INTRODUCTION
The increasing prevalence of protein allergenicity
to cow’s milk has driven the food industry towards the
design, supply, and production of new plant-based milk
alternatives. Studies on the creating of formulations with
sensory acceptability that are suitable for vegetarian
diets have rapidly increased in recent years. In addition,
fermented products made from plant-derived milk
instead of fermented products from animals’ milk have
become of interest [1].
One of the plant-based alternatives to cow’s milk
is lupine milk. Lupine (Lupinus albus L.) is a plant
belonging to the Lupinus species of the Papilioneceae
(Legumineceae; butterfly-flowered) family. Lupine
is used as a soy alternative in such products as bread,
biscuits, cakes, pasta, confectionery, and soy sauce.
Besides, due to its antioxidant content, lupine is also
used in high-quality vegetable oil, gluten-free flour,
emulsifying agents, and alternative fermented products
[2, 3].
Today, lupine attracts attention as functional food
because it is rich in protein, minerals, vitamins, oleic
acid, fiber and other valuable components, as well as
because its antioxidant capacity. Lupine seeds contain
significant amounts of polyphenols, carotenoids,
phytosterols, tocopherols, and alkaloids, as well
as peptides with antioxidant, antimicrobial, anticarcinogenic,
and anti-inflammatory activities [4].
Lupine milk is obtained from lupine grains. The
protein value of lupine milk is 4.90 g/100 g, the fat
content is 5.00 g/100 g, the total dry matter ratio
is 11.20 g/100 g, and the pH is 6.30 [1, 2, 5]. Lupine
milk characteristics make it suitable to produce dairy
products such as set yogurt, probiotic yogurt, and
cheese [6].
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In addition to the use of lupine milk itself, there are
studies on the use of its components in the production of
dairy products. For example, lupine proteins were used
in ice-cream production and yielded ice cream with high
sensory properties [7].
Some studies, have reported that fermented
products can be produced from lupine milk, but since
yogurt starter cultures cannot use carbohydrates in the
composition of lupine milk, it is necessary to enrich this
milk by adding disaccharides, increasing thereby the
activity of starter cultures [6].
The growth of lactic acid bacteria in artificial
environments (other than milk) is difficult, but they
can easily grow in most plant-based media/substrates.
Additionally, lactic acid bacteria have been found to
rapidly increase acidity (decrease in pH) in plantderived
environments to a point where other competitive
organisms cannot thrive.
Krunglevičiūtė et al. have reported that acidity
(hence the growth of lactic acid bacteria) in a fermented
product made using lupine milk depends on the lupine
variety and amino acid levels [8]. However, it has
been stated that lupine milk has a unique nutritional
composition and may support the increase in the
number and survival of lactic acid bacteria in fermented
products.
Egg white protein powder is obtained as a result
of drying the egg white protein by the traditional
drying method. Pasteurized egg white protein powder
is produced by drying egg white protein by the
conventional spraying method [9]. In our research, egg
white protein powder was used to increase dry matter
level.
The aim of this study was to obtain a yogurt-like
product based on lupine milk as an alternative to cow’s
milk and evaluate its microbiological, physico-chemical,
and sensory attributes.
STUDY OBJECTS AND METHODS
Raw lupine (Lupinus albus L.), pasteurized egg
white protein powder (Alfasol®), JOINTEC VB530
lyophilized culture, lactose, and maltose were obtained
from Ödemiş (Turkey-İzmir), Kimbiotek Chemical
Substances Inc. (Istanbul-Turkey), CSL laboratory
(Strade per Merlino, 3,26839, Italy), and Sigma-Aldrich,
respectively.
Production of lupine milk. Lupine milk was
extracted from seeds by the method illustrated in Fig. 1.
Production of non-dairy yogurt-like product.
In the study, plant-based yogurt-like products were
obtained by from lupine milk with functional
properties, egg white protein powder, and disaccharides
(lactose and maltose) with the following fermentation
with Lactobacillus bulgaricus + Streptecoccus
thermophiles (Table 1).
Lupine milk was aliquoted into three equal
batches to obtain three samples, namely milk without
saccharides (control), milk with lactose (0.5%, w/v), and
milk with maltose (0.5%, w/v). Lactose and maltose
were added into the milk, the batches were pasteurized
separately at 85°C for 20 min, and cooled to 50–55°C.
