Review
Microbial fermented
tea e a potential
source of natural
food preservatives
Haizhen Moa, Yang Zhub,* and
Zongmao Chenc
a
Department of Food Science, Henan Institute of
Science and Technology, 453003 Xinxiang,
Henan Province, China
b Leheup, & Cordier, 1998). Because no microorganisms
Food and Bioprocess Engineering Group, Wageningen
University and Research Centre, P.O. Box 8129, 6700
EV, Wageningen, Netherlands (Tel.: D31 343 538103;
fax: D31 343 538405; e-mail: yang.zhu@wur.nl)
c
Chinese Tea Research Institute, Chinese Academy of
Agricultural Sciences, 310008 Hangzhou, Zhejiang
Province, China
Antimicrobial activities of microbial fermented tea are much
less known than its health beneficial properties. These anti-
microbial activities are generated in natural microbial fermen-
tation process with tea leaves as substrates. The antimicrobial
components produced during the fermentation process have
shown inhibitory effects against several food-borne and patho-
genic bacteria. With the trend of increasing use of natural and
biological preservatives in food products, natural antimicrobial
agents from microbial fermented tea may offer an innovative
and interesting measure for such applications. However,
a breakthrough in this field can only be realised after several
critical aspects are clarified and further studied. Only then,
the application of these potential, novel and natural antimicro-
bial substances from microbial fermented tea can be industri-
alized. The present review describes some unique microbial
fermentation of tea and the antimicrobial activities formed
during the fermentation process. Moreover, future needs in
research and development of these antimicrobial compounds
from microbial fermentation of tea are discussed for potential
industrial applications.
* Corresponding author.
Introduction
Tea is one of the most consumed beverages worldwide.
In fact, there are mainly three different kinds of tea. In China
and many Southeast Asian countries, green tea is preferred.
In Western countries and the rest of the world, the consump-
tion of black tea is the highest. Another kind of tea is Kom-
bucha, which is a sweet-sour tea beverage made actually
from tea extract supplemented with sugar and fermented
with yeast and acetic-acid bacteria. Fermentation process
is necessary to produce black tea and Kombucha. Black
tea can be further subdivided into naturally oxidized tea
(although the term of fermentation is often used), and micro-
bial fermented tea. Fermentation process for producing most
black teas is actually an oxidation process catalyzed by
enzymes that are originally present in tea leaves (Fowler,
are involved in this kind of enzyme-oxidized black tea,
the use of the term fermentation is obviously not completely
correct. Microbial fermentation in the production of black
tea, however, is found to be carried out solely in certain Chi-
nese tea assortments. Therefore, this kind of tea is little
known outside China, whereas Kombucha is exceptionally
well known because of its increasing interests in the West.
Recently, several microbial fermented teas got noticed in
the Western world, probably not only because of trade ex-
pansions between China and the West, but also because of
several health beneficial claims associated with microbial
fermented tea. A few studies reveal that extracts from mi-
crobial fermented teas contain natural antimicrobial com-
ponents that have inhibitive effect on several food-borne
pathogen and spoilage bacteria (Greenwalt, Ledford, &
Steinkraus, 1998; Mo, Xu, Yan, & Zhu, 2005; Sreeramulu,
Zhu, & Knol, 2000; Sreeramulu, Zhu, & Knol, 2001;
Steinkraus, Shapiro, Hotchkiss, & Mortlock, 1996).
Application of natural antibacterial agents has been in-
creasingly noticed as a novel trend in biological preserva-
tion of foods in recent years (Schillinger, Geisen, &
Holzapfel, 1996). Although plants can provide a vast source
of natural preservatives, many potential biological preserva-
tives are originated from traditional food fermentations. The
most well known example is probably the production of
nisin by lactic acid bacteria. Some potential and unexplored
food fermentation processes are certainly new possibilities
for innovative applications in the modern society.
This article reveals the background information of micro-
bial fermented Puer tea and Fuzhuan brick-tea, as well as
fermented tea beverage Kombucha. The importance of their
antibacterial effects on several food-borne bacteria are given
in detail. Potential applications and research needs of these
microbial fermented teas are also critically discussed. The
antimicrobial effects of tea catechins or polyphenols have
been intensively described previously elsewhere (An et al.,
2004; Bandyopadhyay, Chatterjee, Dasgupta, Lourduraja,
& Dastidar, 2005; Cushnie & Lamb, 2005; Friedman,
Henika, Levin, Mandrell, & Kozukue, 2006; Hamilton-Miller,
1995; Hirasawa & Takada, 2004; Kim, Ruengwilysup, &
Fung, 2004; Si et al., 2006; Taguri, Tanaka, & Kouno, 2004;
Yamamoto, Matsunaga, & Friedman, 2004; Yilmaz, 2006)
and so this topic is not included here.
