Journal of Food Science - 2022 - Miranda - Kombucha A Review of Substrates Regulations Composition and Biological

17503841, 2022, 2, Downloaded from https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.16029 by UFAL - Universidade Federal de Alagoas, Wiley Online Library on [22/02/2023].
See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Received: 17 July 2021 Revised: 3 December 2021 Accepted: 12 December 2021
DOI: 10.1111/1750-3841.16029
CONCISE REVIEWS & H YPOTH ESES IN FOOD SCIENCE
Kombucha: A review of substrates, regulations,

composition, and biological properties
Jeniffer Ferreira de Miranda1 Larissa Fernandes Ruiz1

Cíntia Borges Silva1 Thais Matsue Uekane1 Kelly Alencar Silva1
Alice Gonçalves Martins Gonzalez1 Fabrício Freitas Fernandes2
Adriene Ribeiro Lima1
1 Department of Bromatology, Pharmacy
School, Fluminense Federal University, Abstract: Kombucha has been gaining prominence around the world and
Niterói, RJ, Brazil becoming popular due to its good health benefits. This beverage is historically
2 Institute of Health Sciences, Paulista
obtained by the tea fermentation of Camellia sinensis and by a biofilm of cellu-
University, Ribeirão Preto, SP, Brazil
lose containing the symbiotic culture of bacteria and yeast (SCOBY). The other
Correspondence substrates added to the C. sinensis tea have also been reported to help kombucha
Adriene Ribeiro Lima, Department of Bro- production. The type as well as the amount of sugar substrate, which is the ori-
matology, Pharmacy School, R. Dr. Mario
Vianna no. 523, Faculdade de Farmácia, gin of SCOBY, in addition to time and temperature of fermentation influence
Niterói, RJ 24241-000, Brazil. the content of organic acids, vitamins, total phenolics, and alcoholic content of
Email: adrienelima@id.uff.br
kombucha. The route involved in the metabolite biotransformation identified in
Funding information kombucha so far and the microorganisms involved in the process need to be fur-
Conselho Nacional de Desenvolvimento ther studied. Some nutritional properties and benefits related to the beverage
Científico e Tecnológico - CNPq; Coorde-
have already been reported. Antioxidant and antimicrobial activities and antidi-
nação de Aperfeiçoamento de Pessoal de
Nível Superior - CAPES; Fundação Car- abetic and anticarcinogenic effects are some of the beneficial effects attributed
los Chagas Filho de Amparo à Pesquisa to kombucha. Nevertheless, scientific literature needs clinical studies to evaluate
do Estado do Rio de Janeiro - FAPERJ,
Grant/Award Number: Project E-26/200
these benefits in human beings. The toxic effects associated with the consump-
930/2017. tion of kombucha are still unclear, but due to the possibility of adverse reactions
occurring, its consumption is contraindicated in infants and pregnant women,
children under 4-years-old, patients with kidney failure, and patients with HIV.
The regulations in place for kombucha address a number of criteria, mainly for
the pH and alcohol content, in order to guarantee the quality and safety of the
beverage as well as to ensure transparency of information for consumers.
KEYWORDS
Camellia sinensis, green tea, black tea, fermented beverage, alternative substrates
1 INTRODUCTION (SCOBY) (Kapp & Sumner, 2019), also known as “tea fun-
gus” (Dutta & Paul, 2019), which consist of a symbiosis of
Kombucha is a slightly sweet and acidic beverage, tradi- acetic bacteria, lactic acid bacteria, and osmophilic yeasts
tionally produced from the fermentation of black or green (Cardoso et al., 2020).
tea (Camellia sinensis), adding sugar by a biofilm of cellu- Kombucha is believed to have originated in northeast-
lose containing the symbiotic culture of bacteria and yeast ern China (Manchuria), where it was valued during the
J. Food Sci. 2022;87:503–527. wileyonlinelibrary.com/journal/jfds © 2022 Institute of Food Technologists

R 503
17503841, 2022, 2, Downloaded from https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.16029 by UFAL - Universidade Federal de Alagoas, Wiley Online Library on [22/02/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
504 KOMBUCHA: A REVIEW
Tsin Dynasty (“Ling Chi”), around 220 BC, for its supposed 2 KOMBUCHA SUBSTRATES
health benefits (Chakravorty et al., 2019). With the expan-
sion of commercial routes, it started migrating to the rest of Black or green tea is traditionally used for the production of
the world. In addition to China, Korea, and Japan, the pop- kombucha. Both black and green tea come from the same
ular practice of consuming kombucha became prevalent in species of plant, C. sinensis, a tree of the botanical fam-
Russia and in other Eastern European countries, emerging ily Theaceae, native to Southeast Asia (Tanaka & Kouno,
in Germany in the 20th century (Dutta & Paul, 2019; Jaya- 2003).
balan et al., 2014). In the 1950s, it reached France and also C. sinensis leaves produce different teas from differ-
North Africa. The habit of consuming fermented beverages ent manufacturing processes. However black, green, and
became acceptable throughout Europe, further increasing oolong teas are the main differentiated types accord-
its popularity in the 1960s. Nowadays, kombucha is sold ing to the degree of fermentation and oxidation of the
worldwide (Jayabalan et al., 2014) and continues to gain leaves (Dutta & Paul, 2019; Sang et al., 2011). Green tea
popularity, especially in Brazil. is produced from fresh leaves of the plant and thus pre-
Although C. sinensis tea is the main substrate used to serve more polyphenol content with a higher concentra-
produce kombucha, there are various studies with other tion of catechins and compounds with functional activ-
raw materials as a substrate to produce this beverage (Emil- ity as compared to other teas produced from C. sinensis
janowicz & Malinowska-Pańczyk, 2019). Fermentation of (Cardoso et al., 2020). Black tea is obtained from aged
kombucha culture on alternative substrates, such as herbal leaves, influenced by the fermentation and oxidation pro-
infusions (Battikh, Bakhrouf & Ammar , 2012), fruit juice cess of catechins that lead to the formation of theaflavins
(Ayed, Ben Abid & Hamdi, 2016), milk (Kanurić et al., and thearubigins, which are the main polyphenolic com-
2018), soy (Tu et al., 2019), unconventional food plant (Mal- pounds present in black tea (Cardoso et al., 2020). Oolong
vaviscus arboreus) (Silva et al., 2021), among others, has tea is produced with unique strategies through strong sun
been reported and they have presented satisfactory results drying and oxidation before torsion. The chemical oxida-
regarding their kinetics and biological properties. The fer- tion process causes changes in the leaves, influencing the
mented beverages produced with these other substrates striking characteristics of teas such as color, flavor, and
were cited in this work as kombucha analogs. aroma (Yanagimoto et al., 2003).
Due to its ease of use, sour flavor, and especially its bene- The high consumption of tea from C. sinensis has been
ficial health-promoting properties, kombucha has become associated with the known functional properties, linking
an attractive drink. Some benefits attributed to kombucha the positive effect due to the presence of antioxidants and
have already been proven in in vitro studies and with ani- phenolic compounds which reduce the risk of diseases
mals, such as oxidative stress control (Srihari, Karthike- (Santos et al., 2018). C. sinensis contains various polyphe-
san, et al., 2013), antimicrobial activity (Battikh et al., 2011), nols, flavonols, catechins, catechin gallates, adenine, caf-
diabetes treatment and prevention (Aloulou et al., 2012), feine, theobromine, theophylline, gallic acids, tannins,
cancer spread reduction (Srihari, Arunkumar, et al., 2013), and gallotannin. Such compounds make this beverage as
and liver function improvement (hepatoprotective) (Wang having high potential antioxidant properties (Aboulwafa
et al., 2013). Kombucha is a beverage rich in substances et al., 2019). Catechins and tannins are the most prevalent
with bioactive properties, especially phenolic compounds. polyphenols. These compounds contribute to the bitter-
These represent the main group of antioxidants present ness, astringency, and sweet taste of tea beverages (Santos
in kombucha and are responsible for the supposed health et al., 2018). In addition, other minor compounds present
benefits of the beverage (Cardoso et al., 2020). in C. sinensis tea, such as amino acids, chloride, oxides,
A considerable increase in the search for foods that carotenoids, lipids, and volatile compounds, may play an
contribute to health promotion has occurred frequently important role in aroma development (Dutta & Paul, 2019).
in recent years and has encouraged the consumption of In addition to its antioxidant activities and the diversity
and development of research on kombucha (Santos-Buelga of bioactive compounds, C. sinensis has other remark-
et al., 2019). In this context, this article aimed to gather data able health benefits such as antimicrobial, antineoplas-
from the main scientific studies on kombucha and make tic, cardioprotective, antidiabetic, and antiobesity activities
them available in the form of this review, focusing on its (Aboulwafa et al., 2019; Santos et al., 2018).
functionality, different substrates and preparation meth-
ods, chemical composition, and its main beneficial contri-
butions. It also aimed to describe the available regulations, 3 KOMBUCHA ANALOGS
especially Brazilian Normative Instruction No. 41 (Brasil,
2019), as well as the practices for obtaining a drink with The use of other products to prepare kombucha has already
quality and microbiological safety. been described in the literature. Examples include herbs
KOMBUCHA: A REVIEW 505
infusions, wax mallow flowers, coffee, oak leaves, euca- tional beverage. All study results indicated that fermenta-
lyptus, bay leaves, fruit juices, milk, and soy products, tion improved the beneficial properties of soy extract, as
which have proven to be good alternatives for fermentation fermentation significantly increased the content of pheno-
with SCOBY kombucha (Emiljanowicz & Malinowska- lic compounds and vitamins. Furthermore, stachyose and
Pańczyk, 2019; Silva et al., 2021). raffinose, two oligosaccharides present in soy, which cause
Herbal infusions have been used over many years as intestinal discomfort, were consumed during fermenta-
health-promoting beverages, and according to literature tion, indicating that the microorganisms present in the fer-
reports, this promoting activity can be enhanced after the mented beverage can produce the enzyme α-galactosidase.
fermentation process with SCOBY kombucha (Emiljanow- All these substrates stand out due to their chemical
icz & Malinowska-Pańczyk, 2019). Battikh, Bakhrouf and composition rich in bioactive compounds, which favor
Ammar (2012) evaluated the antimicrobial activity of kom- various physiological effects, in addition to potential bio-
bucha produced from black tea analogs such as T. vul- logical effects associated with antioxidant, antimicrobial,
garis L. (thyme), R. officinalis (rosemary), F. vulgare (fen- anti-inflammatory, gastroprotective, and anticarcinogenic
nel), and M. piperita (mint) and found higher activity, activities (Rocha-Guzmán et al., 2019). Therefore, studies
mainly against Candida spp., compared to their respec- to investigate beverages fermented with kombucha SCOBY
tive unfermented beverages as well as to black tea kom- prepared from different substrates have been stimulated,
bucha Četojević-Simin et al. (2012) and Velićanski et al. mainly to develop new beverages with more attractive
(2014) showed that the infusion of lemon balm (Melissa properties (Emiljanowicz & Malinowska-Pańczyk, 2019).
officinalis L.) fermented with kombucha culture showed
higher antimicrobial and antioxidant activity compared to
the unfermented infusion. Achillea millefolium (Yarrow), 4 PREPARATION METHODS
a medicinal plant, was used by Vitas et al. (2018) for fer-
mentation with kombucha SCOBY and they found good According to Villarreal-Soto et al. (2018), there are sev-
antimicrobial and antioxidant activity as well as antipro- eral factors which may influence kombucha fermentation,
liferative activity against human tumor cells. such as temperature, pH, oxygen, substrate, sugar con-
Ayed and Hamdi (2015) used prickly pear juice (Opun- centration, SCOBY origin, container geometry, and fer-
tia ficus-indica) as an alternative substrate to develop a mentation time. Any variation in these factors can inter-
new beverage with enhanced nutritional properties and fere with sensory characteristics, chemical properties, and
found increased antioxidant activity throughout fermenta- nutritional quality, and can influence biological activities.
tion, and antimicrobial activity mainly against Escherichia The amount of substrate, sugar, and symbiotic kom-
coli. Ayed, Ben Abid and Hamdi (2016) developed a kombucha culture used in fermentation varies according to
bucha SCOBY-fermented beverage from grape juice that the region of preparation, and several preparation condi-
had better sensory acceptance and functional properties tions have already been reported in the literature (Tables 1
compared to the unfermented grape beverage after 6 days and 2). Generally, as observed in the studies presented in
of fermentation. Salak fruit juice fermented with kom- Tables 1 and 2, tea or infusions can be prepared in a wide
bucha SCOBY presented important antidiabetic proper- range of concentrations ranging from approximately 1.0 to
ties such as plasma glucose reduction and regeneration of 100 g/L. Sucrose can be used in a concentration of 1–20%,
pancreatic β cells of diabetic rats (Zubaidah et al., 2018). as the main source of carbon for the growth of microor-
Zubaidah et al. (2019) found that the activity of fermented ganisms. SCOBY and the liquid from the previous fer-
salak fruit juice in reducing oxidative stress, blood glucose, mentation, in concentrations around 0.25–10% and 3–30%,
and lipid profile was comparable to the activity of met- respectively, should be used as starter culture for fermen-
formin. tation. The addition of the preceding fermented liquid has
Vázquez-Cabral et al. (2017) investigated the antioxi- the function of helping the activation of acetic bacteria due
dant activity and anti-inflammatory effects of a kombucha to the decrease in pH (Leal et al., 2018).
SCOBY-fermented beverage produced from the infusion of Fermentation occurs, ideally, at a temperature close to
oak leaves (Quercus sp.) and concluded that the beverage 25◦ C, in a period of 7–14 days. During the fermentation
showed good antioxidant property due to its rich compo- period, a new microbial biofilm develops on the surface
sition of phenolics (Rocha-Guzmán et al., 2019), as well as of the tea. Then, the recently formed culture is removed
important anti-inflammatory activity due to the decrease and can be stored immersed in the fermented beverage.
of pro-inflammatory cytokines, TNF-alpha and IL-6, and Kombucha’s flavor changes during fermentation and gives
reduced nitric oxide production. a final, sweet, sparkling taste. Fermentation must be inter-
Xia et al. (2019) used kombucha SCOBY for fermentation rupted after 14 days, as after this period, the drink may have
of water-soluble soybean extract to develop a new func- a sour vinegar taste (Jayabalan et al., 2014).
506
TA B L E 1 Different preparation conditions of kombucha in Camellia sinensis substrate

