ArticlePDF Available

Viili as Fermented Food in Health and Disease Prevention: A Review Study


Abstract and Figures

Viili is a unique fermented milk product from Nordic countries, particularly popular in Finland. Viili contains a symbiotic cluster of viable abundant probiotics, including lacto bacteria, fungus and yeasts, has a pleasant sharp taste and good diacetyl aroma linked to a stringy texture and can be consumed easily with spoon because of its semi-solid structure. The slime-forming Lactococcus lactis subsp. cremoris in viili produce phosphate-containing hetero polysaccharides, one of the main characteristics that distinguish viili from other fermented milk products. The symbiotic characteristics of viili and its Exopolysaccharides (EPS) provide various benefits from promoting intestinal probiotics interaction to anti oxidative, anti-inflammation, immunomodulation and antitumor activities. In addition, the tripeptides Isoleucine-Proline-Proline (IPP) and Valine-Proline-Proline (VPP) from the fermentation inhibit Angiotensin-Converting Enzyme, which reduces the blood pressure and the incidents of cardiovascular diseases. All studies show that multi microbes fermented viili not only provides probiotics, also a large number of other beneficial factors to Nordic population, and possibly to other regions of population as well.
Content may be subject to copyright.
©2016 Pearl Research Journals
Viili as Fermented Food in Health and Disease Prevention: A
Review Study
Cheng Luo* and Shanggui Deng
Accepted 6 July 2016
1School of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, China.
Viili is a unique fermented milk product from Nordic countries, particularly popular in Finland. Viili contains a
symbiotic cluster of viable abundant probiotics, including lacto bacteria, fungus and yeasts, has a pleasant
sharp taste and good diacetyl aroma linked to a stringy texture and can be consumed easily with spoon
because of its semi-solid structure. The slime-forming Lactococcus lactis subsp. cremoris in viili produce
phosphate-containing hetero polysaccharides, one of the main characteristics that distinguish viili from other
fermented milk products. The symbiotic characteristics of viili and its Exopolysaccharides (EPS) provide
various benefits from promoting intestinal probiotics interaction to anti oxidative, anti-inflammation,
immunomodulation and antitumor activities. In addition, the tripeptides Isoleucine-Proline-Proline (IPP) and
Valine-Proline-Proline (VPP) from the fermentation inhibit Angiotensin-Converting Enzyme, which reduces the
blood pressure and the incidents of cardiovascular diseases. All studies show that multi microbes fermented
viili not only provides probiotics, also a large number of other beneficial factors to Nordic population, and
possibly to other regions of population as well.
Key words: Viili, Fermentation, Exopolysaccharides, Probiotics, Lactococcus lactis, Fungus, Yeasts, Antioxidants,
Immunomodulation and Anti-carcinogenic activity.
*Corresponding author. E-mail:
Viili is a unique fermented thick milk product
without curds from Nordic countries, but it is believed to
be originally from Sweden, however, now found in
Finland, where it is largely considered as a national
treasure. This domestic fermented milk is typically
consumed at breakfast and is also a popular snack food
among children and elders in Finland. The product is
consumed alone or with cereals, muesli or fruit. A
traditional way of eating viili is by mixing it with cinnamon
and sugar (Leporanta, 2003). Its unique stickiness is due
to phosphate- containing EPS produced by the slime-
forming Lactococcus lactis subsp. cremoris. Viili EPS has
been claimed to exhibit several health benefits including
antioxidant, anti-inflammatory, anti-carcinogenic, anti-
aging and immunomodulatory activities (Kitazawa et al.,
1991, 1992, 1993). Viili was historically made on farms in
large wooden buckets, and with cooking utensils at home.
The industrial manufacture of viili began in Finland in the
1950s. Today, this product constitutes an important
fermented milk product in Finland. Annual consumption
stands at more than 4.5 kg/capita (Leporanta, 2003) even
though the average consumption is actually declined
because of wider range of milk products in market
(Alatossava et al., 2013).
It is believed that the term viili originated from the
Swedish word “fil” (Leporanta, 2003). However, the
tradition and skill of making viili has been passed from
generation to generation for many centuries. Viili is
mainly consumed in Finland, Sweden, Denmark, and, in
less extent, in Norway. Nowadays, there are different viili
starter providers, most of them from U.S. Since viili is
always a starter, and extremely easy to be cultured at
home, there are quite a large number of viili lovers
worldwide. In the last several years we have studied
macrophage RAW 264.7 cells to observe phagocytosis,
the release of NO, and the gene expression of iNOS, IL-6
Journal of Agricultural Science and Food Technology
Vol. 2 (7), pp. 105-113, August, 2016
ISSN: 2465-7522
Luo and Deng 106
Figure 1.Commercial viili, A: viili made from whole milk, B: Structure and consistency of semi-solid viili, with the
permission of Valio Limited.
Figure 2. Viili production at home. A: starter of viili and the milk. B: Approximately 10 to 20% inoculation of viili to pasteurized milk (V/V). C:
Fermentation at Room temperature for 16 to 24 h, the container need to be covered with 2 to 3 layers of textile and stand at 20 to 30°C. D: An
ready viili should contain ropy polysaccharides, should also form a velvet surface layer if whole milk was applied fermented viili. E: One of the way
to comsume viili with sugar and cinnamon (Leporanta, 2003).
and IL-as well as the morphology change at presence
of viili exopolysaccharides (VEPS). We also studied
VEPS influence on tumor antigen MAGEA10 and
inflammatory COX-2 genes expression in cancer cells to
investigate the potential possible roles of VEPS in human
immunity. To understand the complex multiple
mechanisms of viili in the daily health and diseases
prevention, we further review our work and analyze
different approaches for possible immune therapy.
For the commercial industrial production of viili (Figure 1)
the milk is separated and standardized to a fat content in
the range of 1.0 to 3.5%. Traditionally, viili has been
made from non-homogenized milk, which is the reason
why a thicker layer of cream forms on its surface. After
standardization, milk is pasteurized and cooled to 20°C.
A mixed culture of L. lactis sub sp. cremoris, L. lactis sub
sp. lactis, and G. candidum is then inoculated and mixed.
