert their preservative effects mainly by producing antimicrobial metabolites such as organic acids (e.g., lactic and acetic acid), hydrogen peroxide, diacetyl, ethanol, and various bacteriocins (Zapaśnik et al., 2022). Additionally, quorum sensing systems in LAB regulate the production of antimicrobial compounds (particularly bacteriocins) in response to cell density and the surrounding microbial community, which ultimately shapes the antimicrobial profile of their postbiotic preparations (Qian et al., 2023). Lactococcus lactis subsp. lactis, for instance, has been shown to produce active nisin-rich cell-free supernatants (CFSs) as postbiotic preparations when co-cultured with pathogens like Salmonella enterica and Listeria monocytogenes, and to increase nisin production by 50% when co-cultured with Yarrowia lipolytica (Ariana and Hamedi, 2017; Abdollahi et al., 2018). A study examining the effectiveness and antimicrobial potential of LAB-derived postbiotics reported that the lyophilized CFSs of three Lactiplantibacillus and Lacticaseibacillus strains exhibited in vitro antibacterial activity against L. monocytogenes and were also effective against its biofilms (Moradi et al., 2019a).

Reuterin, a powerful postbiotic with strong anti-pathogenic effects, is produced by Limosilactobacillus reuteri during glycerol fermentation under anaerobic conditions (Rodrigues et al., 2021). Reuterin has demonstrated antibacterial activity against E. coli, L. monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus, with particularly strong inhibition observed against the latter two pathogens (Niamah et al., 2023). In another study by Lin et al. (2020), the CFSs of Limosilactobacillus reuteri, isolated from the gut microbiota of fecal samples collected from dairy cows, exhibited inhibitory activity against certain Salmonella serovars (S. Enteritidis, S. Typhimurium, and S. Dublin) as well as Mycobacterium avium subsp. paratuberculosis. The use of LAB, particularly reuterin-producing Limosilactobacillus reuteri, and their metabolites such as reuterin for biopreservation has gained increasing interest in recent years as a natural alternative to enhance food safety and extend product shelf life.

Another noteworthy benefit of postbiotics beyond their antibacterial qualities is their antiviral potential. Postbiotics have been shown to exhibit antiviral effects against enveloped viruses in in vitro studies. A study investigating the in vitro antiviral activity of postbiotics derived from Lactiplantibacillus plantarum and Limosilactobacillus fermentum against herpes simplex virus type 1 (HSV-1) showed that several postbiotic preparations significantly inhibited viral replication and reduced virus adsorption to host cells, with the highest selective index observed for a cell-free supernatant from L. plantarum (Vilhelmova-Ilieva et al., 2022). A study examining the in vitro antiviral activity of Lactobacillus plantarum, Lactobacillus amylovorus, and Enterococcus hirae against enterovirus isolates (Echovirus 7, E13, and E19) found that these LAB strains exhibited marked antiviral effects against E7 and E19 but not against E13. According to the study results, LAB in broth suspension showed stronger antiviral activity than their corresponding CFSs and bacterial pellets, and the pre-treatment approach was more effective than post-treatment, with L. amylovorus AA099 displaying the highest activity against E19 (Sunmola et al., 2019). A study evaluating the potential adjuvant therapeutic effect of Lactobacillus plantarum Probio-88 postbiotics against SARS-CoV-2 reported that CFSs containing plantaricin E and F significantly inhibited viral replication in vitro and showed high binding affinity to SARS-CoV-2 helicase, suggesting that these postbiotic peptides may block viral RNA binding and replication (Rather et al., 2021). Additionally, Kim et al. (2018) investigated the antiviral effects of exopolysaccharides (EPSs) produced by Lactiplantibacillus plantarum LRCC5310, isolated from the traditional Korean fermented food kimchi, against rotavirus infection. The study showed that these EPSs inhibited attachment of human rotavirus Wa strain to host cells in vitro and, when orally administered in a mouse model, reduced viral replication and shedding, shortened the duration of rotavirus-induced diarrhea, and helped preserve intestinal epithelial integrity.