Previously
Substrate fermented Fermentation Temperature
Substrate concentration Sucrose SCOBY beverage time (days) (±3◦ C) Study objectives Reference
Black tea and 12 g/L 100 g/L 3% 10% 18 24◦ C Evaluate the content of organic acids and Jayabalan et al.,
Green tea polyphenols in kombucha made from 2007
green tea, black tea, and residues
generated from tea production.
Black tea 1.5 g/L 70 g/L* *Same – 10% 14 22◦ C Compare black tea kombucha sweetened Malbaša et al.,
measure in sugar with sucrose and with three types of 2008
beet molasses sugar beet molasses.
Black tea 12 g/L 100 g/L 2,5% – 12 28◦ C To investigate and compare the Aloulou et al.,
hypoglycemic and antilipidemic effects 2012
of black tea and black tea kombucha in
alloxan-induced diabetic rats.
Black tea and 12 g/L 100 g/L 3% 10% 15 24◦ C Get an overview of the biochemistry of Kallel et al., 2012
green tea kombucha fermentation and address
the impact of frequent kombucha
consumption on starch digestion.
Black tea 4,5 g/L 100 g/L 2,5% 10% 14 28◦ C To evaluate the antiangiogenic effect of Srihari,
kombucha on angiogenesis Arunkumar,
stimulators/regulators (gene et al., 2013
expression of human inducible factor 1,
vascular endothelial growth factor,
interleukin-8, cyclooxygenase-2, and
matrix metalloproteinases) in human
androgens (cell line independent of
prostate cancer).
Black tea 5 g/L 100 g/L – 10% 21 28◦ C Evaluate the dynamics of yeasts and Chakravorty
bacterial community in the biofilm and et al., 2016
kombucha beverage, and evaluate
changes in the fermentation
biochemistry and antioxidant property
of kombucha.
Black tea 12 g/L 100 g/L 3% 10% 14 30◦ C To investigate the role of kombucha in Ismaiel et al.,
reducing patulin produced by three 2016
strains of toxigenic fungi (Penicillium
expansum, Talaromyces purpureogenus,
and Acremonium implicatum).
(Continues)
KOMBUCHA: A REVIEW
TA B L E 1 (Continued)
Previously
◦
Black tea and 2 g of green 100 g/L 5% 6% 10 30 C Select and optimize the conditions of the Neffe-Skocińska
KOMBUCHA: A REVIEW
Green tea tea/L + 4 g of 25◦ C fermentation process and evaluate et al., 2017
black tea 20◦ C their influence on microbiological,
sensory, and physicochemical changes,
with an emphasis on determining the
sugar, alcohol, and organic acids
content of kombucha.
Black tea and 10 g/L 100 g/L 2% 3% 21 20◦ C Characterize the microbiota involved in De Filippis
Green tea 30◦ C kombucha fermentation and explore et al., 2018
the effects of two fermentation
temperatures on the microbiota, to
define which conditions can be applied
to increase the production of gluconic
and glucuronic acid.
Black tea and 8 g/L 80 g/L 3% 10% 14 27◦ C Compare the microbiological and Gaggìa et al.,
Green tea chemical composition and antioxidant 2019
activity of kombucha produced from
Rooibos infusion with kombucha made
from green and black tea.
Black tea 10 g/L 70 g/L 4% – 21 25◦ C Evaluate the impact of vessel geometry by Villarreal-Soto
examining the dynamic and bioactive et al., 2019
properties of fermentation products
from two vessels with different
surface/height rates, but with the same
surface/volume rate.
Black tea Green 2 g/L 100 g/L – 10% 15 Room To investigate the antioxidant and Kaewkod et al.,
tea Oolong tea temperature antibacterial properties (E. coli 2019
O157:H7, Shigella dysenteriae,
Salmonella typhi, E. coli, and Vibrio
cholera) from kombuchas obtained
from different types of C. sinensis,
including black tea, green tea, and
oolong tea, and the degree of toxicity it
exhibits against colorectal cancer cell
line.
(Continues)
507
508
Previously
◦
Black tea and 12 g/L 50 g/L 3% 10% 10 25 C To investigate the profile of phenolic Cardoso et al.,
Green tea compounds in kombucha produced 2020
with green tea and black tea,
characterizing its chemical and
microbiological composition, as well as
determining its antioxidant profile,
antibacterial activity (S. aureus, S.
typhimurium, E. coli, and L.
monocytogenes), cytotoxic and
antiproliferative.
Black tea 10 g/L 100 g/L 3% 10% 7 25◦ C Determine the properties of the Ben Taheur
kombucha microbiota and isolate the et al., 2020
active bacteria and yeasts in the
biodegradation of AFB1.
Black tea 5 g/L 30 g/L – – 7 22◦ C Determine the chemical composition, Ivanišová et al.,
antioxidant activity, antimicrobial 2020
activity (Candida krusei, Candida
glabrata, Candida albicans, Candida
tropicalis, E. coli, and Haemophilus
influenzae) and sensory profiles of
kombucha.
Black tea Green 8 g/L 100 g/L – – 14 28◦ C Analyze the antioxidant properties and Jakubczyk et al.,
tea White tea content of the drink prepared from , 2020
Red tea black, green, white, and red teas.
Green tea 3 g/L 10 g/L 4% – 14 28◦ C To study the action of symbiotic Najafpour et al.,
kombucha culture in removing Hg2+ , 2020
As3+ , Pb2 +, Cd2+ , and Cr6+ from
kombucha drink, and to evaluate the
effect of four parameters influencing
metal biosorption (initial dosage of
SCOBY, content of tea, sugar content
and water hardness).
– Not presented.
KOMBUCHA: A REVIEW
TA B L E 2 Different preparation conditions of kombucha analogs
Previously
KOMBUCHA: A REVIEW
Infusions and extracts

Infusions: Thyme 10 g/L 20 g/L 1% – 21 Room To investigate the Battikh,
(Thymus vulgaris temperature antimicrobial activity Bakhrouf
L.), lemon verbena against S. epidermidis, S. &
(Lippia citriodora), aureus, Mi. luteus, E. Ammar ,
rosemary coli, P. aeruginosa, S. 2012
(Rosmarinus typhimurium, Listeria
officinalis), fennel monocytogenes, and
(Foeniculum yeasts associated with
vulgare) and cases of candidiasis.
peppermint
(Mentha-piperita)
Lemon balm (Melissa 5 g/L 70 g/L – 10% 7 28◦ C To investigate the Velićanski
officinalis L.) antioxidant activity et al., 2014
infusion (against hydroxyl
radicals and DPPH) and
antibacterial activity
against P. aeruginosa,
Proteus sp., E. coli, C.
freundii, Enterobacter
cloacae, Salmonella sp.,
S. saprophyticus, S.
equorum, Bacillus sp., L.
monocytogenes e L.
innocua)
Coffee (Coffea sp.) 10 g/L 100 g/L 3% – 7 24◦ C To investigate the Watawana,
infusion and antioxidant potential of Jayawar-
instant coffee coffee beverages dena &
fermented with a Waisun-
symbiotic kombucha dara ,
culture and to evaluate 2015
the inhibitory capacity of
coffee starch hydrolase.
(Continues)
509
510
Previously
◦
Infusions: River red 10 g/L 10 g/L 0.25% 10% 7 25 C Determine the chemical Gamboa-
gum (Eucalyptus composition, consumer Gómez
camaldulensis) acceptance, antioxidant, et al., 2016
and mexican bay and
leaf (Litsea angiotensin-converting
glaucescens) enzyme inhibitory
activities.
Oak (Quercus 10 g/L 100 g/L 2.5% 10% 7 25◦ C To investigate the Vázquez-
resinosa, Quercus antioxidant activity and Cabral
arizonica, Quercus anti-inflammatory et al., 2017
convallata) leaves effects evaluating the
infusion modulation capacity of
TNF-alpha and IL-6
derived from
macrophages.
Yarrow (Achillea 1.13 g/500 ml and 35 g/500 mL – 10% 7 25◦ C Evaluate the antioxidant, Vitas et al.,
millefolium L.) 2.26 g/500 ml antimicrobial, and 2018
flowers infusion antiproliferative activity.
and yarrow
flowers subcritical
water extracts
Rooibos (Aspalathus 8 g/L 80 g/L 3% 10% 14 27◦ C Compare the Gaggìa et al.,
linearis) infusion microbiological 2019
composition, chemical,
and antioxidant activity
of kombucha made from
Rooibos infusion with
kombucha made from
green and black tea.
(Continues)
KOMBUCHA: A REVIEW
KOMBUCHA: A REVIEW
Previously
◦
Wax mallow 5 g/L 50 g/L 2.5 % 1% 14 24 C Evaluate the Silva et al.,
(Malvavicus physicochemical 2021
arboreus) flowers characteristics,
infusion microbiological quality,
safety, toxicity,
antioxidant activities
and antimicrobial
properties, and compare
with kombucha made
from green tea.
Fruits and juices
Sour cherry (Prunus 1L 6 g/L 8.7% 1% 94 18◦ C Develop a sour cherry juice Yavari et al.,
cerasus) juice 8 g/L 14 27◦ C fermented beverage 2010
10 g/L 37◦ C using symbiotic
kombucha culture and
improve glucuronic acid
production by
combining sucrose
content, time, and
fermentation
temperature.
Cactus pear (Opuntia 300 ml – 3% 10% 12 30◦ C Make a kombucha using Ayed &
ficus-indica) juice cactus pear juice as a Hamdi,
substrate, and elucidate 2015
the relationship between
fermentation time and
beverage composition.
(Continues)
511
512
Previously
◦
Red grape (Vitis 500 ml – 3% 10% 12 30 C To investigate the effects of Ayed, Ben
Vinifera L.) juice fermentation time on Abid &
the polyphenol content, Hamdi ,
total anthocyanin 2016
content, and antioxidant
activity; evaluate the
sensory properties of the
drink, and the
antimicrobial activity
against S. aureus,
S.epidermidis, E. faecalis,
B. cereus, K. pneumoniae,
P. aeruginosa, and E. coli.
Snake fruit (Salak Fruit:water, 1:1 w/w 100 g/L 10% – 14 room Compare in vivo Zubaidah
suwaru) juice temperature antidiabetes activity of et al., 2019
snake fruit Kombucha,
black tea Kombucha,
and metformin.
Milk and soy
Milk 300 ml – – 10% 8 h, 15 min 42◦ C To investigate the kinetics Kanurić
13 h, 40 min 37◦ C of milk-sugar (lactose) et al., 2018
transformation during
milk fermentation by
symbiotic kombucha
culture under five
different pH and two
different temperatures.
Soymilk Water:dry bean ratio, – – 5% 4 days 28◦ C Use kombucha as a starter Xia et al.,
8:1, w/w 3 days 37◦ C culture for the 2019
fermentation of soymilk
to develop a novel
functional soymilk
beverage.
(Continues)
KOMBUCHA: A REVIEW
Previously
KOMBUCHA: A REVIEW

