INTRODUCTION
Microorganisms have long been widely used in the
food industry: in winemaking and beverage production,
dairy production, silage making and fermentation of
plant materials, as well as in seafood fermentation [1–17].
Products of animal origin make up a significant
part of total food consumption. Like any products, they
undergo biochemical transformations during production
and are exposed to microorganisms during long-term
storage [18–19].
Literature describes various methods for producing
foods under the influence of microorganisms contained
in animal raw materials. One of such methods – aging
– is mainly used to produce dry-cured products. This
process involves curing raw materials and semi-products
under certain conditions to expose them to a gradual,
and sometimes fairly long, effect of the microorganisms’
enzyme systems. As a result, the main food components
transform and develop certain flavoring characteristics
[20–22].
Today, formulators forcibly introduce starter cultures
of microorganisms into raw materials to reduce process
time. In particular, for food systems based on animal
raw materials, they use Staphylococcus (St. xylosus,
St. carnosus), Lactobacillus (L. pentosus, L. plantarum,
L. sakei, L. curvatus), and Pediococcus (P. pentosaceus,
P. acidilactici) [23–26]. Microorganisms can
produce different effects and form a wide variety
of flavors [27–28].
For convenience, microorganisms are commonly used
as lyophilized solids, with culture cells deposited on the
surface of a solid carrier, usually sugars. For products of
animal origin, 1–2 g of a freeze-dried culture containing
(1–10)×1012 CFU/100 g sucrose is usually added per every
5 or 10 kg of raw material [21, 29].
Although the action of microbiological cultures
on various food materials is fully described, scientific
literature lacks systematic data on the quantitative
changes in the most important minor components of
animal materials (individual amino acids, fatty acids,
and monosugars) under the influence of L. planta-rum
and S. carnosus.
We aimed to identify the effects of microorganism
cultures on protein, lipid, and carbohydrate components
of animal-based food systems.
STUDY OBJECTS AND METHODS
Our objects of the study included a model food
system – a mixture of beef muscle tissue Longissimus
dorsi and pork fat tissue Telae adipem with 2% sodium
chloride (75:25%) homogenized in a Buchi Mixer B400
blender (Switzerland), as well as a model protein. The
model food system contained 18.5% protein, 23% fat,
2.5% carbohydrates, and 54% moisture.
The model protein was obtained by a 6 h extraction
at 25°C of Longissimus dorsi of Bubulae beef with a 5%
sodium chloride solution followed by desalting on G25
and freeze-drying [30]. The isolated protein was 93%
pure and had a 6% moisture.
Lactobacillus plantarum ATCC 8014 (LP) and
Staphylococcus carnosus ATCC 51365 (SC) were added
in an amount of 1×107 CFU/g of raw material. We used
preparations of culture on freeze-dried sucrose in an
amount of 2×1010 CFU/g.
The model mixture was treated as follows. First,
animal raw materials were kept in salt for 24 h at
2 ± 2°С. Then, we introduced starter cultures and packed
the mixture in plastic bags to keep in the chamber for
5 days at 2–4°С, relative humidity (W) 85%, and an air
flow speed of 0.1 m/s. Further treatment was carried
out during 5 days (15°C, W 82%) and 10 days (12°C,
W 75%). The control sample was kept in salt for 24 h at
2 ± 2°C.
The model protein was treated with starter cultures
in a 2% sodium chloride solution (hydromodule 1:5)
under similar conditions at pH 7.0.
To measure the proteolytic activity, we placed
a 1% casein solution in 0.05 M Tris-PO4 buffer
(pH 7.0) into two tubes (5 mL in each) and added
10 mL of distilled water to the first tube and 1 mL of
a 1×1010 CFU/mL enzyme solution or 1 mL of the test
solution to the second tube. After a 10-min exposure
at 37°C, we added 5 Ml of a 10% trichloroacetic acid
solution to the test samples, filtered them through a
0.45 μm filter, and measured the optical densities of
the transparent solutions against the control at 280 nm.
The proteolytic activity (units/mg) was calculated as
A = (D280 sample – D280 control)/10·g, where g is the
nominal enzyme concentration in the test sample. The
standard unit of peptidase activity is the amount of
enzyme required to release free amino acids during
proteolytic decomposition. It is equivalent to a change
in the absorption rate of the test solution (0.001D280) per
minute at 37°C and pH 7.0 [31].