The formation of biofilms by pathogens such as L. monocytogenes, S. aureus, Campylobacter jejuni, Bacillus cereus, and Yersinia enterocolitica in food processing environments poses a serious problem in terms of food safety. Biofilms usually contain multiple microorganisms within a polysaccharide or protein matrix, and this structure increases the resistance of bacteria to antimicrobials (Henriques and Fraqueza, 2017; Dhivya et al., 2022). Some studies focused on the potential of postbiotics to prevent biofilm formation and eliminate existing biofilms. Postbiotics work against biofilm mostly by reducing virulence factors, disrupting quorum sensing systems, and suppressing twitching motility of bacteria (Khani et al., 2024). For example, postbiotics derived from Lacticaseibacillus casei 431, Lactobacillus acidophilus LA5, and Ligilactobacillus salivarius have been shown to reduce L. monocytogenes biofilms on polystyrene surfaces, likely due to the presence of organic acids and other antimicrobial metabolites (Moradi et al., 2019a). By inhibiting the production of biofilms or breaking up existing ones, pure teichoic acids extracted from Lactobacillus strains can have an inhibitory effect on pathogens such Streptococcus mutans, S. aureus, and Enterococcus faecalis (Ahn et al., 2018; Jung et al., 2019). However, the anti-biofilm impact of postbiotics may be limited by environmental conditions, particularly pH and temperature; therefore techniques like postbiotic encapsulation or integration into packaging films are recommended to overcome these restrictions (Khani et al., 2024). Overall, studies on the effect of postbiotics in removing biofilms have shown promising results, and the use of postbiotics is considered an innovative solution to limit and destroy biofilm formation of bacteria in the food industry. Building on their demonstrated antimicrobial potential, the following section outlines some of the most commonly studied postbiotic components in relation to food safety and pathogen control. An overview of these components, highlighting their producer strains, target pathogens, and observed mechanisms, is presented in Table 1.

Table 1

Table 1. Summary of postbiotic components, producer strains, target pathogens, and observed antimicrobial/antiviral effects discussed in this review.

8.1 Cell-free supernatants

CFS refers to the liquid portion of a culture medium that remains after the removal of microbial cells, such as bacteria or yeast. It is typically obtained by centrifuging the culture medium after microbial growth, allowing for the collection of the CFS. This CFS contains bioactive metabolites produced by the microorganisms (Scarpellini et al., 2022). Generally, LAB-derived CFS consists of a mixture of low molecular weight (<1 kDa) compounds (hydrogen peroxide, reuterin, organic acids, carbon dioxide, and diacetyl) and medium molecular weight (1 kDa- 10/20 kDa) compounds (bacteriocins and bacteriocin-like substances) (Kapustian et al., 2018; Nataraj et al., 2020). For instance, Moradi et al. (2019a) reported the CFS of Ligilactobacillus salivarius, Lacticaseibacillus casei 431, and Lactobacillus acidophilus LA5 contains a variety of metabolic byproducts, including alcohol, sugars, peptides, phenols, hydrocarbons, SCFA, organic acids, and amino acids and benzoic acids. Researchers highlighted that the CFS from these probiotic strains exhibits significant antibacterial and antibiofilm properties, particularly against L. monocytogenes. CFSs derived from LAB have been shown to exert inhibitory effects against both Gram-positive and Gram-negative bacteria (Rwubuzizi et al., 2025). These effects are mediated through mechanisms such as increasing membrane permeability, disrupting cell wall synthesis, and disturbing metabolic balance, ultimately leading to cell death (Bajpai et al., 2016). For instance, Gram-negative E. coli tends to be more susceptible to postbiotics due to its sensitivity to acidic environments and antimicrobial agents, whereas Gram-positive S. aureus may exhibit greater resistance owing to its thick peptidoglycan layer (Dejene et al., 2021). Similarly, Khodaii et al. (2017) reported that CFSs obtained from Lactobacillus and Bifidobacterium species exhibited antibacterial activity by inhibiting the in vitro invasion of enteroinvasive E. coli strains into enterocytes. These effects were associated with reduced pathogen adhesion, enhanced epithelial barrier function, and upregulation of protective gene expression, supporting the potential use of LAB CFSs as safe antimicrobial alternatives (Khodaii et al., 2017; Mani-López et al., 2022). However, the antimicrobial activity and overall functionality of postbiotics can be influenced by several factors, including food composition, pH, moisture content, temperature, oxygen and light exposure, and storage conditions (Patil et al., 2019). Optimal postbiotic activity has been reported within a pH range of 4 to 9, with acidic or alkaline conditions negatively impacting their effectiveness (Prabhurajeshwar and Chandrakanth, 2017). In addition, temperature fluctuations can also reduce their antimicrobial potency. A study found that while mild inactivation was observed at temperatures below 100 °C during postbiotic production, higher temperatures, such as 121 °C, caused significant changes in the chemical profile of postbiotics (Sun et al., 2023). Furthermore, a study investigating the thermal stability of CFS derived from Lacticaseibacillus casei demonstrated that it remained stable and retained its metabolic activity for up to 15 minutes at 100 °C (Mirnejad et al., 2013). Safari et al. (2019) reported that postbiotics from Lactobacillus, Bifidobacterium, and Acetobacter strains inactivated by heat, paraformaldehyde, or ozone showed varying antibacterial effects, with paraformaldehyde-treated Levilactobacillus crustorum being most effective against S. mutans (70% reduction), while E. coli was more resistant, highlighting the importance of inactivation method selection.