◦
Soy whey 100 ml – 10% – 8 days 28 C To investigate the Tu et al.,
feasibility of 2019
transforming soy whey
into a new functional
beverage using a
kombucha culture, and
to evaluate changes in
the metabolic process,
antimicrobial activity
(against S. aureus, E.
coli, and Bacillus subtilis)
and antioxidant capacity.
Camellia sinensis tea added to alternative substrates
Green tea (Camellia 3,5 g/L green tea 65 g/L 4% – 16 28◦ C Evaluate the Shahbazi
sinensis) + spices + physicochemical, et al., 2018
(cinnamon, 3,5 g/L spices sensory, and
cardamom and antimicrobial properties
Shirazi thyme) (against S. aureus, B.
cereus, S. typhimurium,
and E. coli).
Green tea (Camellia 5 g/L green tea 70 g/L – 10% 30 28◦ C Evaluate bioactive Uțoiu et al.,
sinensis) + bee + compounds, 2018
pollen 50 g/L bee pollen fermentation kinetics
(dynamics of lactic acid
bacteria in SCOBY,
quantify organic acids
and soluble sugars),
cytotoxic and antitumor
effects in vitro
– Not presented.
513
When the fermentation period ends, the resulting drink kombucha drinks was published. This document provides
may be filtered to remove SCOBY residues and made to information about kombucha quality requirements such
undergo a second fermentation. This process, in turn, takes as maximum limits for microbiological contaminants and
2–3 days in a closed container whose objective is to car- chemical contaminants (heavy metals, pesticide residues),
bonate (production of carbon dioxide) kombucha, with production handling and hygiene, as well as analytical
a possibility of adding or not ingredients such as fruits parameters (alcohol content, acidity in acetic acid, acidity
and/or herbs for flavor. After production, kombucha must in lactic acid, and total sugar) (Uganda National Bureau of
be stored in a refrigerated environment at 4◦ C to improve Standards, 2018). The Centre of Disease Control of British
its sensory aspect and reduce the fermentation speed, as Columbia (BCCDC), Canada, has a document with rec-
the bacteria and yeasts are still viable in the beverage (Aus- ommendations for a food safety plan for the commer-
tralian Government Department of Health, 2019). cial kombucha-making process. The document provides
Another very important factor to be considered when comprehensive information on the main biological and
preparing kombucha is the use of sanitized utensils, as well chemical risks and potential hazards associated with kom-
as working in clean areas and following good manufactur- bucha production, with emphasis on monitoring pH and
ing practices, in order to avoid unwanted contamination ethanol content, establishing critical limits for both param-
(Watawana et al., 2015). eters. According to the BCCDC plan, pH levels should not
be below 2.5 and alcohol levels should not exceed 1% or
increase during the shelf life of the product (Centre of Dis-
5 KOMBUCHA REGULATIONS ease Control, 2020).
Kombucha Brewers International (KBI), a trade associ-
Today kombucha is sold as obtained from different sub- ation that globally represents the commercial kombucha
strates and in various flavors and continues to gain pop- beverage category, provides a kombucha code of practice,
ularity in the worldwide potentially functional beverage which aims to create a food safety and quality standard
market (Villarreal-Soto et al., 2018). In Brazil, the kom- for kombucha producers, ensuring more transparency of
bucha Identity and Quality Standard (PIQ) is regulated information to consumers so that they can make safer
by Normative Instruction (IN) No. 41 of September 17, choices. KBI has a certification and seal program that
2019, from the Ministry of Agriculture, Livestock and offers a high level of food safety, indicating that the product
Supply (MAPA), which monitors definitions, composi- follows clear manufacturing standards. The KBI code of
tion (mandatory and optional ingredients), classifications, practice describes the definitions of processing and ingre-
description of labeling, prohibitions, and analytical param- dients, the manufacturing processing steps with process
eters. These analytical parameters include pH, in the range flowchart, hygiene and labeling requirements, and kom-
of 2.5–4.2, alcoholic grading of kombucha without alcohol bucha analytical profile – chemical and microbiological
(up to 0.5%) and with alcohol (0.6–0.8%), volatile acidity standards (Kombucha Brewers International, 2021).
(30–130 mEq/L) and pressure (atm 20◦ C) in the kombucha In 2017, KBI, in cooperation with members and US sen-
added with CO2 (1.1 to 3.9 atm) (Brasil, 2019). In addi- ators, gathered to draft a kombucha law. In this proposal,
tion, IN No. 41/2019 prohibits, on its labeling, the use of kombucha would be defined as a beverage that "(a) is fer-
expressions such as "craft, homemade, familiar, live drink, mented only by a symbiotic culture of bacteria and yeast,
probiotic drink, ancient drink, elixir, elixir of life, ener- (b) contains no more than 1.25% alcohol by volume, (c) is
gizing, invigorating, special, premium," among others that sold or offered for sale as kombucha, and (d) is derived
attribute characteristics of superlative qualities and func- from sugar, malt or malt substitute, tea or coffee and no
tional properties not yet approved in specific legislation more than 20% of other healthy ingredients.” However, the
(Brasil, 2019). “Kombucha Law” has not yet been approved in the United
In international regulatory terms, the National Environ- Sates (S. 926/HR 1961) (Ransom & McDermott, 2020).
mental Health Association (NEHA) published in 2013, in Considering the microbiological safety of kombucha, pH
the Journal of Environmental Health (JEH), a guideline is the most important parameter in the fermentation of
with “Procedural Guidance and Risk Analysis in Kom- this drink, as the organic acids formed are considered to be
bucha Fermentation”, under the Model Code of Food mainly responsible for functional activities, especially the
Administration and Medicines as stated by the FDA (Food antimicrobial activity (Villarreal-Soto et al., 2018). Acidic
and Drug Administration [FDA], 2009), which covers food (pH between 4.0 and 4.5) and very acidic (pH < 4.0) foods
safety plan and risk analysis based on critical control are microbiologically safe, since pathogenic microorgan-
points (HACCP) throughout the entire kombucha produc- isms are hardly able to develop under these conditions
tion chain. In 2018, a draft standard from Uganda that spec- (Coban, 2019). According to available regulations, to be
ifies requirements and sampling and testing methods for considered safe, kombucha drink must have a pH range
between 2.5 and 4.2. Based on FDA Food Code Model rec- well as creats a physical barrier that protects against UV
ommendations, a pH ≤ 4.2 is considered the critical limit, radiation, allow moisture retention and prevent dehydra-
and if a pH does not reach this value within 7 days, the cul- tion. Bacterial cellulose synthesis includes carbon source
ture may be contaminated and should be discarded. Pre- transport, uridine diphosphate-glucose (UDPG) synthe-
suming that a pH value is less than 2.5, the addition of sis via glucose-6-phosphate and glucose-1-phosphate, and
fresh tea is recommended to reach adequate levels (Num- glucose chain polymerization by cellulose synthase. An
mer, 2013). advantage of this form of cellulose production is that the
Under the NEHA guideline, other methods to ensure bacteria grow rapidly under controlled conditions and can
beverage safety include pasteurizing the product, which produce glucose, ethanol, sucrose, and glycerol (Cacicedo
also prevents overproduction of alcohol and carbon diox- et al., 2016; Villarreal-Soto et al., 2018).
ide; addig the preservatives 0.1% sodium benzoate and The acetic acid produced by acetic bacteria stimulates
0.1% potassium sorbate, and finally keeping it refrigerated yeasts to produce ethanol, and ethanol, in turn, can be use-
(Nummer, 2013). ful for the growth of acetic bacteria and then produce more
acetic acid (Gomes et al., 2018; Jayabalan et al., 2014; Leal
et al., 2018). Moreover, autolysis of yeast cells also releases
6 FERMENTATION PROCESS nutrients that stimulate the growth of important bacteria.
Most microbial species excrete metabolic products that can
Several factors can influence the diversity of kombucha stimulate or inhibit the growth of other species, establish-
microbiota, that is, geographic location, weather condi- ing interactions that need to be further explored (Gomes
tions, local species of bacteria and yeasts, substrate, fer- et al., 2018; Leal et al., 2018; Villarreal-Soto et al., 2018).
mentation time, and temperature (Jayabalan et al., 2014; Some of the products formed during the fermentation
Watawana et al., 2015). Nevertheless, studies show that process have antimicrobial activity against pathogenic bac-
the microbiota present in kombucha is predominantly teria, providing protection to the beverage and, conse-
composed of acetic bacteria from Acetobacteraceae fam- quently, can be considered natural preservatives, which
ily, with the dominant genera Komagataeibacter, Aceto- contribute to increase its shelf life (Dutta & Paul, 2019).
bacter, and Gluconobacter, and yeasts, mainly from genera
Brettanomyces and Zygosaccahromyces (Marsh et al., 2014;
Villarreal-Soto et al., 2020). Lactic acid bacteria have also 7 CHEMICAL COMPOSITION
been isolated, but to a lesser extent (De Filippis et al., 2018;
Marsh et al., 2014). Kombucha presents a wide variety of chemical com-
The fermentation process consists of a symbiotic asso- pounds, including substrate polyphenols; sugars such as
ciation of yeasts and bacteria organized in a cellulosic sucrose, glucose, and fructose; organic acids such as acetic,
matrix. Yeasts, through the enzyme invertase, catalyze the gluconic, glucuronic, citric, L-lactic, malic, tartaric, mal-
hydrolysis of sucrose into fructose and glucose, producing onic, oxalic, succinic, and pyruvic; fiber; ethanol; amino
ethanol via a glycolytic pathway. On the other hand, acetic acids; biogenic amines; purines; pigments; lipids; proteins;
bacteria produce acetic acid from the oxidation of ethanol, essential elements such as copper, iron, manganese, nickel,
through the enzymes alcohol dehydrogenase and aldehyde and zinc; vitamins B1, B2, B6, B12, and C; carbon dioxide,
dehydrogenase. At the end of the process, water and car- antibiotic substances, d-acid-saccharic-1, 4-lactone (DSL);
bon dioxide are also found as final products. In addition, and hydrolytic enzymes (Kapp & Sumner, 2019).
they are also capable of converting glucose into gluconic Chemical compounds present in kombucha are pro-
and glucuronic acid (Gomes et al., 2018; Leal et al., 2018). duced by SCOBY and originate from the substrate itself,
During the process, fructose is used less and can be found which can have their structure modified and be trans-
after fermentation (Nummer, 2013). formed into new compounds during fermentation. The
Bacteria and yeasts added to the beverage for fermen- exact biotransformation route of all metabolites already
tation are responsible for a cellulosic film formation that identified in kombucha and the microorganisms involved
makes up SCOBY. When using sucrose as a main source of still need further studies. However, several nutritional
carbon, acetic bacteria, especially Komagataeibacter xyli- properties and benefits related to the bioactive potential
nus, produce a cellulose network, which enhances the of the beverage have already been reported (Dutta & Paul,
association formed between bacteria and yeast, producing 2019). Main compounds identified during the fermenta-
a thick membrane with a gelatinous aspect (Gomes et al., tion process of C. sinensis tea and alternative substrates
2018; Leal et al., 2018). This produced cellulose film allows are shown in Tables 3 and 4, respectively. The study of the
microorganisms to remain in the liquid near the interface composition and properties of kombucha is crucial, as it
to obtain the necessary oxygen for their metabolism, as makes it possible to understand its kinetics. However, it is
516
TA B L E 3 Compounds identified in fermentation process of Camellia sinensis tea

