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Review

Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha

by
Tara Budimac
,
Aleksandra Ranitović
*,
Olja Šovljanski
,
Dragoljub Cvetković
and
Ana Tomić
Faculty of Technology Novi Sad, University of Novi Sad, 21000 Novi Sad, Serbia
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(8), 434; https://doi.org/10.3390/fermentation11080434
Submission received: 20 June 2025 / Revised: 21 July 2025 / Accepted: 28 July 2025 / Published: 29 July 2025
(This article belongs to the Special Issue Fermentation and Bioactive Potential of Kombucha and Bee-Derived Compounds for Health and Wellness)
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Abstract

Kombucha is a fermented tea beverage of Asian origin, widely consumed due to its functional properties; yet, it typically lacks sufficient levels of probiotic micro-organisms to be classified as a probiotic product. This review analyzes the occurrence of lactic acid bacteria (LAB) in kombucha, reporting that concentrations rarely exceed 4–5 log CFU/mL and often decline during fermentation or storage. Strategies to enhance probiotic viability, including the use of robust LAB strains and encapsulation technologies, are critically evaluated. Notably, encapsulation using pea and whey protein has been shown to sustain LAB levels above 6 log CFU/mL during fermentation and up to 21 days under refrigerated storage for whey protein. Fortified kombucha beverages with probiotic strains have also been shown to possess enhanced functional and health-promoting benefits compared to traditional control samples. Despite promising approaches, inconsistencies in microbial survival and regulatory constraints remain key challenges. Future research should focus on the optimization of delivery systems for probiotic cultures, identification of kombucha-compatible LAB strains and standardized protocols to validate probiotic efficacy in real-world beverage conditions.

1. Kombucha

Kombucha is a fermented beverage originating from Asia, which has recently gained considerable popularity in Western countries due to its reported therapeutic properties, including antimicrobial, antioxidant, anticancer and anti-inflammatory effects (Figure 1). Its official origin traces back to northeastern China, specifically the Manchurian region, where it was embraced during the Tsin Dynasty for its detoxifying and energizing benefits. Historical records indicate that in 414 BC, a physician named Kombu introduced fermented tea from Korea to Japan, using it to treat digestive ailments of Emperor Inkyo, hence the name “Kombucha” or “Kombu tea”. Over time, the beverage has been known by various other names, such as tea fungus, Kargasok tea, Manchurian mushroom, Haipao, etc. [1,2]. With the expansion of trade routes, kombucha was first introduced to Russia and then to Eastern Europe, appearing in Germany around the 20th century. During World War II, Germany introduced this drink, and in the 1950s, it reached France and north Africa, where its consumption became quite popular [1]. Years later, Swiss researchers stated that consumption of kombucha was just as beneficial as yogurt due to the presence of acids that promote the growth of beneficial bacteria in the gut. Since then, the popularity of the beverage has grown, and it has been introduced to the market with a variety of new flavors. Today, kombucha can be purchased in retail stores around the world, as well as the tea mushroom itself for preparation [3].
Traditionally, kombucha was produced by fermenting sweetened black or green tea (Camellia sinensis L.), although other types of teas can also be used. The presence of tea in the fermentation medium supplies micro-organisms with nitrogen, which is essential for their metabolic processes. The primary nitrogen sources in tea come from purines such as caffeine and theophylline. Consequently, some studies suggest that teas beyond Camellia sinensis L. species, provided they contain caffeine, such as yerba mate (Ilex paraguariensis), mate tea and even coffee, can be used for kombucha production [4]. However, other research indicates that the kombucha symbiotic culture can also thrive on caffeine-free substrates, implying that the micro-organisms present in kombucha are capable of utilizing alternative nitrogen sources. As a result, kombucha variants have been produced from infusions like mint, lemon balm, jasmine [5], echinacea, Rtanj tea [6], hibiscus and many others [1]. If sucrose is used as the primary source of carbon for fermentation, acetic acid will be the dominant metabolite produced [7].
The fermentation of tea, which lasts from a few days to two weeks, is the product of a symbiotic consortium of bacteria and yeasts embedded in a cellulose film. This cellulose film is called the symbiotic consortium of bacteria and yeast (SCOBY) [1]. In addition, lately, researchers have used other substrates that partially or completely replace tea in the production of kombucha beverages, such as fruit juices, vegetables, spices and herbs, flowers, as well as sugar substitutes, such as honey, molasses, coconut sugar, etc. In this way, products with altered chemical content and enhanced functional properties are created [8].
The chemical composition of kombucha is variable both quantitatively and qualitatively. The presence and concentrations of certain chemical compounds in the beverage itself depend on the micro-organisms used in the fermentation, the fermentation parameters (time and temperature), the concentration of sucrose and other carbon nutrient sources, the type of tea used and the analytical methods used for quantification. However, some components, such as organic acids, vitamins, polyphenols and amino acids, have been shown to be predominantly present [7].
Numerous characteristics of this beverage have been noticed that possess a beneficial effect on health, and due to its simple and safe preparation, it has gained great popularity among consumers around the world. It has been experimentally proven that kombucha has four main characteristics that are necessary for certain biological activities. Namely, it has detoxifying properties, the property of protection against damage caused by free radicals, energizing abilities and the ability to improve immunity [9]. Although it may seem that many of the reports available are based on personal experiences and testimonies, more recently, scientists have provided evidence to explain the therapeutic effects of the beverage [4]. However, most of these benefits have only been studied in experimental models, in the absence of scientific evidence in humans [10].
In addition to its antioxidant, antimicrobial and anticancer properties, kombucha is often marketed as a probiotic beverage, although the number of probiotic micro-organisms present in the final product is frequently inconsistent and unstable, as many strains fail to remain viable throughout the sometimes unfavorable conditions of fermentation and storage [11]. To be officially classified as a probiotic product, a beverage must contain at least 6–7 log CFU/mL of viable probiotic micro-organisms at the point of consumption [12]. This highlights the need for the supplementation of known probiotic strains that can survive throughout the production and shelf life of kombucha. One promising approach to address this issue is encapsulation, a technology already widely and successfully used in other food matrices [13]. However, when it comes to kombucha, this area remains relatively unexplored, with only a limited number of studies published to date.
The worldwide kombucha market was estimated at USD 4.26 billion in 2024 and is projected to reach around USD 5.9 billion by 2029 [14], growing at a compound annual growth rate of approximately 13–19% [15]. Over 1300 new kombucha variants launched globally in 2023, including hard, low- or no-sugar versions. Overall, kombucha has evolved into a robust, fast-growing functional beverage sector with strong growth, wide product variety and increasing global consumer appeal. This growth is driven by the increasing popularity of functional beverages and the trend toward organic and natural products [14,15].
In addition to historical and commercial perspectives, the production and labeling of kombucha beverages, particularly those claiming probiotic properties, are increasingly influenced by national and international regulatory frameworks [16]. In Canada, the Center of Diseases Control of British Columbia (BCCDC) provides a food safety document for kombucha production, covering chemical parameters, ingredients and safety claims to ensure controlled manufacturing. More recently, federal guidelines have been introduced to assess recipe safety, potential hazards, alcohol content and labeling. These guidelines and labeling offer recommendations for hygiene, pH control, packaging, strain safety, alcohol content and product validation for human consumption [17]. Brazil is the only country with specific kombucha legislation, established by Norm Instruction No. 41 from the Ministry of Agriculture, Livestock and Supply. This legislation outlines the definition, classification and labeling requirements for kombucha, as well as the analytical standards, permitted and optional ingredients and rules for production and marketing, as well as addition of probiotic strains. It also specifies mandatory label information, acceptable alcohol limits and the analytical parameters that must be monitored during production [16,17].

