Kombucha

Rasu Jayabalan, National Institute of Technology, Rourkela, Odisha, India

Radomir V Malbasa, University of Novi Sad, Novi Sad, Serbia
Muthuswamy Sathishkumar, R&D Division, Eureka Forbes Ltd., Bangalore, Karnataka, India
Ó 2016 Elsevier Inc. All rights reserved.

Introduction 1
History 2
Preparation of Kombucha Tea 2
Composition of Kombucha Culture 2
Reported Biomolecules Present in Kombucha Tea 3
Kombucha Fermentation on Unusual Substrates 4
Reported Health Benefits of Kombucha 4
Toxic Effects of Kombucha 5
Kombucha is Nontoxic 7
Applications of Cellulose Pellicle 7
Conclusion 7
Acknowledgments 8
References 8

Introduction

Kombucha or kombucha tea locally refers to fermented refreshing beverage which tastes slightly sweet and acidic. It is consumed
worldwide for its unproved health benefits and it can be made locally in home with the kombucha culture brought from kombucha
drinkers without having much technical expertise on fermentation. Kombucha particularly refers to the microbial culture used to
ferment the tea infusion and kombucha tea refers to the tea infusion fermented with kombucha culture. However, the terminologies
kombucha and kombucha tea locally refers to the tea infusion fermented with kombucha culture. In reality, kombucha culture is
a biofilm made by symbiotic association of yeasts and acetic acid producing bacterial group which is otherwise called as ‘tea fungus.’
The name tea fungus was given to the biofilm since it appears like a fungal mat grown under static conditions. When this biofilm
grown in cooled tea broth added with sugar, it forms a cellulosic pellicle layer which is a characteristic feature of acetic acid bacteria.

Tea fungus

Fermented tea broth

Figure 1 Kombucha black tea having fermented broth and tea fungus. Reproduced with prior permission, Jayabalan, R., Malbasa, R.V., Loncar,
E.S., Vitas, J.S., Sathishkumar, M., 2014. A review on kombucha teadmicrobiology, composition, fermentation, beneficial effects, toxicity, and tea
fungus. Compr. Rev. Food Sci. Food Saf. 13, 538–550.

Reference Module in Food Sciences http://dx.doi.org/10.1016/B978-0-08-100596-5.03032-8 1

2 Kombucha

Hence, on completion of fermentation, fermented tea will have two parts, namely cellulosic layer which floats on the surface and
liquid broth which tastes sour (Figure 1). In general, black tea and white sugar are the substrates for the preparation of kombucha
tea. Green tea kombucha is also becoming popular due to the awareness on the health benefits of green tea polyphenols among
consumers. Very little or no clinical evidence available for the health benefits claimed for kombucha.

History

The available literature says that kombucha was initially used in East Asia for its health effects. During Tsin Dynasty (‘Ling Chi’),
about 220 BC, it was used in northeast China (Manchuria) for its energizing and detoxifying properties. Then it was introduced
to Japan by a physician named Kombu in 414 AD to treat the digestive troubles of the Emperor Inkyo. The name kombucha would
have arrived after this incident by adding the Japanese word for tea, ‘cha’ with physician Kombu (Kombu þ Cha). From Japan, the
popularity of kombucha expanded to Russia through trade routes with the name ‘Mo-Gu.’ In Russia, it was called in several names
(Jsakvasska, Kambucha, Japonskigrib, Cainii kvass, Cainiigrib). After getting popularized in other eastern European countries, kom-
bucha appeared in Germany (as Heldenpilz, Kombuchaschwamm) during twentieth century. During World War II, kombucha was
again introduced into Germany and then to France in 1950s followed by France-dominated North Africa. The habit of drinking
kombucha tea in European countries made shortages of tea leaves and sugar which shows the widespread acceptance of the kom-
bucha tea. It gained popularity in Italy (called ‘Funkochinese’) after World War II. During 1960s, Switzerland researchers reported
that habit of drinking kombucha was beneficial as similar as to drinking yogurt. This statement increased the popularity of kom-
bucha tea dramatically. The popularity of kombucha had also expanded due to its beneficial effects on human health and its easy
method of preparation in home with little scientific knowledge. Today, different flavors of kombucha and kombucha culture can be
purchased through online Web sites or from retail food stores worldwide. Gunther W. Frank published an electronic journal of
kombucha which is currently available worldwide in 30 different languages (Dufresne and Farnworth, 2000; Jayabalan et al., 2014).