Total phenolics Fermentation
Substrate Organic acid Vitamins and minerals (GAE*) time (days) References
Black tea Acetic acid 4.69 g/L – – 18 Jayabalan et al., 2007
Glucuronic acid 1.71 g/L
Lactic acid 0.18 g/L
Acetic acid 8.0 g/L – 1.12 g GAE/L 15 Kallel et al., 2012
Acetic acid 16.57 g/L – – 21 Chakravorty et al., 2016
Acetic acid 1.83 g/L – – 14 Ismaiel et al., 2016
Citric acid 2.77 g/L
Acetic acid 9.18 g/L – 67.20 mg GAE/g 14 Gaggìa et al., 2019
Acetic acid 6 to 15.1 g/L – 36 to 221 mg GAE/g 21 Villarreal-Soto et al., 2019
Acetic acid 11.15 g/L Vitamin C 0.70 g/L – 15 Kaewkod, Bovonsombut &
Glucuronic acid 1.58 g/L Tragoolpua, 2019
Succinic acid 3.05 g/L
Acetic acid 3.6 g/L – 1.09 mg GAE/mL 10 Cardoso et al., 2020
Acetic acid 1.55 g/L Iron 0.31 mg/L 412 mg GAE/L 7 Ivanišová et al., 2020
Tartaric acid 0.23 g/L Manganese 1.57 mg/L
Citric acid 0.05 g/L Zinc 0.53 mg/L
Acetic acid 9.08 g/L – 206 mg GAE/L 14 Jakubczyk et al., 2020
Green tea Acetic acid 8.36 g/L – – Jayabalan et al., 2007
Acetic acid 5.4 g/L – 1.08 g GAE/L 15 Kallel et al., 2012
Acetic acid 7.65 g/L – 64.20 mg GAE/g 14 Gaggìa et al., 2019
Acetic acid 10.42 g/L Vitamin C 0.61 g/L – 15 Kaewkod, Bovonsombut &
Glucuronic acid 1.37 g/L Tragoolpua, 2019
Acetic acid 3.2 g/L – 0.70 mg GAE/mL 10 Cardoso et al., 2020
Acetic acid 9.14 g/L – 320 mg GAE/L 14 Jakubczyk et al., 2020
(Continues)
KOMBUCHA: A REVIEW
important to emphasize that the presence of these com-
Neffe-Skocińska et al., 2017

pounds depends on the cultivation substrate, sugar con-
Kaewkod, Bovonsombut &

centration, fermentation process time and temperature, as
Jakubczyk et al., 2020

Jakubczyk et al., 2020
Malbaša et al., 2008
Malbaša et al., 2008

well as the microorganisms present in kombucha SCOBY.
Tragoolpua, 2019
Any change in fermentation conditions can affect the final
product composition (Jayabalan et al., 2014; Villarreal-Soto
References
et al., 2018).
During the fermentation process, kombucha’s pH value
decreases due to the production of organic acids (Jayabalan
et al., 2014). Jayabalan et al. (2007) observed an increase
Fermentation
in acetic acid production up to the 15th day of fermenta-

time (days)
tion, reaching a maximum of 9.5 g/L in green tea kom-

bucha. In the case of l-glucuronic acid, the same authors
observed a maximum concentration of 2.3 g/L, in black
10
14
14
14
14
15
tea kombucha, on the 12th day of fermentation. Likewise,
Chakravorty et al. (2016) observed an increase in acetic and
Total phenolics
279.1 mg GAE/L
228.1 mg GAE/L
gluconic acids concentration in green tea kombucha. The
concentration of both acids increased with time and
(GAE*)
reached 16.57 and 7.36 g/L, respectively, after 21 days of fer-

mentation. Cardoso et al. (2020) reported that acetic acid
–
was the most-produced organic acid, both in green and

black tea kombucha, reaching a concentration of 3 g/L
after 10 days of fermentation.
0.60 g/L
Velićanski et al. (2014), when evaluating the antiox-

Vitamins and minerals
idant and antimicrobial activity of beverage fermented

with kombucha SCOBY, from the infusion of lemon balm
(Melissa officinalis L.), also found an increase in the pro-
duction of organic acids, especially acetic acid, which
Vitamin C
reached a maximum 8.12 g/L after 7 days of fermentation.

Soy whey, a product obtained from tofu production, was
used by Tu et al. (2019) as a substrate for kombucha pro-
–
–
–
duction, and they observed that pH value and organic acid

g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
g/L
content changed throughout fermentation. After 8 days

of fermentation, acetic and glucuronic acid concentration
reached 5.77 and 132.81 g/L, respectively. Malbaša et al.
(2008) used sugar beet molasses as an alternative carbon
0.040 to 0.044
0.040 to 0.063
0.029 to 0.030
0.071 to 0.086
source in the production of black tea kombucha and evi-

0.46 to 0.47
0.16 to 0.40
0.21 to 0.28
1.42 to 1.65
denced that there was a greater drop in pH values and, con-

0.053
10.48
sequently, higher production of organic acids compared to

0.07
9.07
0.53
9.13
the beverage prepared with sucrose. Total acids concentra-

tion ranged from 0.9 to 3.1 g/L and can be explained by the
chemical composition of the beet molasses itself, which
includes fatty acids, lactic acid, and nonvolatile organic
Glucuronic acid
Glucuronic acid
Organic acid
Quinic acid
acids. However, acetic acid production was much lower

Lactic acid
Lactic acid
Oxalic acid
Acetic acid
Acetic acid
Acetic acid
Acetic acid
Acetic acid
Acetic acid
Malic acid
Citric acid
compared to sucrose in the same fermentation stage. Due

(Continued)
to the presence of some components of molasses, it is possi-

ble that the activity of acetic bacteria has decreased, or the
acetic acid generated has been converted into other com-
pounds.
– Not presented.
Green tea
In the human body, vitamins and minerals have ben-

Oolong tea
Substrate
TA B L E 3
White tea
Black tea
Black tea
Black tea
eficial effects on health and are responsible for normal

Red tea
metabolic pathways and different physiological functions

+
(Vitas et al., 2018). Studies have reported that essential

518
TA B L E 4 Compounds identified in fermentation process of kombucha analogs

Total phenolic Fermentation time
Substrate Organic acids Vitamins (GAE*) (days) References
Lemon balm (Melissa officinalis L.) Acetic acid 8.12 g/L – 850.3 µg GAE/L 7 Velićanski et al.,
infusion 2014
Coffee (Coffea sp.) infusion and – – 14.2 to 19.8 mg 7 Watawana,
instant coffee GAE/g Jayawardena &
Waisundara ,
2015
Oak (Quercus resinosa, Quercus Glucuronic acid 0.048 to 0.060 g/L – – 7 Vázquez-Cabral
arizonica, Quercus convallata) et al., 2017
leaves infusion
Yarrow (Achillea millefolium L.) Acetic acid 8.12 to 15.10 g/L Vitamin C 0,48 a 1,14 g/L – 7 Vitas et al., 2018
flowers infusion and yarrow Oxalic acid 2.53 to 3.45 g/L
flowers Formic acid 1.51 to 2.33 g/L
subcritical water extracts Succinic acid 0.16 to 0.30 g/L
Malic acid 0.10 to 1.59 g/Lg/L
Citric acid 0,04 to 0,07
Rooibos (Aspalathus linearis) Acetic acid 4.89 g/L – 40.89 mg GAE/g 14 Gaggìa et al., 2019
infusion Gluronic acid 2.87 g/L
Wax mallow (Malvavicus arboreus) Acetic acid 8.80 g/L – 124.6 mg GAE/L 14 Silva et al., 2021
flowers infusion
Sour cherry (Prunus cerasus) juice Glucuronic acid 132.5 g/L – – 14 Yavari et al., 2010
Cactus pear (Opuntia ficus-indica) Gluconic acid 6.91 g/L – 857 mg GAE/L 15 Ayed & Hamdi,
juice 2015
Snake fruit (Salak suwaru) juice – – 535.59 mg GAE/L 14 Zubaidah et al.,
2019
Soymilk Lactic acid 5.25 g/L Thiamin 51.67 µg/g – 3 Xia et al., 2019
Acetic acid 2.75 g/L Riboflavin 145.59 µg/g
Glucuronic acid 1.23 g/L Vitamin C 1676.43 µg/g
Citric acid 0.12 g/L Vitamin E 335.99 µg/g
Malic acid 0.09 g/L
(Continues)
KOMBUCHA: A REVIEW
Total phenolic Fermentation time
KOMBUCHA: A REVIEW
Substrate Organic acids Vitamins (GAE*) (days) References

Soymilk Lactic acid 1.72 g/L Thiamin 48.51 µg/g – 4 Xia et al., 2019
Acetic acid 6.74 g/L Riboflavin 44.62 µg/g
Glucuronic acid 2.74 g/L Vitamin C 1586.95 µg/g
Citric acid 0.33 g/L Vitamin E 287.01 µg/g
Malic acid 0.14 g/L
Soy whey Acetic acid 5.77 g/L – – 8 Tu et al., 2020

