FACTORS AFFECTING SYNERESIS IN YOGHURT: A REVIEW

Rekha Rania*, V.Unnikrishnana, Dharaiya, C.N.b and Bhopal Singhc

Yoghurt is fermented milk product and syneresis (Whey separation) is a textural defect in it.
During manufacturing yoghurt various factors affects its body and texture like composition, heat
treatment, homogenisation lactic culture and incubation condition, acidity etc. Cow and buffalo
compositional differences affect quality of yoghurt. Increasing amount of protein increase gel
firmness and decreases syneresis. Homogenisation affect product quality, higher viscosity and
firmness observed when homogenisation performed after heat treatment.Degree of denaturation
of whey protein is major factor affecting syneresis and yoghurt texture. Incubation temperature
(42-43oC) is also important factor affecting syneresis, lower the temperature, longer the time for
curd setting and higher temperature leads to more syneresis in yoghurt. Use of contamination
free lactic culture is desired condition for yoghurt manufacture. Type of acidification kinetics,
either slow or fast affects the protein network formation and whey separation.
Key words:Yoghurt, syneresis, composition, culture, heat treatment, temperature

INTRODUCTION
Yoghurt is an important fermented milk
product, obtained by the lactic fermentation
of milk. Yoghurt is gaining in popularity due
its acceptability for the consumers as well as
their nutritional properties and potentially
beneficial effects in human health. Good
quality yoghurt is firm and smooth
consistency with a sweet aroma and pleasant
taste. Syneresis (Whey separation) is a
textural defect in yoghurt. Rearrangement of
the network of casein micelles is the main
cause of syneresis. Due to shrinkage of gel,
water will expel from the network which
leads to syneresis.
The composition of the base milk, the
fermentation process and the post
fermentation treatment, all have
considerable influence on the texture and
wheying-off in yoghurt. Adequate firmness
without syneresis is essential for top quality
yoghurt.
Composition
The amount of total solids in the base milk,
to a large extent, determines the physical
properties of the final yoghurt product.
Syneresis from yoghurt is caused by a
broken coagulum due to protein content
below 3.4 %, a low fat content and a high
mineral content of the milk, heating of the
coagulum during incubation or thereafter
(Kroger, 1976).
The compositional variation between cow
and buffalo milk influences the quality of
the curd. The curd prepared from buffalo
milk is firmer than the curd obtained from
cow milk. The higher curd tension of buffalo
milk curd has been attributed to the higher
levels of SNF (Ganguli and Menon, 1971)
and calcium (Sindhu and Roy, 1973; Yadav
and Singh, 1973). Higher rate of syneresis of
rennet curd prepared from buffalo milk was
observed (Ganguli, 1974). This may hold
true for lactic curd from buffalo milk.
There are several methods to fortify yoghurt

a
Dairy Chemistry and Bacteriology section, NDRI, Bangalore-560030 *Email: verma.rekha@gmail.com
b
Assistant Professor, SMC College of Dairy Science, AAU, Anand-388110
c
M. Tech. Scholar, NDRI, Karnal-132001

Indian J. Dairy and Biosci., 23, 2012

Syneresis in Yoghurt - Review
for improvement of body and texture
(Kulkarni et al., 1990). These include
evaporation of base milk to the desired dry
matter level, reverse osmosis, addition of
skimmed milk powder or the addition of
evaporated milk.
Protein
The protein content is the most important
factor which influences the rheological and
physical properties of yoghurt. Increasing
the amount of protein increases gel
firmness and decreases syneresis. The
formation of the coagulum and hence the
consistency/viscosity of the product is
directly proportional to the level of protein
present. Casein is more effective in this
regard than whey proteins (Molder et al.,
1983). The formation of gel is dependent on
the functional properties of the casein
fraction.
According to Snoeren et al. (1982), milk
casein is able to immobilise as much as 2.82
g water per g protein and shows a
voluminosity of 3.57 ml/g. Corresponding
hydration values for undenatured and
denatured whey proteins are 0.32 g and 2.34
g water per g protein, respectively. Parnell-
Clunies (1985) reported lower water holding
capacity and protein hydration in yoghurt
from unheated milk than heated milk.