2. Lactic Acid Bacteria in Kombucha as Part of Natural Microbiota

Symbiotic consortium of bacteria and yeast (SCOBY) or “tea fungus” mostly consists of acetic acid bacteria (AAB) and osmophilic yeast species. The term was coined by Len Porzio in the 1990s to distinguish the new culture formed in kombucha from its previous culture [18]. The term “symbiosis” is mostly used in the literature, and although there is no evidence that these micro-organisms depend on each other, as is the case in symbiosis, they nevertheless live in a common culture [2]. The exact microbial composition of SCOBY is determined by the geographical and climatic conditions of cultivation, as well as local species of wild yeasts and bacteria, and can differ widely depending on the location [19]. SCOBY micro-organisms can be found both in the fermented liquid (kombucha) and the biofilm (pellicle). The bacteria and yeasts that are added to the beverage are responsible for the formation of a cellulose film, or “mat” [20], which often resembles a surface mold or mushroom, which is where the common name tea fungus comes from [21]. The official, scientific name of this biofilm is Medusomyces gisevii [5] (Figure 2). The microbial composition of kombucha is still a very active field and the target of many studies. A major obstacle in reaching a consensus on the structure of this microbial community, however, comes from the fact that most fermentations are carried out without adhering to and using a specific methodology and that, essentially, no two microbial inoculums are the same [22].
Given that the fermentation of kombucha is usually carried out at the household level, where ingredients and recipes vary greatly, the variety of micro-organisms obtained from this drink is very wide. In general, the yeast population outnumbers the bacteria present in this beverage [23]. Yeast cells in the culture medium are usually deposited in liquid but also get trapped in the pellicle. The yeast component of SCOBY varies more between the studies than AAB, but usually, the genera Zygosaccharomyces and Saccharomyces are found to dominate the culture [24]. Further, Candida, Kloeckera/Hanseniaspora, Torulaspora, Pichia, Brettanomyces/Dekkera, Saccharomyces, Lachancea, Saccharomycoides, Schizosaccharomyces, Kluyveromyces [25], Mycotorula, Mycoderma, Torula and Torulopsis [26] were also detected.
The bacteria that dominate the kombucha starter culture are producers of acetic and gluconic acids and belong to the genus Acetobacter and Gluconobacter [1]. Previous studies have shown that the main bacterial genus present in the tea fungus is Gluconacetobacter, which is present in >85% of the total bacterial population for most samples. When looking at the entire fermentation medium, 86–99% of Gluconobacter genera were found [27]. Similar results were obtained by Watawana et al. [28], who fermented coconut water with tea fungus and found the genus Gluconacetobacter as the main one with a relative percentage of 85.6%. In addition to these genera, the tea fungus and kombucha mainly contain species belonging to the genera Acetobacter and Komagataeibacter. In particular, Komagataeibacter xylinus is the most characteristic micro-organism present in the tea fungus and is thought to be responsible for the production of the cellulose pellicle. Sometimes, filamentous fungi, or mold, can also be found, but they are considered harmful and undesirable micro-organisms, as they are associated with the production of mycotoxins and adverse health effects [29].
Lactic acid bacteria (LAB) can also occasionally occur in SCOBY and kombucha itself, but they are generally not an essential part of the kombucha microbial ecosystem, and their number is often inconsistent. However, in certain industrial beverage fermentations, these bacteria have been found in higher concentrations, mainly the species Lactobacillus nagelii and Oenococcus oeni, which are known to be acid-resistant [29]. Although some studies show that these bacteria are present in small numbers in kombucha, in the first days of fermentation, an increase in their number is observed, but then, there is a gradual decrease in the concentration of the population. Therefore, it is possible that inoculation of a tea infusion with a starter culture from a previous fermentation, which lasts longer than 14 days, may lead to the absence of LAB during the new process [30]. Table 1 provides an overview of samples where LAB were detected or not detected in unsupplemented, pure kombucha beverages or pellicles as part of natural microbiota (i.e., without addition of probiotic cultures).
Marsh et al. [31] reported that Gluconobacter was the predominant bacterial species isolated in 85% of samples, and Lactobacillus, with abundance above 30%, were identified as the second dominant isolated species. The data showed that lactobacilli are more prevalent in kombucha than Lactococcus, and their numbers increased by the end of fermentation in comparison to Lactococcus. Bogdan et al. [32] also studied the possible probiotic potential of five LAB isolated from kombucha. All isolates were identified as Pediococcus pentosaceus species and were screened for bacteriocin, with three of the five strains being positive. In addition, the isolates have also been tested for resistance to bile salts. Among these five isolates, one strain showed the greatest potential as a probiotic because it was the only one that showed both the ability to produce bacteriocin and resistance to bile salts. This strain was further studied for its adhesion potential for Caco-2 intestinal cells, and the authors proposed a probiotic effect. The authors concluded that kombucha could have probiotic potential if these bacteria were found in sufficient quantities.
Fabricio et al. [33] found that Proteobacteria were the most dominant phyla in all the tested samples, but Lactobacillaceae were also present in the fermentation liquid, with 19.9% of relative abundance. The genus Liquorilactobacillus was also found in kombucha, representing 13% of the bacterial population in liquid and less than 1% in biofilm. Another LAB identified in the initial liquid was Ligilactobacillus, with 6.9% of relative abundance. Yang et al. [34] sampled nine commercial kombucha beverages for microbial identification and found that although LAB were not commonly used in the fermentation of kombucha to enhance its biological function or to be added as probiotics, Lactobacillus was dominant in four of the nine samples.
The species Oenococcus oeni is mainly associated with the fermentation of green tea [22]. Coton et al. [35] found that two LAB strains, Oneococcus oeni and Lactobacillus nagelii, present in kombucha during green tea fermentations, were correlated with the production of lactic acid, and their concentration increased during fermentation. In addition to the Lactobacillus strains, Weissella spp. was also isolated in some instances [36,37].
Although LAB are present in some studies both in tea fungus and in the kombucha itself (Table 1), their numbers are often inconsistent, and they are not present in sufficient quantities to confer probiotic effects; therefore, kombucha still cannot be considered a probiotic product. To achieve potential health effects, probiotic food and beverages are required to contain a recommended minimum level of 6–7 log CFU/mL or g of probiotic bacteria at the time of consumption [38].
Table 1. A review of studies detecting/not detecting the presence of LAB in pure, unsupplemented kombucha samples (as part of the natural microbiota).
Table 1. A review of studies detecting/not detecting the presence of LAB in pure, unsupplemented kombucha samples (as part of the natural microbiota).
LABMethodIsolated from Kombucha/PellicleOriginSample Type (Commercial, Laboratory, Household)Reference
LactobacillusDNA amplification and high-throughput sequencingKombuchaCanada, Ireland, UK, USACommercial[31]
LactococcusDNA amplification and high-throughput sequencingKombuchaCanada, Ireland, UK, USACommercial
Lactobacillus and LactococcusDNA amplification and high-throughput sequencingPellicleCanada, Ireland, UKCommercial
Collinsella, Enterobacter,
Weissella, Lactobacillus		Next-generation sequencing and data analysisKombuchansLaboratory[36]
ndMorphological features followed by DNA amplification and high-throughput sequencingPellicleUkraineCommercial[39]
Oenococcus oeni, Lactobacillus nagelii, L. satsumensisM13-PCR genetic profile clustering and sequence analysisKombuchaFranceCommercial[35]
Limosilactobacillus fermentumDNA amplification and high-throughput sequencingKombuchaChinaHousehold[40]
Five isolates belonging to the Lactobacilli groupMolecular identificationKombuchaRomaniaCommercial[32]
LactobacillusDNA amplification and high-throughput sequencingKombucha and pellicleChinaCommercial[41]
LactobacillaceaeMolecular identificationKombucha and pellicleUKCommercial[42]
ndDiversity sequencingPellicleUSA [27]
Lactobacillus plantarumCarbohydrate fermentation pattern followed by molecular identificationPellicleChinaCommercial[43]
ndMetagenomic analysesKombucha and pellicleFranceCommercial and household[44]
ndAmplicon sequencing and shotgun metagenomicsKombuchaTurkeyHousehold[45]
LactobacillusMetabarcoding analysesKombucha and pellicleUSACommercial[46]
ndMetagenomics analysisKombuchaAustraliaCommercial[47]
Weissella spp., Lactobacillus rhamnosus, Lactobacillus acidophilusDNA amplification and high-throughput sequencingKombuchaIranCommercial[38]
ndPhenotypic characterization followed by RNA sequence analysesKombuchaNew ZealandCommercial[23]
Liquorilactobacillus and LigilactobacillusMetagenetic analysisKombuchaBrazilCommercial[33]
Lactobacillus nagelii and L. maliShotgun metagenomics analysisKombuchaUSACommercial[34]
nd—not detected, ns—not specified.