The mixture is then packed in a cup and to an incubator
for ripening, where viili fermentation is conducted for 20 h
at 20°C. After fermentation, viili is cooled (6°C)
throughout the transportation and sale shelf. Viili has a
stringy/ropy texture at optimal temperature of 28°C. The
shelf life of viili is around 3 weeks. There are a wide
range of different types of viili on the market, including
products with different fat contents, with reduced lactose
and flavoured varieties. Viili is also made commercially
from homogenized milk without mould growing on the
surface and from milk other than cow’s milk, but
traditional viili is still the most popular in the Finnish
region (Leporanta, 2003). However, the consumption of
viili has been variable and versatile by multiple food
cultures in Figure 2. Commercial viili carries living
microorganisms, that make it easy for anyone to
reproduce anywhere, at home, farm, or industry scale
without infiltration. This procedure is not possible with
almost all other yogurt, because they do not carry living
microorganisms or they are protected by a patent.
Probiotics are microorganisms that are believed to
provide health benefits when consumed. Introduction of
the concept is generally attributed to the Nobel Prize Élie
Metchnikoff, who in 1907 suggested that the dependence
of the intestinal microorganisms on the food makes it
possible to adopt measures to modify the intestinal
microbiota and to replace the harmful microorganisms by
the useful ones. In addition, probiotics secrete a large
number of vitamins, or effective peptides. However, the
profile of probiotics for each individual was actually
established in a quite early age (Bäckhed et al., 2015).
Normally probiotics in viili are bacteria, fungus and yeasts
that provide positive health benefits, especially for
digestive system. Viili presents high population of
symbiotic bacteria, yeasts and moulds that act
synergically. Viili contains several lactic acid bacteria
(LAB) including L. lactis sub sp. cremoris, L. lactis sub
sp. Lactis biovar. diacetylactis, and Leuconostoc
mesenteries subsp. cremoris. Among them, the slime-
forming L. lactis subsp. cremoris produces a
characteristic phosphate-containing heteropolysacharide
or EPS. These LAB strains are also responsible for lactic
acid fermentation, as well as citrate-based aroma
formation. In addition, traditional viili cultures also contain
yeast strains such as Kluveromycesmarxianus and Pichia
fermentans. Because milk mainly consists of casein
protein, or peptides, so the role of yeasts in viili
fermentation is minimum, but in the end of fermentation,
or as the lactose or glucose increase, small amount of
ethanol can be produced by yeast, this is particularly the
case for overdue viili, which can be smelled by olfactory
system. However, the effect of the yeasts on the viili as
symbiosis is not clear, it is probably that CO2 by the
fermentation of yeast may act as growth factors for
synergistic effects beside the roles of stripping H2S
(Butzke and Park, 2011), this is particularly the case for
yogurt production.
In addition, it is also believed that yeasts in viili may
provide the product’s unique flavour and induce the LAB
to produce more EPS (Wang et al., 2008). However,
yeast is frequently excluded in today’s viili industry to
keep alcohol free (personal communication with Kalle
Leporanta, 2016).
Viili contains at least one kind of fungus: Geotrichum
candidum. White filamentous yeast- like mould, which
forms a velvet-like surface at the end of viili fermentation.
G. candidumis a pathogen of many plants, but it is well
known for its role as a probiotic microorganism present in
viili and cheese fermentation processes. It can grow at
temperatures ranging from 5 to 38°C, with an optimal
growth at around 25°C, and at a large pH interval from 3
to 11, with an optimal pH value at 5.0 to 5.5. Besides the
long lag phase, the generation time of G. candidum is 66
min in liquid culture at 30°C, being one of the shortest
among eukaryotes, with final counts lower than 106
thallus forming units (tfu)/g (Boutrou and Gueguen, 2005;
Hudevoca et al., 2009). G. candidum creates a velvet-like
creamy surface on viili and it is also involved in the
ripening process of various cheeses. Typically during the
J.Agric. Sci. Food Technol. 107
manufacture of viili, the fermentation takes place in the
package and lasts about 18 to 20 h at 18 to 20°C
(Roginski, 2002).
The bacteria strains used for viili manufacturing produce
EPS which give viili a ropey, gelatinous consistency and
a pleasantly mild taste resulting from lactic acid
fermentation (Fondén et al., 2006). L. lactis subsp.
Cremoris in viili produces phosphate-containing hetero
polysaccharides, named viilian. The composition of the
EPS secreted by L. lactiss subsp. Cremoris consists of 3
to 47% protein and 29 to 85% carbohydrates (Macura
and Twonsley, 1984; Nakajima et al., 1990). Viili EPS has
a molecular weight of about 2000 kDa and it is mainly
composed of D-glucose, D-galactose, L-rhamnose, and
phosphate, with a repeating unit of “→4-β- Glcp-(1→4)-β-
D-Galp (1→4)-β-D-Glcp-(1→”, and groups of α-L-Rhap
and α -D-Galp-1-p attached to each side of Galp
(Nakajima et al., 1990; 1992, Higashimura et al., 2000;
Sletmoen et al., 2003). The composition and sugar
components of EPS are sub-strains and medium
The EPSs secreted by L. lactis subsp. Cremoris SBT
0495, ARH53, ARH74, ARH 84, ARH 87, and B30 are
composed of repeating units of galactose, glucose, and
rhamnose with a phosphodiester structural element
(Nakajima et al., 1990; Yang et al., 1999). In contrast, the
EPS produced by L. lactis subsp. Cremoris H414 is a
homopolymer consisting of galactose with a branched-
pentasaccharide repeating unit (Gruter et al., 1992).
Marshall et al. (1995) reported that L. lactis subsp.
Cremoris strain LC33 was able to generate two different
EPSs. One of them contains glucose, galactose,
rhamnose, glucosamine, and phosphate. The other was
composed of galactose, glucose, and glucosamine with
branched terminal galactose moieties. The slime material
obtained from L. lactis subsp. Cremoris SBT0495
supernatant in whey permeate medium consisted of 42%
carbohydrate and 21% protein with large amount of
mannans (Yang et al., 1999). Additionally, L. lactis
produced more EPS on glucose than on fructose as the
sugar substrate, although the transcription level of the
epsgene cluster was independent of the sugar source
(Looijesteijn et al., 1999).