The materials were treated with L. plantarum and
S. carnosus in a 1:5 ratio: 1 g of the enzyme preparation
per 5 kg of the formulation and 1 mg of the preparation
per 5 g of animal protein.
The content of amino acids was determined on
a Biotronic 6001 amino acid analyzer (Germany) by
distribution chromatography after acid hydrolysis of
proteins [31].
Free amino acids were determined after protein
precipitation by adding 10% trichloroacetic acid,
followed by neutralization with a 10M sodium hydroxide
solution to pH 2.0 and filtration through a Millipore
membrane filter with a pore diameter of 0.22 μm. Then,
the filtrate was diluted in a buffer solution (pH 2.2). To
quantify individual amino acids, we compared the peak
areas in the aminogram obtained with the Winpeak
Eppendorf-Biotronic integration system (Germany)
by analyzing a standard mixture of amino acids that
contains 2.5 μmol of each amino acid in 1 ml of the
solution [31].
Fatty acids and chemical components responsible
for the product’s taste and aroma were determined by
chromatography-mass spectrometry [21, 31].
The components were analyzed on a 7890A gas
chromatograph with a 5975C VLMSD mass selective
detector (Agilent Technologies, USA) using a modified
Folch method. In particular, a 1 g sample was subjected
to a mixture of 10 mL chloroform and 10 mL methanol
in the presence of a 1% KCl solution for 24 h to
dissolve the lipid components. The extract was filtered
through paper. After removing the excess solvents by
evaporation to dryness, the residue was subjected to acid
hydrolysis to obtain methyl esters of fatty acids, which
were analyzed by gas chromatography.
A 0.01 g amount of lipids was treated with 3 mL
of a 15% solution of acetyl chloride in methanol at
100°C for 2 h. Then, the mixture was neutralized
by KOH (1.25 mL) in СН3ОН to pH 5.0–6.0. A few
minutes after adding 3 mL of a saturated aqueous NaCl
solution and 3 mL of hexane, we took for analysis 0.2 μl
from a clear hexane layer containing methyl esters
of fatty acids. Chromatography was performed on a
30 m × 0.32 mm × 0.5 mkm HP-Innowax capillary
column under the following conditions: the column
temperature in the thermostat increasing from 100°C
to 260°C at a rate of 10°C/min; injector temperature
250°C, detector temperature 300°C; hydrogen flow
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from the generator at 35 cm3/min; nitrogen flow at
20 cm3/min; flow division 1:100; analysis time
30 min; injection of 1 μl of the sample. A NIST08 MS
Library was used to measure the content of isomers,
an automatic search and identification program for gas
chromatography-mass spectrometry with a probability of
peaks correlation above 65%.
The content of free fatty acids was determined by an
acid-base titration of the samples according to the acid
number. In 2 mg KOH/g of the product, it corresponded
to 1% of the mass content of free fatty acids [31].
The composition of free carbohydrates was analyzed
using a BioLC chromatographic system, including a
GS50 gradient pump, an ED50 electrochemical detector,
an EG50 eluent generator with 10mN NaOH, and an
LC25 chromatographic thermostat with a CarboPac
PA20 column (Dionex, Germany). The content of free
carbohydrates was determined in aqueous extracts of
0.01 g of the sample in 100 ml of demineralized HPLCgrade
water filtered through a 0.45 μm filter at 25°C.
The water retention capacity was determined by
a standard method, recording bound moisture under
load [31].
Our study used casein, tris (hydroxymethyl)
aminomethane (tris), phosphoric acid, sodium chloride,
sodium hydroxide, potassium hydroxide, and Sephadex
G-25 (Sigma, USA). As standards of amino acids, we
used a solution of mixed individual amino acids in a
molar concentration of 2.5 μmol/ml (Supelco, USA):
glycine, alanine, valine, leucine, isoleucine, proline,
phenylalanine, tyrosine, methionine, cysteine, aspartic
acid, glutamic acids, lysine, arginine, histidine, serine,
and threonine.
