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Postbiotics in Food and Pharmaceutical Industry
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This entry is adapted from the peer-reviewed paper 10.3390/foods11193094

The use of postbiotics as functional ingredients in foods offers several benefits during the industrial handling and commercialization of food products, including the ability to be added to some foods that are thought to be detrimental to the survival of probiotics. This helps to grow the market for functional foods. Practically speaking, because their viability is not necessary for either large-scale production or consumption, postbiotics are more stable and secure for food and pharmaceutical uses than the living bacteria they are produced from. A change in processing conditions or scaling that can result in structural modifications and variations in the physiological function of postbiotics may make it difficult to produce postbiotics on a large scale in the industry. This is true even though the production of these compounds is possible at the laboratory scale or under small-scale conditions. 

postbiotics bioactivity lactic acid bacteria health-promoting

1. Biological Activities of Postbiotics

1.1. In Vitro Studies of Bioactivities

Postbiotic therapies are not known to have any adverse effects like inflammation and so on, during clinical and experimental procedures. Indulged with its safer dosages and known chemical structures it can be proposed as a safe alternative to probiotics engrossment. Owing to their exclusive competence in the prevention and treatment of certain diseases and improvement of health status, there is an increased use of these postbiotics in veterinary, medical, and food applications. Among the different properties exhibited by postbiotics, the anticancerous activity of these metabolites is promising and of medical significance. Being a multifactor disease encompassing the irregular or uncontrolled growth of cells, the traditional methods of treatment results in resistance to chemotherapy, the toxicity of the system, and even the reoccurrence of cancer growth [1]. Mainly the involvement of postbiotics in cancer treatment is associated with their capacity to connect with the host immune cells, activating certain signaling routes, which results in an upsurge of inherent immune system reaction and reduction in inflammation [2]. In vitro analysis of exopolysaccharide metabolites from the postbiotic strain of L. plantarum depicts the prospect of the convention of these metabolites in functional food supplements and their use as antitumor drugs [3]. Explorative studies on the effects of metabolites showed that there is dosage and period dependency in results of the inhibition of liver cancer, gastric cancer, and colon cancer cells. Moderate antitumor activity was observed in the case of liver cancer cells and a significant inhibition ratio of around 61% and 88.34% against gastric and colon cancer cells. Similarly, metabolites of six postbiotic strains of L. plantarum also exhibited strain, time, and dosage-dependent cytotoxicity against different cancerous cell lines with an inhibition concentration of about 50% [4]. There was no validated toxicity of normal human breast cells; red blood cells of humans, dogs, rabbits, chickens; and of mice splenocytes. Further clinical studies are required to optimize the delivery approaches, design suitable carriers, and assess the potential for postbiotics following oral treatment as well as the underlying processes of cell death for their uses in vivo.
Additionally, in vitro revisions of postbiotic-constituent mechanisms against the oxidative components is evidently an unresearched area which entails more consideration and research. Chang et al. [5] revise the significance of Lactiplantibacillus plantarum strains as a potential antioxidative source. A positive connection was drawn between the organic acid production and intracellular components like pyrrole derivatives among postbiotics metabolites and accomplished oxidative inhibitory activity. In vitro evaluations of multifunctional properties of postbiotic strains discloses the antioxidant activities of intracellular contents and cellular fractions