3. Kombucha with the Addition of Probiotic Cultures

As stated, the probiotic strains in kombucha are usually absent or remain in low concentrations, mainly after storage. Harrison et al. [48] examined samples of 39 retail non-alcoholic (<0.5%), low-alcohol (>0.5%) and alcoholic kombucha samples to conclude whether the claims of probiotic properties were supported. Interestingly, only 6.3% of non-alcoholic and 10% of low-alcohol kombucha products exceeded 106 CFU/mL, the limit that would deliver at least a billion cells in one package and thus provide an adequate probiotic dose. None of the sampled alcoholic kombucha beverages exceeded this limit. The most abundant communities of bacteria were Glucanobacter and Bacillus. To a small extent, the probiotic species Lactobacillus and the yeast S. Cerevisiae var. boulardii were recognized. The deficiency of the Lactobacillus species can be explained by the poorer survival of these bacteria in kombucha.
An alternative strategy for obtaining kombucha with pronounced probiotic properties may be to add well-known and studied live probiotic strains at the beginning or during the fermentation process or to a finalized product in the post-fermentation process, as has been shown in several studies to date (Figure 3). Table 2 presents a summary of studies where probiotic strains have been added to kombucha, as well as their survivability potential. For example, Al-Dulaimi et al. [49] developed two kombuchas using traditional black tea and kombucha fermented skim milk, composed of a starter probiotic culture, including two species of Lactobacillus (delbruekii and fermentum). Investigating the beneficial effects of these samples on some physiological and biochemical parameters in rats, the authors confirmed that animals treated with these beverages had reduced concentrations of total lipid profile, ALT, AST, ALP hepatic enzymes and serum glucose compared to the control group. Reduced serum glucose levels may be related to the activity of probiotic bacteria because they use glucan compounds for fermentation. In conclusion, kombucha mixed with skim milk that includes LAB species could enhance the functional properties of these beverages.
In another study, kombucha made from coffee was developed to which two probiotic strains were added: Lactobacillus rhamnosus and Lactobacillus casei. After fermentation and removal of the biofilm, the strains were added to the coffee-fermented kombucha, and a control sample without probiotic strains was produced. The authors assessed the survival of probiotic strains in kombucha and the viability under simulated stomach and intestinal conditions. The concentrations of these probiotic bacteria in kombucha were above the minimum recommended concentration for food products at the end of storage. The addition of probiotic strains changed the proportion of micro-organisms from Acetobacteraceae to Lactobacillaceae. The results were very interesting and showed that the viability of L. casei in kombucha remained without significant changes at the end of storage, which made it possible to suggest that this strain survives in the gastric and intestinal environment. The good survivability could be due to relatively high pH values (in most cases above 3.5) compared to other studies [50].
On the other hand, in a study conducted by Fu et al. [51], estimating the survival of specific strains in kombucha after 14 days of refrigerated storage did not show such positive results. The authors developed a green tea kombucha inoculated with a 5% starter culture composed of Saccharomyces cerevisiae, Komagataeibacter sp. and Lactobacillus plantarum in a 1:1:1 ratio. In the end, they concluded that the concentration, when cooled to 4 °C, of AAB decreased moderately during ten days of storage, while the survival rate of LAB was only 0.98% on the eighth day of storage, and their number decreased significantly after two days of storage. These results confirm the importance of the selection of micro-organisms contained in the starter culture and their influence on the final probiotic characteristics of the beverage.
Cvetković et al. [52] added five wild strains of LAB to black tea kombucha to assess their viability, contribution to the functional characteristics of the beverage, as well as interactions with tea fungus micro-organisms. The strains were added on the second day of cultivation to avoid the osmotic stress to which cells would have been exposed at the beginning due to the high concentration of sucrose in the medium. After fermentation, the product was stored at 4 °C for 10 days. The results showed that the addition of these strains did not affect the physiological activity of the tea fungus but contributed to a significant increase in the content of lactic acid in the beverage, which has certain functional properties. In terms of the viability of these strains, Lactobacillus hilgardii has shown the best results both during fermentation and during the storage period. Such results can be explained by the origin of this wild strain, which is the only one isolated from sourdough. Strains isolated from traditionally fermented foods are known to have enhanced metabolic activities compared to micro-organisms used as industrial starter cultures. The authors concluded that all of the above should result in the production of a beverage with increased functional and probiotic properties, with additional studies.
Similar results in terms of biological activity of a beverage with added strains of Lactobacillus paracasei were obtained by Lee et al. [53]; however, the viability of these bacteria during and after fermentation has not been examined. The authors concluded that in samples inoculated with LAB, there was a significantly higher production of acetic and glucuronic acid compared to control samples. Also, the antibacterial activity against E. coli was slightly higher compared to the control sample. Antioxidant and anti-inflammatory properties have been proven and pronounced. These results are consistent with the results of Nguyen et al. [54], who added strains of LAB isolated from kefir (L. plantarum and L. casei) during the fermentation of kombucha. In this research, there was also an improved production of glucuronic acid as well as D-saccharic acid 1,4–lactones (DSL), which are one of the most important carriers of the functional properties of kombucha. In addition, the improved antibacterial and antioxidant activity of the beverage has been confirmed with the addition of the aforementioned strains.
Yang et al. [55] confirmed this in their study, where they examined the interactions between Gluconacetobacter sp. A4, one of the most common AAB in tea fungus, and LAB strains isolated from kefir. The results suggested that LAB stimulated the production of DSL molecules by Gluconacetobacter sp. A4 in the range from 4.86% to as much as 86.70%. The metabolites of these bacteria, xylitol and acetic acid, had a positive effect on the reproduction of AAB and the production of this functional compound; therefore, it was concluded that the combination of these bacteria in tea fungus would be optimal for the fermentation of kombucha with enhanced functional properties. However, the survivability of added LAB strains was not examined in this study.
Some manufacturing companies do not pasteurize kombucha on an industrial scale to maintain the beverage’s natural microbiome, in such a way that keeping the product raw would allow the development of a prebiotic or probiotic product. On the other hand, some companies continue with the pasteurization process in their production and subsequently add recognized probiotic strains, such as Bacillus coagulans, Saccharomyces boulardii and Lactobacillus rhamnosus, to the final formulation of kombucha. It is important to note that this strategy of adding probiotics is beneficial depending on the country in which the sale is regulated. In some countries, such as Brazil, the Normative Guideline for Quality and Identity Standards for Kombucha does not allow the addition of strains after fermentation but only at the beginning of the process itself [51].
There is not much research available that has successfully produced a kombucha beverage that meets the requirements of a probiotic product with the addition of live probiotic strains due to the large drop in the concentration of these bacteria during fermentation and storage of the product. Majid et al. [56] successfully produced a blue pea tea kombucha (Clitoria ternatea L.) that can be considered a probiotic product by adding Lactiplantibacillus plantarum subsp. plantarum Dad-13 strain, where the amount of probiotic strain after fermentation and storage for 28 days was 6.26 log CFU/mL. The strain was added before fermentation, which lasted 8 days at room temperature. These results may be dependent on the type of tea used in comparison to traditional black tea, since survivability remained high, although the pH values decreased noticeably.
As shown in Table 2, the survivability of live LAB added to kombucha beverages varies depending on the pH levels, the point in time when the strains were added, as well as the substrate used for preparation, but usually, the numbers do not remain stable, especially during storage, because of unfavorable conditions during kombucha fermentation and storage, such as low pH values.
Table 2. Overview of strains, viable counts and survivability of probiotic bacteria added to kombucha.
Table 2. Overview of strains, viable counts and survivability of probiotic bacteria added to kombucha.
LAB StrainInitial Number (log cfu/mL)Survivability (Days)Final Number(log cfu/mL)pH Range (Initial–Final)Reference
Lactobacillus plantarum883.43nr–3.15[51]
Lactobacillus plantarum6.8134.394.16–2.78[57]
L. plantarum Lb-37.3812.024.52–3.01[52]
L. plantarum Lb-47.5432.954.30–3.03
L. plantarum Lb-57.1331.904.29–3.06
Lactiplantibacillus plantarum subsp. plantarum6286.263.32–2.68[56]
Lactiplantibacillus plantarum subsp. plantarum78 (not selected for storage analyses)8.323.34–2.72
Lactobacillus plantarum and Lactobacillus casei *nrnrnrnr[54]
Lactobacillus casei8.7215~8.723.56–3.46[50]
Lactobacillus paracasei *3nrnrnr[53]
Lactobacillus rhamnosus8.50157.903.56–3.48[50]
Lactobacillus rhamnosus6.122.844.16–3.06[57]
Lactobacillus brevis and Lactobacillus fermentum~96<1 to 3nr–3[58]
Lactobacillus fermentum7.181<14.43–3.05
Lactobacillus hilgardii8.1677.184.43–3.16[52]
Lactobacillus sp., Lactococcus lactis subsp. and Leuconostoc sp. *7.30nrnr5.10–2.60[55]
nr—not reported. * Survivability data not provided in the original study; functional characteristics reported instead. Bold rows indicate cases where the final LAB count exceeded the minimum threshold required for probiotic products.