Preparation of Kombucha Tea

Sugared tea decoction (infusion of black tea or green tea leaves or powder) is fermented with kombucha culture (tea fungus) for 2–
14 days. Fermentation changes the sugared tea into a refreshing drink with increased nutritive and medicinal properties. The
amount of sugar, tea leaves, or power and kombucha culture will differ from region to region based on the likings of the kombucha
drinkers. The standard procedure is given as in Scheme 1. Fermentation for 1 week will change the taste and flavor of sugared black
tea to slightly sweet and acidic sparkling flavor and longer fermentation results in vinegar like taste. It was reported that traditional
recipes use 50 g sucrose/l concentration for a long time and this sugar concentration provides the optimum concentration of
ethanol and lactic acid. Longer the fermentation higher the acid production by kombucha culture which may pose potential risks
to the consumers and hence an optimum fermentation time must be standardized by the consumer for pleasant flavor and taste
(Jayabalan et al., 2014).

Composition of Kombucha Culture

Lindau gave the formal botanical name ‘Medusomyces gisevii’ to symbiotic association of yeast and acetic bacteria. Local people used
to call it as ‘tea fungus’ or ‘tea mushroom.’ Scientifically, kombucha is neither a fungus nor a mushroom. These names are wrongly
given by the local people due to its similarity with mold growing on an undisturbed medium or with the upper portion of the mush-
room. Acetic acid bacteria synthesize cellulose fibers which make a network and appear like a surface mold. The microbes which
make the kombucha are not same throughout the world. Based on the region and the availability of the bacteria and yeast which
can grow together in the symbiotic association, the composition differs from region to region. Abundantly available prokaryotes in
kombucha belong to the bacterial genera Acetobacter and Gluconobacter. Presence of Acetobacter xylinum is reported by several
researchers. Acetobacter xylinum produces cellulose fibers as its secondary metabolite which floats on the surface of fermenting
tea broth. During fermentation, the bacterial and yeast cells will attach to the cellulosic network and embed in the layer to
make it thick. Acetobacter xylinum, Acetobacter pasteurianus, Acetobacter aceti, and Gluconobacter oxydans were reported as predomi-
nantly available acetic acid bacteria in kombucha. Gluconoacetobacter sp. A4 was identified from a preserved kombucha and the
bacteria was found to produce D-saccharic acid-1,4-lactone (DSL). Acetobacter intermedium sp. nov., which was a new species in genus
Acetobacter was also isolated and characterized from kombucha. Nitrogen fixing Acetobacter nitrogenifigens sp. nov., and nitrogen
fixing and cellulose producing Gluconoacetobacter kombuchae sp. nov., were also isolated from kombucha. Recently, it was found
that the bacteria in kombucha samples received from Canada, Ireland, United States, and United Kingdom belong to Gluconoace-
tobacter and Lactobacillus species and Acetobacter was the least dominant group in those samples (Jayabalan et al., 2014).
Yeasts in kombucha culture belong to genus Brettanomyces/Dekkera, Candida, Koleckera, Mycotorula, Mycoderma, Pichia, Saccharo-
myces, Schizosaccharomyces, Torulospora, and Zygosaccharomyces. Genus Brettanomyces includes Brettanomyces intermedius, Brettanomyces
bruxellensis, and Brettanomyces claussenii. The reported species in the genus Candida includes Candida famata, Candida guilliermondii,
Candida obtusa, Candida stellate, C. guilliermondii, Candida colliculosa, Candida Kefyr, and Candida krusei. Saccharomyces was identified
 Kombucha 3

50 g sucrose is added to 1 L water during boiling and stirred in.

