Kombucha as a Fermented Beverage

Kombucha, a fermented tea drink from Asia, is gaining popularity for its potential health
benefits. According to Esatbeyoglu et al (2022), kombucha possesses antimicrobial, antioxidant,
antidiabetic, and anti-carcinogenic properties. These properties and chemistry are attributed to
the SCOBY (Symbiotic Culture Of Bacteria and Yeast). The SCOBY is the one responsible for
transforming the sweetened solution into kombucha with the process of breakdown of sugars to
produce organic acids such as acetic acid as well as creating the beneficial compounds of
kombucha (Dartora et al. 2023; Nyhan et al. 2022). The microbial community within SCOBY
stems from the symbiotic culture of beneficial microorganisms. However, further studies are
needed to fully understand the mechanisms by which these microbial products contribute to
human health. As with any food or drink, it is important to consume kombucha in moderation as
part of a balanced diet.

Microbial Advantages of Kombucha

Kombucha’s fermentation, from the metabolic activities of its microbial community, is

responsible for its distinctive taste and potential health benefits. Acetic acid, produced during
acetic acid fermentation contributes to the characteristic sour taste and acts as a preservative by
lowering the pH, inhibiting the growth of spoilage organisms and pathogens (Greenwalt et al.
1998).

Ethanol (Alcohol), generated during alcoholic fermentation, by yeast species like

Saccharomyces , although present only in trace amounts, plays a vital role in forming acetic acid
and contributes to the beverage's antimicrobial properties (Teoh et al., 2004). Carbon dioxide
(CO2), also produced during yeast fermentation, adds effervescence and enhances the sensory
experience of kombucha (Villarreal-Soto et al., 2018). Gluconic acid, produced by the oxidation
of glucose by acetic acid bacteria such as Gluconobacter, contributes to the mild acidic taste and
has potential health benefits, including improving mineral absorption (Malbasa et al., 2008).
Additionally, lactic acid, produced during lactic acid fermentation by lactic acid bacteria (LAB)
such as Lactobacillus, enhances the beverage's acidity and probiotic content, supporting gut
health (Battikh et al., 2013).

Kombucha's unique flavor profile and potential health benefits are further enriched by
various flavor compounds like esters, ketones, and aldehydes, resulting from complex microbial
interactions during fermentation (Chen & Liu, 2000). The beverage is also rich in polyphenols
and antioxidants, primarily derived from the tea used in brewing. By leveraging these microbial
advantages, kombucha not only offers a distinctive taste and sensory experience but also presents
numerous health-promoting properties, making it a popular choice among health-conscious
consumers.

Aside from the increase in organic acids and alcohol content of kombucha, certain
metabolites are also produced. During the fermentation process in kombucha, there is an increase
in phenols which also increase the antioxidant activity of the beverage (Srihari & Satyanarayana,
2012). Kombucha fermentation leads to the synthesis of water-soluble B-complex vitamins,
including B1, B2, B6, and B12, which are essential for energy metabolism and maintaining
various physiological functions and are attributed to the metabolic activities of the yeast and
bacteria present in the SCOBY(Srihari & Satyanarayana 2012). Despite the inhibiting factors of
kombucha beverage against pathogenic bacteria, spoilage still occurs.

Intrinsic and Extrinsic Factors

To combat against pathogenic microorganism, and certain spoilage microbes, kombucha
has its characteristic intrinsic features that serves as a barrier with that purpose. One factor of
kombucha is its relatively low pH (2.5 to 4) which inhibits the growth of most spoilage
microorganisms (Içen et al., 2023) due to its high level of acidity. Aside from pH, the water
composition of the drink itself is essential as water is considered to be an initial growth medium
for yeast and bacteria— this is before the fermentation process. In making kombucha, water that
met a baseline standard of potability should be opted to use. With that, the microbial content also
plays an important role in the production of kombucha. The microbes innate kombucha are acetic
acid bacteria, lactic acid bacteria (LAB), and yeasts. These microorganisms comprise the
SCOBY as mentioned priorly (Tran et al., 2020). In fact, some of these mentioned microbes have
been discovered to have an antimicrobial activity against many pathogenic microorganisms.
Temperature is a critical extrinsic factor in the fermentation and storage of Kombucha
because it significantly influences the beverage's overall quality and safety. Higher temperatures
accelerate the acidification process, as observed by Loncar et al. (2006), where fermentation at
30°C resulted in faster sugar consumption compared to 22°C. This is due to the increased
metabolic activity of the yeast and bacteria at higher temperatures, leading to quicker
fermentation. Conversely, lower temperatures slow down microbial activity, resulting in a slower
fermentation process. The temperature also affects the flavor and stability of Kombucha; higher
temperatures can lead to over-fermentation and increased acidity, while lower temperatures help
maintain the desired balance of acids and other compounds, extending the beverage's shelf life.
Proper temperature control during both fermentation and storage is essential to prevent spoilage
and ensure the safety and consistency of the final product.

