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Експериментальні статті
97
Експериментальні статті
Organic acids play an important role in wine
biotechnology because of the taste im par ted by
the concentration, of the different organic
acids. Organic acids in wine include tartaric,
malic, citric, lactic, acetic, succinic and others.
Concentration of organic acids varies depending
on different factors such as temperature, pH,
concentration of oxygen and carbon dioxide.
Wine taste depends mainly on the ratio of
tartaric acid to malic one. If the ratio of these
acids is about 2 or less, the wine will not be har-
monious and will have a sour aftertaste. On the
contrary, wine stock with the best flavor and
bouquet will be obtained at a ratio of tartaric to
malic acid equal coefficient 3 and more.
Some organic acids have their own specific
taste and aroma. For example citric acid gives
freshness to wine, succinic acid has salty-bit-
ter taste, and malic acid gives the taste of
green apples. However, malic acid may exhibit
different flavours depending on the concen-
trations of ethanol, tannic acids, sugar, aro-
matic and mineral substances. Making quanti-
tative changes in the ratio of these compounds
makes it possible to achieve the tastes of dif-
ferent harmonious acidity [1, 2].
Buffering also plays an important role in
formation of taste that results mostly from
potassium ions in malic acid and, to a lesser
extent, from small ions of Ca
2+
, Mg
2+
, and Na
+
.
In sparkling wines, acidic harmony is con-
trolled by concentrations of sugar and carbon
dioxide, but in dry wines it is controlled by
concentrations of organic acids [3, 4].
Interaction of organic acids and alcohol,
with their degradation products (example of
tartaric acid degradation) play an important
role in forming of organoleptic properties of
wine stocks, which is very important in wine
biotechnology.
Organic acids commence to be accumulated
in the grapes at the moment when grapes begin
to accumulate and concentrate sugar. At the
moment of reduced accumulation of organic
acids, titratable acidity decreases sharply [5, 6].
During fermentation of grape must with
high titratable acidity, decreasing acidity
indicates the completion of fermentation. The
amount of tartaric and malic acids decrease,
however the amount of citric and succinic
acids increase, which is natural. Lactic acid is
formed, which is impossible to find in the ripe
grapes. Wine stocks at a high pH have a sour
taste. Therefore, strict adherence to deadlines
in harvesting grapes will be a key to successful
formation of optimal concentration of organic
UDC 634.8:632.93:661.74:663.2
ORGANIC ACIDS CONCENTRATION
ORGANIC ACIDS CONCENTRATION
IN WINE STOCKS AFTER
IN WINE STOCKS AFTER
Saccharomyces cerevisiae
Saccharomyces cerevisiae
FERMENTATION
FERMENTATION
Key words: organic acids, grape varieties, Saccharomyces cerevisiae.
V. N. Bayraktar Odesa Mechnikov National University, Ukraine
Email: vogadro2007@rambler.ru
Received 24.08.2012
The biochemical constituents in wine stocks that influence the flavor and quality of wine are investi-
gated in the paper. The tested parameters consist of volume fraction of ethanol, residual sugar, phenolic
compounds, tartaric, malic, citric, lactic, acetic acids, titratable acidity and volatile acids. The wine stocks
that were received from white and red grape varieties Tairov`s selection were tested. There was a correla-
tion between titratable acidity and volatile acids in the wine stocks from white and red grape varieties.
High correlation was also found between lactic and acetic acids, between volatile acids, acetic acid and
sugar. It was determined that wine stocks with a high concentration of ethanol originated from those yeast
strains of Saccharomyces cerevisiae, in a fermented grape must of high speed of enzyme activity. The taste
of wine stocks correlated with the ratio of tartaric to malic acid. Analysis showed significant differences
between the varieties of white and red wine stocks in concentrations of organic acids, phenolic compounds,
residual sugar, and volume fraction of ethanol. Positive correlation was indicated for both studied groups
for volatile acids and acetic acid, tartaric, malic, lactic acids and total sugar. Prospective yeast cultures
with high productivity of alcohol (ethanol) were selected for winemaking biotechnology.
BIOTECHNOLOGIA ACTA, V. 6, No 2, 2013
98
acids for preparation of high-quality wines
with excellent taste and aroma.