Figure 1 Preparation of lupin milk from the seeds of Lupinus albus L.
Clean lupine seeds (1 kg)
Boiling 1:3 seeds/water (30 min)
Removing bitterness from the seeds
Heat treatment (20 min at 30°C)
Dehulling
Shredding (5 min)
Adding water (10 g/100 mL) so that the lupine dry matter is 10% and pH adjustment (6.5–7.0)
Homogenization
Cooling to room temperature
Storage at +4°C
Filtration
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After adding egg white protein powder (3%, w/v) to
each batch, they were homogenized for 3–5 min with
an Ultra Turrax Blender at 1200 rpm. Then, the samples
were cooled to 42–43°C and the starter culture (3%, w/v)
was added. Thus, we prepared three yogurt-like samples
based on the lupine milk samples. The samples were
incubated at 42–43°C and pH 4.60 for three days. On
days 1, 7, 14, and 28 of storage at 4°C, we performed
physico-chemical, rheological, microbiological, and
sensory analyses.
Physico-chemical analyses. Dry matter was
determined by the gravimetric method. The ash
content was determined using AOAC methods. The
fat content was determined by Gerber’s method. pH
value was detected using an SS-3 Zeromatic (Beckman
Instruments Inc., California, USA) pH meter, and
titration acidity (% lactic acid) was determined
according to the alkali titration method. The protein
value was determined by the Kjeldahl method according
to AOAC in milk and yogurt-like product samples [10].
Viscosity was determined with a digital viscometer
(V100003/FungiLabAlpha), and syneresis was
determined as described by Kieserling et al. [11, 12].
The texture properties were determined with a
texture analyzer (HDPL2/CEL5/TA-XT Plus), and the
carbohydrate content was determined with an Atago
Polax×2L polarimeter (Japanese).
Determination of fatty acid composition. Each
homogenized sample was extracted according
to the Gerber method to obtain oil as described
in [14], fatty acid methyl esters were prepared
according to AOCS and investigated using gas
chromatography [15]. We used a Supelco SP-
2380 (Supelco Inc., Bellefonte, USA) fused a silica
capillary column (60×0.25 mm i.d., 0.2 mm film
thickness) and a Hewlett-Packard gas chromatographer
(model 6890) with a flame ionizing detector.
Injection volume: 1 μL; oven temperature: 4°C/min
100°C to 220°C; injector and detector temperature:
300°C; carrier gas: helium; and flow rate: 1 mL/min.
Fatty acid methyl esters were detected in the lupine milk
and yogurt-like samples on day 1 of storage.
Microbiological analysis. MRS-agar (Merck,
Germany) was used to count L. bulgaricus. All
the samples under study were subjected to anaerobic
incubation at 42°C for 3 days on MRS-agar.
L. bulgaricus count was determined as CFU/g [16].
An M17 agar medium containing lactose was used for
S. thermophiles counting. The incubation of the planted
Petri dishes was carried out under aerobic conditions
at 37°C for 72 h. The typical colonies formed at the end
of the incubation were counted [17].
Sensory tests. Sensory analysis was performed by
10 trained panelists on days 1, 7, 10, 14, and 21 according
to Jovanović et al. [18].
Statistics. The samples were examined in three
repetitions and two replications. SPSS version
15 (IBM SPSS Statistics) statistical analysis package
program was used. The data considered significant
according to the analysis of variance (ANOVA) was
tested at the P < 0.05 level using the Duncan multiple
comparison test.
RESULTS AND DISCUSSION
Physico-chemical properties. In the study, dry
matter in lupine milk was determined as 10.02%, fat
3.58%, protein 5.05%, carbohydrates 2.59 g/100 g,
titration acidity (°SH) 0.131, pH value 6.38, ash 1.3%,
and viscosity 3.52 cP (20°C). The physico-chemical
properties of the yogurt-like samples based on lupine
milk are given in Table 2.