Results on firmness and syneresis of yoghurt
with a modified whey protein/casein ratio
are somewhat contradictory. There is,
however, a tendency towards less viscous,
softer gels for products with an increased
whey protein content compared to products
showing similar protein content on casein
basis (Jelen et al., 1987; Morris et al., 1995).
In any case, it is suggested to limit the
increase in the whey protein/casein ratio due
to potential off-flavour effects (De Boer,
1996).
Fat
Partial replacement of the native fat globule
membrane with other milk proteins, as in
the case of homogenisation allows fat
globules to be incorporated into the gel by
cross linking them to the matrix (van Vliet et
al., 1982). Large fat globules as present in
unhomogenised milk may decrease firmness
of fermented products by interrupting the gel
network of casein particles (Aguilera and
Kessler, 1988). The firmness values for
whole yoghurt are lower than skimmed
yoghurt (Salvador and Fiszman, 2004). If
the milk has been homogenised, most of the
surface coat of fat globules consists of
miceller casein (Tamime and Marshall,
1997). On acidification, these globules will
participate in the aggregation, hence in
building the network. This implies that the
effective concentration of casein has been
greatly increased thereby enhancing
firmness and stability against whey
separation.
After homogenising whole milk at different
conditions, Plock et al. (1992) found a linear
increase in gel firmness with decreasing fat
globule diameter. The covering of the
homogenisation-induced enlarged fat
globule surface area with fragments of milk
proteins leads to the development of a
secondary fat globule membrane, which is of
great importance for the characteristics of
fermented dairy products (Schkoda, 1999).
The texture-enhancing capability of fat in
yoghurt (Lucey et al., 1998; Patrignani et
al., 2007) is related to the ability of
homogenized fat globules to participate in
the gel network formation (Aguilera &
Kessler, 1988; Sodini et al., 2004) and
consequently strengthen the yoghurt gel
structure (Lucey et al., 1998). The effects of
incorporating the fat globules into the gel
network also depend on the globule size and,
consequently, the homogenisation
procedure. A higher efficiency is achieved
by multiple stage homogenisation at low
Indian J. Dairy and Biosci., 23, 2012
Syneresis in Yoghurt - Review
homogenisation pressures (Tamime and
Robinson, 1999).
According to Samuelson and Christiansen
(1978), increased amounts of milk fat
globule membrane material i.e.
phospholipids and proteins in the skim
phase, improve the water holding capacity of
coagulum. A change in water holding
capacity of the milk proteins tends to
influence syneresis (Grigorov, 1966a).
The effect of homogenisation depends on
the layout of the production process. Higher
viscosity and firmness values for products
were observed where homogenisation was
performed after heat treatment (Kulkarni et
al., 1990). Casein micelles incorporated in
the secondary fat globule membrane built
during homogenisation will be covered with
the whey proteins. Their denaturation during
subsequent heating might prevent the active
contribution of the fat phase to the gel
properties (Kulkarni et al., 1990 and
Tamime and Marshall, 1997).
Homogenisation after heat treatment causes
casein micelles to be torn apart by surface
active forces while new fat globules are
being formed. On acidification hydrophobic
interactions then produce more stable
protein network (Kessler, 1997). Schkoda
(1999) stated that heat treatment preceding
homogenisation causes a higher load to the
membrane due to the aggregation of
denatured whey proteins to each other and to
casein. On the other hand, heating after
homogenisation leads to partial aggregation
of membrane proteins to each other,
resulting in the formation of aggregates of
linked fat globules.
Heat treatment
Heating of milk is an essential processing
variable in yoghurt manufacture. For
manufacturing of yoghurt, milk is normally
homogenized (15–20 MPa) and heat-treated
to reduce microbial load, to increase the
yoghurt stability, consistency and
body/texture, and also to decrease whey
separation during storage (Lucey, 2004).
Patel and Chakraborty, (1988) also reported
heat treatment of milk usually improves its
value as a medium for starter organisms and
other lactic acid bacteria and also reduces
the curd tension
Functional properties of whey proteins
become more apparent after heating milk
and they begin to become denatured above
60oC. Heating increases voluminosity and
water binding capacity of whey proteins
(Snoeren et al., 1982) and decreases their
solubility. When milk is heated to such an
extent that most of the serum proteins is
denatured, the fermented product is much
firmer. Also, whey separation tends to be
less (Staff, 1998). In commercial yoghurt
production, temperature-time profiles
usually ranging from 80-85oC for 30 min to
90-95oC for 5 min are applied (Lucey and
Singh, 1998). β-lactoglobulin plays a central
role in interactions and complex forming
reactions with itself, with α-lactalbumin and
with κ-casein. Interaction of whey proteins
with casein increases the hydrophobicity of
the micellar surface and favours gelation
(Jaros and Rohm, 2003).