of Lactobacillus casei and Bacillus coagulans strains [6]. Studies depicted a cellular antioxidant activity (CAA units expressed as quercetin equivalents) that ranged from 22.7–42.9 CAA units in the case of Lactobacillus casei and 43.1–56.9 CAA units in the case of Bacillus coagulans. Reports on exploratory studies on the effectiveness and anti-microbial potential of postbiotic strains were confirmed by several authors. The potential for postbiotics obtained from three different strains of Lactobacillus spp. in preventing the colonization of pathogenic organisms was reported by Moradi et al. [7]. Residual antimicrobial activity of over 50% was observed in the case of all three strains of LAB species with significant reductions in the case of the biofilm formation of Listeria monocytogenes. The concept of controlled pathogenic growth is corrected to the postbiotic secretion of different substances like bacteriocins, hydrogen peroxide, organic acids and so on, which hinder microbial growth. Prevention of microbial growth is dependent on certain factors including the type of postbiotic substances involved, contact time, the process of the carnage of prebiotic strain, and concentration. Alongside the reported variance in the inhibitory activity among different species or strains, there is a possibility of difference in repressive nature according to the killing method of prebiotic strain. The inhibitory effect of Acetobacter pasteurianus and Lactobacillus crustorum strains were found to be affected by the killing method while examining the effectiveness in the prevention of growth of Streptococcus mutans and Escherichia coli [8]. Among the two different strains, S. mutans were found to more sensitive to postbiotics than E. coli, exhibiting a total of 70% diminution in viability. Among the different methods of carnage involved, the heat-killed bacteria to some extent was ineffective against the populations of E. coli. Formaldehyde killed method was found to be effective against the populations of E. coli and S. mutans, with more than a 50% decrease in viability. With little knowledge of the disparity in mechanisms concerning varied results, there should be thorough studies on various factors shaping the efficacy of the process. The antiviral potential of these postbiotics is well thought out as a supplementary benefit to their antibacterial properties. The strains of Lactobacillus amylovorus, Lactobacillus plantarum, and Enterococcus hirae exhibited good antiviral properties against the recovered enterovirus isolates [9]. The antiviral effects of these strains are proposed to different mechanisms including the production of hydrogen peroxide and organic acids and competitive exclusion. Postbiotic metabolites like amino acids, bacteriocins, SCFAs, peptidoglycan-derived muropeptide, vitamins and so on, promote and enhance the anti-inflammatory functions in the human gut. Clinical trials on the anti-inflammatory potential of postbiotics of the Bifidobacterium longum strain showed that it was effective in controlling the acute inflammatory response and gut disruption by activating the pathways related to distinctive immune response [10]. The study explains that the potential of the heat-treated probiotic strains was in agreement with the gut-barrier protective capacity and anti-inflammatory potential of the live strain. As with every functional activity of postbiotics, the effectiveness of metabolites in controlling the inflammatory condition is dependent on several factors, especially the type of postbiotic strain. Lin et al. [11] evaluated the dependency of metabolite efficiency with the strains involved. Multi-strain postbiotics was exhibiting a superior anti-inflammatory potential compared to single-strain postbiotics and efficiency of postbiotic metabolites is closely dependent on the choice of strains integrated together. The results explain that the multi-strain probiotics named as probiotic extracts of four strains—number 1, PE0401 which is an amalgamation of four probiotic strains including Lactobacillus salivarius, Lactobacillus plantarum, Lactobacillus acidophilus, and Bifidobacterium longum in 1:1:1:1 ratio was effective against all the other mixed forms of the metabolites.