The acidity in the samples with lactose and maltose
was higher than that in the samples without sugars
during storage, which was associated with maltose and
lactose added to the lupine milk. The acidity increase
in the samples with maltose during storage was higher
than that in the products with lactose. The relationship
between the increase in acidity and the sugar addition/
type added to the samples was significant (P < 0.05).
Bintsis has reported that lactic acid bacteria develop
better especially in the presence of glucose and some
other sugars (sucrose, maltose), which cause higher
acidity increase [19]. Our research results were found
to be compatible with the literature, and depending
on the glucose ratio, the highest acidity increase was
determined in the yogurt-like product with maltose.
Additionally, the results regarding the increase in acidity
were found to be compatible with studies by Ozcan et al.
who stated that the viability of lactic acid bacteria
in plant-based yogurt-like products increased, thus
increasing the acidity of the product [20].
Dry matter in all the samples decreased by the end
of storage. The highest decrease was determined in
the samples without sugars, while the decrease in the
samples with lactose and maltose was found to be close
to each other. However, the dry matter decrease in the
products with maltose was found to be lower than that
in the samples with lactose. The relationship between
the type of sugar used in the production of non-dairy
Table 1. Experimental design
Lactobacillus bulgaricus +
Streptecoccus thermophilus
Egg white protein powder Lactose Maltose
Lupine milk (control) 3% 3%
Lupine milk with lactose 3% 3% 0.5%
Lupine milk with maltose 3% 3% 0.5%
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yogurt-like products from lupine milk and dry matter
was significant (P < 0.05).
The fat content decreased in all the samples by the
end of storage, but this decrease was not significant
(P > 0.05). We revealed that during storage fat amounts
in the samples with maltose and lactose were higher
than in the samples without sugars. This situation is
associated with a more pronounced syneresis in the
yogurt-like products without sugars compared to the
other samples. In this study, the relationship between the
fat content and the type of sugar added to the samples
and the amount of egg white protein powder was found
to be significant (P < 0.05).
Protein values decreased in all the samples during
storage, and the highest protein hydrolysis had the
samples with maltose, followed by the samples with
lactose and the samples without sugars. Among the
samples, protein hydrolysis is associated with egg
white protein powder added to lupine milk to increase
the protein content and dry matter in lupine milk. The
relationship between the sugar type added to milk and
the addition of egg white protein powder and protein
hydrolysis was significant (P < 0.05). In the study, the
relationship between the type of sugar added to lupine
milk and viscosity and syneresis was found to be
significant (P < 0.05).
Al-Saedi et al. stated that yogurt-like products can be
produced from lupine milk, with shortened fermentation