As standards of fatty acids, we used a solution of
mixed C6–C24 fatty acid methyl esters in methylene
chloride with a mass concentration of 10 mg/mL
(Supelco, USA): caproic (C6:0), octanoic (C8:0),
decanoic (C10:0), decenoic (C10:1), undecanoic (C11:0),
dodecanoic (C12:0), tridecanoic (C13:0), tetradecanoic
(C14:0), cis-9-tetradecenoic (C14:1), pentadecanoic
(C15:0), cis-10-pentadecenoic (C15:1), hexadecanoic
(C16:0), cis-9-hexadecenoic (C16:1), heptadecanoic
(C17:0), cis-10-heptadecenoic (C17:1), octadecanoic
(C18:0), cis-9-octadecenoic (C18:1n9c), trans-9-
octadecenoic (С18:1n9t), cis-9,12-octadecadienoic
(С18:2n6), cis-6,9,12-оctadecatrienoic (С18:3n6), cis-
9,12,15-оctadecatrienoic (С18:3n3), nonadecanoic
(С19:0), eicosanoic (С20:0), Cis-9-eicosenoic (С20:1n9),
cis-11,14,17-eicosatrienoic (C20:3n3), cis-8,11,14-
eicosatrienoic (C20:3n6), cis-11,14,17-eicosatrienoic
(C20:3n3), cis-5,8,11,14-eicosatetraenoic (C20:4n6),
eicosapentaenoic (C20:5n3), heneicosanoic (C21:0),
docosanoic (C22: 0), cis-13-docosenoic (C22:1n9),
cis-13,16-docosadienoic (C22:2n6), cis-7,10,13,16,19-
docosapentaenoic (C22:5n3), cis-4,7,10,13,16,19-
docosahexaenoic (C22:6n3), tricosanoic (C23:0),
tetracosanoic (C24:0), and cis-15-tetracosenoic (C24:1).
As carbohydrate standards, we used arabinose
(Ara, C5H10O5, D-(−)-arabinose ≥ 99%, A3131 Sigma),
galactose (Gal, C6H12O6, D-(+)-galactose ≥ 99%, G0750
Sigma-Aldrich), glucose (Glc, C6H12O6, D-(+)-glucose
≥ 99.5%, G8270 Sigma), xylose (Xyl), mannose (Man,
C6H12O6, D-(+)-mannose from wood, ≥ 99% M2069
Sigma), fructose (Fru, C6H12O6, D-(−)-fructose ≥ 99%,
F0127 Sigma), sucrose (Sug, C12H22O11, α-D-glucose-
(1→2)-β-D-fructose, sucrose ≥ 99.5% S9378 Sigma),
ribose (C5H10O5, D-(−)-ribose ≥ 99% R7500 Sigma),
lactose (Lac, C12H22O11·H2O, β-D-galactose-(1→4)-α-Dglucose,
α-Lactose monohydrate reagent grade L3625
Sigma-Aldrich), aqueous solutions in a concentration of
0.001 mg/mL.
RESULTS AND DISCUSSION
Animal-based products have a protein content of 10
to 25% [18, 29]. Fresh raw materials of animal origin
usually contain from 0.001 to 0.01% of free amino acids,
and their content increases with prolonged storage due to
internal enzyme systems. We determined the amino acid
composition of the model protein and the meat system
protein before and after introducing Lactobacillus
plantarum and Staphylococcus carnosus (Table 1). We
found that the starter cultures significantly increased
the total content of free amino acids both in the model
protein and in the formulation. In the formulation,
their content increased to 2.0 ± 0.1%, 2.2 ± 0.1%,
and 2.8 ± 0.1% after using Lactobacillus plantarum,
Staphylococcus carnosus, and an equimolar mixture of
Lactobacillus plantarum and Staphylococcus carnosus,
respectively.
With the same total concentration of introduced
cultures at 4×109 CFU/kg, the mixture of Lactobacillus
plantarum and Staphylococcus carnosus increased the
rate of protein hydrolysis to free amino acids by 30–40%
(P > 0.96), compared to their separate action.
To determine the treatment time, we observed
changes in the release of free amino acids and the
correlating values of water retention (Fig. 1). We found
Figure 1 Content of free amino acids (curves 1, 4) and water
retention (curves 2, 3) vs. treatment time with Lactobacillus
plantarum and Staphylococcus carnosus (1) and without
them (4)
Amino acids, g/100 g
Time, wk.
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that the optimal treatment time was three weeks. This
time ensured optimal quality parameters of the product,
including water retention (Fig. 1).