Microbial fermentation and antimicrobial effect of
Puer tea, Fuzhuan brick-tea and Kombucha
Both Puer tea and Fuzhuan brick-tea are microbial fer-
mented black teas, whereas Kombucha is a fermented drink
of tea extract supplemented with sucrose and fermented
with yeasts and acetic-acid bacteria. All three teas have
shown obvious antibacterial effects.
Fermentation process and antimicrobial
characteristics of Puer tea
Puer tea is a unique Chinese microbial fermented tea ob-
tained through indigenous tea fermentation where micro-
organisms are involved in the manufacturing process. A
schematic description of Puer tea production is shown in
Fig. 1. This tea is different from common black tea that un-
dergoes a natural oxidation process through enzymes orig-
inally existing in tea leaves (Fowler et al., 1998). The
process of Puer tea manufacture is characterized by solid-
state fermentation with natural flora as inoculums of which
the fungus Aspergillus niger plays a key role (Mo et al.,
2005; Xu, Yan, & Zhu, 2005). The processing of Puer tea
is quite different from that of black tea, although they are
both called fermented teas. During black tea processing,
fresh tea leaves are rolled and cut before drying so that
Collecting of fresh tea leaves
Drying Steaming treatment
Wetting and cooling Ripening
Pasteurization Loose Puer tea product
Steaming and pressing Drying
Fig. 1. Schematic description of Puer tea manufacture.
tea polyphenols in tea leaves come into contact with the
tea polyphenol oxidases and then oxidized in the conse-
quent fermentation process. During the Puer tea fermenta-
tion process, fresh leaves are fixed by heat in a drum to
inactivate polyphenol oxidases. The fixed leaves are then
rolled and partially dried. The partially dried leaves are
piled up in humid conditions (moisture content w40% at
25e60  C) for a few weeks, during which the tea polyphe-
nols are more intensively oxidized by the action of micro-
organisms and environmental oxygen than in black tea
fermentation process, resulting in low concentrations of
tea polyphenols and tea catechins (Liang, Zhang, & Lu,
2005).
Puer tea has not only a unique flavour but also several
health beneficial properties, such as suppressing fatty acid
synthase expression (Chiang et al., 2006), acting as an in-
hibitor of lipid and nonlipid oxidative damage and also
exhibiting metal-binding ability, reducing power, and scav-
enging effect for free radicals (Duh, Yen, Yen, Wang, &
Chang, 2004; Jie et al., 2006). Kuo et al. (2005) reported
that Puer tea and oolong tea can lower the levels of triglyc-
eride more significantly than that of green tea and black tea,
whereas Puer tea and green tea are more efficient than
oolong tea and black tea in lowering the level of total
cholesterol. Liang et al. (2005) reported that Puer tea sup-
presses the genotoxicity induced by nitroarenes, lowers the
atherogenic index and increases HDL (high density lipo-
protein) e total cholesterol ratio. Rather recently, antimuta-
genic and antimicrobial activities of Puer tea were also
reported (Wu, et al., 2007). Although Puer tea has a history
of production and consumption for hundreds of years, few
studies can be found on its microbiological properties,
especially during the fermentation process.
Mo et al. (2005) have done a microbiological analysis on
samples from the indigenously fermented Puer tea. Micro-
bial counting and identification revealed that A. niger was
the dominating microorganism during the fermentation.
Antimicrobial activity of fermentation samples shows
Inactivation of enzymes Dispersion of tea blocks
Grading Natural inoculation and fermentation
Evaluation and grading
Inspection and packaging
Pressed tea product Storage and ripening
Inspection and packaging
inhibitory effect on several food-borne bacteria, including
spore-forming bacteria Bacillus cereus, Bacillus subtilis,
Clostridium perfringens and Clostridium sporogenes. The
antimicrobial activity increases with the course of the fer-
mentation. This implies that certain metabolites of A. niger
growing on tea leaves have the feature of inhibiting certain
food-borne bacteria. As the fermentation process will
decrease the content of tea polyphenols and catechins, the
increase in antimicrobial activity of Puer tea results obvi-
ously in the formation of antimicrobial components other
than tea polyphenols and catechins, or in their conversion
to other new compounds.