In addition to environmental conditions and inactivation methods, the effectiveness of postbiotics also depends significantly on the type of strains used and the delivery matrices applied. For instance, Beristain-Bauza et al. (2016) evaluated the antibacterial activity of postbiotics incorporated into calcium caseinate films derived from Lacticaseibacillus rhamnosus NRRL B-442 at concentrations of 6, 12, and 18 mg/mL. At 18 mg/mL, the postbiotics exhibited strong antimicrobial effects against S. aureus, E. coli, L. monocytogenes, and S. Typhimurium. Similarly, the application of postbiotics in food packaging has shown promising results. In a subsequent study, the use of postbiotics from Latilactobacillus sakei NRRL B-1917 in beef cube packaging led to significant reductions in L. monocytogenes and E. coli during refrigerated storage (Beristain-Bauza et al., 2017). Moreover, in vitro assays have demonstrated that strain-specific postbiotic activity can also extend to cellular-level protection. Lactiplantibacillus plantarum-derived postbiotics showed notable antibacterial effects, while CFSs from Bifidobacterium and Lactobacillus cultures were reported to inhibit the invasion of enteroinvasive E. coli strains into enterocytes, potentially by enhancing epithelial barrier function and reducing pathogen adherence (Aghebati-Maleki et al., 2021).

CFS has also demonstrated promising anti-biofilm properties, offering a potential strategy to combat biofilm-associated infections. Biofilms are notoriously resistant to conventional antimicrobials, contributing significantly to persistent infections and treatment failures. Owing to its amphiphilic chemical composition, LAB-derived CFS can interfere with biofilm formation or disrupt established biofilms. For instance, CFS from Lactobacillus spp. has been shown to inhibit biofilm development and eradicate pre-formed biofilms of Cronobacter sakazakii and L. monocytogenes (Singh et al., 2020). Likewise, pH-neutralized CFSs from strains such as Lactiplantibacillus plantarum, Helvetilactobacillus helveticus, Pediococcus acidilactici, and Enterococcus faecium have significantly reduced biofilms formed by S. aureus and E. coli (Cui et al., 2018). In addition, CFS has been reported to be effective against biofilms formed by multidrug-resistant pathogens, including P. aeruginosa, S. aureus, and E. coli, highlighting its potential role in addressing antimicrobial resistance challenges (Zamani et al., 2017; Dey et al., 2019). Collectively, these findings suggest that LAB-derived CFS can inhibit bacterial adhesion and biofilm formation on both biotic and abiotic surfaces. Despite their promising antimicrobial properties, certain by-products found in CFSs, such as D-lactic acid and biogenic amines, may raise safety concerns that could limit their practical application in foods (Obis et al., 2019; Moradi et al., 2020; Kwon et al., 2021). Other issues such as excessive acidification and sensory changes, potential cytotoxic or immunogenic effects of proteinaceous components, and the presence of antibiotic resistance determinants in the producing strains also need to be carefully evaluated before LAB-derived CFSs can be broadly applied in foods. Therefore, careful strain selection and postbiotic characterization are crucial for ensuring the safety and effectiveness of CFS-based applications.

8.2 Peptides and proteins

Food antimicrobials are various compounds or substances used as preservatives to slow down microbial spoilage in food. They inhibit the growth of microorganisms by inhibiting or inactivating spoilage and pathogenic microorganisms, helping to extend shelf life and maintain food quality (Niaz et al., 2019). Antimicrobial peptides and proteins are components of the immune defense and act as biological molecules that protect against microorganisms (Zhang and Gallo, 2016). Antimicrobial peptides, which are short chains of amino acids, destroy bacterial pathogens primarily by disrupting microbial membranes or inhibiting vital processes like macromolecular synthesis (Waghu and Idicula-Thomas, 2020). These peptides are mainly categorized as ribosomal and non-ribosomal: ribosomal peptides, such as bacteriocins, are synthesized by ribosomes and typically show bactericidal or bacteriostatic activity against related strains (Makarova et al., 2019; Wegh et al., 2019), while non-ribosomal peptides are produced enzymatically and can modulate immune responses, inhibit enzymes, or prevent biofilm formation (Hernández-Granados and Franco-Robles, 2020; Li et al., 2022). Their antimicrobial action includes disrupting cell walls, forming membrane pores that cause leakage, and damaging internal cell structures (Rad et al., 2021). For example, peptides produced by E. faecium WEFA23 have been shown to significantly inhibit L. monocytogenes through interactions with its cell wall proteins (He et al., 2019). Similarly, peptides synthesized by E. coli Nissle 1917 have been reported to inhibit S. enterica by targeting and damaging its cell wall (Forkus et al., 2017). In addition, antimicrobial peptides from strains like B. subtilis and E. coli have shown efficacy in suppressing microbial growth and prolonging food shelf life, highlighting their value not only in infection control but also in food preservation (Osés et al., 2015).