1.2. In Vivo Studies of Bioactivities

Studies on postbiotics have a known level of adherence to the characteristics of prebiotic pattern and environment. However, it does not degenerate the efficiency of postbiotics in fostering their antimicrobial, antidiabetic, antiproliferative, antioxidant and anticancer roles in living organisms. An advantage of postbiotics over other chemical preservatives and antibiotics is their effectiveness against pathogenic bacteria solving the possibility of reducing food spoilage issues. The antimicrobial properties of postbiotics are dependent on factors like the type of the target microorganism, postbiotic concentration, and the nature of the source prebiotics involved. Probiotic stimulation achieved due to the presence of Lactobacillus spp. in mice helped in reducing the infectivity and total activity of E. coli preventing intestinal inflammation in a broader perspective [12]. The incidence of postbiotics has been found to elevate the level of cytokines arbitrated habitually by pathogenic recognition receptors inducing immune homeostasis against the targeted organism. Correspondingly, the efficacy of Lactobacillus spp. against L. monocytogenes in both in vivo and in vitro studies was reported by Nakamura et al. [13]. Inhibition of activity of around 90% was made possible by the postbiotic orientation in live tissues. Theories on the effectiveness of these populations revolve either around artificially reduced nutrient availability or pH decreases indicating the presence of LAB bacteria. Additionally, the antiviral activity of postbiotics is a significant diversion that requires deeper thought. Postbiotics derived from populations of Lactobacillus plantarum were effective against SARS-CoV-2 infection by modifying the immune system and attacking the virus, as discussed by Anwar et al. [14]. Plant-derived probiotics were helpful in controlling the pathogenicity of the Coronavirus 2019 (COVID-19) by affixing itself with the spike glycoprotein which otherwise shows a possibility of binding with angiotensin 2 converting enzyme (ACE2) causing infection. 
Defensive mechanisms of postbiotics constituents, especially against the oxidative properties of different elements, were studied and reported by several authors. Dietary postbiotics of L. plantarum strain reduced the serum lipid peroxidation and boosted the serum and ruminal fluid antioxidant activity with augmented hepatic antioxidant enzymes [15]. Postbiotics of the indicated strain augment the antioxidant activities by exhibiting a high resistance to hydrogen peroxide and elevated rates of scavenging activity against superoxide, hydroxyl, and DDPH free radicals beholden to the presence of metabolites including EPS, lipoteichoic acid, and cell-surface proteins [16]. The incorporation of postbiotics as an oral supplement in broilers positively influenced the antioxidant properties enhancing the activity of enzymes like glutathione peroxidase and decreasing heat-stress markers. The likelihood of the introduction of postbiotic strains of Lactiplantibacillus plantarum was accounted for as an antibiotic substitute and natural antioxidant source in feeding patterns [17]. In vivo studies of postbiotic metabolites require a better understanding of the delivery and reaction mechanisms, signaling pathways, and of the direct effects of constituents on the living cells. With limited information on these scales, in vivo experimental research on the anticancer potential of postbiotics is still in its infancy. Chen et al. [18] elucidate the efficacy of metabolites of Clostridium butyricum in modulating and inhibiting intestinal tumor development in mice. The likelihood of the suppression of tumor growth is retracted to the efficiency of butyrate-producing bacteria in Wnt signal modulation and gut microbiota. More experiential learning are needed to optimize and extend the application ranges of this postbiotic potential. Postbiotics are known to enhance the immune system of beings by varying and strengthening the microflora of the intestinal tract and thereby preventing inflammatory issues in the body Rad et al. [19]. LAB are known for modulating the immune responses in normal conditions which has been evaluated by Menard et al. [20] in mice. The study depicts the clear correlation between the anti-inflammatory effect of postbiotics of Bifidobacterium breve and Streptococcus thermophilus strains in an inflammatory context and their immune stimulatory effects. Strains of Lactobacillus bulgaricus and Streptococcus thermophilus are found to yield health effects in mice by preventing the advance of colitis a chronic inflammatory disease [21].

1.3. Infection Prevention

Postbiotics are defined as the preparation of inactive microorganisms and/or their parts that benefits the host’s health [22]. A novel method for obtaining probiotic bacteria’s positive effects without their potential drawbacks is the use of postbiotics. Despite the rarity of probiotic-related infections and the ongoing debate over whether probiotics could express virulence factors or pass antibiotic-resistant genes to pathogenic bacteria [23][24], using postbiotics might avoid those problems because the bacteria are rendered inactive by heat, high pressure, sonication, or ionizing radiation [25]. Numerous studies claim that postbiotics can enhance physiological functions [26] and prevent and treat diseases including gastroenteritis, respiratory tract infections, and enteric infections as well as conditions like gut barrier dysfunction [27][28][29][30]. Some postbiotics can compete for receptors needed by some pathogenic bacteria, seal the intestinal barrier, change the expression of host genes, or modify the local environment to have direct antimicrobial effects [31].

2. Potential Applications in Food and Pharmaceutical Sector

Functional foods that contain probiotics, prebiotics, and postbiotics have drawn a lot of interest from researchers, manufacturers, and consumers in recent years. A significant amount of postbiotic research is currently focused on not only accurately defining their mechanisms of action but also creating novel functional food and preventative medication formulations for improving host health [32]. There are a wide variety of food products with bioactive substances like probiotics, dairy, and non-dairy products already on the market [33] to suit the nutritional needs of customers with diverse dietary preferences, such as people who are allergic to milk proteins, lactose intolerant, and vegetarians [32]. Since postbiotics are stable across wide temperatures and pH ranges, it is possible to add foods and ingredients before thermal processing without compromising their functionality. This could give producers some technical and financial advantages [32]. Postbiotics can be used in delivery systems like functional foods and/or pharmaceutical products because their adequate amount can be controlled during production and storage conditions, where survival is not the main determining factor.