4. Encapsulation of Probiotic Bacteria as a Promising Method for Kombucha

Encapsulation emerges as one of the main strategies for protection and enhancement of the survivability and viability of probiotic strains during unfavorable conditions of fermentation, transportation, desiccation, storage and gastrointestinal transit, such as low pH values, temperature, oxygen etc., thus ensuring product stability and timely release of probiotic strains at appropriate sites, such as the small intestine, in an adequate amount [59]. Encapsulation refers to a physicochemical or mechanical process of entrapping nutrients, therapeutic compounds or probiotics (called core materials, active agents, internal phase, etc.) in various materials (called coating membrane, carrier, wall material, shell or matrix) to produce particles with diameters of a few nanometers, micrometers or millimeters [60,61]. The survival of encapsulated probiotic cells will depend on several factors, including the strain, carrier material and concentration, the number of initial cells, encapsulation method, etc. [13,62].
Encapsulation of active substances into carrier materials can be achieved through various methods, such as spray drying, spray cooling or freezing, extrusion coating, fluidized bed coating, lyophilization, coacervation, emulsion methods and many others. The choice of an appropriate encapsulation process is influenced by numerous factors, such as the physical and chemical characteristics of the core and carrier materials, as well as the intended application within the food matrix [63]. The selection of carrier materials is very important, especially when used in food products or processes. They should be food-grade, biodegradable and must have a generally recognized as safe (GRAS) status for consumption [64]. To microencapsulate probiotic micro-organisms, materials such as polysaccharides (gum arabic [65], pectin [66], maltodextrin [67], alginate, inulin [68], chitosan, xanthan [62]), proteins (whey protein [13], gelatin, legume proteins [69]) and dietary fibers (resistant starch, maize starch) are commonly used [62].
The effectiveness of encapsulation has been well demonstrated in various food matrices, particularly in fruit juices and yogurt, which often share similar fermentation conditions with kombucha, including low pH and the presence of organic acids. Several studies have reported increased viability of probiotic strains during storage and gastrointestinal transit when encapsulated [70,71]. The survivability is generally higher in encapsulated cells compared to free cells, especially in acidic environments, such as fruit juices [72]. In addition, encapsulated probiotics have shown positive effects on the sensory and functional properties of the final products [73]. Table 3 summarizes recent applications of encapsulated probiotic strains in different food matrices.
Table 3. Review of the application of encapsulated probiotic strains in different food matrices.
Table 3. Review of the application of encapsulated probiotic strains in different food matrices.
Probiotic Bacterial StrainCarrier MaterialEncapsulation MethodFood ProductReference
Lactococcus lactis supsb. lactis 303 CFELiposomesMicrofluidizationCheddar cheese ripening[74]
Lactobacillus acidophilus and Bifidobacterium bifidumSodium alginateInternal gelationGrape juice[72]
Enterococcus faeciumSodium alginateExtrusionSour cherry juice[75]
Lactobacillus caseiSodium alginateVibration technologyPineapple, orange and raspberry juice[76]
Lactobacillus casei and Lactobacillus acidophilusSodium alginate and cocoa powderLyophilizationChocolate[77]
Lacticaseibacillus rhamnosusSodium alginateIonic gelation following emulsification process as pretreatmentApple juice and yogurt[78]
Lactobacillus gasseriSodium alginateEmulsification and extrusionApple juice[79]
Lactobacillus rhamnosus and Lactobacillus plantarumWhey protein isolateConventional and multilayer emulsionYogurt[80]
Lactobacillus plantarumWhey proteinLyophilizationApple juice[81]
Given the success of encapsulation in enhancing probiotic viability in juices and other acidic matrices, it could be assumed that similar strategies could be effective in kombucha fermentation. However, despite the growing interest in functionalizing kombucha with probiotics, the application of encapsulated probiotic strains in kombucha remains underexplored. Figure 4 illustrates the general concept of probiotic encapsulation for kombucha enrichment, highlighting key stages, such as the selection of probiotic strains, encapsulation methods and carrier materials.
To date, only a few studies have investigated the use of encapsulated probiotics in kombucha. In one study, the survivability of encapsulated Lactobacillus plantarum ATCC 14917 was evaluated using three different carrier materials and the conventional emulsification method during kombucha fermentation. The viability of these LAB strains was also monitored throughout the storage period of the final product. Among the tested carriers, pectin showed the highest protective potential, with viable LAB cells detected in the beverage until the end of fermentation and for up to 4 days of storage, maintaining levels around 4 log CFU/mL. In contrast, with the other two carrier materials, LAB cells were only detectable until the third day of fermentation, after which they were no longer present. Although pectin outperformed the other carriers in preserving cell viability, the overall LAB counts remained below the minimum recommended threshold for probiotic products (6–7 log CFU/mL). These findings underscore the need for further research to improve encapsulation strategies and enhance probiotic survival in kombucha [82].
In another study, Budimac et al. [13] examined five different carrier materials—pectin, inulin, maltodextrin, pea protein and whey protein—for the encapsulation of Lactobacillus rhamnosus ATCC 53103 using the lyophilization method. This same strain had previously been tested as free cells added at the start of kombucha fermentation, where its viability drastically decreased and became undetectable by the end of the fermentation period [52]. Among the tested carriers, pea protein and whey protein showed the most promising protective effects during preliminary trials that assessed survival in different pH environments, leading to their selection for further testing in kombucha fermentation. During this phase, both whey and pea proteins showed good potential for protection of LAB cells until the end of fermentation, since their numbers were 6.51 logCFU/mL and 7.71 logCFU/mL for pea and whey proteins, respectively. However, whey protein was also noticed to positively influence the fermentation process, which is why, consequently, it was selected for use during the storage period as well. After 21 days of storage, only a slight, statistically insignificant decline in cell count was observed, from 7.71 log CFU/mL to 7.43 log CFU/mL, remaining well above the recommended minimum for probiotic products. The kombucha enriched with whey protein–encapsulated LAB was further evaluated for its functional properties and sensory profile. In addition to maintaining a higher concentration of viable bacteria, this probiotic-enriched kombucha exhibited enhanced health-promoting effects, and the sensory characteristics remained comparable to the control sample without added probiotics, indicating strong consumer acceptability.
Table 4 summarizes the very few available studies on the use of encapsulated probiotics in kombucha fermentation.
Table 4. Review of the application of encapsulated probiotic strains in kombucha.
Table 4. Review of the application of encapsulated probiotic strains in kombucha.
Probiotic Bacterial StrainCarrier MaterialEncapsulation MethodFood ProductReference
Lactobacillus plantarumPectin, inulin and mixture of maltodextrin and glucoseEmulsionKombucha[82]
Lactobacillus rhamnosusPectin, inulin, maltodextrin, pea protein, whey proteinLyophilizationKombucha[13]
While encapsulation has proven beneficial in multiple food matrices, its application in kombucha is still in the early stages. Future research should aim to optimize the encapsulation techniques and carrier materials specifically for kombucha fermentation in order to fully realize its potential as a probiotic functional beverage. Although certain conditions in other food matrices where encapsulated probiotic strains have been successfully applied may resemble those in kombucha, future research should also consider specific differences, particularly the complex interactions between the SCOBY micro-organisms and the added LAB strains, as well as their potential interactions with the encapsulation carriers.