Lactoferrin is an iron-binding glycoprotein found in mammalian secretions such as milk and colostrum, known for its antimicrobial activity (Xu et al., 2022). By binding to iron and reducing its availability for bacterial growth, lactoferrin exhibits both bacteriostatic and antibacterial properties against a wide range of microorganisms. It serves as a natural antimicrobial agent in various food products, including meat, dairy, seafood, baked goods, and beverages, helping to preserve them and extend shelf life (Niaz et al., 2019; Tavassoli et al., 2024). Another important antimicrobial protein is lactoperoxidase, a heme protein commonly found in milk, particularly in high concentrations in colostrum. It is an animal-derived peroxidase present in human and animal secretions such as saliva and tears (Zhang and Rhim, 2022). The antimicrobial agents produced by lactoperoxidaase have been reported to prevent milk spoilage, thereby preserving its microbiological quality (Lara-Aguilar and Alcaine, 2019). Additionally, lysozyme is a widely distributed antimicrobial protein found in various biological tissues, cells, and body fluids. It is most abundant in egg white and is also present in tears, saliva, human milk, and mucus (Nawaz et al., 2022). In one study, lysozyme peptides derived from chicken egg white lysozyme exhibited only 11% of the original lysozyme’s lytic activity. However, at a concentration of 100 μg/mL, these peptides completely inhibited Bacillus species responsible for food contamination (Abdou et al., 2007). Such naturally occurring antimicrobial proteins have long been used as bio-preservatives in food products due to their broad-spectrum activity and compatibility with natural formulations (Moradi et al., 2020). Following this approach, recent studies have turned attention to postbiotics, which are bioactive compounds produced by LAB and other beneficial microorganisms, as promising antimicrobial agents against a wide range of foodborne pathogens.

8.3 Bacteriocins

Bacteriocins are ribosomal antimicrobial peptides synthesized by both Gram-positive and Gram-negative bacteria and are generally effective against same or closely related bacteria, with bactericidal or bacteriostatic capabilities (Wegh et al., 2019). These peptides show their lethal activity by altering membrane permeability and are typically highly effective even at nanomolar concentrations (Cotter et al., 2013). The antimicrobial mechanisms of bacteriocins include suppressing spore production, pore formation in pathogenic cell membranes, and membrane lysis (Wang et al., 2018). For instance, bacteriocins generated by Lactiplantibacillus plantarum LPL-1 isolated from fish have been found to limit L. monocytogenes by creating pores in bacterial membranes, while Lactobacillus taiwanensis bacteriocins act against E. coli and S. Gallinarum by disrupting protein structures essential to cell viability (Kim et al., 2020).

Bacteriocins are natural antibacterial chemicals that have been consumed through fermented foods for thousands of years and are currently generating interest for their application as food biopreservers. Bacteriocins, particularly those produced by LAB, are commonly used to preserve foods such as cheese, meat, and vegetables (Drider et al., 2016). LAB-derived bacteriocins are the most studied bacteriocin group due to their food preservative effects. Among these, nisin stands out as the most studied bacteriocin. Moreover, nisin is the first bacteriocin approved by the FDA and it is frequently used as a food preservative in various countries (Chen and Hoover, 2003; Surati, 2020). Nisin is well known for its effectiveness against Gram-positive bacteria, exhibiting an inhibitory effect on both food spoilage bacteria and foodborne pathogens including L. monocytogenes, S. aureus, B. cereus and C. botulinum (Ibrahim, 2019). Except nisin, there are many other bacteriocins such as enterocin, leukocin, pediocin, sukacin, and mycocin derived from Enteroccoccus, Leuconoctoc, Pediococcus, Staphlylococcus, and Debaryomyces spp. respectively (Zhang et al., 2020). Enterocin A, B, and P have been shown to bind to gelatin and suppress the development of L. monocytogenes, S. aureus, and B. cereus (Ibarguren et al., 2015). Lactisin is another bacteriocin with significant antimicrobial potential against various pathogenic microorganisms. Lactisin 3147 and Lactisin 481 exhibit broad-spectrum antimicrobial activity and are considered as safe (GRAS) for food biopreservation (Hugo and Hugo, 2015). Lactisin 3147, produced by Lactococcus lactis, has been reported to help maintain the quality of cheddar cheese and inhibit L. monocytogenes growth on mold-ripened cheese surfaces, reducing its count by 3 log CFU/mL (Lahiri et al., 2022).