Fermentation process and antimicrobial characteristics
of Fuzhuan brick-tea
Fuzhuan brick-tea is another kind of microbial fer-
mented tea uniquely found in China (Xu, Mo, Yan, &
Zhu, 2007). A schematic description of Fuzhuan brick-tea
production is given in Fig. 2.
In a similar manner to studying Puer tea, we analyzed
the microbiological composition and tested the antimicro-
bial activity of extracts from the indigenously fermented
Fuzhuan brick-tea (unpublished data). Microbial counting
and identification revealed that Aspergillus spp., Penicil-
lium spp. and Eurotium spp. were the main microorganisms
isolated from the samples during fermentation and Euro-
tium spp. was the dominating fungus during the fermenta-
tion. Antibacterial tests of extracts of fermented tea
showed inhibitory effect on several food-borne bacteria,
including spore-forming bacteria B. cereus, B. subtilis,
C. perfringens and C. sporogenes. The antibacterial activity
increased with the course of the fermentation. This implies
that certain metabolites of the fungi growing on tea leaves
had the feature of inhibiting certain food-borne bacteria.
Yao, Tan, Zhang, Su, & Wei (1998) developed a bulk
acoustic wave bacterial growth sensor to study inhibitory
effects of tea by continuous monitoring of disturbances in
Proteus spp. growth in the aqueous extracts of various
teas, including Fuzhuan brick-tea. The kinetic parameters,
such as asymptote, maximum specific growth rate, lag
time, and generation time were accurately estimated by us-
ing the growth response model. This model characterizes
antimicrobial properties of the tested teas. All the parame-
ters were changed via the inhibitory effects by tea. These
inhibitory effects have also been examined by using the
pour plate count technique for validation of the model.
Collecting of fresh tea leaves Enzyme inactivation and grinding
Mixing Steaming Natural oxidation
Fungal fermentation Drying
Fig. 2. Schematic description of Fuzhuan brick-tea manufacture.
Both results show that, in addition to the antimicrobial
properties of tea polyphenols and catechins, the inhibitory
effects are probably due to the metabolites produced during
the fermentation processing.
Fermentation process and antimicrobial characteristics
of Kombucha
Kombucha is a slightly sweet-sour flavoured tea bever-
age, obtained by fermentation of sweetened boiled tea
with a mixed culture of yeasts and acetic-acid bacteria
(Aidoo, Nout, & Sarkar, 2006). A flowchart of Kombucha
preparation is given in Fig. 3. Kombucha originated in
northeast China (former Manchuria region) and later spread
to Russia and the rest of the world. Kombucha is also fre-
quently called ‘‘tea fungus’’ in the literature, although there
is actually no fungus involved in the fermentation (Benk,
1988; Liu, Hsu, Lee, & Liao, 1996; Mayser, Fromme,
Leitzmann, & Grunder, 1995; Sievers, Lanini, Weber,
SchulerSchmid, & Teuber, 1996; Steinkraus et al., 1996).
This beverage reportedly has a number of health benefits,
for example, against metabolic disease, arthritis, psoriasis,
constipation, indigestion, and hypertension, but there are
few solid scientific evidences available yet for its efficacy
(Dufresne & Farnworth, 2000; Ernst, 2003; Greenwalt,
Steinkraus, & Ledford, 2000; Hartmann, Burleson, Holmes,
& Geist, 2000; Pauline et al., 2001; Ram et al., 2000). By
virtue of the numerous health-promoting aspects reported
and the easy and safe preparation of this beverage at
home, it has gained popularity as other traditional beverages.
Kombucha is a symbiotic growth of bacteria (Acetobacter
xylinum, Acetobacter xylinoides, Bacterium gluconicum)
and yeast strains (Schizosaccharomyces pombe, Saccharo-
mycodes ludwigii, Saccharomyces cerevisiae, etc.) cultured
in a sugared tea (Chen & Liu, 2000; Chu & Chen, 2006;
Greenwalt et al., 1998; Teoh, Heard, & Cox, 2004). The ex-
act microbiological composition also depends on the source
of inoculums of the tea fermentation. Growth patterns of
these microorganisms during the fermentation process of
Kombucha are not well documented. Cellulose produced
during the fermentation by A. xylinum appears as a thin
film on top of the sugared tea broth where the cell mass of
bacteria and yeasts is attached. This fungus-like mixture of
microorganisms and cellulose is likely why Kombucha is
also called ‘‘tea fungus’’. Glucose liberated from sucrose is
metabolized for the synthesis of cellulose and gluconic
acid by Acetobacter strains. Fructose is metabolized into
Grading and bagging
Softening with steam Cooling and
brick-making
Packaging & storage
 Black tea Sucrose and glucose
Water Boiling Take out tea Cooling
Inoculation with tea fungus Fermentation Product
Fig. 3. Schematic description of Kombucha manufacture.
ethanol and carbon dioxide by yeasts. Ethanol is oxidized to
acetic acid by Acetobacter strains.