5. Beneficial Effect of Probiotic Kombucha

According to the World Health Organization (WHO/FAO), probiotics are “living microorganisms that, when used in adequate amounts, confer health benefits to the host” [83]. The most common probiotic bacteria belong to the group of lactic acid bacteria (LAB), particularly Lactobacillus and Bifidobacterium, although certain non-LAB species and yeasts such as Saccharomyces boulardii are also used [4]. Documented benefits include the alleviation of lactose intolerance, immune system modulation and reduction in digestive infections and even precancerous lesions [84,85].
Due to the health benefits mentioned, there has been an increased demand for probiotic products over the last decade, which has boosted a sharp increase in the development of novel foods and supplements containing probiotics for the consumer market [86]. Fermented probiotic beverages are often produced with the involvement of diverse microbial consortia, such as the kombucha culture, which exemplifies mutual metabolic interactions between prokaryotic and eukaryotic organisms, including various bacteria and yeasts. Numerous studies have highlighted that the micro-organisms present in kombucha exhibit probiotic potential, with reports indicating the presence of over 50 distinct probiotic strains in this beverage [87]. The bacteria and yeasts found in kombucha function as probiotics, while the cellulose pellicle supports the growth of beneficial gut micro-organisms [2]. Additionally, kombucha provides short-chain fatty acids and other bioactive metabolites, such as organic acids, catechins, DSL etc., which contribute to immune system enhancement, microbiota balance and improved gastrointestinal health in humans [4].
The addition of specific LAB strains has been shown to further enhance the functional potential of kombucha. For example, several Lactobacillus strains have demonstrated beneficial effects, such as increased production of lactic acid, DSL and glucuronic acid, as well as reduction in serum glucose levels [49,52,55]. Probiotic kombucha enriched with encapsulated L. rhamnosus showed significant antidiabetic, antihypertensive and cholesterol-lowering activity, along with preserved sensory qualities compared to the control sample [13].
While there is substantial evidence supporting the health benefits of probiotics, much of the research has been conducted on individuals with pre-existing health conditions. Data on the effects of probiotic consumption in healthy populations remain limited and less conclusive [86]. Furthermore, many promising results observed in experimental models have yet to be validated through clinical trials. Taken together, kombucha shows strong potential as a probiotic functional beverage, especially when enriched with encapsulated probiotic strains. However, further research is needed to optimize the encapsulation strategies, understand microbial interactions and validate functional claims through clinical studies in diverse populations.

6. Conclusions and Future Perspectives

Kombucha continues to gain popularity as a functional beverage, but its characterization as a true probiotic product remains premature. The review highlights that although LAB are occasionally present in kombucha, their concentrations are typically too low and inconsistent to meet the accepted thresholds for probiotic efficacy. This microbial instability, both during fermentation and throughout storage, remains one of the most critical barriers to probiotic kombucha development. Moreover, the acidic nature of the kombucha matrix, although central to its sensory and antimicrobial properties, poses a major physiological challenge for probiotic survival. Most LAB strains are poorly adapted to persist in such environments, particularly over prolonged storage, and the rapid decline in CFU counts following fermentation further limits probiotic function. Encapsulation has emerged as a promising solution for improving probiotic stability in kombucha, but the research to date remains fragmented. Few studies directly address kombucha-specific encapsulation workflows, and those that do often lack methodological consistency, long-term viability data or industrial scalability assessments. Most techniques remain confined to lab-scale experiments with limited commercial relevance.
A path forward will require a coordinated effort to overcome these limitations. First, acid-tolerant probiotic strains with demonstrated compatibility with SCOBY consortia must be identified and validated. Second, the encapsulation methods should be optimized and standardized, particularly regarding materials suitable for beverage matrices under acidic stress. Third, shelf-life studies and real-world testing across different kombucha formulations are urgently needed. Finally, industrial translation will depend not only on microbiological success but also on regulatory clarity. Some regions still restrict the addition of probiotics following fermentation, which significantly narrows the production options. In short, kombucha holds clear promise as a probiotic carrier but only if its microbial, technological and regulatory challenges are met with targeted, multidisciplinary solutions. Without these, claims of probiotic benefit will remain speculative at best.