Potential Food Spoilage

Microbial interactions in kombucha, often described as “symbiotic”, involve acetic acid

bacteria, lactic acid bacteria, and yeast coexisting stably. These microorganisms play a role in
metabolizing the tea and sugar mixture to produce organic acids like acetic acid, contributing to
the kombucha’s taste and acidity (Wang et al., 2022). During fermentation, the formation of the
pellicle and the drop of pH serves as a protective mechanism against contamination by external
spoilage agents such as molds (May et al. 2019). However, factors related to the fermentation
process and contamination by harmful microorganisms such as Staphylococcus aureus,
Escherichia coli, Salmonella spp., and Listeria monocytogenes can influence spoilage in the
food, especially in home-made kombucha (Greenwalt et al. 1998; Sreeramulu et al. 2000).
Home-made kombucha has been a subject of interest due to its antimicrobial properties. Despite
kombucha’s antimicrobial properties, attributed to components such as acetic acid, polyphenols,
gluconic acid, and lactic acid, and antibiotics produced during fermentation, improper
fermentation conditions or contamination during the fermentation process can lead to spoilage
(Wang et al. 2022).
Acetic acid bacteria are the most characterized bacteria in kombucha fermentation as it plays a
role in converting the ethanol produced by yeast into acetic acid. However, Acetic Acid Bacteria
(AAB) of the genera Acetobacter and Gluconobacter have been deemed as the spoiling agent of
many alcoholic products (Bishop, 2022). Prolonged fermentation causes excessive amounts of
acetic acid that makes kombucha overly acidic and undesirably sour that is unpalatable.
Excessive acetic acid also changes the text texture of kombucha creating a slimy feel to the
drink. Kombucha in this state is not necessarily spoiled but its overly sour taste and slimy texture
makes it undesirable for most consumers. Along with the potential food spoilage microbes, it is
also important to identify the extrinsic and intrinsic factors that play a role in the spoilage
process of kombucha.

Potential Health Risk

Kombucha has gained popularity due to its potential health benefits. However, despite its
therapeutic effects, concerns have been raised regarding its safety and potential health risks,
especially in immuno-compromised individuals (Miranda et al. 2022). One of the risks
associated with kombucha consumption is its potential to contain high levels of alcohol, as it can
pose health risks in children and pregnant individuals (Chan et al. 2020). The variability in
alcohol levels can be influenced by fermentation time, temperature, and the composition of the
symbiotic culture used in the fermentation process (Suprayogi et al. 2023).
add more…