5 g tea leaves is added and stirred for 5 minutes.

Tea leaves are filtered and tea is left to cool to room temperature (approximately 20
to 22 ºC)

Cooled sugared tea broth is inoculated with 24 g of kombucha culture (tea fungus)
and poured into a sterilized glass bottle of 1 to 1.5 L capacity.

0.2 L of previously fermented kombucha tea is added to lower the initial pH of tea
so that undesirable microorganisms would not grow during fermentation

The preparation is covered with a paper towel or cheese cloth to keep the insects
(Drosophila fruit flies) away

The preparation is incubated at 20 to 22ºC (the optimal temperature of incubation

ranges from 18 to 26ºC) in dark and away from sunlight.

Newly formed daughter culture will be visible above the old tea fungus in 3 to 4
days of incubation and will start to float and form a clear thin gel-like membrane
(biofilm) across the available surface. The tea starts to smell fermented and gas
bubbles appear from the carbonic acid production

During the course of fermentation, the old tea fungus will sinks to the bottom of the
tea broth and remains at its original volume

After 10-14 days, the newly formed tea fungus which appears as a disc of 2 cm
thickness is removed with a spoon and kept in small volume of fermented tea under
refrigeration. The remaining beverage is filtered and stored in capped bottles under
refrigeration and consumed as chilled beverage without any heating

Scheme 1 Standard procedure to prepare kombucha tea. Reproduced from Jayabalan, R., Malbasa, R.V., Loncar, E.S., Vitas, J.S., Sathishkumar,
M., 2014. A review on kombucha teadmicrobiology, composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr. Rev. Food Sci.
Food Saf. 13, 538–550.

by several researchers and the genus includes Saccharomyces cerevisiae and Saccharomyces bisporus. Schizosacchromyces genus was found
to have Schizosaccharomyces pombe and Zygosaccharomyces was identified as Zygosaccharomyces rouxii, Zygosaccharomyces bailii, and
Zygosaccharomyces kombuchaensis sp. n. Apart from these yeast species, Sacchromycodes ludwigii, and S. pombe were also reported.
The following yeast species were also reported: Torula, Torulopsis, Torulaspora delbrueckii, Mycoderma, Pichia, Pichia membranefa-
ciens, Kloeckera apiculata, and Kluyveromyces africanus (Jayabalan et al., 2014).

Reported Biomolecules Present in Kombucha Tea

Biochemical investigation carried out by several researchers around the world has revealed the presence of several biomolecules in
kombucha tea. The composition of kombucha is given as Table 1. All the investigations were carried out in static conditions. The
chemical composition of kombucha cannot be similar throughout the world due to the different metabolic pathways found in
different genus of bacteria and yeast. The heterogeneity in chemical composition can be complemented by the utilization of
different amount of sugar and black tea, different amount of kombucha tea broth used to start the fermentation, and different
lengths of fermentation time. Glucose and fructose is produced through invertase action on sucrose. Majority of fructose is con-
verted to ethanol by yeasts through glycolysis. Gluconic acid is produced from glucose by acetic acid bacteria and they produce ace-
tic acid from ethanol oxidation. pH reduction during the fermentation period is due to the production of various organic acids
(Dufresne and Farnworth, 2000). The chemical composition of kombucha beverage depends on the substrate in which it is
4 Kombucha

Table 1 Chemical composition of kombucha tea

Organic acids Sugars Vitamins Metals and inorganic salts Others

Acetic, gluconic, glucuronic, Sucrose, B1, B2, B6, Manganese, iron, nickel, copper, 14 amino acids, biogenic amines, purines,
citric, L-lactic, malic, glucose, B12, and C zinc, lead, cobalt, chromium, pigments, lipids, proteins, some hydrolytic
tartaric, malonic, oxalic, and fructose fluoride, chloride, bromide, enzymes, ethanol, antibiotically active matter,
succinic, pyruvic, usnic iodide, nitrate, phosphate, and carbon dioxide, phenol, tea polyphenols,
sulfate D-saccharic acid 1,4-lactone (DSL)

Reproduced with prior permission, Jayabalan, R., Malbasa, R.V., Loncar, E.S., Vitas, J.S., Sathishkumar, M., 2014. A review on kombucha teadmicrobiology, composition,
fermentation, beneficial effects, toxicity, and tea fungus. Compr. Rev. Food Sci. Food Saf. 13, 538–550.

cultivated, time and temperature of the fermentation, microbial profile of the kombucha culture, and the method used for analysis
(Table 2).