time and the increased amount of the starter culture
(especially probiotic microorganisms) [6]. In this respect,
our research results are compatible with the literature.
Table 2 Physicochemical properties in yogurt-like samples based on lupine milk (n = 3)
Storage, days Milk without sugars Milk with lactose Milk with maltose
Dry matter, % 1 12.82 ± 1.44aA 13.00 ± 1.14aB 13.00 ± 1.13aB
7 12.25 ± 1.26aA 12.75 ± 1.06aB 12.85 ± 1.69aC
14 10.69 ± 1.33aA 11.36 ± 1.46aB 11.65 ± 1.47aC
28 10.44 ± 1.67aA 11.03 ± 1.57aB 11.26 ± 1.33aC
Viscosity, cP 1 899.00 ± 5.11aA 941.00 ± 8.21aB 1021.00 ± 9.25aC
7 1056.00 ± 8.23bA 1154.00 ± 8.36bB 1163.00 ± 8.73aC
14 1274.00 ± 9.63bA 1566.00 ± 9.45bB 1621.00 ± 8.91bC
28 1663.00 ± 9.74bA 1841.00 ± 9.98bB 2047.00 ± 9.95bC
Syneresis, g 1 9.52 ± 1.01aA 8.67 ± 1.06aB 8.55 ± 1.02aC
7 12.25 ± 2.06aA 11.36 ± 1.03AB 11.22 ± 1.11aC
14 13.49 ± 1.12aA 12.41 ± 1.07aB 12.10 ± 1.53aC
28 15.95 ± 2.07aA 14.65 ± 2.06aB 14.24 ± 2.54aC
pH 1 4.60 ± 1.22aA 4.58 ± 1.29aB 4.56 ± 1.11aC
7 4.57 ± 0.81aA 4.42 ± 1.06aB 4.39 ± 1.46bC
14 4.45 ± 0.63aA 4.29 ± 1.21aB 4.25 ± 1.89bC
28 4.41 ± 0.78bA 4.19 ± 1.63bB 4.16 ± 1.42bC
Acidity (%LA), °SH 1 0.912 ± 0.120aA 0.938 ± 0.100aB 0.944 ± 0.550aC
7 0.988 ± 0.220aA 1.045 ± 0.650aB 1.095 ± 0.630bC
14 1.039 ± 0.350bA 1.121 ± 0.750bB 1.133 ± 0.710bC
28 1.044 ± 0.630bA 1.128 ± 0.430bB 1.139 ± 0.390bC
Fat, % 1 3.55 ± 0.41aA 3.57 ± 0.66aB 3.57 ± 0.51aB
7 3.12 ± 0.96aA 3.36 ± 0.60aB 3.38 ± 0.62aB
14 2.75 ± 0.25aA 3.19 ± 0.82aB 3.22 ± 0.84aB
28 2.35 ± 0.57aA 2.88 ± 0.74aB 2.93 ± 0.78aB
Protein, %
1 5.03 ± 0.91aA 5.00 ± 0.82aA 4.98 ± 0.52aB
7 4.62 ± 0.93aA 4.45 ± 0.67aA 4.33 ± 0.87aB
14 4.22 ± 0.67bA 4.16 ± 0.88aA 4.02 ± 0.74aB
28 4.06 ± 0.76bA 3.86 ± 0.50bA 3.75 ± 0.46bB
Carbohydrates, % 1 2.57 ± 0.92 aA 3.51 ± 0.80 aB 3.48 ± 0.91 aB
7 2.54 ± 0.82 aA 2.85 ± 0.85 aB 2.44 ± 0.72 aC
14 2.12 ± 0.49 aA 1.57 ± 0.63 aB 1.25 ± 0.56 aC
28 1.95 ± 0.57 aA 0.92 ± 0.22 aB 0.84 ± 0.21 aC
Ash, % 1 0.55 ± 0.09 aA 0.57 ± 0.09 aA 0.57 ± 0.03 aA
7 0.31 ± 0.08 aA 0.41 ± 0.07 aA 0.43 ± 0.07 aA
14 0.21 ± 0.08 aA 0.33 ± 0.09 aA 0.34 ± 0.06 aA
28 0.16 ± 0.07 aA 0.29 ± 0.02 aA 0.30 ± 0.02 aA
a,b,c different letters on the same column are statistically significant (P < 0.05)
A,B,C different letters on the same line are statistically significant (P < 0.05)
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In this study, the carbohydrate content decreased in
all the samples during storage. The highest decrease
was detected in the yogurt-like products with maltose,
following by the samples with lactose. The relationship
between the acidity increase and sugar addition/type
and carbohydrate content was found to be significant
(P < 0.05).
The ash values of all the samples decreased during
storage, with the highest decrease in the sample without
sugars, while the values for the products with lactose
and maltose did not differ significantly. There was no
significant difference between the samples in terms of
the ash level (P > 0.05).
Fatty acid composition. Saturated fatty acids,
monounsaturated fatty acids, and polyunsaturated fatty
acids in the yogurt-like products made from lupine
milk were detected as 14.17, 52.3, and 9.86 g/100 g,
respectively (Table 3).