Formic acid 0.78 g/L
Citric acid 1072 g/L
Green tea (Camellia sinensis) + Acetic acid 2.67 g/L – 0.582 mg 16 Shahbazi et al.,
cinnamon Glucuronic acid 1.15 g/L GAE/mL 2018
Green tea (Camellia sinensis) + Acetic acid 1.28 g/L – – 16 Shahbazi et al.,
cardamon Glucuronic acid 0.11 g/L 2018
Green tea (Camellia sinensis) + Acetic acid 1.13 g/L – – 16 Shahbazi et al.,
Shirazi thyme Lactic acid 0.09 g/L 2018
Green tea (Camellia sinensis) + bee Acetic acid 3.51 g/L – – 30 Uțoiu et al., 2018
pollen Glucuronic acid 2.26 g/L
– Not presented.
519
minerals content (Cu, Fe, Mn, Ni, and Zn) increased as Unfermented wax mallow flower infusions showed 50.9 µg
a result of kombucha metabolic activity (Jayabalan et al., GAE/ml, while the beverage fermented produced from
2014). The most commonly found vitamins in kombucha the 14-day fermented wax mallow flower infusion showed
are those from the B complex (B1, B2, B6, and B12) and C, 124.6 µg GAE/ml, representing a 145% increase in total phe-
and both have a high antioxidant activity due to their redox nolic compound content. Therefore, we believe that com-
potential (Malbaša et al., 2011; Vitas et al., 2018). plexed phenolic compounds in fermented beverages may
Malbaša et al. (2011) investigated the influence of starter be subject to degradation in the acidic environment and by
cultures isolated from the symbiotic kombucha culture on the enzymes released by bacteria and yeasts present in the
the antioxidant activity of green and black tea kombucha symbiotic culture, increasing their total content (Jayabalan
and found that vitamins C and B2 content increased over et al., 2008).
the fermentation period; this made it possible to correlate
antioxidant activity of the beverage to the amount of vita-
min C. Vitas et al. (2018) also observed an increase in vita- 8 ALCOHOLIC CONTENT
min C content in all kombucha samples produced from A.
millefolium flowers, ranging from 0.48 to 1.14 mg/L. How- Ethanol is produced during the fermentation process, even
ever, the authors found that, when comparing their results if the beverage undergoes a second fermentation by yeast
with other studies, C. sinensis kombuchas have higher vita- (Jayabalan et al., 2014). Also, if fermentation is not prop-
min C content, reinforcing the idea that different sub- erly controlled, ethanol production can be increased (Aus-
strates can influence the beverage final composition. tralian Government Department of Health, 2019). Sugar
Jayabalan et al. (2007) have observed changes in the concentration, storage conditions (Talebi et al., 2017), time
content of organic acids and polyphenols in tea and (Chakravorty et al., 2016), and fermentation temperature
kombucha, prepared from green tea, black tea, and tea- are factors that can influence ethanol concentration in the
manufacturing residues during 18 days of fermentation. drink (Neffe-Skocińska et al., 2017).
The authors have distinguished an increase in organic The maximum limit for ethanol for nonalcoholic drinks,
acids concentration and degradation of the epicatechin iso- including kombucha beverages, is 0.5% (v/v) (Brasil, 2019;
mers in the fermentation process. Catechins in tea during Tax and Trade Bureau, 2016; Food Standards Australian
kombucha fermentation were found to be degraded. All New Zealand, 2016). Also, if a product has an alcohol con-
catechins studied were degraded by day 9 of kombucha fer- tent above 0.5% (v/v), it must be classified as an alcoholic
mentation and showed a marked increase on day 12. This beverage (Brasil, 2019; Tax and Trade Bureau, 2016; Food
result can be explained by the biotransformation of cate- Standards Australian New Zealand, 2016) and is subject to
chins by enzymes excreted by the microorganisms present important federal and state regulations (Talebi et al., 2017).
in kombucha culture, consequently increasing epicatechin According to the document published by NEHA (Num-
isomer concentration. Similarly, Kallel et al. (2012) also mer, 2013) and by Australian Government Department of
showed an increase in the concentration of phenolic com- Health (2019), in recent years, some kombucha beverages
pounds in black and green tea kombucha after 2 weeks have had an alcohol content above 0.5% (v/v) and this
of fermentation. On day zero of fermentation, the con- information has not been displayed on the labels of com-
centration of total phenolics was 0.78 g gallic acid equiv- mercial kombucha beverages. Table 5 shows the ethanol
alents (GAE)/L in green tea kombucha and 1.01 g GAE/L content in kombucha and kombucha analogs as described
in black tea kombucha. After 15 days of fermentation, the in several studies.
green and black tea kombuchas presented 1.08 and 1.12 g Kallel et al. (2012), when evaluating the biochemistry
of GAE/L of total phenolics, respectively. In the study of green and black tea kombucha fermentation, evidenced
by Bhattacharya et al. (2013), the total content of phe- that ethanol concentration increased during kombucha
nolics and flavonoids increased by approximately 27.2% fermentation, faster in black tea kombucha compared to
and 75%, respectively, after 14 days of black tea fermen- green tea kombucha. On the 15th day of fermentation,
tation with kombucha intercropping. Ayed and Hamdi ethanol concentration was approximately 0.2% and 0.3% in
(2015) also observed increased phenolic compound content green and black tea kombucha, respectively.
over the fermentation period in beverage fermented and In the study conducted by Chakravorty et al. (2016) on
produced from prickly pear juice (O. ficus-indica), show- black tea kombucha, ethanol concentration increased with
ing highest values on day 9 of fermentation. Silva et al. time, reaching a maximum value of 0.028% on the 7th day
(2021) also found an increase in the content of total phe- of fermentation, followed by a decrease to about 0.0073%
nolic compounds when they produced beverage fermented after 21 days of fermentation. This decrease in ethanol con-
with kombucha SCOBY from infusion of wax mallow flow- centration was related to its use by acetic bacteria in the
ers (Malvavicus arboreus), an unconventional food plant. production of acetic acid.
TA B L E 5 Ethanol content in kombucha and kombucha analogs