Kombucha Fermentation on Unusual Substrates

Usual substrates for the preparation of kombucha are black or green tea decoction sweetened with 5–8% sucrose. Kombucha
fermentation on unusual substrates like Coca-Cola, red wine, white wine, vinegar, Jerusalem artichoke extract, milk, honey, recon-
stituted whey, Echinacea, Mentha, etc., was reported. Kombucha fermentation had also been carried out by using unusual substrates
like Gugija and Omija teas, cheese whey, tea waste material, lemon balm, mulberry tea, Japanese green tea, jasmine tea, oolong tea,
sage, thyme, peppermint, sugar beet molasses, cherry juice, sweetened stinging nettle and winter savory extracts, milk and cactus
pear juice (Jayabalan et al., 2014; Ayed and Hamdi, 2015). Researchers found that the composition of the beverage made out of
fermenting these unusual substrates with kombucha resulted in different composition than the traditional kombucha beverage.

Reported Health Benefits of Kombucha

Kombucha drinkers claim several health benefits for consuming kombucha regularly. But, none of these benefits are proved with
human models. Reports on experimental animals concluded that kombucha is a potential antimicrobial, antioxidant, hepatopro-
tective, and anticancer agent.
Following are the reported effects of kombucha from tea drinkers’ testimony and Russian researchers (Dufresne and Farnworth,
2000).
Detoxify the blood
Reduce cholesterol level

Table 2 Predominant components in kombucha tea at the end of the fermentation on sugared black tea infusion

Component Initial Fermentation Fermentation

Component content (g l 1) sucrose (%) Black tea temperature ( C) time (days)

Acetic acid 8 10 2 bags 24  3 60

4.69 10 12 g l1 24  3 18
Glucuronic acid 0.0031 5 1.5 g l1 28 21
0.0026 7 1.5 g l1 28 21
0.0034 10 1.5 g l1 28 21
1.71 10 12 g l1 24  3 18
Gluconic acid 39 10 2 bags 24  3 60
Glucose 179.5 7 1.5 g l1 28 21
24.59 7 1.5 g l1 28 21
12 10 2 bags 24  3 60
Fructose 76.9 7 1.5 g l1 28 21
5.40 7 1.5 g l1 28 21
55 10 2 bags 24  3 60
Remained sucrose 192.8 7 1.5 g l1 28 21
11 10 2 bags 24  3 60
2.09 7 1.5 g l1 28 21

Reproduced with prior permission, Jayabalan, R., Malbasa, R.V., Loncar, E.S., Vitas, J.S., Sathishkumar, M., 2014. A review on kombucha teadmicrobiology,
composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr. Rev. Food Sci. Food Saf. 13, 538–550.
 Kombucha 5