Rheological properties. Syneresis and viscosity
values are given in Table 2, and the texture characteristic
are given in Table 4. Consistency values of the samples
were found to be significant in terms of sugar type and
storage time interaction (P < 0.05). During storage,
the stability (hardness) of the curd increased and the
effect of storage was significant (P < 0.05). Hardness,
flexibility, gumminess, and chewiness increased, while
stickiness decreased in all the samples. The hardness
determined in the sample with maltose was higher than
that in the other samples. This situation is associated
with lower syneresis, higher viscosity increase, lower
dry matter decrease compared to the other samples, and
higher increase in acidity (decrease in pH value) during
storage. The hardness of the samples with lactose was
higher than that in the samples without sugars. The
difference between the samples in terms of syneresis
Table 3 Fatty acid composition in yogurt-like products based on lupine milk
Fatty acid, g/100 g Milk without sugars Milk with maltose Milk with lactose
Oleic Acid (C18:1) 49.00 ± 1.13 49.10 ± 1.05 49.10 ± 1.05
Linoleic Acid (C18:2) 23.41 ± 1.21 23.40 ± 1.07 23.40 ± 1.05
Palmitic Acid (C16:0) 7.33 ± 1.03 7.35 ± 1.13 7.34 ± 1.05
Gadoleic Acid (C20:1) 3.46 ± 1.12 3.47 ± 1.15 3.46 ± 1.05
Stearic Acid (C18:0) 1.62 ± 0.26 1.62 ± 0.04 1.62 ± 0.13
Arachidic Acid (C20:0) 2.85 ± 0.65 2.86 ± 0.70 2.85 ± 0.55
Miristic Acid (C14:0) 0.49 ± 0.01 0.48 ± 0.03 0.48 ± 0.06
Pentadecanoic Acid (C15:0) 0.21 ± 0.02 0.20 ± 0.01 0.21 ± 0.02
Lauric Acid (C12:0) 0.050 ± 0.001 0.050 ± 0.004 0.050 ± 0.003
Table 4 Texture changes in yogurt-like products based on lupine milk during storage
Indicator Storage, days Milk without sugars Milk with lactose Milk with maltose
Hardness, N 1 0.32 ± 0.01aA 0.33 ± 0.02aB 0.34 ± 0.05aC
7 0.36 ± 0.09aA 0.37 ± 0.07bB 0.40 ± 0.03bC
14 0.37 ± 0.01bA 0.40 ± 0.06bB 0.43 ± 0.08cC
28 0.38 ± 0.02cA 0.42 ± 0.01cB 0.48 ± 0.05cC
Adhesiveness 1 0.04 ± 0.02aA 0.05 ± 0.03aA 0.05 ± 0.01aA
7 0.03 ± 0.01aA 0.03 ± 0.01aA 0.04 ± 0.02aA
14 0.02 ± 0.01aA 0.03 ± 0.01aA 0.03 ± 0.01aA
28 0.01 ± 0.01aA 0.02 ± 0.01aA 0.02 ± 0.01aA
Springiness, mm 1 4.00 ± 0.52aA 4.20 ± 0.94aB 4.63 ± 0.99aC
7 4.11 ± 0.83aA 4.29 ± 0.63aA 5.16 ± 0.66aB
14 4.36 ± 0.64aA 4.62 ± 0.80bB 5.55 ± 0.83bC
28 4.58 ± 0.84bA 4.81 ± 0.91bA 5.87 ± 0.80bB
Gumminess, g 1 41.05 ± 1.12aA 72.56 ± 1.27aB 79.47 ± 1.22aC
7 45.63 ± 1.23aA 78.56 ± 2.24aB 83.47 ± 2.88aC
14 49.22 ± 1.66aA 82.10 ± 1.96bB 85.33 ± 2.10bC
28 52.11 ± 1.35bA 85.45 ± 2.67cB 87.22 ± 2.66cC
Chewiness, mJ 1 0.14 ± 0.05aA 0.86 ± 0.11aA 0.92 ± 0.30aA
7 0.16 ± 0.04aA 1.12 ± 0.26aA 1.21 ± 0.14aA
14 0.21 ± 0.03aA 1.82 ± 0.49aA 1.88 ± 0.47aA
28 0.35 ± 0.13aA 2.02 ± 0.28aA 2.09 ± 0.66aA
a,b,c different letters on the same column are statistically significant (P < 0.05)
A,B,C different letters on the same line are statistically significant (P < 0.05)
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and the relationship between syneresis and sugar
addition/type was significant (P < 0.05).
During storage, syneresis and dry matter decrease in
the yogurt-like products with maltose were lower than in
the samples without sugars, while those in the samples
with lactose were lower than in the samples without
sugars. The relationship between increased acidity and
syneresis was found to be significant (P < 0.05).