2.1. Potential Role of Postbiotics in the Food Industry

Postbiotics can be composed of bacterial lysates with cell surface proteins, bacterial enzymes, peptides, metabolites (produced by bacteria such as teichoic acids), neuropeptides (derived from peptidoglycans, polysaccharides), and lower organic acids like lactic acid [32][34][35]. Fermentation is the most prevalent postbiotic source in the food industry. The presence of postbiotics can be found naturally in several milk-based and other products like kefir, kombucha, yogurt, and pickled vegetables [33]. The producer strains mostly include strains of Lactobacillus and Bifidobacterium but may also include Streptococcus, Akkermansia muciniphila, Eubacaterium hallii, Faecalibacterium, Saccharomyces boulardii and can be used to extract the postbiotics in situ [34][36]. EPS are extracellular biopolymers that microbes produce or secrete during the course of their growth. They differ greatly in both the amount of branching they exhibit—from linear molecules to highly branched molecules and the monosaccharide content of these molecules [37]. EPS produced by LAB, such as Lactobacillus rhamnosus, plays a significant role in dairy products and can enhance the physicochemical and sensory qualities of food-based products [34]. Similarly, another study found that the postbiotic supernatant from Lactobacillus plantarum YML007 can be a possible bio-preservative, extending the shelf life of soybeans by up to two months [38]. Additionally, a number of bacteriocins have been isolated, characterized, and may have potential industrial uses. The microbial strain and culture conditions will determine their isolation and characterization. They must first be biologically inactive before being altered to become active [39]. Nisin, which is known as a preservative in several foods (infant formula, canned soups, dairy products), can be produced by Lactococcus lactis subsp. Lactis [40]. Several studies have also focused on the use of enzymes instead of probiotic bacteria to achieve specific effects. Postbiotic enzymes such as purified phytases from Bifidobacterium pseudocatenulatum and Bifidobacterium longum spp. infantis, for example, decreased the amount of phytates in cereal combinations and elevated myoinositol triphosphate levels [41]. Vitamin enrichment in food products has also been reported by several authors [42][43]. Increasing vitamin B as a result of fermentation is a very common strategy applied in cereal grains. Cereal grains are rich in vitamin B. However, during milling or heat processing, these vitamins are lost. Cereal fermentation and LAB pre-treatment increase the amount of bacteria that can produce the vitamins B1, B2, B3, B9, B11, and B12. In vitro levels of total lysine, protein fractions, sugars, soluble dietary fiber, and bioavailability of Ca, Fe, and Zn were all dramatically increased by the LAB fermentation of cereals. Additionally, wheat could produce antihypertensive angiotensin I-converting enzyme-inhibitory peptides, γ-aminobutyric acid, and antioxidant peptides by LAB fermentation [44].
Another novel approach to probiotic usage also involves removing some potentially toxic food ingredients during probiotic-induced fermentation in addition to supplementing food with postbiotics. A study by Sarno et al. [45] reported that the amount of toxic gliadin peptides is reduced in celiac patients by fermentation with the probiotic bacteria Lactobacillus paracasei CBA L74 [45]. With the aforementioned in mind, the use of foods as a delivery vehicle for postbiotics appears to be a field with many prospects but also some challenges.