It is also reported that the fermentation process induces
the synthesis of B complex of vitamins and folic acid
(Bauer-Petrovska & Petrushevska-Tozi, 2000). The pH
value of Kombucha decreases during the fermentation pro-
cess following the increase in organic acid content (Blanc,
1996; Sievers et al., 1996). The resultant low pH and pres-
ence of antimicrobial metabolites reduce the competition of
other bacteria, yeasts and filamentous fungi.
The antimicrobial activity of Kombucha was tested
against a number of pathogenic microorganisms (Sreeramulu
et al., 2000). Staphylococcus aureus, Shigella sonnei, Escher-
ichia coli, Aeromonas hydrophila, Yersinia enterolitica,
Pseudomonas aeruginosa, Enterobacter cloacae, Staphylo-
coccus epidermis, Campylobacter jejuni, Salmonella enteriti-
dis, Salmonella typhimurium, B. cereus, Helicobacter pylori,
and Listeria monocytogenes were found to be sensitive to
Kombucha. According to the literature on Kombucha, acetic
acid is considered to be responsible for the inhibitory effect
towards a number of microbes tested (Greenwalt et al.,
1998; Sreeramulu et al., 2000; Steinkraus et al., 1996). How-
ever, Sreeramulu et al. (2000, 2001) found that Kombucha
exerts antimicrobial activities against E. coli, S. sonnei,
S. typhimurium, S. enteritidis, and C. jejuni, even at neutral
pH and after thermal denaturation of Kombucha. This finding
suggests the presence of antimicrobial compounds other than
acetic acid or large proteins in Kombucha.
Sreeramulu et al. (2001) further characterized the anti-
microbial compounds in Kombucha. During the fermenta-
tion process of 14 days, several metabolites were analyzed
every two days in Kombucha. Levels of acetic acid and
gluconic acid were found to increase with fermentation
time. No lactic acid or ethanol was detected. Systematic
investigation of the antimicrobial activity in Kombucha
confirmed the presence of antimicrobial compounds other
than organic acids or proteins (enzymes) produced during
fermentation, or the tannins originally present in the tea
broth. Synergistic activity of antimicrobial components
and low pH due to the increase of organic acids probably
strengthen the inhibitory effect of Kombucha.
Challenges
Although several antimicrobial activities were found in
the extracts of Puer tea and Fuzhuan brick-tea samples
during the fermentation process, as well as Kombucha,
critical arguments still remain.
Antimicrobial components generated by microbial
fermentation in tea
The first question is whether tea catechins or polyphe-
nols have played the role of inhibition of microbial
growth. This doubt can be excluded by the fact that the
antibacterial activity increases with the fermentation
time; namely, the longer the fermentation is, the more an-
tibacterial activity the sample has. This implies that the
activity comes from fungal metabolism and not from the
originally present catechins or polyphenols of tea leaves.
Nevertheless, Chou, Lin, and Chung (1999) reported that
conventional fermentative processed teas have decreased
antimicrobial activity with the fermentation time and
this implies that the originally present tea catechins or
polyphenols lose their antimicrobial activity during the
enzymatic oxidation. In their study, B. subtilis, E. coli,
Proteus vulgaris, Pseudomonas fluorescens, Salmonella
spp. and S. aureus were used to test the antimicrobial ac-
tivity of various tea extracts. Among the six test micro-
organisms, P. fluorescens was the most sensitive to the
extracts, whereas B. subtilis was the least sensitive. In
general, antimicrobial activity decreased when the extent
of tea fermentation increased. The antimicrobial activities
of tea extracts with different extents of fermentation var-
ied with testing microorganisms. Extract of green tea, the
unfermented or non-oxidized tea, showed the strongest
antimicrobial activity followed by the partially fermented
tea products such as Longjing, Tieh-Kuan-Ying, Pao-
chung, and oolong teas. On the other hand, black tea,
the completely fermented (oxidized) tea, showed the least
antimicrobial activity. It was also noted that extracts of
oolong tea prepared in summer exhibited the strongest an-
timicrobial activity, followed by those prepared in spring,
winter and fall (Chou et al., 1999). It must be pointed out
that none of the teas used by these authors is microbial
fermented tea although the term of fermentation is often
used for black teas (Fowler et al., 1998). This indicates
the possibility that fungus involved in microbial tea fer-
mentation contributes to the formation of antimicrobial
metabolites other than tea catechins or polyphenols that
are originally present in unfermented or green tea leaves.