Organic acids in wine are of technological
importance and have different dissociation con-
stants. Most of strong organic acids in grapes
and wine is tartaric acid — (K
dis
=1.3× 10
–3
),
next is citric acid (K
dis
=8.4×10
–4
), malic acid
(K
dis
=3.95×10
–4
) , lactic acid (K
dis
=1.4 ×10
–4
),
succinic acid (K
dis
=7.4×10
–5
), acetic acid
(K
dis
=1.8×10
–5
) [7, 8]. Organic acids in wine
play an important role in the taste and quality
of wines. Because a low content of organic acids
resulting in insufficient acidity, wine loses its
fullness and roundness of taste and becomes
characterless and expressionless [9–11].
Each type of wine must correspond to its
optimum acidity. Tingling acidity typical of
sparkling wines, up to incompletely fermented
or freshly fermented grapes must contain car-
bon dioxide, which is produced during fermen-
tation. Organic acids preserve the wine from
bacterial diseases. Under acidic conditions the
redox processes proceed slower, slowing down
the maturation of wine, and prevents iron or
iron-phosphate turbidity as well. Organic
acids influence the bouquet of wine, and form
complex esters with alcohols. A special role in
the maturation of wines belongs to tartaric
acid, which is converted to dioxy-fumaric acid
and restores the taste of wine [12, 13].
The aim of this work is to establish correla-
tions between quantitative contents of organic
acids and the quality of the resulting wine fer-
mented on yeast culture isolated from diffe -
rent industrial varieties in the Tairov`s selec-
tion. To achieve this goal, the following tasks
were undertaken.
To determine the concentrations of the
organic acids in wines made from different
industrial varieties of Tairov`s selection it was
needed to determine among them tartaric, cit-
ric, malic, lactic and acetic acids. Based on
these determinations it would be easier to
determine the quality of investigated wines
using the ratio between tartaric and malic
acids.
Materials and methods
Samples of wine from different industrial
grape varieties were collected after comple ting
fermentation with different yeast cultures of
Saccharmyces cerevisiae during the vintage
season from the vineyard of the Ukrainian
Tairov`s Research Institute of Viticulture and
Oenology, located in the Odessa region of
Ukraine. The total number of the species
selected for the research was thirteen. The fol-
lowing industrial grape varieties were selected
for the research:
– white wine grape varieties: Aromatic,
Odessa`s Muscat, Opalovy (Opaline), Ovidio -
polskij, Selena, Sukholimansky, Zagrey.
– red wine grape varieties: Charivny (Ma -
gic), Illychevsky Early, Odessa`s Black, Odes -
sa`s Pearl, Ruby Jubilee, Tairov`s Ruby.
Yeast strains:
I. Laboratory yeast cultures isolated from
grape must from the vineyard of the Ukrai -
nian Tairov`s Research Institute of Viticul tu -
re and Oenology.
Yeast culture isolated from white wine
grape varieties:
*Y-3444; *MAFF-230106. Saccharo my ces
cerevisiae isolated from grape must of the
variety Aromatic;
Y-3445; MAFF-230107. Saccharomyces
cerevisiae isolated from grape must of the
variety Odessa`s Muskat;
Y-3441; MAFF-230103. Saccharomyces
cerevisiae isolated from grape must of the
variety Opalovy (Opaline);
Y-3442; MAFF-230104. Saccharomyces
cerevisiae isolated from grape must of the
variety Ovidiopolskij;
Y-3439; MAFF-230101. Saccharomyces
cerevisiae isolated from grape must of the
variety Selena;
Y-3440; MAFF-230102. Saccharomyces
cerevisiae isolated from grape must of the
variety Sukholimansky;
Y-3443; MAFF-230105. Saccharomyces
cerevisiae isolated from grape must of the
variety Zagrey.
The designated yeast culture numbers were
derived from those deposited in the MAFF
Collection, National Institute of Agrobio lo gi -
cal Sciences, Genbank of Japan.
* MAFF — Culture Collection of microor-
ganisms Ministry of Agriculture, Forestry
and Fisheries, Tsukuba, Ibaraki, Japan.
Yeast culture isolated from red wine grape
varieties:
Y-3438; MAFF-230100. Saccharomyces
cerevisiae isolated from grape must of the
variety Charivny (Magic);
Y-3448; MAFF-230110. Saccharomyces
cerevisiae isolated from grape must of the
variety Illychevsky Early;
Y-3447; MAFF-230109. Saccharomyces
cerevisiae isolated from grape must of the
variety Odessa`s Black;
Y-3446; MAFF-230108. Saccharomyces
cerevisiae isolated from grape must of the
variety Odessa`s Pearl;
Експериментальні статті
99
Y-3437; MAFF-230099. Saccharomyces
cerevisiae isolated from grape must of the
variety Ruby Jubilee;
Y-3436; MAFF-230098. Saccharomyces
cerevisiae isolated from grape must of the
variety Tairov`s Ruby.