Reduce atherosclerosis by regeneration of cell walls

Reduce blood pressure
Reduce inflammatory problems
Alleviate arthritis, rheumatism, and gout symptoms
Promote liver functions
Normalize intestinal activity, balance intestinal flora, cure hemorrhoids
Reduce obesity and regulate appetite
Prevent/heal bladder infection and reduce kidney calcification
Stimulate glandular systems
Protect against diabetes
Increase body resistance to cancer
Have an antibiotic effect against bacteria, viruses, and yeasts
Enhance the immune system and stimulate interferon production
Relieve bronchitis and asthma
Reduce menstrual disorders and menopausal hot flashes
Improve hair, skin, and nail health
Reduce an alcoholic’s craving for alcohol
Reduce stress and nervous disturbances, and insomnia
Relieve headaches
Improve eyesight
Counteract aging
Enhance general metabolism
Reported beneficial effects of kombucha tea in research papers are listed out in Table 3 with appropriate references. Antimicro-
bial activity of kombucha is due to the presence of higher concentration of acetic acid, catechins, and unidentified proteins and
compounds produced during fermentation. Kombucha inhibits the growth of Aeromonas hydrophila, Agrobacterium tumefaciens,
Bacillus cereus, Campylobacter jejuni, Candida albicans, Escherichia coli, Entamoeba cloacae, Helicobacter pylori, Pseudomonas aeruginosa,
Salmonella choleraesuis serotype Typhimurium, Salmonella typhimurium, Salmonella enteritidis, Shigella sonnei, Staphylococcus aureus,
Staphylococcus epidermis, Vibrio parahaemolytica, and Yersinia enterocolitica (Jayabalan et al., 2014).
Kombucha has been found to have higher antioxidant activity than the original black tea or green tea taken for fermentation.
Antioxidant activity of kombucha was tested by several researchers using both in vitro test and cell line models (Hoon et al.,
2014). Presence of tea polyphenols, ascorbic acid, DSL, low molecular weight components, and biotransformed compounds
produced from tea polyphenols by the enzymes of bacteria and yeast during fermentation period have been attributed for antiox-
idant potential of kombucha. Kombucha has been studied for hepatoprotective property against several liver toxicants and carcin-
ogens and had been proved using animal models like albino rats and mice. Hepatoprotection by kombucha tea is ascribed to its
antioxidant potential and its ability to support the detoxification process in liver. It was concluded that DSL produced by Glucona-
cetobacter sp. A4 in kombucha might be the responsible component for the hepatoprotective property of kombucha tea shown
against acetaminophen induced toxicity. In a recent review about the health promoting aspects of kombucha, Vina et al. (2014)
proposed that kombucha can be a fermented functional beverage for health prophylaxis and is due to detoxification, antioxidation,
energizing potencies, and promotion of depressed immunity. In 1951, Central Oncological Research Unit and Russian Academy of
Sciences in Moscow conducted a population study in Russia based on personal observations and testimonials from kombucha
drinkers about the anticancer property of kombucha and concluded positively. Several research studies used cancer cell lines to
study the antiproliferative and antiangiogenic activities of kombucha prepared from conventional and unconventional sources.
Higher concentration of tea polyphenols and their biotransformed products formed during fermentation were attributed for the
anticancer property of kombucha tea (Dufresne and Farnworth, 2000; Jayabalan et al., 2014).

Toxic Effects of Kombucha

While kombucha is celebrated for its numerous health benefits, there are some evidences for the toxic effects connected with intake
of kombucha. Centers for Disease Control and Prevention reported two cases of mysterious illness due to kombucha consumption.
Some drinkers reported for dizziness and nausea. Kombucha is not suggested for pregnant and lactating women. Anthrax Bacillus
was identified in kombucha fermented in unsanitary conditions. Four patients consumed kombucha were documented with
nausea, vomiting, jaundice, allergic reactions, head and neck pain. Lead poisoning was documented in people who were consuming
kombucha for 6 months which was brewed in ceramic pot. Lead poisoning might be due to the elution of lead from the ceramic pot
by acids in kombucha. Toxic nature of kombucha is not acceptable due to the small number of reports available, while the kom-
bucha is spreaded all over the world. The symptoms observed due to kombucha consumption might be due to the fermentation in
unhygienic conditions and presence of pathogenic microorganisms which cannot be sensed by the kombucha drinkers (Jayabalan
et al., 2014).
Table 3 Biological activities of kombucha tea reported through studies with experimental animals and cell lines