The relationships between the viscosity, sugar ratio,
egg white protein powder, and acidity increase in the
samples under study were significant (P < 0.05). We
revealed that the viscosity increased in all the samples
during storage, with the highest increase in the product
with maltose, while the lowest increase was in the
samples without sugars. Egg white protein powder in the
amount of 3% increased the viscosity of all the samples
during storage.
The highest acidity and viscosity increase was
determined in the samples with maltose, followed by
the samples with lactose and the samples without sugars.
The acidity (4.56 pH) and viscosity (1021 cP) values
determined in the product with maltose on day 1 of
storage were higher than those in the other samples. In
the following days of storage, the increase in acidity was
higher than in the other samples (Table 2). Accordingly,
hardness and viscosity also increased.
The texture properties of the samples showed
similar changes during storage. We determined that the
rheological properties determined in the samples with
maltose and lactose during storage were similar to those
for the product without sugars but more acceptable. It
was observed that the yogurt-like products without
sugars were similar to the yogurt gel but had a more
watery (yogurt-like beverage) consistency compared to
the other samples. During storage, the increase in acidity
was lower, the syneresis was higher, and viscosity was
lower in the samples without sugars.
The rheological properties of curd in fermented milk
products develop depending on the composition of milk,
applied temperature, pH, soluble Ca++ ratio, and other
factors (such as casein micelle size, various interactions,
etc.). The increase in acidity decreases syneresis and
increases protein hydrolysis, hardness, more soluble
calcium, and in turn, viscosity [21].
In this study, we detected good physico-chemical
and rheological properties of the fermented products
produced with the addition of maltose and lactose.
It was associated with high protein content and
saccharide derivatives in the composition of lupine
milk. Krunglevičiūtė et al. reported that the acidity of
the fermented product made using lupine milk and the
development of lactic acid bacteria are related to the
lupine variety and amino acid level [8].
Sensory evaluation. Sensory evaluation revealed
good sensory properties of the yogurt-like products with
maltose and lactose (Fig. 2.) During storage, the samples
with the disaccharides got close scores in terms of
structure and consistency. This situation was associated
with the scantiness in syneresis and an increase in
acidity, viscosity, and hardness during storage. The
relationship between the increase of storage time and
structure and consistency was found to be significant
(P < 0.05).
In the study, the samples with maltose and lactose
were found to be close to yogurt with a prolonged
storage time, which is a classical fermented product,
in terms of structural properties. Additionally, the
structure and consistency of these samples were
more similar to the classical fermented product
(yogurt) with no lupine flavor or with weak one during
storage. The texture and consistency in the samples
without sugars were found to be less viscous and the
panelists concluded that they could be considered as
yogurt. Apparently, the level of fat in the samples with
disaccharides during storage also influenced the taste of
the product.
Microbiological analysis. Changes in L. bulgaricus
and S. thermophilus amounts in the yogurt-like products
obtained from lupine milk are given in Fig. 3. The
samples with maltose and lactose demonstrated the
increased growth and activity of starter cultures.
In the production of non-dairy yogurt-like products
from lupine milk, the relationship between the addition
of sugar and the growth of starter cultures was found to
be significant (P < 0.05). In the samples with maltose
and lactose, syneresis decreased during storage which
had a positive effect on the development of starter
cultures.
In the products without sugars, the L. bulgaricus and
S. thermophilus growth was weaker compared to those
with sugars. However, this situation did not appear as a
problem in the production of yogurt-like products in the
study. On the contrary, it strengthened the opinion that
the composition of lupine milk is a suitable raw material
to produce non-dairy yogurt-like products. Different
studies have reported that lupine proteins effectively
maintain the viability of starter cultures in different
Figure 2 Sensory properties of yogurt-like products based on
lupine milk (1), lupine milk with maltose (2), and lupine milk
with lactose (3)
.0
.0
consisteny
taste odor
acceptability
color
.0
.0
1
.0
.0
.0
.0
2 3
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products based on lupine milk and can protect the starter
cultures by wrapping them like a capsule [22].
Our research results were found to be compatible
with studies that stated that starter cultures show
better growth in the presence of some sugars (such
as glucose and maltose) [19]. It has been reported
that lupine milk contains carbohydrates at the level of
2.83 g/100 g, including different types of carbohydrates
(galactose and arabinose) [23, 24]. In this work, the
maximum growth of L. bulgaricus and S. thermophilus
(8 log10 CFU/mL) in the starter cultures on day 14
in the samples with maltose was associated with the
adaptation (prolongation of the lag+ phase) and growth
phase of starter cultures in the presence of maltose.