Author Contributions

Conceptualization, T.B. and A.R.; methodology, A.R. and T.B.; validation, A.T. and O.Š.; formal analysis, A.R., T.B. and O.Š.; investigation, O.Š. and A.T.; resources, A.R. and T.B.; writing—original draft preparation, T.B.; writing—review and editing, A.R. and O.Š.; visualization, A.T.; supervision, D.C.; project administration, D.C. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Ministry of Science, Technological Development and Innovation of the Republic of Serbia under the following grant numbers: 451-03-136/2025-03/200134, 451-03-137/2025-03/200134 and 451-03-136/2025-03/200051.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Coelho, R.M.D.; de Almeida, A.L.; do Amaral, R.Q.G.; da Mota, R.N.; de Sousa, P.H.M. Kombucha. Int. J. Gastron. Food Sci. 2020, 22, 100272. [Google Scholar] [CrossRef]
  2. Šovljanski, O.; Budimac, T.; Tomić, A.; Cvetković, D.; Ranitović, A. Kombucha As a Functional Beverage Rich in Phenolic Compounds. In Natural Products; Springer: Berlin/Heidelberg, Germany, 2025; pp. 1–42. [Google Scholar]
  3. Martins, H.F.; de Oliveira Santos, L.T.S.; de Carvalho, G.B.M.; Martinez, E.A. Kombucha: A literature review. Cuad. Educ. Y Desarro. 2023, 15, 11175–11197. [Google Scholar] [CrossRef]
  4. Watawana, M.I.; Jayawardena, N.; Gunawardhana, C.B.; Waisundara, V.Y. Health, wellness, and safety aspects of the consumption of kombucha. J. Chem. 2015, 2015, 591869. [Google Scholar] [CrossRef]
  5. Martínez Leal, J.; Valenzuela Suárez, L.; Jayabalan, R.; Huerta Oros, J.; Escalante-Aburto, A. A review on health benefits of kombucha nutritional compounds and metabolites. CyTA-J. Food 2018, 16, 390–399. [Google Scholar] [CrossRef]
  6. Cetojević-Simin, D.D.; Bogdanovic, G.M.; Cvetkovic, D.D.; Velicanski, A.S. Antiproliferative and antimicrobial activity of traditional Kombucha and Satureja montana L. Kombucha. J. BUON 2008, 133, 395–401. [Google Scholar]
  7. Bishop, P.; Pitts, E.R.; Budner, D.; Thompson-Witrick, K.A. Chemical composition of kombucha. Beverages 2022, 8, 45. [Google Scholar] [CrossRef]
  8. Emiljanowicz, K.E.; Malinowska-Pańczyk, E. Kombucha from alternative raw materials–The review. Crit. Rev. Food Sci. Nutr. 2020, 60, 3185–3194. [Google Scholar] [CrossRef]
  9. Jayabalan, R.; Waisundara, V.Y. Kombucha as a functional beverage. In Functional and Medicinal Beverages; Academic Press: Cambridge, MA, USA, 2019; pp. 413–446. [Google Scholar]
  10. Bortolomedi, B.M.; Paglarini, C.S.; Brod, F.C.A. Bioactive compounds in kombucha: A review of substrate effect and fermentation conditions. Food Chem. 2022, 385, 132719. [Google Scholar] [CrossRef]
  11. Vargas, B.K.; Fabricio, M.F.; Ayub, M.A.Z. Health effects and probiotic and prebiotic potential of Kombucha: A bibliometric and systematic review. Food Biosci. 2021, 44, 101332. [Google Scholar] [CrossRef]
  12. Maia, M.S.; Domingos, M.M.; de São José, J.F.B. Viability of Probiotic Microorganisms and the Effect of Their Addition to Fruit and Vegetable Juices. Microorganisms 2023, 11, 1335. [Google Scholar] [CrossRef]
  13. Budimac, T.; Pezo, L.; Šovljanski, O.; Cvetković, D.; Cvanić, T.; Vučetić, A.; Ranitović, A. An Optimal Probiotic Carrier: Multiple Steps Toward Selection and Application in Kombucha. Fermentation 2025, 11, 256. [Google Scholar] [CrossRef]
  14. Kim, J.; Adhikari, K. Current Trends in Kombucha: Marketing Perspectives and the Need for Improved Sensory Research. Beverages 2020, 6, 15. [Google Scholar] [CrossRef]
  15. Market Growth Reports. Kombucha Beverage Market Report. 2024. Available online: https://www.marketgrowthreports.com/market-reports/kombucha-beverage-market-100707 (accessed on 17 June 2025).
  16. De Oliveira, P.V.; da Silva Júnior, A.H.; de Oliveira, C.R.S.; Assumpção, C.F.; Ogeda, C.H. Kombucha benefits, risks and regulatory frameworks: A review. Food Chem. Adv. 2023, 2, 100288. [Google Scholar] [CrossRef]
  17. Batista, P.; Penas, M.R.; Pintado, M.; Oliveira-Silva, P. Kombucha: Perceptions and Future Prospects. Foods 2022, 11, 1977. [Google Scholar] [CrossRef] [PubMed]
  18. Soares, M.G.; de Lima, M.; Schmidt, V.C.R. Technological aspects of kombucha, its applications and the symbiotic culture (SCOBY), and extraction of compounds of interest: A literature review. Trends Food Sci. Technol. 2021, 110, 539–550. [Google Scholar] [CrossRef]
  19. Cvetković, D.D.; Markov, S.L.; Velićanski, A. Antimicrobial activity of kombucha made from Rtanj tea. Hem. Ind. 2025, 59, 248–253. [Google Scholar] [CrossRef]
  20. De Miranda, J.F.; Ruiz, L.F.; Silva, C.B.; Uekane, T.M.; Silva, K.A.; Gonzalez, A.G.M.; Fernandes, F.F.; Lima, A.R. Kombucha: A review of substrates, regulations, composition, and biological properties. J. Food Sci. 2022, 87, 503–527. [Google Scholar] [CrossRef]
  21. Greenwalt, C.J.; Steinkraus, K.H.; Ledford, R.A. Kombucha, the fermented tea: Microbiology, composition, and claimed health effects. J. Food Prot. 2000, 63, 976–981. [Google Scholar] [CrossRef]
  22. Chakravorty, S.; Bhattacharya, S.; Bhattacharya, D.; Sarkar, S.; Gachhui, R. Kombucha: A promising functional beverage prepared from tea. In Non-Alcohol. Beverages; Woodhead Publishing: Cambridge, UK, 2019; pp. 285–327. [Google Scholar]
  23. Wang, B.; Rutherfurd-Markwick, K.; Zhang, X.-X.; Mutukumira, A.N. Kombucha: Production and Microbiological Research. Foods 2022, 11, 3456. [Google Scholar] [CrossRef]
  24. Laavanya, D.; Shirkole, S.; Balasubramanian, P. Current challenges, applications and future perspectives of SCOBY cellulose of Kombucha fermentation, J. Clean. Prod. 2021, 259, 126454. [Google Scholar] [CrossRef]
  25. Villarreal-Soto, S.A.; Beaufort, S.; Bouajila, J.; Souchard, J.-P.; Taillandier, P. Understanding Kombucha Tea Fermentation: A Review. J. Food Sci. 2018, 83, 580–588. [Google Scholar] [CrossRef]
  26. Jayabalan, R.; Malbaša, R.V.; Lončar, E.S.; Vitas, J.S.; Sathishkumar, M. A Review on Kombucha Tea–Microbiology, Composition, Fermentation, Beneficial Effects, Toxicity, and Tea Fungus. Compr. Rev. Food Sci. Food Saf. 2014, 13, 538–550. [Google Scholar] [CrossRef]
  27. St-Pierre, D.L. Microbial Diversity of the Symbiotic Colony of Bacteria and Yeast (SCOBY) and Its Impact on the Organoleptic Properties of Kombucha. Master’s Thesis, The University of Maine, Orono, ME, USA, 2019. Available online: https://digitalcommons.library.umaine.edu/etd/3063/ (accessed on 12 June 2025).
  28. Watawana, M.I.; Jayawardena, N.; Gunawardhana, C.B.; Waisundara, V.Y. Enhancement of the antioxidant and starch hydrolase inhibitory activities of king coconut water (Cocos nucifera var. aurantiaca) by fermentation with Kombucha “tea fungus”. Int. J. Food Sci. Technol. 2016, 51, 490–498. [Google Scholar] [CrossRef]
  29. Laureys, D.; Britton, S.J.; de Clippeleer, J. Kombucha Tea Fermentation: A Review. J. Am. Soc. Brew. Chem. 2020, 78, 165–174. [Google Scholar] [CrossRef]
  30. Antolak, H.; Piechota, D.; Kucharska, A. Kombucha Tea-A Double Power of Bioactive Compounds from Tea and Symbiotic Culture of Bacteria and Yeasts (SCOBY). Antioxidants 2021, 10, 1541. [Google Scholar] [CrossRef] [PubMed]
  31. Marsh, A.J.; O’Sullivan, O.; Hill, C.; Ross, R.P.; Cotter, P.D. Sequence-based analysis of the bacterial and fungal compositions of multiple kombucha (tea fungus) samples. Food Microbiol. 2014, 38, 171–178. [Google Scholar] [CrossRef]
  32. Bogdan, M.; Justine, S.; Filofteia, D.C.; Petruta, C.C.; Gabriela, L.U.Ț.Ă.; Roxana, U.E.; Florentina, M.; Camelia Filofteia, D.; Călina Petruța, C.; Gabriela, L. Lactic acid bacteria strains isolated from Kombucha with potential probiotic effect. Rom. Biotechnol. Lett. 2018, 23, 13592–13598. [Google Scholar]
  33. Fabricio, M.F.; Mann, M.B.; Kothe, C.I.; Frazzon, J.; Tischer, B.; Flôres, S.H.; Ayub, M.A.Z. Effect of freeze-dried kombucha culture on microbial composition and assessment of metabolic dynamics during fermentation. Food Microbiol. 2022, 101, 103889. [Google Scholar] [CrossRef]