Moreover, the results of Yao et al. (1998) also confirm
that the antimicrobial effect of microbial fermented
Fuzhuan brick-tea is not catechins or polyphenols that
are actually fully oxidized during the process.
Thermo-stable characteristics of antimicrobial
components
The second question is whether the antimicrobial com-
pounds are large proteins or enzymes that may have the in-
hibitory activity. This question is answered by the fact that
sample extraction with hot water has wider antibacterial
spectrum and stronger activity, and this confirms that the
antimicrobial compounds are unlikely large proteins or en-
zymes, because they are otherwise inactivated through
a thermal treatment. Sakanaka, Juneja, and Taniguchi
(2000) studied the inhibitory action of tea polyphenols to-
wards the development and growth of bacterial spores.
Among the tested Bacillus bacteria, tea polyphenols show
antibacterial effects towards Bacillus stearothermophilus,
which is a thermophilic spore-forming bacterium. The
heat resistance of B. stearothermophilus spores is reduced
by the addition of tea polyphenols. Clostridium thermoace-
ticum, an anaerobic spore-forming bacterium, also exhibits
reduced heat resistance of its spores in the presence of tea
polyphenols. Epigallocatechin gallate, a main component
of tea polyphenols, shows strong activity against both
B. stearothermophilus and C. thermoaceticum. The heat re-
sistance of these bacterial spores is more rapidly decreased
by the addition of tea polyphenols at high temperatures.
This phenomenon also indicates that the potential antimi-
crobial agents produced by microbial fermentation in tea
are thermo-stable compounds, at least stable at 80  C dur-
ing the extraction time needed. With this thermo-stable fea-
ture, this kind of natural preservatives can be used in food
products that will go through pasteurisation and short-time
heat treatment without losing the inhibitory activity. This
can be a very interesting measure to control thermophilic
spore-forming bacteria.
Influences of extract concentration
The third question to answer is whether the conclusion
is solid enough that the extract has no inhibitory effect on
yeasts and fungi. In fact, Mo et al. (2005) used only one
extract concentration of the antimicrobial compounds for
the test. Therefore, it is critical whether a higher concen-
tration of the extracts will have inhibitory effect on yeasts
and fungi. Other challenges might arise when a higher
concentration of the fungal antimicrobial metabolites
will be applied as natural preservatives in food products.
First, the flavour of such natural preservatives might
have impact on the food products concerned. These natu-
ral preservatives should be desirably colourless and taste-
less so that they will not bring about any off flavour
troubles. Second, whether there are interactions between
these natural preservatives and nutritional components in
the concerned food products. Ideally, these natural preser-
vatives should not bring about any anti-nutritional effects.
Davis (1990) described the anti-nutritional effect of caf-
feine and Eggum, Pedersen, and Jacobsen (1983) reported
the anti-nutritional effect of tannins. Lule and Xia (2005)
indicated in a review on phenolic compounds that several
anti-nutritional effects can happen despite the positive
effect of these compounds. However, little is known if
microbial fermented tea still has the anti-nutritional effect
on other food components after weeks of fungal fermen-
tation and oxidation. This, however, can be easily tested
in future studies.
Perspectives and future research needs of microbial
fermentation in tea
It is clear that microbial fermentation of tea results in
certain metabolites that have inhibitory effects against
a number of bacteria. However, before these antimicrobial
metabolites can be used as natural preservatives in food
products, several critical issues must be clarified.
The responsible microorganisms for microbial
fermentation in tea
First, it is necessary to confirm the responsible fungi,
yeast and bacteria for the production of inhibitive compo-
nents. Although certain microorganisms are claimed to be
responsible for the antimicrobial activity in Puer tea, Fuz-
huan brick-tea and Kombucha, little is known about the fer-
mentation of these teas by a pure culture of the claimed
microorganisms. When a pure culture of the claimed micro-
organism is applied in the fermentation on the same sub-
strate, and can generate the same inhibitory effect against
certain microorganisms, we can be sure that these claimed
microorganisms are really responsible for the activity. Fur-
thermore, the confirmation of the responsible role of the
claimed microorganisms will stimulate and promote the
standardization of the tea fermentation process and improve
the industrial efficiency.