Pediococcus acidilactici BAMA 15, isolated from the traditional “naniura” dish, demonstrated antibacterial activity against E. coli and S. aureus, with a stronger inhibitory effect on S. aureus. The study also highlighted that Gram-negative bacteria tend to be more resistant than Gram-positive bacteria (Nasution et al., 2023). Another study examined the bacteriocins derived from Lactiplantibacillus plantarum, E. faecalis, and Lactobacillus delbrueckii subsp. lactis isolated from cheese, butter, and strained yogurt. A bacteriocin cocktail at 100 AU/mL concentration led to significant reductions in B. cereus, L. monocytogenes, and S. aureus counts in milk, whereas an increase in pathogen levels was observed in the control group (Kaya and Simsek, 2019). On the other hand, bacteriocins from LAB have also been investigated on Gram-negative bacteria. Probiotic LAB isolated from kiwi fruit pulp were tested against Staphylococcus, Pseudomonas, and E. coli. The results showed inhibition against Gram-positive bacteria but no effect on Gram-negative bacteria, emphasizing that the outer membrane of Gram-negative bacteria provides resistance against bacteriocins (Kamaliya et al., 2023). This suggests that while not all bacteriocins lead to direct membrane lysis, many still disrupt membrane integrity to levels that can cause cellular death depending on the target strain and bacteriocin type (Wang et al., 2018; Kim et al., 2020). However, studies have shown that bacteriocins demonstrating strong in vitro antimicrobial activity may not always exhibit the same efficacy in real food matrices. For instance, Hacıomeroglu and Cufaoglu (2024) observed antibacterial activity of bacteriocins against P. aeruginosa in vitro, but no significant reduction was achieved in a milk model, emphasizing the importance of evaluating bacteriocin performance in complex food environments.

Despite their limited efficacy against Gram-negative bacteria, bacteriocins have gained attention in food safety and therapeutic contexts due to their low toxicity, high stability, and target specificity. Their use as starter cultures in food processing has also been linked to enhanced fermentation, improved preservation, and better organoleptic qualities (Carneiro et al., 2024). However, interactions with food matrices or degradation in complex environments may reduce their effectiveness. One proposed strategy is incorporating bacteriocins into food packaging films to maintain their activity and stability (Silva et al., 2018b).

Beyond food preservation, bacteriocins hold promise as natural alternatives to antibiotics, offering new strategies to combat antimicrobial resistance (Cotter et al., 2013; WHO, 2017). Effective resistance management strategies include combining bacteriocins with other bacteriocins with differing modes of action (Hols et al., 2019), with traditional antimicrobials (Mathur et al., 2017), or with bacteriophages (Rendueles et al., 2022), and using bioengineering to design peptides less prone to resistance (Field et al., 2019). Moreover, bacteriocins have been shown to inhibit biofilm formation, disrupt quorum sensing, and reduce bacterial virulence factors, expanding their potential applications in antimicrobial therapy and biocontrol (Mathur et al., 2018; Zhang et al., 2023). Yet, to fully realize their potential, challenges such as optimizing their incorporation into foods, ensuring stability in diverse matrices, and improving production scalability must be addressed. Continued research into bioengineering, resistance minimization, and cost-effective production is essential for broader application.

8.4 Exopolysaccharide

EPSs are long-chain, high molecular weight polymers composed of branched, repeating units of sugars or sugar derivatives that are mostly generated by LAB. They are classified into two types based on their chemical composition: homopolysaccharides and heteropolysaccharides. Homopolysaccharides include just one type of monosaccharide unit (e.g., cellulose, levan, curdlan, pullulan, dextran), whereas heteropolysaccharides contain many monosaccharide units (e.g., xanthan, gellan, galactan, kefiran) (Chaisuwan et al., 2020). In the food industry, LAB-derived EPSs are commonly used for improving the texture, taste, flavor, and shelf life of fermented foods. Lactococcus, Leuconostoc, Streptococcus, Pediococcus and Bifidobacteria are essential microbes in the formation of EPS in fermented dairy products such as milk, yogurt, curd, cheese, buttermilk, and sour cream (Roca et al., 2015). Additionally, the antimicrobial and antioxidant properties of LAB-derived EPSs are being intensively investigated (Nehal et al., 2019; Liu et al., 2019; Xiao et al., 2020). A study on the in vitro antibacterial activity of a novel EPS (EPS-Ca6) from Lactobacillus sp. Ca6 revealed strong effects against Salmonella spp. and Micrococcus luteus, with inhibition zones of 14 mm and 10 mm, respectively. However, no significant activity was detected against L. ivanovii, S. aureus, B. cereus, and E. coli (Trabelsi et al., 2017). Furthermore, the antimicrobial potential of EPS was demonstrated by findings showing that EPS produced by Weissella confusa MD1 exhibited strong inhibitory effects against S. aureus, L. monocytogenes, S. enterica and S. Typhi (Lakra et al., 2020).