According to Fig. 1, a joint use of the bacterial
cultures ensured the maximum concentration of free
amino acids in two weeks. In addition, the water
retention capacity of the food system – an important
technological indicator of product quality – was almost
twice as high as when the process lasted a month
without using bacterial cultures.
The proteolytic activity with respect to the model
and food system proteins showed a synergistic effect of
L. plantarum and S. carnosus on the protein components
(Table 2). Their joint use increased the efficiency of
hydrolytic decomposition of the system proteins leading
to a release of free amino acids (Table 1).
Table 3 shows the composition of fatty acids in the
animal-based food system and changes in their contents
under the influence of L. plantarum and S. carnosus.
At the initial stages, we found a decrease in the
contents of lower C6–C10 and unsaturated fatty acids,
especially essential omega-3 acids (α-оctadecatrienoic
C18:3, eicosapentaenoic C20:5, and docosapentaenoic
C22:5). However, there was an increase in cis-11,14,17-
eicosatrienoic С20:3 acid that is important for proper
nutrition of mammals. Three weeks of treating the
food system with L. plantarum and S. carnosus led to
a decrease in unsaturated fatty acids and an increase
in saturated acids by 1–5% (P > 0.95). Similar changes
in contents of saturated and unsaturated fatty acids are
usually observed for animal-based products subjected to
long-term storage at low temperatures [22, 23].
We did not evaluate the direct lipolytic activity of
L. plantarum and S. carnosus in the presence of
synthetic substrates commonly used for this purpose.
However, Table 3 shows that a combined action of the
cultures on the system led to a more efficient breakdown
of animal fats and a release of free fatty acids, compared
to their individual action. Yet, this effect was not
expressed clearly.
Thus, the action of L. plantarum and S. carnosus
on the fat components of the food system not only
transformed the fatty acid composition, but also, to a
much greater extent, increased the amount of free fatty
acids.
Fig. 2 shows a kinetic curve of changes in the
content of free fatty acids as a result of treatment with
L. plantarum and S. carnosus. Longer treatment time
led to a higher mass fraction of free fatty acids in all
cases. We found no differences in the kinetics of free
fatty acids formation under individual or joint action of
the cultures, apparently due to their comparable lipolytic
activity.
Table 1 Amino acid composition of model and food system proteins and the content of free amino acids after treatment with
Lactobacillus plantarum and Staphylococcus carnosus
Name
of amino acid
Model
protein
Food system proteins Model protein Food system proteins
g/100g product g/100g protein LP SC LP+SC LP SC LP+SC
Asparagine 7.98 1.43 7.24 0.14 0.04 0.17 0.19 0.17 0.16
Glutamine 16.4 2.95 15.9 0.81 0.31 0.54 0.87 0.73 0.56
Serine 1.59 0.28 1.52 0.01 0.01 0.10 0.03 0.13 0.13
Histidine 3.46 0.62 3.38 0.08 0.01 0.08 0.08 0.08 0.14
Glycine 2.24 0.40 2.19 0.08 0.01 0.12 0.11 0.13 0.11
Threonine 5.91 1.06 5.94 0.04 0.02 0.06 0.03 0.07 0.09
Arginine 7.80 1.40 7.77 0.10 0.05 0.13 0.12 0.13 0.12
Alanine 3.63 0.65 3.54 0.18 0.04 0.13 0.16 0.15 0.15
Tyrosine 3.98 0.71 3.76 0.04 0.01 0.09 0.05 0.07 0.12
Cysteine 1.18 0.21 1.15 0.02 0.01 0.04 0.01 0.02 0.04
Valine 5.56 1.00 5.48 0.07 0.01 0.08 0.07 0.06 0.09
Methionine 3.35 0.60 3.41 0.02 0.02 0.04 0.01 0.02 0.05
Phenylalanine 4.55 0.81 4.53 0.01 0.01 0.12 0.02 0.04 0.09
Isoleucine 4.93 0.88 4.91 0.01 0.02 0.14 0.02 0.08 0.13
Leucine 8.57 1.54 8.49 0.03 0.04 0.12 0.03 0.07 0.13
Lysine 10.6 1.91 10.5 0.22 0.17 0.58 0.16 0.17 0.54
Proline 3.10 0.55 3.06 0.03 0.03 0.08 0.03 0.06 0.12
Tryptophan 1.32 0.23 1.31 0.04 0.01 0.03 0.01 0.02 0.03
Ʃ 96.2 17.23 94.1 1.93 0.82 2.65 2.00 2.2 2.8
LP – Lactobacillus plantarum; SC – Staphylococcus carnosus
Table 2 Proteolytic activity of Lactobacillus plantarum
and Staphylococcus carnosus in relation to proteins of animalbased
food systems, units/mg
Name LP SC LP+SC
Model protein 12 14 17
Food system protein 6 4 8
LP – Lactobacillus plantarum; SC – Staphylococcus carnosus
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The synergistic effect of L. plantarum and
S. carnosus on the fat components of the animalbased
food system did not manifest reliably, since the
amounts of free fatty acids in the final products were
approximately the same.