Fermentation
Substrate Ethanol content (%) time (days) References
Black tea 0.3 15 Kallel et al., 2012
0.007 21 Chakravorty et al., 2016
0.58 14 Gaggìa et al., 2019
1.4 21 Villarreal-Soto et al., 2019
0.49 10 Cardoso et al., 2020
0.4 7 Ivanišová et al., 2020
2 14 Jakubczyk et al., 2020
Green tea 0.2 15 Kallel et al., 2012
0.4 14 Gaggìa et al., 2019
1.4 21 Villarreal-Soto et al., 2019
0.7 10 Cardoso et al., 2020
2.75 14 Jakubczyk et al., 2020
Black tea + green tea 0.69 to 1.1 10 Neffe-Skocińska et al., 2017
White tea 3 14 Jakubczyk et al., 2020
Red tea 3 14 Jakubczyk et al., 2020
Red grape (Vitis vinifera L.) juice 0.29 12 Ayed, Ben Abid & Hamdi ,
2016
Rooibos (Aspalathus linearis) 0.11 14 Gaggìa et al., 2019
infusion
Soymilk 0.062 3 Xia et al., 2019
Soymilk 0.075 4 Xia et al., 2019
When analyzing different temperature conditions (20, to stabilize alcohol production. Refrigeration reduces the
25, and 30◦ C) during the fermentation of green and fermentation speed of viable microbial cells in the bever-
black tea blends with symbiotic kombucha culture, Neffe- age (Australian Government Department of Health, 2019).
Skocińska et al. (2017) found that after 10 days of fermen- Villarreal-Soto et al. (2019) have assessed the geometry
tation, at all temperatures analyzed, ethanol content was impact of two vessels with different surface/height ratios
higher than 0.5% (v/v), reaching a maximum of 1.1% at on black tea kombucha’s dynamic and bioactive properties.
25◦ C. In this study, ethanol production was faster in the vessel
Talebi et al. (2017) evaluated ethanol concentration in with the largest surface area, with a maximum production
kombucha samples of equal validities and batches over a of 32.8 g/L (4.1% v/v) after 11 days of fermentation, com-
period of 60 days under different temperature conditions pared to the vessel with the smallest surface area, which
using the gas-phase chromatography method. In all sam- produced 23.1 g/L (2.9% v/v) of ethanol after 16 days. How-
ples analyzed, the alcohol content ranged from 1.2% to 2% ever, after 21 days of fermentation, the final ethanol con-
(v/v), in disagreement with the Alcohol and Tobacco Tax centration was almost the same in both vessels, reaching a
and Trade Bureau, US government regulation (Tax and concentration of about 11 g/L (1.4% v/v).
Trade Bureau, 2016). The ethanol content of kombucha
bottles kept at 22◦ C increased significantly after 7 days
and reached a maximum level of 1.57% after 14 days, fol- 9 BIOLOGICAL PROPERTIES
lowed by a slight decline after 21 days. This result can be
explained by the production of carbon dioxide, mainly in Regular consumption of kombucha is believed to have
the secondary fermentation, which occurs after the bever- health benefits. Given its long history of consumption,
age is bottled. Carbon dioxide accumulation inhibits the especially as a health-promoting beverage, kombucha has
conversion of ethanol to acetic acid in the case of closed continually been explored for its beneficial properties
containers (Nummer, 2013). On the other hand, in bottles (Dutta & Paul, 2019). Kombucha protective activities can
stored at 4◦ C, the amount of ethanol began to rise grad- mainly be attributed to the presence of organic acids and
ually for 14 days, and remained virtually unchanged dur- phenolic compounds that are produced as a result of
ing the period tested, indicating that refrigeration helped microbial fermentation (Bhattacharya et al., 2016).
Studies have shown antioxidant activity of beverages fer- Many scientific reports prove that kombucha can exert
mented with SCOBY kombucha (Jayabalan et al., 2008; antimicrobial activity against pathogenic bacteria and
Silva et al., 2021). This ability to effectively fight free radi- fungi. This is due to both compounds present in the sub-
cals can be explained by the high content of phenolic com- strates and the metabolites produced by the bacteria and
pounds, especially flavonoids, present in the substrates yeasts present in the SCOBY kombucha that inhibit the
used in their production (Bhattacharya et al., 2013). About growth of microorganisms with potential pathogenicity
127 phenolic compounds (70.2% flavonoids, 18.3% pheno- (Silva et al., 2021; Battikn et al., 2011).
lic acids, 8.4% other polyphenols, 2.3% lignans, and 0.8% Bhattacharya et al. (2016) evaluated the antimicrobial
stilbenes) were identified in green and black tea bever- activity of black tea kombucha against enteric bacteria at
ages fermented by kombucha culture for a period of 10 different time intervals (0, 7, 14, and 21 days). The results
days (Cardoso et al., 2020). In addition, the kombucha showed that kombucha had better antibacterial activity
starter microbial culture (Malbaša et al., 2011) and the after 14 and 21 days of fermentation, showing bacterio-
fermentation time (Chu & Chen, 2006; Puspitasari et al., static and bactericidal activity against enterohemorrhagic
2017) can also influence the antioxidant capacity of the E. coli (O157:H7), Vibrio cholerae, Shigella. flexneri, and
beverage. Salmonella. typhimurium.
Complications related to diabetes and cancer may be Vitas et al. (2018) used infusions and extracts from A.
associated with increased oxidative stress, which, in turn, millefolium L. flowers for beverage fermented with kom-
can be suppressed due to the presence of bioactive com- bucha SCOBY production, and examined its antimicrobial
pounds, which have antioxidant properties. Such evi- activity against Staphylococcus aureus, Klebsiella pneumo-
dence may justify the alleged antidiabetic and anticarcino- niae, E. coli, Proteus vulgaris, P. mirabilis, Bacillus subtilis,
genic kombucha effects (Bhattacharya et al., 2013; Srihari, Candida albicans, and Aspergillus niger. The drink pre-
Arunkumar, et al., 2013). sented antimicrobial activity against all microorganisms
Srihari, Karthikesan, et al. (2013) found that there was tested after 7 days of fermentation.
a reduction in glycosylated hemoglobin (HbA1c) and an Furthermore, kombucha can also contribute to the
increase in plasma insulin, hemoglobin, and tissue glyco- improvement of liver function (Wang et al., 2013), immune
gen levels in diabetic rats induced by streptozotocin. The system, and gastrointestinal functions (Kozyrovska et al.,
rats had consumed 6 mg/kg of body mass of lyophilized 2012; Watawana, et al., 2015), can possess detoxifying
black tea kombucha over 45 days of treatment. A sig- action (Nguyen et al., 2015) and anti-inflammatory proper-
nificant reduction in enzymatic activities of glucose-6- ties (Vázquez-Cabral et al., 2017), and help reduce choles-
phosphatase, fructose-1, 6-bisphosphatase, and hexoki- terol levels and blood pressure (Jayabalan et al., 2016).
nase, the main enzymes responsible for the control of gly- According to literature data, there are many benefits
colysis and gluconeogenesis in liver and muscle, was also attributed to kombucha, however, it is important to empha-
observed, indicating that kombucha has a potential to act size that the studies are carried out predominantly in ani-
at various points in the glucose regulatory pathways (Sri- mals, or in vitro in cell cultures of animals or humans,
hari, Karthikesan, et al., 2013). Bhattacharya et al. (2013) with no evidence yet to support these benefits to human
also observed a potential antidiabetic effect of black tea health (Kapp & Sumner, 2019). Still, despite not being able
kombucha in alloxan-induced diabetic rats. The blood glu- to receive any official claims about its health effects, kom-
cose level was reduced by approximately 56.4% after 14 days bucha can be considered a high-value food product in a
of treatment with 150 mg/kg body weight of lyophilized healthy diet (Jayabalan et al., 2014).
kombucha extract.
Kombucha antiproliferative actions have been proven
in several human tumor cells such as HeLa cells (epithe- 10 TOXIC AND ADVERSE EFFECTS
lial carcinoma of the cervix), HT-29 (adenocarcinoma of
the colon), MCF-7 (breast adenocarcinoma) (Četojević- The mechanisms associated with the adverse effects of
Simin et al., 2012), Hep-2 (human laryngeal epidermoid kombucha are still unclear, but microbiological composi-
carcinoma) (Deghrigue et al., 2013; Uțoiu et al., 2018), tion, storage conditions, chemical composition of the tea,
PC-3 (prostate cancer) (Srihari, Arunkumar, et al., 2013), and host factors may contribute to the effects. In almost
RD (human rhabdomyosarcoma-derived cell line), Hep2c all cases of toxicity related to kombucha consumption, the
(HeLa-derived human cervical carcinoma-derived cell individuals already had a previous illness or some predis-
line) (Vitas et al., 2018), A549 (pulmonary adenocarci- position factor (Leal et al., 2018).
noma), HCT8 (human ileocecal colorectal adenocarci- Despite the evidence on kombucha’s therapeutic effects,
noma), and Caco-2 (colorectal adenocarcinoma) (Cardoso there are concerns about its toxicity and the possi-
et al., 2020). ble damage it could cause to health, particularly for
immune-compromised individuals (Sinir et al., 2019). Several metabolic reactions occur during the fermenta-
Some individuals reported dizziness, nausea, vomiting, tion process leading to the formation of secondary metabo-
allergic reactions, jaundice, headache, and neck pain after lites. Most metabolites originate from the substrate used
consuming kombucha (Jayabalan et al., 2014). for their production. Many of these metabolites have
Kole et al. (2009) recorded a case of acute renal fail- their structure changed and are transformed into new
ure with lactic acidosis and hyperthermia, in a 22-year- compounds during fermentation. The main compounds
old HIV-positive man, after 15 hours of ingesting 1 L of formed in the beverage are organic acids, ethanol, sugars,
kombucha. Holbourn and Hurdman (2017) reported a case vitamins, and minerals. Among these compounds, ethanol
of lactic acidosis in an asthmatic woman who consumed has been the focus of attention. According to the literature,
kombucha daily for several months and showed improve- many kombuchas produced have presented an ethanol
ment after discontinuing consumption. A case of hepato- content above 0.5% (v/v), which, according to legislations,
toxicity has also been reported in a woman with a history must be considered as an alcoholic beverage.
of diabetes and hypothyroidism who consumed kombucha The variety and amount of raw material, as well as car-
for a month (Gedela et al., 2016). For pregnant women and bon source, starter culture, fermentation time and temper-
children under 4-years-old, the consumption of kombucha ature, pH, and oxygen are factors that can affect fermen-
is contraindicated as a safety parameter due to the ethanol tation, contributing to beverages with different chemical
content, which may represent a chemical hazard for these properties, which will consequently affect their biological
groups (Centre for Disease Control, 2020). activity. Several therapeutic and health-promoting prop-
Consumption of kombucha has also been associated erties have been investigated in kombucha over the ages.
with lead poisoning. Jayabalan et al. (2014) mentions a Various beneficial effects in in vitro experiments and in
case of symptomatic lead poisoning in a couple who con- animal models have been confirmed, but there is still no
sumed kombucha fermented in a ceramic jar. There was clinical evidence for kombucha biological activities in
evidence that the organic acids present in the drink release humans. Toxic effects related to kombucha consumption
the lead from the enamel used in the ceramic jar, highlight- need further investigation and study, and, as a preven-
ing the importance of the way of preparation, the utensils tive measure, it should not be recommended for individ-
used, and good manufacturing practices, as the drink can uals with pre-existing diseases, pregnant women, nursing
become harmful (Jayabalan et al., 2014). mothers, and children under the age of 4.
According to the U.S. Centers for Disease Control and Reiterating the given data, kombucha can be considered
Prevention (CDC), a daily kombucha consumption of a potentially expanding product for use and applications
110 mL for healthy individuals poses no health risk. How- in food industries, which makes research on this product
ever, possible risks may be related to excessive drinking or even more important. Investigating the microbial diver-
consumption by individuals with pre-existing health prob- sity involved, as well as the interactions, chemical compo-
lems (Nummer, 2013). Due to the possibility of generating sitions, and "influences" on biological activities allows us
toxic reactions in some individuals, the consumption of to obtain better information about its functionality. These
kombucha is contraindicated for nursing mothers and considerations have driven the development of this work,
pregnant women, children under 4-years-old, patients with the aim to better understand the kombucha fermenta-
with renal insufficiency, and patients with HIV (Centre tion process through the gathering of important scientific
for Disease Control, 2020; Kole et al., 2009; Leal et al., data published over the years.
2018).
AC K N OW L E D G M E N T S
The authors received financial support from Fundação de
11 CONCLUSION Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ,
Brazil), through Project E-26/200 930/2017. We are also
Kombucha is a beverage traditionally obtained by the fer- grateful to the Coordenação de Aperfeiçoamento de Pes-
mentation technique of C. sinensis tea, with sugar added, soal de Nível Superior (CAPES, Brazil) and to the Conselho
by a SCOBY. Due to its supposed health benefits, this Nacional de Desenvolvimento Científico e Tecnológico
drink has gained popularity, has been perpetuated in sev- (CNPq, Brazil) for their continued support.
eral countries, and is currently consumed and sold world-
wide. In addition to C. sinensis, the use of herbal infusions, AU T H O R CO N T R I B U T I O N S
coffee, fruit juices, milk, and soy, associated or not with Jeniffer Ferreira de Miranda: data curation; inves-
green or black tea for fermentation with kombucha culture tigation; visualization; writing – original draft; writ-
has also been reported, and shown to be good alternatives, ing – review and editing. Larissa Fernandes Ruiz:
mainly due to their chemical composition. investigation; visualization; writing – original draft;
writing – review and editing. Cíntia Borges Silva: inves- Ayed, L., & Hamdi, M. (2015). Manufacture of a beverage from cactus
tigation; visualization; writing – original draft; writing – pear juice using “tea fungus” fermentation. Annals of Microbiol-
review and editing. Thais Matsue Uekane: conceptual- ogy, 65(4), 2293–2299. https://doi.org/10.1007/s13213-015-1071-8
Battikh, H., Bakhrouf, A., & Ammar, E. (2012). Antimicrobial effect of
ization; supervision; visualization; writing – original draft;
kombucha analogues. LWT— Food Science and Technology, 47(1),
writing – review and editing. Kelly Alencar Silva: con-
71–77. https://doi.org/10.1016/j.lwt.2011.12.033
ceptualization; supervision; visualization; writing – orig- Battikh, H., Chaieb, K., Bakhrouf, A., & Ammar, E. (2011). Antibac-
inal draft; writing – review and editing. Alice Gonçalves terial and antifungal activities of black and green kombucha teas.
Martins Gonzalez: resources; visualization; writing – Journal of Food Biochemistry, 37(2), 231–236. https://doi.org/10.
review and editing. Fabrício Freitas Fernandes: con- 1111/j.1745-4514.2011.00629.x
ceptualization; visualization; writing – review and editing. Ben Taheur, F., Mansour, C., Ben Jeddou, K., Machreki, Y., Kouidhi,
Adriene Ribeiro Lima: conceptualization; investigation; B., Abdulhakim, J. A., & Chaieb, K. (2020). Aflatoxin B1 degrada-
tion by microorganisms isolated from kombucha culture. Toxicon,
project administration; supervision; visualization; writing
179(2020), 76–83. https://doi.org/10.1016/j.toxicon.2020.03.004
– original draft; writing – review and editing. Bhattacharya, D., Bhattacharya, S., Patra, M. M., Chakravorty, S.,
Sarkar, S., Chakraborty, W., Koley, H., & Gachhui, R. (2016).
CONFLICT OF INTEREST Antibacterial activity of polyphenolic fraction of kombucha
The authors declare that they have no known compet- against enteric bacterial pathogens. Current Microbiology, 73(6),
ing financial interests or personal relationships that could 885–896. https://doi.org/10.1007/s00284-016-1136-3
have appeared to influence the work reported in this paper. Bhattacharya, S., Gachhui, R., & Sil, P. C. (2013). Effect of kombucha,
a fermented black tea in attenuating oxidative stress mediated tis-
sue damage in alloxan induced diabetic rats. Food and Chemical
ORCID
Toxicology, 60, 328–340. https://doi.org/10.1016/j.fct.2013.07.051
Jeniffer Ferreira de Miranda https://orcid.org/0000- Brasil. (2019). Estabelece o Padrão de Identidade e Qualidade da
0002-3749-9381 Kombucha em todo o território nacional (Instrução Normativa
Larissa Fernandes Ruiz https://orcid.org/0000-0001- No. 41, de 17 de Setembro de 2019). Ministério da Agricultura,
6344-3317 Pecuária e Abastecimento. Diário Oficial da União, Brasilia.
Cíntia Borges Silva https://orcid.org/0000-0002-4391- https://www.in.gov.br/en/web/dou/-/instrucao-normativa-n-41-
6807 de-17-de-setembro-de-2019-216803534
Cacicedo, M. L., Castro, M. C., Servetas, I., Bosnea, L., Boura, K.,
Thais Matsue Uekane https://orcid.org/0000-0001-
Tsafrakidou, P., Dima, A., Terpou, A., Koutinas, A., & Castro,
7239-9274
G. R. (2016). Progress in bacterial cellulose matrices for biotech-
Kelly Alencar Silva https://orcid.org/0000-0003-2239- nological applications. Bioresource Technology, 213, 172–180.
4787 https://doi.org/10.1016/j.biortech.2016.02.071
Alice Gonçalves Martins Gonzalez https://orcid.org/ Cardoso, R. R., Neto, R. O., dos Santos D’Almeida, C. T., do Nasci-
0000-0001-6789-502X mento, T. P., Pressete, C. G., Azevedo, L., Martino, H. S. D. L.,
Fabrício Freitas Fernandes https://orcid.org/0000- Cameron, C., Ferreira, M. S. L., & de Barros, F. A. R. (2020).
0003-2375-7455 Kombuchas from green and black teas have different phenolic
profile, which impacts their antioxidant capacities, antibacterial
Adriene Ribeiro Lima https://orcid.org/0000-0003-
and antiproliferative activities. Food Research International, 128,
2653-066X 108782. https://doi.org/10.1016/j.foodres.2019.108782
Centre for Disease Control. (2020). Food safety assessment
REFERENCES of kombucha tea recipe and food safety plan: British
Aboulwafa, M. M., Youssef, F. S., Gad, H. A., Altyar, A. E., Al-Azizi, Columbia. Environmental Health Services, (March), 1–14.
M. M., & Ashour, M. L. (2019). A comprehensive insight on the http://www.bccdc.ca/resource-gallery/Documents/Educational%
health benefits and phytoconstituents of Camellia sinensis and 20Materials/EH/FPS/Food/kombucha1.pdf
recent approaches for its quality control. In Antioxidants, 8(10), Četojević-Simin, D. D., Velićanski, A. S., Cvetković, D. D., Markov, S.