However, this effect could not be detected in the
samples with lactose added. On the contrary, the
development of starter cultures in the samples with
lactose was particularly high (7–8 log10 CFU/mL) until
day 7 of storage, and decreased to 6–7 log10 CFU/mL
after day 14 of storage.
L. bulgaricus levels in the products with maltose
were found to be lower than those in the samples with
lactose between days 1 and 7 of storage. However, at
days 14 and 28 of storage, L. bulgaricus levels in the
samples with maltose were found to be higher than those
in the samples with lactose. S. thermophilus levels, on
the other hand, were found to be lower in the yogurt-like
product with maltose until day 7 of storage (including
day 7) and higher after day 7 than those in the products
with lactose.
These results show that S. thermophilus was
effective, also it is associated with an increased glucose
concentration in the medium as a result of reaching a
higher level of S. thermophilus than of L. bulgaricus.
We determined relationships between the glucose
ratio and bacterial growth, between the bacterial
growth and acidity increase, and between acidity
increase and hardness, viscosity, and syneresis. These
results were found to be compatible with Bintsis [19].
L. bulgaricus and S. thermophilus were determined in
the samples without sugars during storage, as well as
the slower growth of acidity on the same storage days
was attributed to the increase in syneresis observed in
those samples. With the increase in syneresis in the
yogurt-like products, the symbiotic relationship between
microorganisms was disrupted, pH development slowed
down or stopped [25].
Elsamani determined that the levels of Bifidobacterium
bifidum and Lactobacillus acidophilus
were preserved and increased in probiotic ice creams
produced from lupine milk on day 30 of storage [26].
The author associated it with the protection of the
proteins found in high levels in the composition of
lupine milk that wrap the probiotics like a capsule.
Figure 3 Number of microorganisms in yogurt-like products based on: lupine milk during storage (a), lupine milk with lactose
during storage (b), and lupine milk with maltose during storage (c)
S. termophilus L. bulgaricus
9.00
Microorganism count,
log10 CFU/mL
7.50
7.00
6.50
6.00
6.50
4.50
4.00
day 1 day 7 day 14 day 28
8.00
8.50
5.00
a b
Microorganism count,
log10 CFU/mL
7.50
7.00
6.50
6.00
6.50
4.50
4.00
day 1 day 7 day 14 day 28
S. termophilus L. bulgaricus
8.00
8.50
9.00
5.00
Microorganism count,
log10 CFU/mL
7.00
6.50
6.00
5.50
5.00
4.50
4.00
3.50
3.00
1.00
1.50
2.00
2.50
day 1 day 7 day 14 day 28
S. termophilus L. bulgaricus
c
384
Kavas N. Foods and Raw Materials. 2022;10(2):377–385
CONCLUSION
In our study, lupine milk was obtained from lupine
with functional properties and egg white protein powder.
Different concentrations of lactose and maltose were
added to lupine milk to obtain a non-dairy yogurt-like
product.
The growth of Lactobacillus bulgaricus and Streptecoccus
thermophiles was weaker in the disaccharidefree
products compared to the samples with maltose
and lactose. The increase in acidity in the samples
with disaccharides during storage (28 days) was higher
than that in the samples without sugars, and this was
associated with maltose and lactose added to lupine
milk.
It has been concluded that yogurt-like samples
produced from lupine milk can be produced due to their
similarity to fermented products (especially yogurt)
of animal origin in terms of physico-chemical and
rheological properties. However, with time, the samples
with maltose and lactose were found to be closer to
classical yogurt in terms of all properties. Sensory
evaluation revealed that the smell and aroma of lupine
were not pronounced. Thus, lupine yogurt-like products
had high sensory properties.
Consequently, yogurt-like products based on lupine
milk can be used as an alternative to fermented products
produced from cow’s milk, and more detailed studies
should be conducted to formulate and optimize lupine
fermented milk products.
CONFLICT OF INTEREST
The authors declare that there is no conflict of
interest.

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