2.2. Potential Role of Postbiotics in Pharmaceutical or Health Industry

Numerous postbiotic molecules have attracted interest because of their unique properties such as their chemical makeup, prolonged storage stability, and capacity to activate various mechanisms regulating inflammation, obesity, hypertension, cardiovascular disease, cancer, and oxidative stress [37]. Postbiotics provide immunomodulatory activity to the host and may represent a safer alternative when the use of live probiotic bacteria is not indicated. A recent review by [46] highlighted several pharmacodynamic features of postbiotics over probiotics which are (1) Invulnerable and immunocompromised individuals have minimal or no risk of bacterial translocation from the stomach lumen to blood; (2) No possibility of acquiring and transferring genes for antibiotic resistance; (3) The extraction, standardization, transportation, and storage is more natural; (4) Additional positive effects may result from cell lysis-induced viability loss; (5) Improved interactions between the epithelial cells and molecules released from the disrupted cells [46].
Nowadays, industries are attempting to incorporate postbiotics into the pharmaceutical product matrix due to their beneficial role in clinical scenarios. For example, CytoFlora® manufactured by BioRay Inc., Laguna Hills, CA, USA is a well-known postbiotic in the pharmaceutical industry. CytoFlora® is composed of cell walls isolated from bacteria some of which include Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus acidophilus DDS-1. CytoFlora has been reported to have several important positive impacts, including preserving intestinal homeostasis, enhancing intestinal dysbiosis, enhancing immunological response, and alleviating symptoms in autistic individuals [32][33][34]. Del-Immune V® from Pure Research Products, LLC, Boulder, CO, USA is another postbiotic pharmaceutical product. It is typically advised to take them with probiotics for therapeutic purposes. A combination of amino acids, muramyl peptides, and DNA fragments from Lactobacillus rhamnosus V make up the Del-Immune V formulation. Studies have shown that Del-Immune V significantly lessens the severity of the gastrointestinal disorder in people with autism spectrum disorder [32][34]. In another example, for the treatment of intestinal active inflammatory diseases, Zakofalk® (a mixture of sodium butyrate and inulin) is frequently advised [32]. A sterile liquid postbiotic medication called Hylak® Forte (Ratiopharm/Merckle, Germany) contains biological metabolites such as organic acids, short-chain fatty acids, and other metabolites. The development of gut-beneficial microbes, regulation of gut environment pH, support for healthy digestion, energy supply for intestinal epithelial cells, control of vitamin K and B balances, and treatment of salmonellosis and intestinal disorders in adults and children with chronic gut disorders are some of the main health effects of Hylak® Forte [32][47].
In addition to the commercial availability of postbiotic products, much attention has been paid to various health benefits that could result in the future production of other drugs. For example, reproductive health and the impact of postbiotics were recently reviewed [48]. They supported the use of postbiotics as a promising tool in a personalized medicine approach for restoring vaginal eubiosis/health. Additionally, it was recommended for both preventative and adjuvant treatment strategies in women with reproductive-related disorders due to their unique features in terms of clinical, technological, and economic aspects [48]. Researchers also reviewed the impact of postbiotics in children younger than 5 years old and recommended heat-killed Lactobacillus acidophilus LB for the management of acute diarrhea based on limited evidence. Additionally, it was also recommended heat-killed Lactobacillus paracasei CBA L74 for the prevention of gastrointestinal and respiratory tract infections based on limited evidence and suggested further need for studies to impart further understanding [30]. Similarly, another research group studied the antibacterial mechanism of indole propionic acid, a gut microbiota metabolite, and reported it as an anti-tubercular agent [49][50]. They discovered that indole propionic acid might imitate the physiological allosteric inhibitor of TrpE, limit tryptophan production in M. tuberculosis, and hence, exhibit antimycobacterial activity after conducting metabolic, chemical rescue, genetic, and biochemical tests [50].
Animal health has also received some attention similar to human health as it offers a promising tool in veterinary research [51][52]. A study reported increased final body weight, total weight gain, and the height of duodenal and ileal villus in broilers when their meals were fortified with postbiotics of Lactobacillus plantarum [52]. In another study, supplementing animal meals with Lactobacillus plantarum postbiotics increased the length of mucosal villi and strengthened the population of beneficial intestinal microbiota, which in turn improved protein digestibility and growth efficiency, and decreased the incidence of diarrhea [34]. Additionally, postbiotic use changed the intestinal microbiota, increased the number of protective bacteria (such as Lactobacillus and Bifidobacterium), and improved the health of the test animals. Postbiotics may be helpful as preventative medications, fermented functional foods, and microbial-free food supplements as supplemental therapy for a number of disorders [34].
In summary, there have been numerous postbiotics introduced into the pharmaceutical, veterinary, and food industries as medications, food, and feed, demonstrating an extraordinary aptitude for the prevention and treatment of specific diseases, boosting animal health status, and generating functional foods. Consequently, postbiotics may be a secure substitute and a novel application approach in the pharmaceutical and/or food industries for fostering and developing health advantages, as well as for preventing diseases and achieving therapeutic goals. However, food and drug product’s safety heavily depends on the safety of the ingredients that make up those products. Therefore, additional research is necessary to define novel postbiotics and analyze their stability and safety criteria in the pharmaceutical and food industries. Clinical trials are also required to establish the ideal postbiotic administration frequency and dosage in a delivery method (pharmaceutical formulations and/or functional postbiotic meals).

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Subjects: Food Science & Technology
Contributors MDPI registered users' name will be linked to their SciProfiles pages. To register with us, please refer to https://encyclopedia.pub/register :
Priyamvada Thorakkattu
,
Anandu Chandra Khanashyam
,
Kartik Shah
,
Karthik Sajith Babu
,
Anjaly Shanker Mundanat
,
Aiswariya Deliephan
,
Gitanjali S. Deokar
,
Chalat Santivarangkna
,
Nilesh Prakash Nirmal
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Update Date: 20 Oct 2022
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