455. https://doi.org/10.3390/antiox8100455 L., Mrdanović, J. Ž., Bogdanović, V. V., & Šolajić, S. V. (2012). Bioac-
Aloulou, A., Hamden, K., Elloumi, D., Ali, M. B., Hargafi, K., Jaouadi, tivity of lemon balm kombucha. Food and Bioprocess Technology,
B., Ayadi, F., Elfeki, A., & Ammar, E. (2012). Hypoglycemic and 5(5), 1756–1765. https://doi.org/10.1007/s11947-010-0458-6
antilipidemic properties of kombucha tea in alloxan-induced dia- Chakravorty, S., Bhattacharya, S., Bhattacharya, D., Sarkar, S., &
betic rats. BMC Complementary and Alternative Medicine, 12(63), Gachhui, R. (2019). Kombucha: A promising functional beverage
1–9. https://doi.org/10.1186/1472-6882-12-63 prepared from tea. In Non-Alcoholic Beverages. https://doi.org/10.
Australian Government Department of Health. (2019). Fermented 1016/b978-0-12-815270-6.00010-4
beverages stakeholder roundtable. Department of Health. http:// Chakravorty, S., Bhattacharya, S., Chatzinotas, A., Chakraborty, W.,
www.foodregulation.gov.au Bhattacharya, D., & Gachhui, R. (2016). Kombucha tea fermen-
Ayed, L., Ben Abid, S., & Hamdi, M. (2016). Development of a bev- tation: Microbial and biochemical dynamics. International Jour-
erage from red grape juice fermented with the kombucha con- nal of Food Microbiology, 220, 63–72. https://doi.org/10.1016/j.
sortium. Annals of Microbiology, 67(1), 111–121. https://doi.org/10. ijfoodmicro.2015.12.015
1007/s13213-016-1242-2
Chu, S. C., & Chen, C. (2006). Effects of origins and fermentation time dant, antimicrobial and sensory properties of kombucha tea bev-
on the antioxidant activities of kombucha. Food Chemistry, 98(3), erage. Journal of Food Science and Technology, 57(5), 1840–1846.
502–507. https://doi.org/10.1016/j.foodchem.2005.05.080 https://doi.org/10.1007/s13197-019-04217-3
Coban, H. B. (2019). Organic acids as antimicrobial food agents: Jakubczyk, K., Kałduńska, J., Kochman, J., & Janda, K. (2020). Chem-
Applications and microbial productions. Bioprocess and Biosys- ical profile and antioxidant activity of the kombucha beverage
tems Engineering, 43(4), 569–591. https://doi.org/10.1007/s00449- derived from white, green, black and red tea. Antioxidants, 9(5).
019-02256-w https://doi.org/10.3390/antiox9050447
De Filippis, F., Troise, A. D., Vitaglione, P., & Ercolini, D. (2018). Dif- Jayabalan, R., Marimuthu, S., & Swaminathan, K. (2007). Changes in
ferent temperatures select distinctive acetic acid bacteria species content of organic acids and tea polyphenols during kombucha tea
and promotes organic acids production during kombucha tea fer- fermentation. Food Chemistry, 102(1), 392–398. https://doi.org/10.
mentation. Food Microbiology, 73, 11–16. https://doi.org/10.1016/j. 1016/j.foodchem.2006.05.032
fm.2018.01.008 Jayabalan, R., Subathradevi, P., Marimuthu, S., Sathishkumar, M., &
Deghrigue, M., Chriaa, J., Battikh, H., & Abid, K. (2013). Antiprolif- Swaminathan, K. (2008). Changes in free-radical scavenging abil-
erative and antimicrobial activities of kombucha tea. African Jour- ity of kombucha tea during fermentation. Food Chemistry, 109(1),
nal of Microbiology Research, 7(27), 3466–3470. https://doi.org/10. 227–234. https://doi.org/10.1016/j.foodchem.2007.12.037
5897/AJMR12.1230 Jayabalan, R., Malbaša, R. V., Lončar, E. S., Vitas, J. S., & Sathishku-
Dutta, H., & Paul, S. K. (2019). Kombucha drink: Production, quality, mar, M. (2014). A review on kombucha tea-microbiology, compo-
and safety aspects. In Production and Management of Beverages. sition, fermentation, beneficial effects, toxicity, and tea fungus.
https://doi.org/10.1016/b978-0-12-815260-7.00008-0 Comprehensive Reviews in Food Science and Food Safety, 13(4), 538–
Emiljanowicz, K. E., & Malinowska-Pańczyk, E. (2019). Kombucha 550. https://doi.org/10.1111/1541-4337.12073
from alternative raw materials–The review. Critical Reviews in Jayabalan, R., Malbaša, R. V., & Sathishkumar, M. (2016). Kom-
Food Science and Nutrition, 0(0), 1–10. https://doi.org/10.1080/ bucha. Reference Module in Food Science, 1–8. https://doi.org/10.
10408398.2019.1679714 1016/b978-0-08-100596-5.03032-8
Food and Drug Administration. (2009). Food code. http://www.fda. Kaewkod, T., Bovonsombut, S., & Tragoolpua, Y. (2019). Efficacy
gov/Food/GuidanceRegulation/RetailFoodProtection/FoodCode/ of kombucha obtained from green, oolong and black teas on
UCM2019396.htm inhibition of pathogenic bacteria, antioxidation, and toxicity on
Food Standards Australian New Zealand. (2016). No-alcoholic bever- colorectal cancer cell line. Microorganisms, 7(12), 1–18. https://doi.
ages and brewed soft drinks. Australian New Zealand Food Stan- org/10.3390/microorganisms7120700
dards Code—Standard 2.6.2. www.legislation.gov.au Kallel, L., Desseaux, V., Hamdi, M., Stocker, P., & Ajandouz, E. H.
Gaggìa, F., Baffoni, L., Galiano, M., Nielsen, D. S., Jakobsen, R. R., (2012). Insights into the fermentation biochemistry of kombucha
Castro-Mejía, J. L., Bosi, S., Truzzi, F., Musumeci, F., Dinelli, G., teas and potential impacts of kombucha drinking on starch diges-
& Di Gioia, D. (2019). Kombucha beverage from green, black and tion. Food Research International, 49(1), 226–232. https://doi.org/
rooibos teas: A comparative study looking at microbiology, chem- 10.1016/j.foodres.2012.08.018
istry and antioxidant activity. Nutrients, 11(1), 1–22. https://doi.org/ Kanurić, K. G., Milanović, S. D., Ikonić, B. B., Lončar, E. S., Iličić,
10.3390/nu11010001 M. D., Vukić, V. R., & Vukić, D. V. (2018). Kinetics of lactose
Gamboa-Gómez, C. I., González-Laredo, R. F., Gallegos-Infante, J. fermentation in milk with kombucha starter. Journal of Food
A., Pérez, M. M. L., Moreno-Jiménez, M. R., Flores-Rueda, A. and Drug Analysis, 26(4), 1229–1234. https://doi.org/10.1016/j.jfda.
G., & Rocha-Guzmán, N. E. (2016). Antioxidant and angiotensin- 2018.02.002
converting enzyme inhibitory activity of Eucalyptus camaldulen- Kapp, J. M., & Sumner, W. (2019). Kombucha: A systematic review
sis and Litsea glaucescens infusions fermented with kombucha of the empirical evidence of human health benefit. Annals of Epi-
consortium. Food Technology and Biotechnology, 54(3), 367–373. demiology, 30, 66–70. https://doi.org/10.1016/j.annepidem.2018.11.
https://doi.org/10.17113/ftb.54.03.16.4622 001
Gedela, M., Potu hakravarth, K. C., Gali, V. L., Alyamany, K., & Jha, Kole, A. S., Jones, H. D., Christensen, R., & Gladstein, J. (2009). A
L. K. (2016). A case of hepatotoxicity related to kombucha tea con- case of kombucha tea toxicity. Journal of Intensive Care Medicine,
sumption. South Dakota Medicine: The Journal of the South Dakota 24(3), 205–207. https://doi.org/10.1177/0885066609332963
State Medical Association, 69(1), 26–28. Kombucha Brewers International. (2021). Kombucha code of practice.
Gomes, R. J., Borges, M. de, F., Rosa, M. de, F., Castro-Gómez, R. J. H., https://kombuchabrewers.org/kombucha-code-of-practice/
& Spinosa, W. A. (2018). Acetic acid bacteria in the food industry: Kozyrovska, N. O., Reva, O. M., Goginyan, V. B., & Devera, J. P. (2012).
Systematics, characteristics and applications. Food Technology and Kombucha microbiome as a probiotic: A view from the perspec-
Biotechnology, 56(2), 139–151. https://doi.org/10.17113/ftb.56.02.18. tive of post-genomics and synthetic ecology. Biopolymers and Cell,
5593 28(2), 103–113. https://doi.org/10.7124/bc.000034
Holbourn, A., & Hurdman, J. (2017). Kombucha: Is a cup of tea good Leal, J. M., Suárez, L. V., Jayabalan, R., Oros, J. H., & Escalante-
for you? BMJ Case Reports 1–2. Aburto, A. (2018). A review on health benefits of kombucha nutri-
Ismaiel, A. A., Bassyouni, R. H., Kamel, Z., & Gabr, S. M. (2016). tional compounds and metabolites. CYTA—Journal of Food, 16(1),
Detoxification of patulin by kombucha tea culture. CYTA— 390–399. https://doi.org/10.1080/19476337.2017.1410499
Journal of Food, 14(2), 271–279. https://doi.org/10.1080/19476337. Malbaša, R., Lončar, E., & Djurić, M. (2008). Comparison of the prod-
2015.1096828 ucts of kombucha fermentation on sucrose and molasses. Food
Ivanišová, E., Meňhartová, K., Terentjeva, M., Harangozo, Ľ., Kántor, Chemistry, 106(3), 1039–1045. https://doi.org/10.1016/j.foodchem.
A., & Kačániová, M. (2020). The evaluation of chemical, antioxi- 2007.07.020
Malbaša, R. V., Lončar, E. S., Vitas, J. S., & Čanadanović-Brunet, J. nandes, F. F., & Lima, A. R. (2021). Kombucha beverage from
M. (2011). Influence of starter cultures on the antioxidant activity non-conventional edible plant infusion and green tea: Charac-
of kombucha beverage. Food Chemistry, 127(4), 1727–1731. https:// terization, toxicity, antioxidant activities and antimicrobial prop-
doi.org/10.1016/j.foodchem.2011.02.048 erties. Biocatalysis and Agricultural Biotechnology, 34(2021), 1–9.
Marsh, A. J., O’Sullivan, O., Hill, C., Ross, R. P., & Cotter, P. D. (2014). https://doi.org/10.1016/j.bcab.2021.102032
Sequence-based analysis of the bacterial and fungal compositions Sinir, G. Ö., Tamer, C. E., & Suna, S. (2019). Kombucha tea: A
of multiple kombucha (tea fungus) samples. Food Microbiology, promising fermented functional beverage. In Fermented Bever-
38(2014), 171–178. https://doi.org/10.1016/j.fm.2013.09.003 ages, 5(2019), 401–432. https://doi.org/10.1016/b978-0-12-815271-3.
Najafpour, A., Rajabi Khorrami, A., Aberoomand Azar, P., & Saber 00010-5
Tehrani, M. (2020). Study of heavy metals biosorption by tea fun- Shahbazi, H., Hashemi Gahruie, H., Golmakani, M. T., Eskandari, M.
gus in kombucha drink using central composite design. Journal of H., & Movahedi, M. (2018). Effect of medicinal plant type and con-
Food Composition and Analysis, 86(2020), 103359. https://doi.org/ centration on physicochemical, antioxidant, antimicrobial, and
10.1016/j.jfca.2019.103359 sensorial properties of kombucha. Food Science and Nutrition,
Neffe-Skocińska, K., Sionek, B., Ścibisz, I., & Kołożyn-Krajewska, 6(8), 2568–2577. https://doi.org/10.1002/fsn3.873
D. (2017). Contenido de ácido y efectos de las condiciones de Srihari, T., Arunkumar, R., Arunakaran, J., & Satyanarayana,
fermentación en las propiedades fisicoquímicas, microbiológ- U. (2013). Downregulation of signalling molecules involved in
icas y sensoriales de bebidas de té de kombucha. CYTA— angiogenesis of prostate cancer cell line (PC-3) by kombucha
Journal of Food, 15(4), 601–607. https://doi.org/10.1080/19476337. (lyophilized). Biomedicine and Preventive Nutrition, 3(1), 53–58.
2017.1321588 https://doi.org/10.1016/j.bionut.2012.08.001
Nguyen, N. K., Nguyen, P. B., Nguyen, H. T., & Le, P. H. (2015). Screen- Srihari, T., Karthikesan, K., Ashokkumar, N., & Satyanarayana,
ing the optimal ratio of symbiosis between isolated yeast and acetic U. (2013). Antihyperglycaemic efficacy of kombucha in strep-
acid bacteria strain from traditional kombucha for high-level pro- tozotocin-induced rats. Journal of Functional Foods, 5(4), 1794–
duction of glucuronic acid. LWT—Food Science and Technology, 1802. https://doi.org/10.1016/j.jff.2013.08.008
64(2), 1149–1155. https://doi.org/10.1016/j.lwt.2015.07.018 Talebi, M., Frink, L. A., Patil, R. A., & Armstrong, D. W. (2017). Exam-
Nummer, B. A. (2013). Kombucha brewing under the food and drug ination of the varied and changing ethanol content of commercial
administration model food code: Risk analysis and processing kombucha products. Food Analytical Methods, 10(12), 4062–4067.
guidance. Journal of Environmental Health, 76(4), 8–11. https://doi.org/10.1007/s12161-017-0980-5
Puspitasari, Y., Nurikasari, M., Palupi, R., Siwi, Y., Surya, S., & Kediri, Tanaka, T., & Kouno, I. (2003). Oxidation of tea catechins: Chemi-
M. H. (2017). Characterization and analysis kombucha tea antioxi- cal structures and reaction mechanism. Food Science Technology
dant activity based on long fermentation as a beverage functional. Research, 9(2), 128–133. https://doi.org/10.3136/fstr.9.128
Journal of Global Research in Public Health, 2(2), 90–96. Tax and Trade Bureau. (2016). Kombucha. U.S. Department of the
Ransom, D., & McDermott, W. (2020). Progress on the kom- Treasury. https://www.ttb.gov/kombucha
bucha act continues in 2020. Kombucha Brewers International, Tu, C., Tang, S., Azi, F., Hu, W., & Dong, M. (2019). Use of kom-
https://kombuchabrewers.org/progress-on-the-kombucha-act- bucha consortium to transform soy whey into a novel func-
continues-in-2020/ tional beverage. Journal of Functional Foods, 52(July 2018). 81–89.
Rocha-Guzmán, N. E., González-Laredo, R. F., Vázquez-Cabral, B. https://doi.org/10.1016/j.jff.2018.10.024
D., Moreno-Jiménez, M. R., Gallegos-Infante, J. A., Gamboa- Uganda National Bureau of Standards. (2018). Kombucha
Gómez, C. I., & Flores-Rueda, A. G. (2019). Oak leaves as a new specification. https://ec.europa.eu/growth/toolsdatabases/tbt/
potential source for functional beverages: Their antioxidant capac- en/search/?tbtaction=search.detail&Country_ID=UGA&num=
ity and monomer flavonoid composition. Functional and Medici- 930&dspLang=en&basdatedeb=01/10/2018&basdatefin=31/10/
nal Beverages, 11(2019), 381–411. https://doi.org/10.1016/B978-0-12- 2018&baspays=&basnotifnum=&basnotifnum2=&bastypepays=
816397-9.00011-X ANY&baskeywords=
Sang, S., Lambert, J. D., Ho, C. T., & Yang, C. S. (2011). The chemistry Uțoiu, Elena, Matei, Florentina, Toma, Agnes, Diguță, Camelia
and biotranformation of tea constituents. Pharmacological, 64(2), Filofteia, Ștefan, Laura Mihaela, Mănoiu, Sorin, Vrăjmașu,
87–99. https://doi.org/10.1016/j.phrs.2011.02.007 Virgil Valeriu, Moraru, Ionut, Oancea, Anca, Israel-Roming,
Santos, J. S., Deolindo, C. T. P., Hoffmann, J. F., Chaves, F. C., Do Florentina, Cornea, Călina Petruta, Constantinescu-Aruxandei,
Prado-Silva, L., Sant’ana, A. S., Azevedo, L., Do Carmo M. A. Diana, Moraru, Angela, & Oancea, Florin (2018). Bee collected
V., & Granato, D. (2018). Optimized Camellia sinensis var. sinen- pollen with enhanced health benefits, produced by fermentation
sis, Ilex paraguariensis, and Aspalathus linearis blend presents with a kombucha consortium. Nutrients, 10(10), 1–24. https://doi.
high antioxidant and antiproliferative activities in a beverage org/10.3390/nu10101365
model. Food Chemistry, 15(254), 348–358. https://doi.org/10.1016/ Vázquez-Cabral, B. D., Larrosa-Pérez, M., Gallegos-Infante, J.
j.foodchem.2018.02.021 A., Moreno-Jiménez, M. R., González-Laredo, R. F., Rutiaga-
Santos-Buelga, C., González-Paramás, A. M., Oludemi, T., Ayuda- Quiñones, J. G., Gamboa-Gómez, C. I., & Rocha-Guzmán, N. E.
Durán, B., & González-Manzano, S. (2019). Plant phenolics as (2017). Oak kombucha protects against oxidative stress and inflam-
functional food ingredients. Advances in Food and Nutrition matory processes. Chemico-Biological Interactions, 272(2017), 1–9.
Research, 90(2019), 183__257. https://doi.org/10.1016/bs.afnr.2019. https://doi.org/10.1016/j.cbi.2017.05.001
02.012 Velićanski, A. S., Cvetković, D. D., Markov, S. L., Tumbas Šaponjac,
Silva, K. A., Uekane, T. M., Miranda, J. F. d., Ruiz, L. F., Motta, J. V. T., & Vulić, J. J. (2014). Antioxidant and antibacterial activity of
C. B. d., Silva, C. B., Pitangui, N. S., Gonzalez, A. G. M., Fer- the beverage obtained by fermentation of sweetened lemon balm
(Melissa offi cinalis L.) tea with symbiotic consortium of bacte- ing and Preservation, 39(6), 2596–2603. https://doi.org/10.1111/jfpp.
ria and yeasts. Food Technology and Biotechnology, 52(4), 420–429. 12509
https://doi.org/10.17113/ftb.52.04.14.3611 Xia, X., Dai, Y., Wu, H., Liu, X., Wang, Y., Yin, L., Wang, Z., Li, X.,
Villarreal-Soto, S. A., Beaufort, S., Bouajila, J., Souchard, J. P., Renard, & Zhou, J. (2019). Kombucha fermentation enhances the health-
T., Rollan, S., & Taillandier, P. (2019). Impact of fermentation con- promoting properties of soymilk beverage. Journal of Functional
ditions on the production of bioactive compounds with anticancer, Foods, 62(2019), 1–13. https://doi.org/10.1016/j.jff.2019.103549
anti-inflammatory and antioxidant properties in kombucha tea Yanagimoto, K., Ochi, H., Lee, K. G., & Shibamoto, T. (2003). Antiox-
extracts. Process Biochemistry, 83(2019), 44–54. https://doi.org/10. idative activities of volatile extracts from green tea, oolong tea, and
1016/j.procbio.2019.05.004 black tea. Journal of Agricultural and Food Chemistry, 51(25), 7396–
Villarreal-Soto, S. A., Beaufort, S., Bouajila, J., Souchard, J., & Tail- 7401. https://doi.org/10.1021/jf030127i
landier, P. (2018). Understanding kombucha tea fermentation: A Yavari, N., Assadi, M. M., Larijani, K., & Moghadam, M. B. (2010).
review. HAL Archives-Ouvertes, 83(3). 580–588. https://doi.org/10. Response surface methodology for optimization of glucuronic acid
1111/1750-3841.14068 production using kombucha layer on sour cherry juice. Australian
Villarreal-Soto, S. A., Bouajila, J., Pace, M., Leech, J., Cotter, P. D., Journal of Basic and Applied Sciences, 4(8), 3250–3256.
Souchard, J. P., Taillandier, P., & Beaufort, S. (2020). Metabolome- Zubaidah, E., Afgani, C. A., Kalsum, U., Srianta, I., & Blanc, P. J.
microbiome signatures in the fermented beverage, kombucha. (2019). Comparison of in vivo antidiabetes activity of snake fruit
International Journal of Food Microbiology, 333(2020), 108778. kombucha, black tea kombucha and metformin. Biocatalysis and
https://doi.org/10.1016/j.ijfoodmicro.2020.108778 Agricultural Biotechnology, 17(2019), 465–469. https://doi.org/10.
Vitas, J. S., Cvetanović, A. D., Mašković, P. Z., Švarc-Gajić, J. V., & 1016/j.bcab.2018.12.026
Malbaša, R. V. (2018). Chemical composition and biological activ- Zubaidah, E., Dewantari, F. J., Novitasari, F. R., Srianta, I., & Blanc, P.
ity of novel types of kombucha beverages with yarrow. Journal of J. (2018). Potential of snake fruit (Salacca zalacca (Gaerth.) Voss)
Functional Foods, 44(2018), 95–102. https://doi.org/10.1016/j.jff. for the development of a beverage through fermentation with the
2018.02.019 kombucha consortium. Biocatalysis and Agricultural Biotechnol-
Wang, Y., Ji, B., Wu, W., Wang, R., Yang, Z., Zhang, D., & Tian, ogy, 13(2018), 198–203. https://doi.org/10.1016/j.bcab.2017.12.012
W. (2013). Hepatoprotective effects of kombucha tea: Identifica-
tion of functional strains and quantification of functional compo-
nents. Journal of the Science of Food and Agriculture, 94(2), 265–
272. https://doi.org/10.1002/jsfa.6245 How to cite this article: de Miranda, J. F., Ruiz,
Watawana, M. I., Jayawardena, N., Gunawardhana, C. B., & Waisun- L. F., Silva, C. B., Uekane, T. M., Silva, K. A.,
dara, V. Y. (2015). Health, wellness, and safety aspects of the Gonzalez, A. G. M., Fernandes, F. F., & Lima, A. R.
consumption of kombucha. Journal of Chemistry, (2015). 1–11.
(2022). Kombucha: A review of substrates,
https://doi.org/10.1155/2015/591869
regulations, composition, and biological properties.
Watawana, M. I., Jayawardena, N., & Waisundara, V. Y. (2015).
Enhancement of the functional properties of coffee through fer- J Food Sci. 87, 503–527.
mentation by “tea fungus” (kombucha). Journal of Food Process- https://doi.org/10.1111/1750-3841.16029