There are different enzymes involved in EPS formation
by L. lactis subsp. Cremoris suchasthose responsible for
carbohydrate metabolism, enzymes leading to
sugar nucleotide synthesis and interconversion,
glycosyltransferases that form the repeating unit attached
to the glycosyl carrier lipid, and translocases and
polymerases that form the polymer. The genes encoding
the enzymes involved in the biosynthesis of EPS are
placed in an EPS plasmid in Lc. lactis subsp. cremoris.
The gene products EpsD, EpsE, EpsF, and
EpsGareglycosyl transferases and are required for the
synthesis of the EPS backbone (Van Kranenburg et al.,
1997, 1999). The presence of the EPS genes on a
plasmid has been suggested to be the cause of EPS
expression instability at higher temperatures and when
there are frequent batch inoculations of starter culture
(Vedamuthu and Neville, 1986; Cerning et al., 1992). It
has been shown that the slime-forming capacity is stable
when the bacteria are grown at17°C but is lost when they
are grown at >30°C(Forsén et al.,1973).
Besides its high nutritional value, viili exhibits positive
benefits for human health. Viili contains high populations
of different probiotic bacteria (108/mL), antihypertensive
effects as well as anti-inflammatory, antioxidant and anti-
carcinogenic activities associated to the presence of viili.
Hence, viili can be considered a traditional natural
functional food. It has been shown that viili EPS help the
interaction with intestine mucusa (Ruas-Madiedo et al.,
2006), but it acts in symbiosis with other microorganism
in viili, multiple metabolites in the same life chain, which
make it safe to consume. The precise mechanism is
unknown, but the risk of developing an infection due to
“pathogen” of G. candidum in connection with its
technological use and consumption of dairy products has
been virtually nil because fewer than 100 cases reported
only between year 1842 to 2006 (Pottier et al., 2007).
Many dietary compounds are known to have health
benefits owing to their antioxidative and anti-inflammatory
properties. In addition to the antioxidant diets, an
intracellular Nrf2/ARE antioxidant pathway has also been
established. That is, there is possible an association of
intracellular anti-ROS's activation via Nrf2/ARE with the
hypoxia, inflammation as well as the promotion of cell
migration and invasion in cellular micro environment, and
extracellular antioxidant's interaction may remain cell
health by balancing the oxidative state (Luo et al., 2011).
Viili and its isolated bacteria have been reported to
possess several health benefits including anti-oxidative
effects (Wang et al., 2014). Endogenous metabolic
processes and exogenous chemicals in the human body
or in a food system are able to produce, in some
circumstances, highly reactive oxygen species that
Luo and Deng 108
are able to oxidize biomolecules, resulting in tissue
damage and cell death. This may lead to inflammation,
diabetes, genotoxicity, cancer, and accelerated aging
(Biswas et al., 2010).
In a recent study that showed the antioxidant capacity of
ursolic acid (UA) increased with concentration from 0 to
100 μg/ml in a dose dependent manner, while antioxidant
power of viili EPS (VEPS) was milder, but clear synergic
interaction was observed from 0 to 50 μg/ml of both
compounds. Significant anti-proliferation of HepG2 cells
with UA and viili EPS at concentration of 200 μg/ml by
MTT assay was observed, however, the cells started
sene scences at 12.5 μg/ml, which indicated multiple
molecular modulation mechanisms rather than an
induction of apoptosis or necrosis. All these tests have
shown a compatibility of herbal UA and foodborne viili
EPS, which means they can be either processed together
for health products or functional food because viili EPS is
an antioxidant synergic with ursolic acid (Liu et al., 2012).
In addition to viili EPS, viili bacteria have been
demonstrated to contain α,α-diphenyl-β-picylhydrazl,
which has a free radical-scavenging effect and Fe2+-
chelating ability (Chiang et al., 2011). The anti-oxidative
nature of viili and its products may help the human body
to reduce oxidative damage. Since milk proteins are
precursors of many different biologically active
compounds, some peptides with free radical-scavenging
activities have been identified in fermented dairy. Viili’s
proteolytic abilities that aid the digestion of the milk
protein into peptides and free amino acids were
characterized in early 1990s (Tan and Konings, 1990;
Alting et al., 1995). The change in antioxidant activity
noted in low-fat cheeses made with viili is probably also
associated with the viable populations of LAB plus the
anti-oxidative peptides.
ACE (EC is known to be associated with
hypertension and congestive heart failure. The enzyme
converts angiotensin I into angiotensin II, the former
being an inert peptide and the latter being a pressor
agent. The enzyme is also responsible for the breakdown
of bradykinin, which is a dilatory peptide. The enzyme is
thus an obvious drug target for the treatment of certain
cardiovascular diseases, including hypertension. LAB is
known to produce inhibitors of the enzyme during
fermentation. Viili containing L. lactis subsp. Cremoris
strain has been demonstrated to have a strong inhibitory
effect on ACE activity (Chiang et al., 2011). The
proteolytic activity of the starters and the rate of
proteolysis seem to play an important role in the inhibitory
activity of these dairy products. The proteolytic system of
L. lactis has been studied, and it consists of a cell
J.Agric. Sci. Food Technol. 109
Figure 3. Tripeptides from viili and many other fermented milk products (Turpeinen et al., 2009).
wall bound proteinase and several intracellular
peptidases (Tan and Konings, 1990; Alting et al., 1995).
Since the high proteolytic activity, viili, as fermented milk
product demonstrates a better ACE inhibitory effects
comparing to other fermented milk products. Research
has mainly focused on IPP + VPP from viili, two lacto
tripeptides that can inhibit the ACE in vitro. In both
Finnish and Japanese subjects with (mild) hypertension,
IPP and VPP in Figure 3 showed their inhibition on ACE
and reduced diastolic blood pressure (DBP) and systolic
blood pressure (SBP) by 7 to 14 mmHg, which is a
statistically significant reduction (Hata et al., 1996; Pins
and Keena, 2006). Moreover, many studies have
reported that the types and concentrations of bioactive
peptides are able to significantly affect the functional
properties of the dairy product (Ong and Shah, 2008a, b).