6
Biological activity Experimental animal/cells Treatment period/dose Parameters studied
1

Kombucha
Hypoglycemic activity Mice 3 days and 1.71 ml kg body weight Blood sugar level
Antioxidative stress against chromate Rat 30 days and 0.6 ml/200 g body weight Plasma and tissue MDA levels, delayed type hypersensitivity response,
GSH, peroxidase, catalase
Longevity Mice 3 years and ad libitum access Longevity, general health, and open-field exploratory behavioral outcomes
Antistress activity against cold and hypoxia Rat 15 days and 1.6, 8.0, and 16 ml kg1 body Plasma/blood MDA and reduced GSH, fecal output
weight
Antioxidative stress against lead Rat 45 days and 1 ml kg1 body weight Lipid peroxidation, creatine phosphokinase, GSH, SOD, GPx, DNA
fragmentation in liver
Prevention of weight loss in diabetics Rats 15 days and different dilutions of kombucha Weight loss
tea (25, 50, 75, and 100%) in place of water
Prevention of postoperative intraabdominal Rats 14 days and 15 ml kg1 of body weight Adhesion intensity score, inflammatory cell reaction, number of adhesion
adhesion formation bands
Protection on chromosomal aberrations Human peripheral 250, 500, and 1000 ml doses Chromosomal aberrations, mitotic index
induced by g-radiation lymphocytes
Protection on nephrotoxicity induced by Rat 2 weeks and 0.1 ml/100 g body weight Lipid peroxidation, oxidative stress
trichloroethylene
Hypocholesterolemic effect Rat 12 weeks and 66 ml kg1 body weight Total cholesterol, low-density and high-density lipoprotein cholesterol
Healing property on indomethancin-induced Mice 7 days and 15 mg kg1 body weight Histopathological and biochemical studies
gastric ulceration
Protection on phenol-induced cytotoxicity Mice 20 days and 1 ml kg1 body weight Micronuclei formation
Protection on mitomycin C- induced genotoxic Chinese hamster cell line 1 h and 0.295, 1.185, 4.75 mg ml1 (dry Chromosome aberrations frequency
effect CHO-K1 weight)
Hypoglycemic and antilipidemic properties Diabetic rats 30 days and 5 ml kg1 body weight a-amylase and lipase in plasma and pancreas, blood glucose
against alloxan
Cytogenic activity Human peripheral blood 1 h and 40 mg ml1 Frequencies of sister chromatic exchange and micronuclei formation
lymphocytes
Protective effects against oxidative stress- Swiss albino male rats 14 days and 150 mg lyophilized powder of Blood glucose, glycated hemoglobin, lipid peroxidation end products,
mediated damages in alloxan-induced kombucha tea/kg body weight protein carbonyl content, glutathione content, antioxidant enzyme
diabetic rats activities
Amelioration of changes in trace element Male Wistar rats 9 weeks and 0.1 ml/100 g body weight/day Iron, zinc, and copper in brain, spleen, and intestine
levels in electromagnetic field-exposed rats
(950 MHz)
Antihyperglycemic effect in streptozotocin- Male albino Wistar rats 45 days and 3, 6, 12 mg of lyophilized solvent Glycosylated hemoglobin, plasma insulin, hemoglobin, and tissue
induced diabetic rats extract of kombucha/kg body weight/day glycogen, glucose-6-phosphatase, fructose-1,6-bisphosphatase and
hexokinase
Attenuation of oxidative damage in Male Wistar rats 57 days and 0.1 ml/100 g body weight/day Malondialdehyde, superoxide dismutase, lactate dehydrogenase,
electromagnetic field-exposed rats aspartate amino transferase, tissue glutathione levels in heart and lung,
(950 MHz) serum total antioxidant capacity
Full thickness wound healing Wistar albino rats 20 days and 0.1 ml kombucha as ointment Angiogenesis, fibroplasias, complete healing and inflammatory cell
form infiltration, histopathological examination
Antihypercholesterolemic activity Male albino Wistar rats 60 days and 2.5 ml/kg body weight/day Total cholesterol, high density cholesterol, low density cholesterol,
triglycerides, alanine transaminase, asparagine transaminase,
homocysteine