  34. Yang, J.; Lagishetty, V.; Kurnia, P.; Henning, S.M.; Ahdoot, A.I.; Jacobs, J.P. Microbial and Chemical Profiles of Commercial Kombucha Products. Nutrients 2022, 14, 670. [Google Scholar] [CrossRef]
  35. Coton, M.; Pawtowski, A.; Taminiau, B.; Burgaud, G.; Deniel, F.; Coulloumme-Labarthe, L.; Fall, A.; Daube, G.; Coton, E. Unraveling microbial ecology of industrial-scale Kombucha fermentations by metabarcoding and culture-based methods. FEMS Microbiol. Ecol. 2017, 93, fix048. [Google Scholar] [CrossRef]
  36. Chakravorty, S.; Bhattacharya, S.; Chatzinotas, A.; Chakraborty, W.; Bhattacharya, D.; Gachhui, R. Kombucha tea fermentation: Microbial and biochemical dynamics. Int. J. Food Microbiol. 2016, 220, 63–72. [Google Scholar] [CrossRef] [PubMed]
  37. Jafari, R.; Naghavi, N.S.; Khosravi-Darani, K.; Doudi, M.; Shahanipour, K. Isolation, molecular and phylogenetic identification of microorganisms from Kombucha solution and evaluation of their viability using flow cytometery. Food Sci. Technol. 2021, 42, e63220. [Google Scholar] [CrossRef]
  38. Zhu, W.; Lyu, F.; Naumovski, N.; Ajlouni, S.; Ranadheera, C.S. Functional Efficacy of Probiotic Lactobacillus sanfranciscensis in Apple, Orange and Tomato Juices with Special Reference to Storage Stability and In Vitro Gastrointestinal Survival. Beverages 2020, 6, 13. [Google Scholar] [CrossRef]
  39. Podolich, O.; Zaets, I.; Kukharenko, O.; Orlovska, I.; Reva, O.; Khirunenko, L.; Sosnin, M.; Haidak, A.; Shpylova, S.; Rohutskyy, I.; et al. The first space-related study of a kombucha multimicrobial cellulose-forming community: Preparatory laboratory experiments. Orig. Life Evol. Biosph. 2017, 47, 169–185. [Google Scholar] [CrossRef]
  40. Zhao, Z.J.; Sui, Y.C.; Wu, H.W.; Zhou, C.B.; Hu, X.C.; Zhang, J. Flavour chemical dynamics during fermentation of kombucha tea. Emir. J. Food Agric. 2018, 30, 732–741. [Google Scholar] [CrossRef]
  41. Tu, C.; Tang, S.; Azi, F.; Hu, W.; Dong, M. Use of kombucha consortium to transform soy whey into a novel functional beverage. J. Funct. Foods 2019, 52, 81–89. [Google Scholar] [CrossRef]
  42. Gaggìa, F.; Baffoni, L.; Galiano, M.; Nielsen, D.S.; Jakobsen, R.R.; Castro-Mejía, J.L.; Bosi, S.; Truzzi, F.; Musumeci, F.; Dinelli, G.; et al. Kombucha Beverage from Green, Black and Rooibos Teas: A Comparative Study Looking at Microbiology, Chemistry and Antioxidant Activity. Nutrients 2019, 11, 1. [Google Scholar] [CrossRef]
  43. Pei, J.; Jin, W.; Abd El-Aty, A.M.; Baranenko, D.A.; Gou, X.; Zhang, H.; Geng, J.; Jiang, L.; Chen, D.; Yue, T. Isolation, purification, and structural identification of a new bacteriocin made by Lactobacillus plantarum found in conventional kombucha. Food Control 2020, 110, 106923. [Google Scholar] [CrossRef]
  44. Villarreal-Soto, S.A.; Bouajila, J.; Pace, M.; Leech, J.; Cotter, P.D.; Souchard, J.P.; Taillandier, P.; Beaufort, S. Metabolome-microbiome signatures in the fermented beverage, Kombucha. Int. J. Food Microbiol. 2020, 333, 108778. [Google Scholar] [CrossRef]
  45. Arıkan, M.; Mitchell, A.L.; Finn, R.D.; Gürel, F. Microbial composition of Kombucha determined using amplicon sequencing and shotgun metagenomics. J. Food Sci. 2020, 85, 455–464. [Google Scholar] [CrossRef]
  46. Harrison, K.; Curtin, C. Microbial Composition of SCOBY Starter Cultures Used by Commercial Kombucha Brewers in North America. Microorganisms 2021, 9, 1060. [Google Scholar] [CrossRef] [PubMed]
  47. Kaashyap, M.; Cohen, M.; Mantri, N. Microbial Diversity and Characteristics of Kombucha as Revealed by Metagenomic and Physicochemical Analysis. Nutrients 2021, 13, 4446. [Google Scholar] [CrossRef] [PubMed]
  48. Harrison, K.; Navarro, R.; Jensen, K.; Cayler, W.; Nielsen, T.; Curtin, C. Live, Probiotic, or Neither? Microbial Composition of Retail-Available Kombucha and “Hard” Kombucha in the Pacific Northwest of the United States. Beverages 2023, 9, 59. [Google Scholar] [CrossRef]
  49. Al-Dulaimi, F.K.; Abd-Alwahab, W.I.; Hasan, A.S. Bioactivity study of kombucha black tea and kombucha with skim milk on some of physiological and biochemical parameters in male albino rats. Int. J. Pharm. Res. 2018, 10, 301. [Google Scholar]
  50. Bueno, F.; Chouljenko, A.; Sathivel, S. Development of coffee kombucha containing Lactobacillus rhamnosus and Lactobacillus casei: Gastrointestinal simulations and DNA microbial analysis. LWT 2021, 142, 110980. [Google Scholar] [CrossRef]
  51. Fu, C.; Yan, F.; Cao, Z.; Xie, F.; Lin, J. Antioxidant activities of kombucha prepared from three different substrates and changes in content of probiotics during storage. Food Sci. Technol. 2014, 34, 123–126. [Google Scholar] [CrossRef]
  52. Cvetković, D.; Ranitović, A.; Savić, D.; Joković, N.; Tomić, A.; Pezo, L.; Markov, S. Survival of wild strains of lactobacilli during Kombucha fermentation and their contribution to functional characteristics of beverage. Pol. J. Food Nutr. Sci. 2019, 69, 407–415. [Google Scholar] [CrossRef]
  53. Lee, S.M.; Lee, J.Y.; Yoo, D.G.; Jeon, Y.B.; Yoon, H.S.; Kim, C.H. Functional characteristics of kombucha fermented with lactic acid bacteria, yeast, and acetic acid bacteria derived from Korea traditional foods. J. Dairy Sci. Biotechnol. 2022, 40, 23–34. [Google Scholar] [CrossRef]
  54. Nguyen, N.K.; Dong, N.T.N.; Nguyen, H.T.; Le, P.H. Lactic acid bacteria: Promising supplements for enhancing the biological activities of kombucha. Springerplus 2015, 4, 91. [Google Scholar] [CrossRef]
  55. Yang, Z.; Zhou, F.; Ji, B.; Li, B.; Luo, Y.; Yang, L.; Li, T. Symbiosis between microorganisms from kombucha and kefir: Potential significance to the enhancement of kombucha function. Appl. Biochem. Biotechnol. 2010, 160, 446–455. [Google Scholar] [CrossRef]
  56. Majid, A.A.; Suroto, D.A.; Utami, T.; Rahayu, E.S. Probiotic potential of kombucha drink from butterfly pea (Clitoria ternatea L.) flower with the addition of Lactiplantibacillus plantarum subsp. plantarum Dad-13. Biocatal. Agric. Biotechnol. 2023, 51, 102776. [Google Scholar] [CrossRef]
  57. Cvetković, D.; Ranitović, A.; Budimac, T.; Šovljanski, O. Examination of Lactobacillus plantarum and Lactobacillus rhamnosus during kombucha fermentation. In Proceedings of the 2nd International Symposium on Biotechnology, Čačak, Serbia, 14–15 March 2024; pp. 409–414. [Google Scholar]
  58. Bromley, A.; Perry, J. Survival of Probiotic Lactobacillus spp. During Kombucha Fermentation. Curr. Dev. Nutr. 2022, 6, 507. [Google Scholar] [CrossRef]
  59. Chávarri, M.; Marañón, I.; Villarán, M.C. Encapsulation technology to protect probiotic bacteria. In Probiotics; IntechOpen: London, UK, 2012. [Google Scholar]
  60. Rama, G.R.; Führ, A.J.; da Silva, J.A.B.S.; Gennari, A.; Giroldi, M.; Goettert, M.I.; Volken de Souza, C.F. Encapsulation of Lactobacillus spp. using bovine and buffalo cheese whey and their application in orange juice. 3 Biotech 2020, 10, 263. [Google Scholar] [CrossRef] [PubMed]
  61. Razavi, S.; Janfaza, S.; Tasnim, N.; Gibson, D.L.; Hoorfar, M. Nanomaterial-based encapsulation for controlled gastrointestinal delivery of viable probiotic bacteria. Nanoscale Adv. 2021, 3, 2699–2709. [Google Scholar] [CrossRef] [PubMed]
  62. Rajam, R.; Subramanian, P. Encapsulation of probiotics: Past, present and future. Beni Suef Univ. J. Basic Appl. Sci. 2022, 11, 46. [Google Scholar] [CrossRef]
  63. Kavitake, D.; Kandasamy, S.; Devi, P.B.; Shetty, P.H. Recent developments on encapsulation of lactic acid bacteria as potential starter culture in fermented foods–A review. Food Biosci. 2018, 21, 34–44. [Google Scholar] [CrossRef]
  64. Serna-Cock, L.; Vallejo-Castillo, V. Probiotic encapsulation. Afr. J. Microbiol. Res. 2013, 7, 4743–4753. [Google Scholar] [CrossRef]
  65. Gul, O.; Atalar, I. Different stress tolerance of spray and freeze dried Lactobacillus casei Shirota microcapsules with different encapsulating agents. Food Sci. Biotechnol. 2019, 28, 807–816. [Google Scholar] [CrossRef]