EPSs also function in the adhesion and protection of bacteria in processes such as bacterial surface interactions and biofilm formation. LAB-derived EPS is recognized as a foreign biomolecule by pathogens, preventing their intracellular entry or activity. One possible in vitro antimicrobial mechanism of LAB-derived EPS involves interfering with biofilm-associated signaling molecules or glycocalyx receptors on pathogen surfaces, thereby disrupting cell-to-cell communication. This interaction inhibits biofilm formation, suppresses pathogen growth, and exerts an antimicrobial effect (Spanò et al., 2016). Wang et al. (2020) reported that EPS produced by Limosilactobacillus fermentum strains consist of glucose, galactose, mannose, and arabinose in varying ratios. Although these EPS exhibited minimal inhibitory effects on the growth of E. coli and S. aureus, they demonstrated significant, dose-independent antibiofilm activity. This effect is likely due to their influence on microbial cell surfaces, prevention of initial attachment, and downregulation of biofilm-related gene expression, with variations attributed to differences in EPS composition and surface charges (Wang et al., 2020). Kim et al. (2009) reported that EPS obtained from Lactobacillus acidophilus exhibited antibiofilm activity against both Gram-positive and Gram-negative pathogens, resulting in 87% to 94% inhibition of E. coli O157:H7 biofilms depending on the biofilm surface. EPS obtained from Lacticaseibacillus rhamnosus strains isolated from human milk exhibited antibacterial effects, with inhibition zones ranging from 12 to 14.3 mm against E. coli and 10 to 13 mm against S. Typhimurium. Additionally, S. Typhimurium biofilm formation was inhibited by 58–71%, demonstrating strong antibacterial activity (Rajoka et al., 2018). These findings collectively highlight the promising role of LAB-derived EPSs as postbiotic compounds with multifunctional bioactivities, particularly in antimicrobial and antibiofilm applications. Their natural origin, structural diversity, and functional versatility position them as valuable biocontrol agents in the development of safer and more sustainable food preservation strategies.

8.5 Organic acids

Organic acids are important postbiotic components due to their antibacterial properties, limiting acid-intolerant pathogens by lowering pH and acting as acidifiers (Chang et al., 2021). They are widely used in the food industry as pH regulators and preservatives. Their antimicrobial mechanism primarily involves reducing environmental pH, causing intracellular anion accumulation, and lowering intracellular pH, ultimately disrupting bacterial survival (Wang et al., 2019). Especially the L and D isomers of lactic acid effectively reduce the effect of pathogens when obtained through bacterial fermentation (Rad et al., 2021). Acetic and lactic acids are critical compounds secreted by Lactiplantibacillus plantarum during postbiotic production and support the growth of the cells (Kareem et al., 2014). Moreover, metabolites such as phenyllactic acid and lactic acid produced by Lactiplantibacillus plantarum CECT-221 have been proven to exhibit strong inhibitory effects against pathogens such as Carnobacterium piscicola, S. aureus, P. aeruginosa, L. monocytogenes and S. enterica (Rodríguez-Pazo et al., 2013).

Low pH and high concentrations of organic acids offer effective protection against food spoilage and the growth of pathogenic organisms (Chang et al., 2021). For instance, biological preservation methods that combine various organic acids have been explored as a promising strategy for developing antibacterial agents for widespread use in the food industry (Rad et al., 2021). In this context, organic acids produced by LAB, Bifidobacteria, and other postbiotic species exhibit dose-dependent antimicrobial activity, particularly against Gram-negative pathogens (Lukic et al., 2017). Additionally, a strong correlation has been observed between the lactic and acetic acid content of postbiotics and their antimicrobial properties (Tejero-Sariñena et al., 2012). However, relying on single organic acids requires specific concentrations, which can increase costs, alter product flavor, and lead to nutrient loss. To enhance bacterial inhibition while minimizing organic acid usage, they can be combined to leverage their synergistic effects against target bacteria. Several studies have investigated the combination of organic acids for more effective bacterial control, particularly in the cleaning and disinfection of meat and fresh vegetables (Wang et al., 2019; Ji et al., 2023). Hu et al. (2019) investigated the effects of tartaric, acetic, lactic, citric, and malic acids secreted by three different Lactiplantibacillus plantarum strains (P1, S11, and M7) on E. coli and Salmonella spp., reporting significant bacterial growth inhibition. Beyond their antimicrobial properties, organic acids also play a crucial role in food biopreservation packaging. Their impact goes further than pathogen inhibition, involving multiple mechanisms such as cell membrane disruption and inhibition of macromolecular synthesis. Bacterial fermentation and combination techniques offer the food industry novel and efficient antibacterial strategies against foodborne pathogens by facilitating organic acid production (Prabhurajeshwar and Chandrakanth, 2017; Moradi et al., 2021).