Thus, the action of L. plantarum and S. carnosus
resulted in not only the hydrolytic decomposition
of fat components, but also in their biochemical
transformation into ultimate chemical structures. The
increased content of saturated acids found in our study
should be considered critically, in light of current trends
in the production of foods with an increased amount of
unsaturated, especially polyunsaturated, fatty acids
of the omega-3 family. This problem should be taken
into account in further development of methods for
producing animal-based foods.
Table 3 Changes in the fatty acid composition of the animal-based food system treated with Lactobacillus plantarum,
Staphylococcus carnosus and their mixture, % of total
Name of fatty acid Initial
mixture
3 weeks 4 weeks without
LP SC LP+SC cultures
Caproic С 6:0 0.06 0.05 nd – –
Octanoic С 8:0 0.17 – – 0.04 –
Decanoic С10:0 1.05 1.24 0.48 0.88 0.65
Decenoic С 10:1 0.14 0.12 0.08 0.07 0.03
Undecanoic C11:0 0.12 0.06 0.06 – 0.02
Dodecanoic С12:0 0.75 0.79 1.28 1.14 1.23
Tridecanoic C13:0 – – 0.07 – 0.08
Tetradecanoic С14:0 2.61 2.18 2.16 2.2 2.34
Cis-9-tetradecenoic С 14:1 0.52 0.21 0.18 0.16 0.15
Pentadecanoic C15:0 – – 0.12 – 0.12
Cis-10-pentadecenoic C15:1 0.29 0.22 0.17 0.16 0.14
Hexadecanoic С16:0 19.3 21. 22.8 22.6 23.1
Cis-9-hexadecenoic С16:1 4.23 2.24 2.55 2.45 3.18
Heptadecanoic С17:0 0.51 – 0.16 0.15 0.33
Cis-10-heptadecenoic С17:1 0.18 0.14 0.1 0.12 0.24
Octadecanoic С18:0 21.6 24.1 22.4 23.9 23.6
Cis-9-octadecenoic С18:1n9c 18.8 19.8 19.2 19.1 18.7
Trans-9-octadecenoic С18:1n9t – – 0.01 – 0.02
Cis-9,12-octadecadienoic С18:2n6 4.56 3.62 4.23 3.56 3.88
Cis-6,9,12-оctadecatrienoic С18:3n6 3.44 3.02 3.17 2.34 2.51
Cis-9,12,15-оctadecatrienoic С18:3n3 0.55 0.44 0.61 0.23 0.43
Nonadecanoic С19:0 0.07 0.16 0.41 0.28 0.25
Eicosanoic С20:0 0.35 0.48 0.49 0.61 0.56
Cis-9-eicosenoic С20:1n9 0.45 0.43 0.47 0.45 0.38
Cis-11,14-eicosadienoic С20:2n6 3.0 4.45 4.13 4.65 3.62
Cis-8,11,14-eicosatrienoic С20:3n6 2.05 3.0 3.23 3.18 3.17
Cis-11,14,17-eicosatrienoic С20:3n3 0.41 0.5 0.38 0.44 0.39
Cis-5, 8, 11, 14-eicosatetraenoic С20:4n6 0.45 – 0.32 0.11 0.24
Cis-5, 8,11,14,17-eicosapentaenoic С20:5n3 0.06 0.02 0.12 0.13 0.07
Heneicosanoic C21:0 0.11 0.11 0.06 – 0.15
Docosanoic С22:0 0.12 – 0.13 0.12 0.16
Cis-13-docosenoic С22:1n9 0.11 0.25 0.2 0.11 0.11
Cis-13,16-docosadienoic C22:2 n6 0.46 0.19 0.11 – –
Cis-7,10,13,16,19-docosapentaenoic С22:5n3 0.05 0.03 – 0.04 0.01
Cis-4,7,10,13,16,19-docosahexaenoic С22:6n3 0.04 0.02 – – 0.03
Tricosanoic С23:0 0.1 0.2 0.16 0.15 0.15
Tetracosanoic C24:0 0.21 – 0.12 – 0.18
Cis-15-tetracosenoic С24:1 – 0.23 0.06 – –
Unidentified fatty acids 13.1 10.7 9.8 10.6 9.7
Mass fraction of free fatty acids 0.01 0.2 0.2 0.3 0.3
nd – not detected
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Table 4 shows the quantitative identification of
free carbohydrates in the food system treated with
L. plantarum and S. carnosus. As we can see, the
content of some sugars (galactose, glucose, fructose)
decreased, while the content of others increased.