Viili and its LAB have been demonstrated to have
immune regulatory effects in vitro and in vivo, including
anti-allergic effects and anti-colitis effects (Huang et al.,
2010). Viili and the viili bacterium L. lactis subsp.
Cremoris TL1 have been shown to induce the production
of the helper cell type I (Th1) cytokine tumor necrosis
factor-α, the proinflammatory cytokine interleukin (IL)-6,
and T regulatory cell (Treg) cytokine IL-10 in vitro, which
suggests that viili may be beneficial and improve the
Th1/Th2 balance. Oral feeding of Lc. lactis subsp.
Cremoris TL1 contained in viili has been shown to
suppress total immunoglobulin IgE and ovalbumin (OVA)-
specific IgE levels in the serum of OVA-sensitized mice.
Suppression of IgE production is an important target
when treating allergies. Additionally, in vivo effects of the
viili bacterium L. lactis subsp. Cremoris TL1 on the
regulation of intestinal physiology have been
demonstrated. This strain is able to ameliorate dextran
sulfate sodium (DSS)-induced colitis as exemplified by a
significant attenuation of the bleeding score and a
reduction in colon shortening. Histological analysis also
showed regeneration and epithelial restitution in the colon
among the animals included in the L. lactis subsp.
Cremoris TL1 treated group.
These findings suggest that the viili isolated strain L.
lactis subsp. Cremoris TL1, has a potential direct anti-
inflammatory activity with respect to epithelial cells and
that this effect may lead to inhibition of neutrophil
accumulation in the mucosal region of the DSS-colitis
mice. It is widely known that the toll-like receptor (TLR)
family plays an important role in host defense through
recognizing bacterial pathogen-associated molecular
patterns that engages anti-inflammatory processes, that
is why colitis caused uncomfortability can be minimized
because the inflammation is kept to a minimum. The
immune regulatory effects of viili probiotic LAB has been
investigated in a swine study, showing an intestinal
immune regulation by probiotic LAB mediated by the TLR
in the gut, which is similar to TLR signalling transduction
pathways through the molecular mechanisms of the
ligands of lipopolysaccharides (LPS) of  in
human (Tohno et al., 2007, 2008). Recently such TLR
family protein was defined as RP105/MD1 complex that
is involved in the immunoregulatory effect of EPS from
Lactobacillus plantarum N14 (Murofushi et al., 2015). By
studying the effects of VEPS in RAW 264.7 macrophages
the phagocytosis was observed by scanning electronic
microscopy (SEM), the titer of NO, IL-6, and IL- was
increased by ELISA, the gene expression of IL-6 and IL-
was enhanced by RT-PCR, the gene and protein
expression of inducible nitric oxide synthase (iNOS) were
also increased by RT-PCR and Western blotting, which
suggest the potential possible roles of VEPS in human
immunity (Figure 4). The cytokines’ induction indicates a
Luo and Deng 110
Figure 4. Viili EPS influence on macrophage RAW 264.7 cells (Wu et al., 2013)
possibility of boosting humoral immunity via T-helper cells
(Wu et al., 2013). A: Protein expression of iNOS in
macrophage RAW264.7 cell with different concentrations
of VEPS by Western blotting. B: Cell morphology of
RAW264.7 under stimulation of viili EPS and LPS by
SEM. a) Control, b) 1 μg/mL of LPS, c–e) with 50, 100,
and 200 μg/mL of viili EPS, respectively. Original
magnification was × 4,000, the original scale of SEM
represents 10 μm in each image (Wu et al., 2013).
Immunotherapy is a kind of treatment of disease by
inducing, enhancing, or suppressing an immune
response through a unique matching HLA typing epitope,
or nonspecific antigenic epitope. Immunotherapies
designed to elicit or amplify an immune response are
classified as activation immunotherapies, while those that
reduce or suppress the immune response are classified
as suppression immunotherapies. MAGE-A antigens
belong to a cluster of cancer/testis antigens (CTA) that
are expressed in tumors but not in normal tissues with
the exception of testis and placenta. Among MAGE-A
antigens, MAGEA10 represents an attractive target for
cancer immunotherapy because its epitopes extensively
elicit cytotoxic T - lymphocyte responses. However, the
suppressive cyto environment for gene expression and
the requirement of specific HLA-alleles presentation have
frequently led to immunotherapy failure. By studying the
genes expression and methylation of MAGE genes,
MAGEA10 was scarcely expressed in cancer patients,
but it has been recently shown that its expression can be
enhanced by viili polysaccharides, which indicate a
possibility of increasing epitopes presentation Figure 5.
A few studies have reported the antitumor activities of viili
Kitazawa et al. (1992) reported that viili and one of the
starter bacteria, L. lactis subsp. Cremoris KVS20,
inhibited the metastasis of Lewis lung carcinoma and
reduced the growth of solid and ascetic forms of
sarcoma-180 in vivo. The antitumor effect of viili might be
due to the increase of cytotoxic activity of the
macrophages stimulated by L. lactis subsp. Cremoris
KVS20. Liu et al. (2012) observed the senescence of
HepG2 cancer cells after treatment with viili EPS, which
supports the idea that viili EPS might have anti-tumor
activity (Figure 6). Moreover, villi EPS induced cancer cell
apoptosis via inhibition of COX-2 gene and protein
expression (Liu et al., 2014). However, comparing to
other polysaccharides, such as coix polysaccharides, the
anti-oxidative power of villi EPS was less competent, and
the induction of antioxidation dependent apoptosis was
less sensitive (Liu et al., 2014; Wang et al., 2014). On
March 25, 2011, the U. S. Food and Drug Administration
(USFDA) approved ipilimumab injection (Yervoy, made
by Bristol-Myers Squibb Company) for the treatment of
unrespectable or metastatic melanoma. Ipilimumab is a
monoclonal antibody that activates the immune system
by targeting CTLA-4, a protein receptor that down
regulates the immune system, thus activating cytotoxic T
lymphocytes (CTL), which increase the opportunities for
fighting against cancer cells.