9 Postbiotic preparations in food applications

Postbiotic preparations have the potential to inhibit pathogenic and spoilage microorganisms in food production and preservation. Several postbiotic components have been shown to efficiently suppress the growth of pathogens and spoilage microorganisms in dairy products, meat, seafood, fruits and vegetables (Sharafi et al., 2023). In food systems, these preparations are most commonly applied as LAB–derived CFSs or CFS-based concentrates, which are widely referred to as postbiotic preparations, even though they typically contain soluble metabolites rather than deliberately inactivated cells in the strict ISAPP sense (Salminen et al., 2021). It should be noted that most food-related studies described below use CFS-based preparations rather than fully ISAPP-compliant postbiotics containing intentionally inactivated cells; however, these preparations are commonly referred to as ‘postbiotics’ in the food technology literature. In most in vitro and in situ food-related studies, these CFS-based preparations are typically obtained by fermenting LAB cultures followed by centrifugation and/or membrane filtration to remove bacterial cells. The resulting supernatant, rich in organic acids, hydrogen peroxide, diacetyl, bacteriocins, and other bioactive metabolites, is then directly applied to the food matrix or incorporated into edible coatings and packaging materials. This strategy has been successfully used in meat, dairy, seafood and fruit–vegetable products, either as liquid CFSs or, more frequently, as concentrated, lyophilized or encapsulated CFS-based preparations, which provide improved stability, easier handling and more consistent antimicrobial effects against foodborne pathogens and spoilage organisms during storage.

In the study of Incili et al. (2022), CFS–based postbiotics obtained from Pediococcus acidilactici, especially when incorporated into chitosan coatings, were reported to effectively control microbial spoilage and foodborne pathogens in vacuum-packaged frankfurters by significantly reducing the counts of E. coli O157:H7, S. Typhimurium, L. monocytogenes, and total viable microorganisms during refrigerated storage. Similarly, a study on the anti-L. activity of L. sakei-derived postbiotics reported that applying CFS to inoculated meat cuts significantly decreased L. monocytogenes counts (up to about 3 log CFU/g) over refrigerated storage, confirming their potential as bioprotective agents in fresh meat systems (Valipour et al., 2024).​ Investigations of reuterin produced by Limosilactobacillus reuteri E81 demonstrated that reuterin-containing preparations exerted strong in situ bioprotective effects in a white cheese model, markedly reducing Gram-negative pathogens such as E. coli and inhibiting molds and yeasts during cold storage (Ispirli et al., 2025). Similarly, application of CFS from Weissella viridescens to stainless steel and polystyrene surfaces, as well as to chilled pork, was shown to inhibit L. monocytogenes biofilm formation on contact surfaces and significantly limit L. growth on meat during refrigerated storage, highlighting the potential of Weissella-derived postbiotics for surface and product decontamination in meat processing (Yang et al., 2022). The application of CFS–based postbiotics from Lacticaseibacillus rhamnosus at a 100 mg/g concentration to chicken meat contaminated with C. perfringens effectively eliminated the pathogen during storage at 6 °C (Hamad et al., 2020). In line with these findings, Moradi et al. (2019a) investigated the antibacterial activity of freeze-dried CFSs derived from Ligilactobacillus salivarius against L. monocytogenes both in vitro and in whole milk and ground meat models. The results demonstrated that L. salivarius CFS effectively inhibited pathogen growth and showed potential as a natural antimicrobial additive in meat and dairy products. However, the study also emphasized the need to combine such natural compounds with hurdle technologies to enhance food safety. In a follow-up study, the application of 35.00 mg/g L. salivarius CFS significantly extended the shelf life of refrigerated ground meat by controlling both microbial and oxidative spoilage, further supporting its use as a food preservative (Moradi et al., 2019b).

The incorporation of antimicrobial compounds into food requires careful consideration, as it can lead to undesirable changes in sensory properties such as taste, texture, and appearance, a challenge observed for both live cultures and inactive components (Isaac-Bamgboye et al., 2024). For instance, a study by Szydłowska and Sionek (2022) showed that the addition of LAB did not significantly alter the sensory qualities of semi-hard cheese and cream, but did affect the sensory characteristics of sour cream. Specifically, the inherent acidity of postbiotic preparations (e.g., cell-free supernatants rich in organic acids) can undesirably lower the pH of the food matrix, leading to texture defects such as protein coagulation or an overly sour taste. Furthermore, certain metabolites or growth media residues may introduce bitterness or off-flavors (Moradi et al., 2020). Studies also indicate that color and appearance changes caused by postbiotic components in opaque and liquid matrices, such as milk, can negatively affect consumer acceptance. Moradi et al. (2019b) evaluated different concentrations of lyophilized CFS from Lactobacillus salivarius in a ground beef model and noted that, although the preparation effectively inhibited microbial growth and oxidation, its intrinsic light-brown color could be undesirable in white and opaque products such as milk, where even slight discoloration may reduce consumer acceptance. Kamaliya et al. (2023) examined the application of Lactobacillus acidophilus CFS in milk and reported that, besides antimicrobial performance, changes in appearance, flavor, and overall sensory scores had to be carefully monitored, because visible deviations from the typical color and aspect of fresh milk negatively influenced panelists’ acceptability ratings. At the same time, it has also been observed that postbiotic applications can contribute to sensory improvement; for example, postbiotics derived from Lactobacillus reuteri were reported to enhance the texture of bread (Jonkuviene et al., 2016). These findings highlight the need for a delicate balance when applying postbiotics, requiring maximization of antimicrobial efficacy while minimizing sensory deterioration. Therefore, the selection of the application method is as critical as the choice of the strain. We propose that emerging technologies, particularly encapsulation and active packaging, offer viable solutions to mitigate these sensory defects. Recent studies suggest that encapsulation techniques can effectively mask the acidity and bitterness of postbiotic metabolites (Aghebati-Maleki et al., 2021), while active packaging films isolate bioactive compounds from the food matrix until the point of action (Moradi et al., 2021). Together, these strategies can help maximize antimicrobial efficacy while preserving desirable sensory attributes, making postbiotics a more feasible option for a broader range of non-fermented foods.