It appears that the decrease was caused by the
consumption of those sugars by the cultures themselves,
whereas the increase was associated with proteolytic
and carbohydrolase activity, leading to the breakdown
of animal polysaccharides. The mass fraction of such
polysaccharides in animal raw materials is 2–3%,
but their decomposition can lead to the formation of
0.1–100 mg% free carbohydrates [18, 23].
According to Table 4, free monosaccharides formed
most intensively under the joint action of L. plantarum
and S. carnosus. It manifested through changes in
the content of galactose, glucose, xylose, and ribose
and through a higher rate of disaccharides (lactose
and sucrose) formation in the food system. In fact, the
amount of free lactose was almost twice as high as when
the cultures were used individually. As a result, the
product acquired a sweetish taste. Thus, L. plantarum
and S. carnosus produced a synergistic effect on the
changes in carbohydrate components of the animalbased
food system.
Further, treating food systems with microcultures
totally changes the chemical composition of organic
substances in raw materials and intermediates or those
substances formed as a result. Table 5 shows changes
in some minor components of the food system in the
presence of L. plantarum and S. carnosus. The mass
spectrometric analysis made it possible to identify over
250 organic compounds which could be considered
as a result of biochemical effects that microorganisms
had on protein, lipid, and carbohydrate components of
the food system. A large amount of those compounds
were derivatives of fatty acids. Table 5 lists the main
substances identified, whose content exceeded 0.001%.
As can be seen in Table 5, the joint use of
L. plantarum and S. carnosus resulted in more
pronounced changes in almost all compounds, compared
to their individual action. To some extent, this result
indicated a synergistic mechanism of action of both
cultures used to treat animal raw materials.
Given that all identified substances can affect the
taste and aroma of final products, varying the use of
starter cultures – both individual and joint – can make
it possible to obtain products with a wide range of
consumer properties [21].
Finally, the biochemical transformations of
cholesterol require our special attention (Table 5). This
substance is a significant component of food systems
based on animal raw materials. Its high content in
products is considered as an unfavorable factor leading
to the development of atherosclerosis. In our case,
the combined action of the cultures led to a more
considerable degradation of cholesterol, which is an
important advantage of using these cultures together.
CONCLUSION
Thus, the joint use of starter cultures, Lactobacillus
plantarum and Staphylococcus carnosus, to treat
animal-based food systems not only increased the yield
of the product, but also had a synergistic effect on the
protein, lipid, and carbohydrate components of the
system. This may allow us to change the component
composition of the system and form the desired
characteristics of the food product.
Figure 2 Changes in the acid number of the animal-based food
system caused by treatment with Lactobacillus plantarum and
Staphylococcus carnosus (1), Staphylococcus carnosus (2) and
without starter cultures (3)
Table 4 Changes in free carbohydrate contents in the animal-based food system caused by Lactobacillus plantarum,
Staphylococcus carnosus and their mixture, % (g/100 g of sample weight)
Name Initial mixture LP LP+SC SC Without starter cultures
arabinose 0.016 0.13 0.33 0.16 0.23
galactose 1.85 0.0025 0.004 0.0018 0.0015
glucose 0.03 0.002 0.003 0.002 0.001
xylose + mannose 0.03 0.008 0.05 0.022 0.028
fructose + sucrose 0.08 0.006 0.13 0.095 0.053
ribose 0.15 0.095 0.19 0.11 0.12
lactose 0 0.08 0.18 0.085 0.12
LP – Lactobacillus plantarum; SC – Staphylococcus carnosus
0.00
0.20
0.40
0 1 2 3 4
Acid number, mg KOH/g
Time, wk.