MDA - Malondialdehyde; SOD - Superoxide dismutase; GSH - Reduced glutathione; GPx - Glutathione peroxidase.
Reproduced with prior permission, Jayabalan, R., Malbasa, R.V., Loncar, E.S., Vitas, J.S., Sathishkumar, M., 2014. A review on kombucha teadmicrobiology, composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr. Rev. Food Sci.
Food Saf. 13, 538–550.
 Kombucha 7

(a)

(b)

Figure 2 (a, b) Scanning electron microscope image of the consortia of yeasts and bacteria in a portion of tea fungus (magnification (a) ¼ 3500
and (b) ¼ 3700). Reproduced with prior permission, El-Taher, E.M., 2011. Kombucha: a new microbial phenomenon and industrial benefits. Afr. J.
Biol. Sci. 7, 41–60; Jayabalan, R., Malbasa, R.V., Loncar, E.S., Vitas, J.S., Sathishkumar, M., 2014. A review on kombucha teadmicrobiology,
composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr. Rev. Food Sci. Food Saf. 13, 538–550.

Kombucha is Nontoxic

There are evidences from authentic sources to prove that kombucha is nontoxic drink if it is prepared with good knowledge and
under hygienic conditions. The U.S. Food and Drug Administration and Kappa Laboratories, Miami, Florida, USA have reported
that kombucha is safe for human consumption. Kombucha did not show any toxic signs in rats after 90 days feeding. Kombucha
did not have any toxicity in rats after feeding three times higher doses than the normal dose for 15 days. Administration of 2.5 ml/kg
body weight/day in male albino Wistar rats for 60 days did not show any liver toxicity and mortality (Jayabalan et al., 2014).

Applications of Cellulose Pellicle

The cellulose pellicle produced by Acetobacter species during fermentation floats on the surface of the tea and attaches the bacterial
and yeast cells (Figure 2(a) and 2(b)). This appearance is similar to the fungal mycelial mat floating on the surface of static cultures
and hence given the name ‘tea fungus’ to kombucha. Dried tea fungus has been applied as biosorbent to remove metal pollutants
like arsenic, chromium, and copper from waste water. Dried tea fungal biomass was utilized as protein supplement in broiler feed.
Tea fungus was studied to be rich in amino acid lysine, crude protein, and crude fiber and the composition have positive correlation
with the fermentation time. Due to its pure form, tea fungal cellulose can be utilized for the production of cellulose-based biochem-
ical and bioethanol. Biocompatibility of tea fungal cellulose on cell line models was also demonstrated and reported that it did not
show any toxic effects (Jayabalan et al., 2014).

Conclusion

At present kombucha is consumed worldwide in different forms sold by several companies and the local people still
drink kombucha fermented in their home. The benefits claimed for kombucha is not similar throughout world due to the variations
8 Kombucha

in fermentation time, amount of sugar and tea, types of tea, and microbial composition of tea fungus. None of the biological effects
claimed for kombucha consumption are studied on human models. Kombucha can be considered as a functional food and health
drink rich in pharmacologically active biomolecules. The identification of the potential biomolecule responsible for the health
benefits and the extracellular enzymes responsible for the biotransformation of tea polyphenols during kombucha fermentation
will change the current opinion about kombucha in future.

Acknowledgments

Author Rasu Jayabalan acknowledges the support given by the National Institute of Technology (Rourkela, Odisha, India), Prof. K. Swaminathan
(Dept. of Microbial Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India), Prof. Sei Eok Yun (Department of Food Science and
Technology, Institute of Agricultural Science and Technology, Chonbuk National University, Jeonju, Republic of Korea), Dr S. Marimuthu (R & D
Center, Parry Agro Industries Ltd, Valparai, Tamil Nadu, India), Department of Science and Technology (DST grant No. SERC/LS-156/2012) and
Department of Biotechnology (DBT grant No. 102/IFD/SAN/2770/2013–2014), Ministry of Science and Technology, Government of India.
Author Radomir Malbasa thanks the Ministry of Education, Science and Technological Development, Republic of Serbia (Grant III-46009).

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and tea fungus. Compr. Rev. Food Sci. Food Saf. 13, 538–550.
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