The degree of denaturation of whey proteins
is a major factor affecting syneresis and
yoghurt texture (Mottar et al., 1989).
Dannenberg and Kessler (1988a, 1988b)
demonstrated the close relationship between
the degree of β-lactoglobulin denaturation
and selected physical properties of yoghurt.
Within a β-lactoglobulin denaturation range
of 60-99%, susceptibility to syneresis
decreased linearly. Increasing heating
temperature and time led to higher
denaturation of β-lactoglobulin and α-
lactalbumin with β-lactoglobulin being more
heat sensitive (Lucey et al., 1997).
Indian J. Dairy and Biosci., 23, 2012
Syneresis in Yoghurt - Review
According to Grigorov (1996), optimum
hydrophilic properties of the proteins and
hence coagulation of the yoghurt milk are
obtained when milk is heated to 85oC for 30
min.
In heated milks, the gel is formed as the
casein micelles gradually increase in size
and form chain matrix. This behaviour
results in an even distribution of the protein
throughout the yoghurt and the aqueous
phase is immobolised within the network;
the resultant coagulum is firm and less
susceptible to syneresis (Tamine and
Robinson, 1999). An investigation of milks
subjected to heat (95oC for 10 min) revealed
filamentous appendages composed of β-
lactoglobulin/ κ-casein. The interaction
appears to involve sulphydryl linkage and,
possibly, the involvement of various salts.
For example, calcium phosphate and citrates
(Harwalkar and Kalab, 1981). These
appendages tend to become “diffuse” after
fermentation, but their presence in the
coagulum of heated milks inhibited micellar
coalescence, giving rise to curds with
reduced tendencies to syneresis (Tamine and
Robinson, 1999).
Lactic cultures and incubation conditions
The use of contamination free lactic culture
in a desired state of activity is an essential
pre-requisite for the manufacture of
fermented milk. The basic role of starter
culture is to supply a large number of
desirable microorganisms in their optimum
state of activity. This is attained in a starter
culture by promoting the growth of culture
microorganisms under optimum conditions
of incubation time and temperature and
nutritional supply of the culture media
(Tamime and Deeth, 1980).
In general, Streptococci and Lactobacilli are
the two major groups of microorganisms
that are used in fermented milk products for
acid production (Davis, 1975; Tamime and
Grig, 1979). The type and activity of
microorganisms present in the starter culture
have a great influence on the quality and
flavour characteristics of the curd (Law,
1981). Increased rate of acid production has
been observed in mixed strains of
Streptococcus thermophilus and
Lactobacillus bulgaricus as compared to
their individual ability (Accolas et al.,
1977). Yoghurt culture produces more acid
in buffalo milk than in cow milk (Thomas et
al., 1966; Singh and Ranganathan, 1978).
The classical yoghurt starter culture is a
mixture of Streptococcus thermophilus and
Lactobacillus bulgaricus, usually in 1:1
ratio. Lactic acid bacteria that produce extra
cellular polysaccharides (EPS) are also used
in the yoghurt industry to improve the
physical stability of the product. The use of
slime producing yoghurt cultures reduces
susceptibility to syneresis (Sehellhaass and
Morris, 1985; Toba et al., 1990). The
bacterial cells synthesize EPS, which may
occur as a capsule intimately associated with
the cell surface, or secreted into the
extracellular environment in the form of
slime. EPS producing starter cultures are
used in fermented milk to improve viscosity,
prevent syneresis, improve consistency and
minimise mechanical damage (Shah, 2002).
Polysaccharide production is normally
highest under aerobic conditions. EPS
produced by yoghurt bacteria give a thicker
texture and higher viscosity, enhance
smoothness, impart desirable body and
texture characteristics and prevent gel
fracture and wheying off (Skriver et al.,
2002).