In a recent study it was shown that viili polysaccharides
were able to induce cancer antigen MAGEA10’s gene
and protein expression, which gives a higher opportunity
for immunotherapy because over expression of cancer
antigen will increase the CTL opportunity to destroy the
cancer cells (Wang et al., 2015). Increasing cancer
antigen expression and presentation is possible to
increase the opportunities of CTL mediated cytotoxicity
against cancer cells (Figure 7). Cancer antigen (epitope)
presented by type 1 of MHC (HLA-A) could be
recognized by TCR of CTL, and eventually led to cancer
cell destruction. The right inserted circle image featured
J.Agric. Sci. Food Technol. 111
Figure 5. Enhancement of MAGEA 10 protein
expression of A549 cells under influence of villi EPS by
Western blotting (Wang et al., 2015).
Figure 6. Inhibitory effects of different compounds on (COX)-2 expression in HepG2 cells. HepG2 cells were treated
with doses of UA (5 μg/ml), villi EPS (50 μg/ml), Astragalus polysaccharides (APS) (50 μg/ml) or combined treatments
for 48 h. (Left): RT-PCR analysis of COX-2 genes. Lane 0, control; lane 1, UA (5 μg/ml); lane 2, VEPS (50 μg/ml); lane
3, APS (50 μg/ml), lane 4, UA+VEPS; lane 5, UA+APS for 48 h. (Right): Quantitative analysis of protein levels with the
same samples. *P<0.05, **P<0.01, compared with the control group (Liu et al., 2014).
Figure 7. Autologous / non autologous immunotherapy with CTL activator (such as ipilimumab) and
cancer antigen presenting promoter (such as viili polysaccharides).
Luo and Deng 112
the MAGEA3 pulsed cancer cells (red) were attacked by
corresponding specific CTL, where non pulsed cancer
cells (green) kept intact (Wang et al., 2014)
The functional health roles from different cultural foods,
such as fermented viili, have been widely recognized.
Foodborne polysaccharides VEPS possess anti-
inflammation and anticancer properties, particularly by
down regulating Interleukins and COX-2 expression,
which may have increased internal oxidation and
triggered apoptosis together with a change in internal
antioxidant response elements, leading to a reduction in
cell proliferation. These specific mechanisms need to be
further studied.
Alatossava T, Li R, Munsch-Alatossava P (2013). From “Viili” Towards
“Termoviili”, a Novel Type of Fermented Milk: Characterization of
Growth Conditions and Factors for a co-culture of Lactobacillus
delbrueckii and Geotrichumcandidum . J. Food Process. Beverages.
1(2): 8
Alting RC, Engels WJM, Schalkwijk S, Exterkate FA (1995). Purification
and characterization of cystathionineβ-lyase from
Lactococcuslactissubsp. Cremoris B78 and its possible role in flavor
development in cheese. Appl. Environ. Microbiol., 61:4037-42.
Bäckhed F, Roswall J, Peng Y, Feng Q, Jia H, Kovatcheva-Datchary P,
Li Y, Xia Y, Xie H, Zhong H, Khan MT, Zhang J, Li J, Xiao L, Al-Aama
J, Zhang D, Lee YS, Kotowska D, Colding C, Tremaroli V, Yin Y,
Bergman S, Xu X, Madsen L, Kristiansen K, Dahlgren J, Wang J
(2015).Dynamics and Stabilization of the Human Gut Microbiome
during the First Year of Life. Cell. Host Microbe.,17 (5): 690-703.
Biswas M, Haldar PK. and Ghosh AK (2010). Antioxidant and free-
radicalscavenging effects of fruits of Dregeavolubilis.J. Natural Sci.
Biol. Med., 1(1): 29-34.
Boutrou R, Guéguen M (2005). Interest in Geotrichumcandidumfor
cheese technology.Int. J. Food Microbiol., 102:1-20.
Butzke CE, Park SK (2011). Impact of fermentation rate changes on
potential hydrogen sulfide concentrations in wine. J. Microbiol.
Biotechnol., 21(5): 519-24.
Cerning J, Bouillanne C, Landon M, Desmazeaud M (1992). Isolation
and characterization of exopolysaccharides from slime-forming
mesophilic lactic acid bacteria.J. Dairy Sci., 75: 692-699.
Chiang ML, Chen HC, Wang SY, Hsieh YL, Chen MJ (2011). Use of
Taiwanese ropy fermented milk (TRFM) and Lactococcuslactissubsp.
cremorisisolated from TRFM in manufacturing of functional low-fat
cheeses. J. Food Sci., 76: M504-M510.
Fondén R, Leporanta K, Svensson U (2006). Nordic/Scandinavian
Fermented Milk Products, in Fermented Milks (ed A. Tamime),
Blackwell Publishing Ltd, Oxford, UK.
doi: 10.1002/9780470995501.ch7
Forsén R, Raunio V, Myllymaa R, Nousiainen R, and Päkkilä M (1973).
Studies on the slime forming group N Streptococcus strains. I.
Differentiation between some lactic streptococcusstrains by
polyacrylamide gel electrophoresisof soluble cell proteins. Acta
Universitatis Ouluensis: Scientiae Rerum Naturalium, 3: l.
Gruter M, Leeflang BR, Kuiper J, Kamerling JP, Vliegenthart JFG
(1992). Structure of the exopolysaccharide produced by
Lactococcuslactissubsp. cremorisH414 grown in a defined mediumor
skimmed milk. Carbohydr. Res., 231: 273-291.
Hata Y, Yamamoto M, Ohni M, Nakajima K, Nakamura Y, Takano T
(1996). A placebo-controlled study of the effect of sour milk on blood
pressure in hypertensive subjects. Am. J. Clin.Nutr., 64(5): 767-71.
Higashimura M, Mulder-Bosman BM, Reich R, Iwasaki T. and Robijn
GW (2000). Solution properties of viilian, theexopolysaccharide from
Lactococcuslactis subsp. cremoris SBT0495. Biopolymers, 54: 143-
Huang IN, Dai TY, Wang SY, Chen MJ (2010). Inhibitory effect of
Taiwanese ropy fermented milk in an ovalbumin-induced allergy
mouse model. J.Dairy Sci.,93 S(1): 807.