Characterization of antimicrobial components
Second, we need to further characterize the antimicro-
bial agents produced by microbial fermentation of tea. Al-
though several characterization attempts have been made to
exclude the possibilities that the antimicrobial agents are
tea catechins, large proteins and organic acids, the accurate
formula of the compounds remains unknown. The identifi-
cation and characterization of the antimicrobial compounds
are further of importance for understanding the metabolic
pathways how compounds in tea leaves are transformed
into antimicrobial agents. When this metabolism is clari-
fied, to overproduce these agents will be feasible.
Standardization and optimization of the process
Third, as for Puer tea and Fuzhuan brick-tea, both are
produced through a solid-state fermentation process; stan-
dardization and optimization are necessary. Solid-state fer-
mentation is described as a process where no free water is
present (Smits et al., 1999). During a solid-state fermenta-
tion process, water will be evaporated to release heat pro-
duced by microbial metabolism and this brings about the
limitation of growth and production, as a result of reduced
water activity. In a typical solid-state fermentation process,
moisture, heat transfer (affecting substrate temperature) and
microbial growth are closely interactive with each other
(Smits, Rinzema, Tramper, Van Sonsbeek, & Knol, 1996;
Smits et al., 1999; Von Meien & Mitchell, 2002). In an un-
controlled traditional indigenous fermentation process such
as Puer tea and Fuzhuan brick-tea manufacture, not only
the natural floras for inoculation, but also the season of
production strongly affect the product quality and consis-
tency. Therefore, a standardized fermentation process will
not only ensure food safety but also the product quality.
Furthermore, during the standardization and optimization
of the process, more insight will be obtained for the meta-
bolic mechanism of the fungi involved, how they produce
antimicrobial metabolites and eventually an overproduction
of these useful natural preservatives can be expected.
References
Aidoo, K. E., Nout, M. J. R., & Sarkar, P. K. (2006). Occurrence and
function of yeasts in Asian indigenous fermented foods. FEMS Yeast
Research, 6(1), 30e39.
An, B. J., Kwak, J. H., Son, J. H., Park, J. M., Lee, J. Y., Jo, C., et al.
(2004). Biological and anti-microbial activity of irradiated green
tea polyphenols. Food Chemistry, 88(4), 549e555.
Bandyopadhyay, D., Chatterjee, T. K., Dasgupta, A., Lourduraja, J., &
Dastidar, S. G. (2005). In vitro and in vivo antimicrobial action of
tea: the commonest beverage of Asia. Biological and Pharmaceu-
tical Bulletin, 28(11), 2125e2127.
Bauer-Petrovska, B., & Petrushevska-Tozi, L. (2000). Mineral and water
soluble vitamin content in the Kombucha drink. International
Journal of Food Science and Technology, 35(2), 201e205.
Benk, E. (1988). Ueber Teepilz-Gaergetraenk. [The tea fungus
fermented drink]. Verbraucherdienst, 33(10), 213e214.
Blanc, P. J. (1996). Characterization of the tea fungus metabolites.
Biotechnology Letters, 18(2), 139e142.
Chen, C., & Liu, B. Y. (2000). Changes in major components of tea
fungus metabolites during prolonged fermentation. Journal of
Applied Microbiology, 89(5), 834e839.
Chiang, C. T., Weng, M. S., Lin-Shiau, S. Y., Kuo, K. L., Tsai, Y. J., &
Lin, J. K. (2006). Pu-erh tea supplementation suppresses fatty acid
synthase expression in the rat liver through down regulating Akt
and JNK signalings as demonstrated in human hepatoma HepG2
cells. Oncology Research, 16(3), 119e128.
Chou, C. C., Lin, L. L., & Chung, K. T. (1999). Antimicrobial activity of
tea as affected by the degree of fermentation and manufacturing
season. International Journal of Food Microbiology, 48(2),
125e130.
Chu, S. C., & Chen, C. S. (2006). Effects of origins and fermentation
time on the antioxidant activities of Kombucha. Food Chemistry,
98(3), 502e507.
Cushnie, T. P. T., & Lamb, A. J. (2005). Antimicrobial activity of
flavonoids. International Journal of Antimicrobial Agents, 26(5),
343e356.
Davis, P. N. (1990). Various effects of caffeine upon the body.
International Clinical Nutrition Review, 10(2), 333e339.
Dufresne, C., & Farnworth, E. (2000). Tea, Kombucha, and health:
a review. Food Research International, 33(6), 409e421.
Duh, P. D., Yen, G. C., Yen, W. J., Wang, B. S., & Chang, L. W. (2004).
Effects of pu-erh tea on oxidative damage and nitric oxide
scavenging. Journal of Agricultural and Food Chemistry, 52(26),
8169e8176.