  66. Tarifa, M.C.; Piqueras, C.M.; Genovese, D.B.; Brugnoni, L.I. Microencapsulation of Lactobacillus casei and Lactobacillus rhamnosus in pectin and pectin-inulin microgel particles: Effect on bacterial survival under storage conditions. Int. J. Biol. Macromol. 2021, 179, 457–465. [Google Scholar] [CrossRef]
  67. Barbosa, J.; Brandão, T.R.; Teixeira, P. Spray drying conditions for orange juice incorporated with lactic acid bacteria. Int. J. Food Sci. Technol. 2017, 52, 1951–1958. [Google Scholar] [CrossRef]
  68. Frakolaki, G.; Kekes, T.; Lympaki, F.; Giannou, V.; Tzia, C. Use of encapsulated Bifidobacterium animalis subsp. lactis through extrusion or emulsification for the production of probiotic yogurt. J. Food Process Eng. 2022, 45, e13792. [Google Scholar] [CrossRef]
  69. Gharibzahedi, S.M.T.; Smith, B. Legume proteins are smart carriers to encapsulate hydrophilic and hydrophobic bioactive compounds and probiotic bacteria: A review. Compr. Rev. Food Sci. Food Saf. 2021, 20, 1250–1279. [Google Scholar] [CrossRef]
  70. Afzaal, M.; Saeed, F.; Saeed, M.; Azam, M.; Hussain, S.; Mohamed, A.A.; Alamri, M.S.; Anjum, F.M. Survival and stability of free and encapsulated probiotic bacteria under simulated gastrointestinal and thermal conditions. Int. J. Food Prop. 2020, 23, 1899–1912. [Google Scholar] [CrossRef]
  71. Zeashan, M.; Afzaal, M.; Saeed, F.; Ahmed, A.; Tufail, T.; Ahmed, A.; Anjum, F.M. Survival and behavior of free and encapsulated probiotic bacteria under simulated human gastrointestinal and technological conditions. Food Sci. Nutr. 2020, 8, 2419–2426. [Google Scholar] [CrossRef] [PubMed]
  72. Mokhtari, S.; Jafari, S.M.; Khomeiri, M. Survival of encapsulated probiotics in pasteurized grape juice and evaluation of their properties during storage. Food Sci. Technol. Int. 2019, 25, 120–129. [Google Scholar] [CrossRef]
  73. Ayivi, R.D.; Gyawali, R.; Krastanov, A.; Aljaloud, S.O.; Worku, M.; Tahergorabi, R.; Silva, R.C.D.; Ibrahim, S.A. Lactic acid bacteria: Food safety and human health applications. Dairy 2020, 1, 202–232. [Google Scholar] [CrossRef]
  74. Nongonierma, A.B.; Abrlova, M.; Kilcawley, K.N. Encapsulation of a Lactic Acid Bacteria Cell-Free Extract in Liposomes and Use in Cheddar Cheese Ripening. Foods 2013, 2, 100–119. [Google Scholar] [CrossRef]
  75. Azarkhavarani, P.R.; Ziaee, E.; Hosseini, S.M.H. Effect of encapsulation on the stability and survivability of Enterococcus faecium in a non-dairy probiotic beverage. Food Sci. Technol. Int. 2019, 25, 233–242. [Google Scholar] [CrossRef]
  76. Olivares, A.; Soto, C.; Caballero, E.; Altamirano, C. Survival of microencapsulated Lactobacillus casei (prepared by vibration technology) in fruit juice during cold storage. Electron. J. Biotechnol. 2019, 42, 42–48. [Google Scholar] [CrossRef]
  77. Hossain, M.N.; Ranadheera, C.S.; Fang, Z.; Ajlouni, S. Impact of encapsulating probiotics with cocoa powder on the viability of probiotics during chocolate processing, storage, and in vitro gastrointestinal digestion. J. Food Sci. 2021, 86, 1629–1641. [Google Scholar] [CrossRef]
  78. Romero-Chapol, O.O.; Varela-Pérez, A.; Castillo-Olmos, A.G.; García, H.S.; Singh, J.; García-Ramírez, P.J.; Viveros-Contreras, R.; Figueroa-Hernández, C.Y.; Cano-Sarmiento, C. Encapsulation of Lacticaseibacillus rhamnosus GG: Probiotic Survival, In Vitro Digestion and Viability in Apple Juice and Yogurt. Appl. Sci. 2022, 12, 2141. [Google Scholar] [CrossRef]
  79. Varela-Pérez, A.; Romero-Chapol, O.O.; Castillo-Olmos, A.G.; García, H.S.; Suárez-Quiroz, M.L.; Singh, J.; Figueroa-Hernández, C.Y.; Viveros-Contreras, R.; Cano-Sarmiento, C. Encapsulation of Lactobacillus gasseri: Characterization, Probiotic Survival, In Vitro Evaluation and Viability in Apple Juice. Foods 2022, 11, 740. [Google Scholar] [CrossRef]
  80. Mahmoodi Pour, H.; Marhamatizadeh, M.H.; Fattahi, H. Encapsulation of different types of probiotic bacteria within conventional/multilayer emulsion and its effect on the properties of probiotic yogurt. J. Food Qual. 2022, 2022, 7923899. [Google Scholar] [CrossRef]
  81. Sun, W.; Nguyen, Q.D.; Sipiczki, G.; Ziane, S.R.; Hristovski, K.; Friedrich, L.; Visy, A.; Hitka, G.; Gere, A.; Bujna, E. Microencapsulation of Lactobacillus plantarum 299v Strain with Whey Proteins by Lyophilization and Its Application in Production of Probiotic Apple Juices. Appl. Sci. 2023, 13, 318. [Google Scholar] [CrossRef]
  82. Budimac, T.; Ranitović, A.; Cvetković, D.; Šovljanski, O.; Tomić, A.; Vučetić, A.; Cvanić, T. Survivability of microencapsulated Lactobacillus plantarum during kombucha fermentation. In Book of Abstracts: 5th International Congress “Food Technology, Quality and Safety–FoodTech 2024”, Proceedings of the 5th International Congress “Food Technology, Quality and Safety–FoodTech 2024”, Novi Sad, Serbia, 16–18 October 2024; University of Novi Sad, Institute of Food Technology: Novi Sad, Serbia, 2024. [Google Scholar]
  83. FAO/WHO. The Food and Agriculture Organization of the United Nations and the World Health Organization Joint FAO/WHO Expert Consultation on the Evaluation of Health and Nutritional Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria FAO/WHO Report No. 10-1-2001. 2001. Available online: https://openknowledge.fao.org/items/db384295-64d9-47e2-b65b-3c918efc5140 (accessed on 1 June 2025).
  84. Asar, R.; Erenler, S.; Devecioglu, D.; Ispirli, H.; Karbancioglu-Guler, F.; Ozturk, H.I.; Dertli, E. Understanding the Functionality of Probiotics on the Edge of Artificial Intelligence (AI) Era. Fermentation 2025, 11, 259. [Google Scholar] [CrossRef]
  85. Mazziotta, C.; Tognon, M.; Martini, F.; Torreggiani, E.; Rotondo, J.C. Probiotics Mechanism of Action on Immune Cells and Beneficial Effects on Human Health. Cells 2023, 12, 184. [Google Scholar] [CrossRef] [PubMed]
  86. Khalesi, S.; Bellissimo, N.; Vandelanotte, C.; Williams, S.; Stanley, D.; Irwin, C. A review of probiotic supplementation in healthy adults: Helpful or hype? Eur. J. Clin. Nutr. 2019, 73, 24–37. [Google Scholar] [CrossRef]
  87. Kozyrovska, N.O.; Reva, O.M.; Goginyan, V.B.; de Vera, J.-P. Kombucha microbiome as a probiotic: A view from the perspective of post-genomics and synthetic ecology. Biopolymers Cell 2012, 28, 103–113. [Google Scholar] [CrossRef]
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Figure 1. Kombucha as a traditional fermented beverage.
Figure 1. Kombucha as a traditional fermented beverage.
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Figure 2. Symbiotic consortium of bacteria and yeast.
Figure 2. Symbiotic consortium of bacteria and yeast.
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Figure 3. Approaches of addition of probiotic strains in kombucha samples.
Figure 3. Approaches of addition of probiotic strains in kombucha samples.
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Figure 4. Encapsulation of probiotic(s) for kombucha addition.
Figure 4. Encapsulation of probiotic(s) for kombucha addition.
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MDPI and ACS Style

Budimac, T.; Ranitović, A.; Šovljanski, O.; Cvetković, D.; Tomić, A. Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha. Fermentation 2025, 11, 434. https://doi.org/10.3390/fermentation11080434

AMA Style

Budimac T, Ranitović A, Šovljanski O, Cvetković D, Tomić A. Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha. Fermentation. 2025; 11(8):434. https://doi.org/10.3390/fermentation11080434

Chicago/Turabian Style

Budimac, Tara, Aleksandra Ranitović, Olja Šovljanski, Dragoljub Cvetković, and Ana Tomić. 2025. "Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha" Fermentation 11, no. 8: 434. https://doi.org/10.3390/fermentation11080434

APA Style

Budimac, T., Ranitović, A., Šovljanski, O., Cvetković, D., & Tomić, A. (2025). Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha. Fermentation, 11(8), 434. https://doi.org/10.3390/fermentation11080434

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