1 2 3
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Table 5 Chemical components in products with Lactobacillus plantarum and Staphylococcus carnosus (n = 6, P = 0.95)
Name CAS No. Relative intensity of
mass spectrum main
signals, units
Probability of mass
spectrometric peak
identification from
the mass spectra
library, %
Raw
material,
mg/kg
Final product, mg/kg Time,
LP SC LP+SC minutes
2-phenyl-4-quinolinol 001144-20-3 221(999), 193(44),
220(183), 222(171),
165(134)
66 nd nd 0.003 0.008 4.123
2,5-diphenyl-oxazole 000092-71-7 221(999), 165(617),
166(388), 892(386),
77(351)
70 nd nd 0.003 0.002 4.174
Cyclobarbital 000052-31-3 207(999), 141(195),
81(189), 67(132),
79(122)
66 0.002 0.003 0.009 0.013 4.237
Nonanal dimethyl acetal 018824-63-0 75(999), 157(97),
41(88), 69(86),
43(83)
78 nd 0.001 0.012 0.013 4.346
8H-dinaphtho[2,3-
b:2’,3’-g]carbazole
003557-50-4 367(999), 368(243),
366(168), 183(106),
184(98)
72 0.001 0.001 0.004 0.01 6.603
Decanoic acid, methyl
ester **
000110-42-9 74(999), 87(585),
55(249), 43(205),
143(198)
96 0.002 0.004 nd 0.006 8.933
7-methoxy-9-(3-methyl-
2-butenyl)-furo[2,3-b]
quinolin-4(9H)-one
018904-40-0 215(999), 283(222),
216(133), 284(42),
172(37)
74* 0.002 0.001 0.003 0.011 10.271
Ethyl-6-amino-4-[pchloroanilino]-
5-nitro-2-
pyridincarbamate
021271-60-3 351(999), 43(432),
353(338), 352(297),
270(208)
88 0.003 0.006 0.003 0.005 10.889
1-hexadecenyl methyl
ester
015519-14-9 71(999), 41(330),
82(270), 43(170),
96(150)
96 nd 0.005 0.423 0.579 14.007
1,1-dimethoxy-9-
octadecene
015677-71-1 31(999), 71(850),
32(730), 29(520),
41(430)
91 nd 0.046 0.061 0.067 15.112
Methyl-1-octadecenyl
ether
026537-06-4 71(999), 82(370),
41(300), 43(210),
68(190)
93 0.009 nd 0.017 0.169 15.252
2-methyl-1H-indole 000095-20-5 130(999), 131(657),
77(131), 103(110),
51(83)
63 0.056 0.108 0.106 0.680 16.970
cis-11-hexadecenal 053939-28-9 55(999), 41(554),
69(436), 67(392),
81(376)
90 0.03 0.027 0.037 0.082 18.298
Cholesterol 000057-88-5 43(999), 55(886),
57(744), 105(686),
86(681)
98 0.331 0.097 0.145 0.065 22.854
nd – not detected
* – not detected at less than 0.001 mg/kg
** – compounds identified as methyl esters by the method
LP – Lactobacillus plantarum; SC – Staphylococcus carnosus
CONTRIBUTION
A.N. Ivankin led the project; he set the research
problem and the objects of study and decided on the
methods. A.N. Verevkin conducted experimental
work with the strains of cultures. A.S. Efremov
developed formulations and determined their
physicochemical parameters. N.L. Vostrikova identified
and experimentally confirmed the combined effect
of the cultures on the protein components of the food
system. A.V. Kulikovskii identified and experimentally
confirmed the microcultures’ effect on the lipid
components, as well as established, through mass
spectrometry, their synergistic action on the main
chemical components of the food system. M.I. Baburina
284
Ivankin A.N. et al. Foods and Raw Materials, 2020, vol. 8, no. 2, pp. 277–285
identified and experimentally confirmed the combined
effect of the cultures on the carbohydrate components of
the food system.
CONFLICTS OF INTEREST
The authors declare that there is no conflict of
interests.

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