Eggum, B. O., Pedersen, B., & Jacobsen, I. (1983). The influence of
dietary tea, coffee and cocoa on protein and energy-utilization of
soybean-meal and barley in rats. British Journal of Nutrition, 50(2),
197e205.
Ernst, E. (2003). Kombucha: a systematic review of the clinical
evidence. Forschende Komplementarmedizin und Klassische
Naturheilkunde. [Research in Complementary and Natural
Classical Medicine], 10(2), 85e87.
Fowler, M. S., Leheup, P., & Cordier, J.-L. (1998). Cocoa, coffee and
tea. In B. J. B. Wood (Ed.), Microbiology of Fermented Foods
(pp. 128e147). London: Blackie Academic & Professional.
Friedman, M., Henika, P. R., Levin, C. E., Mandrell, R. E., &
Kozukue, N. (2006). Antimicrobial activities of tea catechins and
theaflavins and tea extracts against Bacillus cereus. Journal of Food
Protection, 69(2), 354e361.
Greenwalt, C. J., Ledford, R. A., & Steinkraus, K. H. (1998). Determi-
nation and characterization of the antimicrobial activity of the
fermented tea Kombucha. LWT - Food Science and Technology,
31(3), 291e296.
Greenwalt, C. J., Steinkraus, K. H., & Ledford, R. A. (2000). Kombu-
cha, the fermented tea: microbiology, composition, and claimed
health effects. Journal of Food Protection, 63(7), 976e981.
Hamilton-Miller, J. M. T. (1995). Antimicrobial properties of tea
(Camellia sinensis L.). Antimicrobial Agents and Chemotherapy,
39(11), 2375e2377.
Hartmann, A. M., Burleson, L. E., Holmes, A. K., & Geist, C. R. (2000).
Effects of chronic Kombucha ingestion on open-field behaviors,
longevity, appetitive behaviours, and organs in C57-BL/6 mice:
a pilot study. Nutrition, 16(9), 755e761.
Hirasawa, M., & Takada, K. (2004). Multiple effects of green tea cat-
echin on the antifungal activity of antimycotics against Candida
albicans. Journal of Antimicrobial Chemotherapy, 53(2), 225e229.
Jie, G. L., Lin, Z., Zhang, L. Z., Lu, H. P., He, P. M., & Zhao, B. L.
(2006). Free radical scavenging effect of Pu-erh tea extracts and
their protective effect on oxidative damage in human fibroblast
cells. Journal of Agricultural and Food Chemistry, 54(21),
8058e8064.
Kim, S., Ruengwilysup, C., & Fung, D. Y. C. (2004). Antibacterial effect
of water-soluble tea extracts on food borne pathogens in laboratory
medium and in a food model. Journal of Food Protection, 67(11),
2608e2612.
Kuo, K. L., Weng, M. S., Chiang, C. T., Tsai, Y. J., Lin-Shiau, S. Y., &
Lin, J. K. (2005). Comparative studies on the hypolipidemic and
growth suppressive effects of oolong, black, pu-erh, and green tea
leaves in rats. Journal of Agricultural and Food Chemistry, 53(2),
480e489.
Liang, Y. R., Zhang, L. Y., & Lu, J. L. (2005). A study on chemical
estimation of pu-erh tea quality. Journal of the Science of Food
and Agriculture, 85(3), 381e390.
Liu, C. H., Hsu, W. H., Lee, F. L., & Liao, C. C. (1996). The isolation
and identification of microbes from a fermented tea beverage,
Haipao, and their interactions during Haipao fermentation.
Food Microbiology, 13(6), 407e415.
Lule, S. U., & Xia, W. S. (2005). Food phenolics, pros and cons: a
review. Food Reviews International, 21(4), 367e388.
Mayser, P., Fromme, S., Leitzmann, C., & Grunder, K. (1995).
The yeast spectrum of the tea fungus Kombucha. Mycoses,
38(7e8), 289e295.
Mo, H. Z., Xu, X. Q., Yan, M. C., & Zhu, Y. (2005). Microbiological
analysis and antibacterial effects of the indigenous fermented Puer
tea. Agro Food Industry Hi-Tech, 16(6), 16e18.
Pauline, T., Dipti, P., Anju, B., Kavimani, S., Sharma, S. K., Kain, A. K.,
et al. (2001). Studies on toxicity, anti-stress and hepato-protective
properties of Kombucha tea. Biomedical and Environmental
Sciences, 14(3), 207e213.