Microbial control
A) Variation in tea leaves
The variety of tea leaves plays a role in microbial regulation that may not be
immediately apparent, as many homebrewers overlook the potential impact of tea leaves
on the microorganisms present in kombucha. However, based on the literature of Al-
Mohammadi et al. (2021), different tea leaves, such as green, oolong, and black tea, can
have inhibitory effects against different pathogenic bacteria. Kombucha made from green
tea has been reported to inhibit various bacteria, including S. epidermidis, S. aureus,
Micrococcus gluteus, E. coli, etc. (Kaewkod et al., 2019). The ability to hinder the
growth of harmful microorganisms is attributed to the bioactive substances present in tea
leaves. These include tea polyphenols, D-saccharic acid-1,4-lactone, and vitamin C,
 responsible for kombucha's antioxidant properties (Zailani & Adnan, 2022). Big
manufacturers recommended that homebrewers always include tea leaves in their
kombucha.
B) Boiling Water for Preparation
In the preparation of kombucha, boiling water plays a crucial role in ensuring the
safety and quality of the beverage. Boiling water before brewing kombucha serves to
eliminate any potential contaminants that could interfere with the fermentation process or
pose health risks to consumers. The high temperature of boiling water, typically 100°C
(212°F), effectively kills most pathogenic microorganisms, including bacteria, viruses,
and protozoa, that might be present in tap water (World Health Organization, 2022). This
step is essential as it creates a sterile environment for the tea and sugar solution, which is
the primary substrate for the kombucha culture (SCOBY). Using boiled water helps
prevent unwanted microbial competition, ensuring that the desired fermentation
microorganisms, primarily acetic acid bacteria and yeast, can thrive without interference
from harmful pathogens (Teoh et al., 2004). By starting with boiled water, home brewers
and commercial producers alike can reduce the risk of contamination and ensure a safe
and consistent product.
C) Pasteurization
Pasteurization, a heat treatment process, is sometimes applied in kombucha
production to control microbial growth and enhance shelf life. This process involves
heating the kombucha to a specific temperature for a set period, sufficient to kill harmful
pathogens and spoilage organisms without significantly altering the beverage's flavor and
nutritional properties (Jay et al., 2005). Typically, pasteurization for kombucha involves
heating the beverage to around 60-70°C (140-158°F) for 15-30 minutes (Roussin, 1996).
While traditional raw kombucha is prized for its probiotic content, pasteurization ensures
microbial safety, particularly important for commercial distribution where longer shelf
life and consistency are desired. This process effectively reduces the risk of
contamination from harmful bacteria like Escherichia coli, Salmonella, and Listeria,
which can occasionally be found in raw or improperly fermented beverages (Jayabalan et
al., 2014). However, it is essential to balance the pasteurization parameters to maintain
 the beneficial properties of kombucha, as excessive heat can degrade sensitive bioactive
compounds and probiotic microorganisms.
D) pH and Acidty

In kombucha, pH and acidity are crucial for controlling microbial growth by creating an
inhospitable environment for pathogenic and spoilage microorganisms. The beverage typically
maintains a pH between 2.5 and 3.5, making it highly acidic. This low pH effectively inhibits the
growth of harmful bacteria and molds. During fermentation, acetic acid, gluconic acid, and lactic
acid are produced, lowers the pH and contributes antimicrobial properties.. These acids act as
natural preservatives, preventing the proliferation of undesirable microorganisms and ensuring
the microbiological safety of kombucha. This method of microbial control is highly effective due
to the stringent environment created by the low pH. However, it is essential to regularly monitor
pH levels during brewing to ensure they stay within the safe range. If the pH is too high, there is
a risk of contamination; if it is too low, it may affect the taste and harm beneficial microbes. In
order to develop an effective method for microbial control in kombucha, suitable techniques for
microbial identification and measurement should be established.

Microbiological Analysis

In the study of Wang et al. (2022), as highlighted in the study of six commercial samples
sold in New Zealand, it revealed the importance of enumerating acetic acid bacteria (AAB) and
yeast. The enumeration process focused on identifying viable AAB and yeast, which are crucial
for understanding the microbial composition of Kombucha at the time of consumption.

According to FDA Circular no. 2013-010 under non-alcoholic beverages, which includes
Kombucha, the required tests are for yeast and molds count, aerobic plate count, and coliforms.
Although Kombucha is not explicitly mentioned in FDA standards, it falls under the broader
category of non-alcoholic beverages, making these tests essential for ensuring product safety and
quality.

Aside from microbial-based tests, chemical characteristics of kombucha are also tested
including the pH, alcohol content, titratable acidity, and Total Soluble Solids (TSS). The pH of
each 50 ml of Kombucha sample was determined using a glass electrode pH meter as well as
the alcohol content using an ebulliometer (Wang et al. 2022). Titratable acidity was determined
using acid - base titration with a standardized 0.1 M sodium hydroxide to pH 8.2 with the use of
phenolphthalein as an indicator in which calculated TA was expressed as a percentage (%) of
acetic acid per gramme of sample(Wang et al 2022). The TSS of Kombucha mainly reflects the
sugar concentration of the beverage, expressed as °Brix which was determined using a
refractometer (Zubaidah et al. 2019).

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