Normally there are greater interactions
among bacteria, EPS and protein at 43oC as
compared to 32oC. The yoghurt is usually
made at 42-43oC and greater interactions
among bacteria and EPS are expected to
occur. It seems that not the amount of
Indian J. Dairy and Biosci., 23, 2012
Syneresis in Yoghurt - Review
polysaccharides but rather the type, charge
and molecular mass of EPS are important for
rheological properties (Laws and Marshall,
2001; Pleijsier et al., 2002; Ruas-Madiedo et
al., 2002).
The conditions of incubation influence the
properties of the final product. Two different
fermentation procedures are usually applied:
the short incubation method and the long
incubation method. By providing the
microorganisms their optimum temperature
range, incubation times of less than 6 h can
be achieved. When incubation should take
place overnight due to technical
requirements, temperature has to be reduced
to around 30°C, leading to a fermentation
time of 10 –12 h to reach the desired end pH
(Staff, 1998). It is generally accepted that
the lower the fermentation temperature, the
longer it takes to reach a certain pH and
therefore firmness, but the final product is
much firmer (Walstra et al., 1999). There
are also reports to the contrary. Lankes et
al. (1998) reported that yoghurt
manufactured at 42°C had higher gel
firmness than the product fermented at
30°C. Similarly, Haque et al. (2001) found a
systematic increase in gel strength with
increase in the temperature of fermentation
from 37°C to 46°C. Any agitation during
incubation leads to whey separation.
Acidity
Lactose fermentation, with lactic acid
production determines the precipitation of
casein particles in yoghurt. Yoghurt milk
will become solid at an acidity of about 0.6
% lactic acid. pH at this point is about 5.3
depending on the type of milk and protein
content (Davis, 1956). High acid yoghurt
may have titratable acidity of above 1.0 %.
A „solidity‟ of gel will begin to be seen at
approximately pH 5.6. Lowering the pH
causes demineralisation of casein micelles
(Lucey and Singh, 1998). Below a pH of 5.5
the casein micelles begin to swell and, as
almost all colloidal calcium phosphate is
dissociated, start to precipitate. This
precipitation leads to sufficient decrease in
voluminosity of casein micelles (Van
Hooydonk et al., 1986) and to the formation
of clusters and chains that link together to
form a gel, composed of a continuous three-
dimensional network with the milk serum
containing whey proteins, lactose and salts
entrapped as liquid phase (Staff, 1998).
Since the protein is most insoluble and has
lowest water binding properties at its iso-
electric point, the yoghurt gel is very
sensitive at this pH (Binanchi Salvadori,
1997; Staff, 1998). Above pH 4.5 a weak
coagulum is obtained (Kroger, 1976). For a
lower pH, the gel is generally firmer,
presumably because the protein-protein
bonds are stronger (Walstra, 1997).
Enhanced acidification to pH values below
4.0 may lead to body and texture defects
such as gel shrinkage and syneresis (Jaros
and Rohm, 2003).
The type of acidification kinetics, either
slow or fast, affects the protein network
formation, including coagulum permeability
and whey separation. A granular, lumpy,
grainy and sandy structure is the result of a
very slow or irregular acidification due the
presence of inhibitory substances in milk or
bacteriophages or too little inoculum or
incompatibility among the strains due to the
production of bacteriocins (Rasic and
Kurman, 1978). However, too rapid
acidification, due to starter inoculum, or
incubation temperatures higher than the
normal ones should be avoided in either the
initial phase of fermentation or during
incubation, as it can lead to dense
aggregation of protein particles, with a
decrease in bound water, causing micelles
dehydration, grainy clot formation and whey
separation which is not reabsorbed by the
Indian J. Dairy and Biosci., 23, 2012
Syneresis in Yoghurt - Review
coagulum in the subsequent phases
(Yuguchi et al., 1989).
CONCLUSION
There is currently considerable growth in
industrialized scale in the economical
importance of yoghurts. In the last few
years, this tendency has driven the
manufacturers to develop and produce
yoghurt with firm body and texture and
having minimum syneresis. Compositional
and processing parameters like solid content,
fat content, protein content, heat treatment,
homogenization, incubation temperature
(42-43oC), pH / acidity etc. affect the
syneresis in yoghurt. While manufacturing
yoghurt these parameters should be taken
into account to decrease syneresis and it will
have positive effect on consumer
acceptance, shelf life and market demand of
product.
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