Fresh fruits and vegetables risk contamination from pathogenic bacteria in manure, irrigation of water, soil, and livestock. Once introduced, pathogens can colonize, persist, and threaten human health, causing economic and social impacts (Alegbeleye et al., 2018). Thus, the antimicrobial efficacy of postbiotic preparations, particularly antimicrobial peptides and LAB-derived CFSs, and their impact on the shelf life of fruits and vegetables have been studied. Antimicrobial peptides produced by Lactiplantibacillus plantarum UTNCys5–4 and Lactococcus lactis subsp. lactis Gt28 have been found effective in preserving fresh-cut fruits by inhibiting pathogen growth. These peptides exhibited bacteriolytic effects on fresh pineapple slices, enhancing food safety (Tenea et al., 2020). Similarly, researchers have evaluated the antimicrobial activity of LAB-derived CFSs from Lactiplantibacillus plantarum Cs and Lactobacillus acidophilus ATCC 314, along with their ability to extend the shelf life of tomato paste. Concentrated CFSs exhibited strong antimicrobial effects against S. aureus, E. coli, Aspergillus niger, and Aspergillus flavus, increasing the shelf life of tomato paste to 25–30 days at room temperature (George-Okafor et al., 2020). Additionally, a combination of CFSs from Lactobacillus brevis WK12 and Leuconostoc mesenteroides WK32 with grape seed extract significantly reduced the growth of native microbial flora, E. coli O157:H7, and L. monocytogenes in ready-to-eat baby leaf vegetables. This natural disinfectant blend has been proposed as a potential alternative to chlorine sanitation for improving microbial safety (Lee et al., 2016). These applications pave the way for alternative strategies that support the use of postbiotic preparations and their associated bioactive version in various food models. The available data suggest that these inactive preparations and compounds not only suppress pathogens but also contribute to extending the shelf life of food products. However, challenges such as variability in composition, limited data from real food matrices, and the absence of standardized production and regulatory criteria still constrain their wider industrial implementation (Salminen et al., 2021).Studies demonstrating the antimicrobial effects of postbiotic preparations in food models are presented in Table 2.

Table 2

Table 2. Applications of postbiotic preparations in food matrices for control of foodborne pathogens.

10 Conclusion

Postbiotics present promising antimicrobial solutions for food safety. Compounds such as bacteriocins, organic acids, enzymes, and peptides have shown effectiveness against both Gram-positive and Gram-negative bacteria by inhibiting pathogen growth and biofilm formation. Their high stability, longer shelf life, and resistance to environmental stressors make them attractive for food preservation. However, their antimicrobial mechanisms are not fully understood, and efficacy may vary with environmental conditions. While in vitro studies on food-related applications are increasing, there remains a pressing need for validation in real food matrices. Moreover, despite the ISAPP consensus definition, there is still no globally harmonized regulatory definition of the term “postbiotic.” The use of varying terminology for similar concepts in the literature contributes to confusion in the industry, complicating the proper classification, labeling, and regulatory evaluation of these products. The absence of standardized production methods, inconsistent dosages, and sensitivity to environmental factors limit the industrial application of postbiotics. Clearer terminology, identification of active compounds, optimized production, and regulatory frameworks are needed to support their effective use in food safety. Future work should focus on elucidating the precise mechanisms of action of postbiotics, optimizing their production and delivery systems, and validating their antimicrobial efficacy in real food matrices. Future inquiries should prioritize well-designed in situ studies in diverse food matrices to bridge the gap between promising in vitro findings and real-world performance, including dose–response relationships and sensory/technological impacts. In parallel, research efforts should aim to elucidate the interactions between food matrix elements and postbiotic bioavailability, alongside confirming the economic viability of industrial-scale production. Emphasis must be placed on advanced delivery systems such as nano-encapsulation, as well as omics-based mechanistic studies that can identify strain-specific active components and validate biomarkers of efficacy, thereby supporting the development of clear regulatory frameworks and harmonized terminology for industry adoption.

Author contributions

AE: Conceptualization, Investigation, Methodology, Resources, Visualization, Writing – original draft. GC: Conceptualization, Investigation, Methodology, Resources, Supervision, Writing – original draft, Writing – review & editing.

Funding

The author(s) declared that financial support was not received for this work and/or its publication.

Conflict of interest

The authors declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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