Hudevoca A, Valik L, Liptakova D (2009). Quantification of
Geotrichumcandidum growth in co-culture with lactic acid bacteria.
Czech J. Food Sci., 27: 8-27.
Kitazawa H, Toba T, Itoh T, Kumano N, Adachi S, Yamaguchi T (1991).
Antitumoral activity of slime-forming, encapsulated
Lactococcuslactisssp. Cremorisisolated from Scandinavian ropy sour
milk viili. Anim. Sci. Technol.,62: 277-283.
Kitazawa H, Yamaguchi T, Miura M, Saito T. and Itoh T (1993). B-cell
mitogen produced by slime-forming encapsulated Lactococcuslactiss
sp. Cremoris isolated from ropy sour milk, viili. J.Dairy Sci.,76: 1514-
Kitazawa H, Yamaguchi T. and Itoh T (1992). B-Cell Mitogenic activity
of slime products produced from slime-forming, encapsulated
Lactococcuslactisssp. cremoris. J. Dairy Sci.,75: 2946-2951.
Leporanta K (2003).Viili and Långfil-Exotic fermented products from
Scandinavia. Valio Foods & Functionals. On line versión.
(Access June 8, 2010).
Ling Liu, Jingkai Z, Meiling Li, Xiaohong Z, Jinlu Z, Zhenjing L, Likui W,
Jihui W, Cheng L (2014). Inhibition of HepG2 cell proliferation by
ursolic acid and polysaccharides via the downregulation of
cyclooxygenase-2. Mol. Med. Rep.,9(6): 2505-2511.
Liu L, Wu J, Zhang J, Li Z, Wang C, Chen M, Wang Y, Sun Y, Wang L.
and Luo C (2012).A compatibility assay of ursolic acid and foodborne
microbial exopolysaccharides by antioxidant power and anti-
proliferative properties in hepatocarcinoma cells. J. Food, Agric.
Environ., 10: 111-114.
Luo C, Urgard E, Vooder T, Metspalu A (2011). The role of COX-2 and
Nrf2/ARE in anti-inflammation and antioxidative stress: Aging and
anti-aging. Med. Hypotheses, 77(2):174-178 .
Macura D, Townsley PM (1984). Scandinavian ropy milk-identification
and characterization of endogenous ropy lactic streptococci and their
extracellular excretion. J. Dairy Sci., 67: 735.
Marshall VM, Cowie EN, Moreton RS (1995). Analysisand production of
two exopolysaccharides from Lactococcuslactis sub sp. Cremoris
LC330. J. Dairy Res., 62: 621-628.
Murofushi Y, Villena J, Morie K, Kanmani P, Tohno M, Shimazu T, Aso
H, Suda Y, Hashiguchi K, Saito T, Kitazawa H (2015). The toll-like
receptor family protein RP105/MD1 complex is involved in the
immunoregulatory effect of exopolysaccharides from Lactobacillus
plantarum N14. Mol. Immunol., 64(1): 63-75.
Nakajima H, Suzuki Y, Kaizu H and Hirota T (1992). Cholesterol
lowering activity of ropy fermented milk. J. Food Sci., 57: 1327-1329.
Nakajima H, Toyoda S, Toba T, Itoh T, Mukai T, Kitazwa H, Adachi S
(1990). A novel phosphophlysaccharide from slime-forming
Lactococcuslactis sub sp. Cremoris SBT 0495. J. Dairy Sci., 73:
Ong L, Shah NP (2008a). Influence of probiotic Lactobacillus
acidophilus and L. helveticuson proteolysis, organic acid profiles, and
ACE-inhibitory activity of Cheddar cheeses ripened at 4, 8, and 12°C.
J. Food Sci., 73: M111-M120.
Ong L, Shah NP (2008b). Release and identification of angiotensin-
converting enzyme inhibitory peptides as influenced by ripening
temperatures and probiotic adjuncts in Cheddar cheeses. LWT Food
Sci. Technol., 41: 1555-1566.
Pins JJ, Keenan JM (2006). Effects of whey peptides on cardiovascular
disease risk factors.J. Clin. Hypertens., (Greenwich). 8(11): 775-82.
Roginski H (2002) Fermented milk: Northern Europe. Encyclopedia of
Dairy Sci., 2: 1034-1041, Academic Press.
Ruas-Madiedo P, Gueimonde M, de los Reyes-Gavilán GC, Salminen S
(2006). Short communication: Effect of exopolysaccharide isolated
from “viili” on the adhesion of probiotics and pathogens to intestinal
mucus. J. Dairy Sci., 89: 2355-2358.
Sletmoen M, Maurstad G, Sikorski P, Paulsen BS, Stokke BT (2003).
Characterization of bacterial polysaccharides: Steps towards single
molecular studies. Carbohydr. Res., 338: 2459-2475.
Tan PST, Konings WN (1990). Purification and characterization of an
aminopeptidase from Lactococcuslactis sub sp. Cremoris Wg2. Appl.
Environ. Microbiol., 56: 526-532.
Tohno M, Shimazu T, Aso H, Kawai Y, Saito T, Kitazawa H (2007).
Molecular cloning and functional characterization of porcine MyD88
essential for TLR signaling. Cell. Mol. Immunol., 4(5): 369-76
Tohno M, Shimazu T, Ueda W, Anzawa D, Aso H, Nishimura J, Kawai Y,
Saito Y, Saito T, Kitazawa H (2007). Molecular cloning of porcine
RP105/MD-1 involved in recognition of extracellular
phosphopolysaccharides from Lactococcuslactisssp. cremoris. Mol.
Immunol., 44: 2566-2577.
Tohno M, Ueda W, Azuma Y, Shimazu T, Katoh S, Wang JM, Aso H,
Takada H, Kawai Y, Saito T, Kitazawa H (2008). Molecular cloning
and functional characterization of porcine nucleotide-binding
oligomerization domain-2 (NOD2). Mol. Immunol., 45(1): 194-203.
Van Kranenburg R, Marugg JD, Van Swam II, Willem J, DeVos WM
(1997). Molecular characterization of the plasmid-encoded eps gene
cluster essential for exopolysaccharide biosynthesis in
Lactococcuslactis. Mol. Microbiol., 24: 387-397.