Ram, M. S., Anju, B., Pauline, T., Prasad, D., Kain, A. K., Mongia, S. S.,
et al. (2000). Effect of Kombucha tea on chromate(VI)-induced
oxidative stress in albino rats. Journal of Ethnopharmacology,
71(1e2), 235e240.
Sakanaka, S., Juneja, L. R., & Taniguchi, M. (2000). Antimicrobial
effects of green tea polyphenols on thermophilic spore-forming
bacteria. Journal of Bioscience and Bioengineering, 90(1), 81e85.
Schillinger, U., Geisen, R., & Holzapfel, W. H. (1996). Potential of
antagonistic microorganisms and bacteriocins for the biological
preservation of foods. Trends in Food Science and Technology,
7(5), 158e164.
Si, W. D., Gong, J., Tsao, R., Kalab, M., Yang, R., & Yin, Y. L. (2006).
Bioassay-guided purification and identification of antimicrobial
 components in Chinese green tea extract. Journal of Chromatog-
raphy A, 1125(2), 204e210.
Sievers, M., Lanini, C., Weber, A., SchulerSchmid, U., & Teuber, M.
(1996). Microbiology and fermentation balance in a Kombucha
beverage obtained from a tea fungus fermentation. Systematic and
Applied Microbiology, 18(4), 590e594.
Smits, J. P., Rinzema, A., Tramper, J., Van Sonsbeek, H. M., & Knol, W.
(1996). Solid-state fermentation of wheat bran by Trichoderma
reesei QM9414: substrate composition changes, C balance,
enzyme production, growth and kinetics. Applied Microbiology
and Biotechnology, 46(5e6), 489e496.
Smits, J. P., Van Sonsbeek, H. M., Tramper, J., Knol, W., Geelhoed, W.,
Peeters, M., et al. (1999). Modelling fungal solid-state fermenta-
tion: the role of inactivation kinetics. Bioprocess Engineering,
20(5), 391e404.
Sreeramulu, G., Zhu, Y., & Knol, W. (2000). Kombucha fermentation
and its antimicrobial activity. Journal of Agricultural and Food
Chemistry, 48(6), 2589e2594.
Sreeramulu, G., Zhu, Y., & Knol, W. (2001). Characterization of
antimicrobial activity in Kombucha fermentation. Acta
Biotechnologica, 21(1), 49e56.
Steinkraus, K. H., Shapiro, K. B., Hotchkiss, J. H., & Mortlock, R. P.
(1996). Investigations into the antibiotic activity of tea fungus/
Kombucha beverage. Acta Biotechnologica, 16(2e3), 199e205.
Taguri, T., Tanaka, T., & Kouno, I. (2004). Antimicrobial activity of 10
different plant polyphenols against bacteria causing food-borne
disease. Biological and Pharmaceutical Bulletin, 27(12),
1965e1969.
Teoh, A. L., Heard, G., & Cox, J. (2004). Yeast ecology of Kombucha
fermentation. International Journal of Food Microbiology, 95(2),
119e126.
Von Meien, O. F., & Mitchell, D. A. (2002). A two-phase model for
water and heat transfer within an intermittently-mixed solid-state
fermentation bioreactor with forced aeration. Biotechnology and
Bioengineering, 79(4), 416e428.
Wu, S. C., Yen, G. C., Wang, B. S., Chiu, C. K., Yen, W. J.,
Chang, L. W., et al. (2007). Antimutagenic and antimicrobial
activities of pu-erh tea. LWT - Food Science and Technology,
40(3), 506e512.
Xu, X. Q., Mo, H. Z., Yan, M. C., & Zhu, Y. (2007). Analysis of
characteristic aroma of fungal fermented Fuzhuan brick-tea by gas
chromatography/mass spectrophotometry. Journal of the Science of
Food and Agriculture, 87(8), 1502e1504.
Xu, X. Q., Yan, M. C., & Zhu, Y. (2005). Influence of fungal fermen-
tation on the development of volatile compounds in the Puer
tea manufacturing process. Engineering in Life Sciences, 5(4),
382e386.
Yamamoto, Y., Matsunaga, K., & Friedman, H. (2004). Protective
effects of green tea catechins on alveolar macrophages against
bacterial infections. Biofactors, 21(1e4), 119e121.
Yao, S. Z., Tan, H. W., Zhang, H., Su, X. L., & Wei, W. Z. (1998). Bulk
acoustic wave bacterial growth sensor applied to analysis of
antimicrobial properties of tea. Biotechnology Progress, 14(4),
639e644.
Yilmaz, Y. (2006). Novel uses of catechins in foods. Trends in Food
Science and Technology, 17(2), 64e71.