Journal of Food Science - 2022 - Miranda - Kombucha A Review of Substrates Regulations Composition and Biological

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Journal of Food Science - 2022 - Miranda - Kombucha A Review of Substrates Regulations Composition and Biological

Uploaded by

Copyright:

Available Formats

17503841, 2022, 2, Downloaded from https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.16029 by UFAL - Universidade Federal de Alagoas, Wiley Online Library on [22/02/2023].

CONCISE REVIEWS & H YPOTH ESES IN FOOD SCIENCE

Kombucha: A review of substrates, regulations,

Jeniffer Ferreira de Miranda1 Larissa Fernandes Ruiz1

1 Department of Bromatology, Pharmacy

J. Food Sci. 2022;87:503–527. wileyonlinelibrary.com/journal/jfds © 2022 Institute of Food Technologists

TA B L E 1 Different preparation conditions of kombucha in Camellia sinensis substrate

Infusions and extracts

Substrate fermented Fermentation Temperature

TA B L E 3 Compounds identified in fermentation process of Camellia sinensis tea

important to emphasize that the presence of these com-

Neffe-Skocińska et al., 2017

Kaewkod, Bovonsombut &

Jakubczyk et al., 2020

Malbaša et al., 2008

in acetic acid production up to the 15th day of fermenta-

tion, reaching a maximum of 9.5 g/L in green tea kom-

reached 16.57 and 7.36 g/L, respectively, after 21 days of fer-

was the most-produced organic acid, both in green and

Velićanski et al. (2014), when evaluating the antiox-

idant and antimicrobial activity of beverage fermented

reached a maximum 8.12 g/L after 7 days of fermentation.

duction, and they observed that pH value and organic acid

content changed throughout fermentation. After 8 days

source in the production of black tea kombucha and evi-

denced that there was a greater drop in pH values and, con-

sequently, higher production of organic acids compared to

the beverage prepared with sucrose. Total acids concentra-

acids. However, acetic acid production was much lower

compared to sucrose in the same fermentation stage. Due

to the presence of some components of molasses, it is possi-

In the human body, vitamins and minerals have ben-

eficial effects on health and are responsible for normal

metabolic pathways and different physiological functions

(Vitas et al., 2018). Studies have reported that essential

TA B L E 4 Compounds identified in fermentation process of kombucha analogs

Substrate Organic acids Vitamins (GAE*) (days) References

Soy whey Acetic acid 5.77 g/L – – 8 Tu et al., 2020

TA B L E 5 Ethanol content in kombucha and kombucha analogs

You might also like