Van Kranenburg, R, Van Swam II, Marugg JD, Kleerebezem M, De Vos,
WM (1999). Exopolysaccharide biosynthesis in Lactococcuslactis
NIZO B40: Functional analysis of the glycosyltransferase genes
involved in synthesis of the polysaccharide backbone. J.
Bacteriol.,181: 338-340.
J.Agric. Sci. Food Technol. 113
Vedamuthu ER, Neville JM (1986). Involvement of a plasmid in
production of ropiness (mucoidness) in milk cultures by
Streprococcuscremoris MS. Appl. Environ. Microbiol., 51: 677.
Wang L, Gao S, Jiang W, Luo C, Xu M, Bohlin L, Rosendahl M, Huang
W (2014). Antioxidative Dietary Compounds Modulate Gene
Expression Associated with Apoptosis, DNA Repair, Inhibition of Cell
Proliferation and Migration. Int. J. Mol. Sci., 15: 16226-16245.
Wang L, XuY, Luo C, Sun J, Zhang J, Lee M-W, Bai A, Chen G, M.
Frenz C.M., Huang W (2015). MAGEA10 gene expression in non
small cell lung cancer and A549 cells, and the affinity of epitopes with
the complex of HLA-A*0201 allelCell. Immunol.,297(1): 10-18.
Wang SY, Chen HC, Liu JR, Lin YC.and Chen MJ (2008). Identification
of yeasts and evaluation of their distribution in Taiwanese kefir and
viili starters.J. Dairy Sci., 91: 3798-3805.
Wu J, Li M, Liu L, An Q, Zhang J, Zhang J, Li M, Duan W, Liu D, Li Z,
Luo C (2013). Nitric Oxide and Interleukins are Involved in Cell
Proliferation of RAW264.7 Macrophages Activated by Viili
Exopolysacchrides. Inflammation, 36 (4): 954-961.
Yang Z, Huttunen E, Staaf M, Widmalm G, Tenhu H (1999). Separation,
purification and characterisation of extracellular polysaccharides
produced by slime-forming Lactococcuslactiss sp. Cremoris strains.
Int. Dairy J., 9: 631-638.
... Viili is a traditional fermented milk product that originated in Scandinavia (Luo & Deng, 2016;Bakry & Campelo, 2018). To make viili, milk is inoculated with a starter and incubated at about 20 • C for 16-24 h. ...
Full-text available
Starter culture of viili contains lactic acid bacteria belonging to Lactococcus lactis. These bacteria secrete large polysaccharides (EPSs) into milk, resulting in a ropy texture of viili. In mouse experiments, a large dose of EPS (5–140 mg/day) has been shown to alleviate severity of artificially induced illness through modulation of the gut microbiota. The present study investigated whether supplementary amounts of EPS affects the gut microbiota of normal mouse. EPS with high glucosamine content (VEPS) was isolated from home-made viili. C57BL/6J male mice fed ordinary diet took 49 ± 1 μg VEPS/day for 28 days by drinking ad libitum tap water containing 8 μg/mL VEPS. The relative abundance of Muribaculum increased significantly by VEPS supplementation. The relative abundance of fecal butyric acid decreased in control mice, and VEPS prevented this decrease. These findings indicated that the gut microbiota can be modulated by a small dose of VEPS.
Full-text available
Fermentation of various food stuffs by lactic acid bacteria is one of the oldest forms of food biopreservation. Bacterial antagonism has been recognized for over a century, but in recent years, this phenomenon has received more scientific attention, particularly in the use of various strains of lactic acid bacteria (LAB). Certain strains of LAB demonstrated antimicrobial activity against foodborne pathogens, including bacteria, yeast and filamentous fungi. Furthermore, in recent years, many authors proved that lactic acid bacteria have the ability to neutralize mycotoxin produced by the last group. Antimicrobial activity of lactic acid bacteria is mainly based on the production of metabolites such as lactic acid, organic acids, hydroperoxide and bacteriocins. In addition, some research suggests other mechanisms of antimicrobial activity of LAB against pathogens as well as their toxic metabolites. These properties are very important because of the future possibility to exchange chemical and physical methods of preservation with a biological method based on the lactic acid bacteria and their metabolites. Biopreservation is defined as the extension of shelf life and the increase in food safety by use of controlled microorganisms or their metabolites. This biological method may determine the alternative for the usage of chemical preservatives. In this study, the possibilities of the use of lactic acid bacteria against foodborne pathogens is provided. Our aim is to yield knowledge about lactic acid fermentation and the activity of lactic acid bacteria against pathogenic microorganisms. In addition, we would like to introduce actual information about health aspects associated with the consumption of fermented products, including probiotics.
Full-text available
The growth dynamics of filamentous fungus G. candidum was studied during the co-cultivation with the commercial lactic acid bacteria (LAB) culture Fresco. The experiments were carried out in milk and on the surface of a milk agar at the temperature ranging from 5 to 37°C. Ratkowsky model was used to describe the relationships of the fungal growth rate to the temperature during both, single and co-cultivation with LAB in milk. Simultaneous growth of LAB affected significantly the growth rate of the filamentous fungus. The growth of G. candidum was in average 39% slower in the co-culture than in the single cultivation. LAB pre-inoculated and growing in the solid medium did not show any significant inhibitory effect on the surface growth of G. candidum at all tested temperature. The precise data describing the growth of this cheese yeast-like fungus, G. candidum, may fill a gap in the field of quantitative food mycology and may be used for predicting its behavior in real conditions.
Full-text available
MAGEA10, a cancer/testis antigens expressed in tumors but not in normal tissues with the exception of testis and placenta, represents an attractive target for cancer immunotherapy. However, suppressive cytoenvironment and requirement of specific HLA-alleles presentation frequently led to immunotherapy failure. In this study MAGEA10 was scarcely expressed in cancer patients, but enhanced by viili polysaccharides, which indicates a possibility of increasing epitopes presentation. Furthermore the correlation of gene expression with methylation, indicated by R(2) value for MAGEA10 that was 3 times higher than the value for other MAGE genes tested, provides an explanation of why MAGEA10 was highly inhibited, this is also seen by Kaplan-Meier analysis because MAGEA10 did not change the patients' lifespan. By using Molecular-Docking method, 3 MAGEA10 peptides were found binding to the groove position of HLA-A(∗)0210 as same as MAGEA4 peptide co-crystallized with HLA-A(∗)0210, which indicates that they could be promising for HLA-A(∗)0201 presentation in immunotherapy. Copyright © 2015. Published by Elsevier Inc.
Full-text available
The gut microbiota is central to human health, but its establishment in early life has not been quantitatively and functionally examined. Applying metagenomic analysis on fecal samples from a large cohort of Swedish infants and their mothers, we characterized the gut microbiome during the first year of life and assessed the impact of mode of delivery and feeding on its establishment. In contrast to vaginally delivered infants, the gut microbiota of infants delivered by C-section showed significantly less resemblance to their mothers. Nutrition had a major impact on early microbiota composition and function, with cessation of breast-feeding, rather than introduction of solid food, being required for maturation into an adult-like microbiota. Microbiota composition and ecological network had distinctive features at each sampled stage, in accordance with functional maturation of the microbiome. Our findings establish a framework for understanding the interplay between the gut microbiome and the human body in early life. Copyright © 2015 Elsevier Inc. All rights reserved.
Oxidation is a chemical reaction that transfers electrons from a substance to an oxidizing agent, which produces harmful free radicals. However, natural antioxidants possess antioxidative properties and exogenous antioxidants are normally capable of inhibiting the oxidation in an organism. The synergy of different antioxidants would reverse the oxidation to a greater extent compared with a single antioxidant. We analyzed ursolic acid (UA) together with foodborne phosphate-containing viili exopolysaccharides (EPS) for antioxidant activities, and anti-proliferation properties of cancer cells. The strong antioxidant capacity of UA was observed at concentrations of 100 μg/ml in ferric reducing antioxidant power test, where mild synergy with viili EPS was visible. Interestingly viili EPS had as strong antioxidative power as UA for the free radical scavenging activity in Fenton assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl-tetrazolium bromide (MTT) assay with hepatoma HepG2 cells, the mild, but significant anti-proliferation by UA and EPS was seen at concentration of 200 μg/ml, a synergic inhibition were particularly seen at 12.5 μg/ml with a ratio of 3:2 EPS and UA. The senescence of HepG2 cancer cells after the combined treatment of UA and viili EPS at 12.5 μg/ml were observed. These data indicated that UA is compatible with North European diets viili, together with potential antioxidative and anti-proliferation complement modulating activities.
Lactococcus lactis strain NIZO B40 produces an extracellular phosphopolysaccharide containing galactose, glucose, and rhamnose. A 40 kb plasmid encoding exopolysaccharide production was isolated through conjugal transfer of total plasmid DNA from strain NIZO B40 to the plasmid-free L. lactis model strain MG1614 and subsequent plasmid curing. A 12 kb region containing 14 genes with the order epsRXABCDEFGHIJKL was identified downstream of an iso-IS982 element. The predicted gene products of epsABCDEFGHIJK show sequence homologies with gene products involved in exopolysaccharide, capsular polysaccharide, lipopolysaccharide, or teichoic acid biosynthesis of other bacteria. Transcriptional analysis of the eps gene cluster revealed that the gene cluster is transcribed as a single 12 kb mRNA. The transcription start site of the promoter was mapped upstream of the first gene epsR. The involvement of epsD in exopolysaccharide (EPS) biosynthesis was demonstrated through a single gene disruption rendering an exopolysaccharide-deficient phenotype. Heterologous expression of epsD in Escherichia coli showed that its gene product is a glucosyltransferase linking the first sugar of the repeating unit to the lipid carrier.
抗腫瘍活性を示したスカンジナビアの粘質酸乳"ヴィリー"中の活性成分を検索する目的の一環として,ヴィリーより莢膜性粘質物産生Lactococcus lactis subsp. cremoris (L. cremoris KVS 20)を分離し,その菌体の抗腫瘍活性をSarcoma-180 (S-180)腹水および固型腫瘍系で検討した.その結果,腫瘍移植後連日9日間の腹腔内投与で両腫瘍系において腫瘍増殖抑制効果が認められた.50mg/kg投与では,腫瘍増殖が有意に抑制され,平均生存日数(WMST)が増加した.一方,L. cremoris KVS 20はin vitroにおいて,S-180腫瘍細胞に対して直接細胞障害活性を示さなかった.これらの結果より,L. cremoris KVS 20の腫瘍増殖抑制効果は宿主免疫の増強を介して誘導されるものと考えられた.
The radioprotective 105 (RP105)/MD1 complex is a member of the Toll-like receptor (TLR) family. It was reported that RP105/MD1 cooperates with the lipopolysaccharide (LPS) receptor TLR4/MD2 complex and plays a crucial role in the response of immune cells to LPS. This work evaluated whether RP105, TLR4 or TLR2 were involved in the immunoregulatory capacities of Lactobacillus plantarum N14 (LP14) or its exopolysaccharides (EPS). EPS from LP14 were fractionated into neutral (NPS) and acidic (APS) EPS by anion exchange chromatography. Experiments with transfectant HEKRP105/MD1 and HEKTLR2 cells demonstrated that LP14 strongly activated NF-κB via RP105 and TLR2. When we studied the capacity of APS to activate NF-κB pathway in HEKRP105/MD1 and HEKTLR4 cells; we observed that APS strongly stimulated both transfectant cells. Our results also showed that LP14 and APS were able to decrease the production of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) in porcine intestinal epithelial (PIE) cells in response to enterotoxigenic Escherichia coli (ETEC) challenge. In order to confirm the role of TLR2, TLR4 and RP105 in the immunoregulatory effect of APS from LP14, we used small interfering RNA (siRNA) to knockdown these receptors in PIE cells. The capacity of LP14 and APS to modulate pro-inflammatory cytokine expression was significantly reduced in PIERP105−/− cells. It was also shown that LP14 and APS were capable of upregulating negative regulators of the TLR signaling in PIE cells. This work describes for the first time that a Lactobacillus strain and its EPS reduce inflammation in intestinal epithelial cells in a RP105/MD1-dependend manner.