Pure yeast cultures were isolated from
grapes and followed by fermentation by using
traditional microbiological methods consis -
ting of inoculation of a sample into a Petri dish
with a few modifications of nutrient selective
agar for yeast isolation and cultivation.
Primary yeast isolation was carried out using
Inhibitory Mold Agar medium (IMA) (Becton
Dickinson Company, USA). Morphological
properties of the yeast culture were analyzed
after the primary yeast culture isolation.
Yeasts were identified by polymerase chain
reaction (PCR) using universal yeast primers.
Then yeast cultures were cultivated on a Wort
Agar medium (Becton Dickinson Company,
USA). Each isolated, yeast culture was
deposited in the NRRL Culture Collection
(National Regional Research Laboratory),
Peoria, USA, in the British National
Collection of Yeast Culture (NCYC), Norwich,
UK and Genbank of Japan.
All biochemical parameters were tested in
the wine stocks followed by fermentation of
the white and red grape varieties Tairov`s
selection.
The volume fraction of ethanol, total sugar
(glucose/fructose), and pH were determined
by the spectroscopy method using the
Bacchus-II spectrometer (Microdom Company,
France).
Concentrations of organic acids: tartaric,
malic, citric, lactic, acetic and phenolic com-
pounds were determined using a method of
liqui d chromatography (Ultimate 3000, Dio -
nex Company, Germany).
Statistical deviation and significance were
evaluated by the Student`s t-test with P —
value: P < 0.1; P < 0.05; P < 0.01. We calcu-
lated Spearman’s rank correlation coefficient
for the tested biochemical parameters between
tested wine stocks from different white and
red grape varieties of the Tairov`s selection.
Each biochemical test was repeated three
times to confirm the exact result. For the
groups of white and red wine grape varieties,
fermentation made with different Saccharo -
my ces cerevisiae yeast cultures dispersion
analysis (ANOVA) was done as well. The dis-
persion analysis (ANOVA) based on the
Fisher`s test (unifactorial model) was applied,
where F- criterion determined whether the re -
levant samples belong to one from general
aggregate and then possible to pool them or
not. Standard deviation was calculated, statis-
tical significance of the difference was evalua -
ted by the Student’s t-test.
Results and discussion
The results of the organic acid study in
wine stocks received from the white and red
grape varieties had specific differences in con-
centrations of organic acids. Results of orga -
nic acids concentrations are given in table 1
for white grape varieties and in table 2 for red
grape varieties. The normal ranges of tested
parameters in wine stocks are given in Table 3.
The study showed that concentration of
ethanol and residual sugar in wine stocks
depends on enzymatic activity of Saccharo -
myces cerevisiae yeast strains which are used
for winemaking. For example, the index of
ethanol produced by yeast cultures in wine
stocks received from white grape varieties has
concentrations of ethanol compared with min-
imal amounts in: Odessa`s Muscat MAFF-
230107 more than 125%, Opalovy (Opaline)
MAFF-230103 more than 122%, Selena
MAFF-230101 more than 112%.
Study of the morphology of Saccharo my -
ces cerevisiae yeast followed by fermentation
illustrate that between wine stocks of white
and red grape varieties there exist some dif-
ferences in level of ethanol production. It`s
more for yeast cultures isolated from white
grape varieties and less (moderate) for yeast
cultures isolated from red grape varieties. All
the yeast cells were stained by Gram method.
Some cells of yeast are large and either roun -
ded or oval shaped (Fig. 1–6).
In wines received from red grape varieties,
the index of ethanol produced in wine stocks
showed maximal concentration of ethanol,
compared with minimal amounts in such
Fig. 1. Morphology of Saccharomyces cerevisiae
yeast culture isolated from white grape variety
«Odessa`s Muscat» MAFF-230107:
the volume fraction of alcohol (ethanol) produc-
tion — 15.55 v/v%; stained by Gram method;
magnification —
×720
BIOTECHNOLOGIA ACTA, V. 6, No 2, 2013
100
Table 1. Organic acid content (g/L) after fermentation and formation of wine stocks from grape varieties
of Tairov`s selection (white wine grape varieties)
Wine
stocks
from
white
grape
vari-
eties
MAFF
Collec -
tion
number
in
Genbank
of Japan
Titrata
ble
acidity
in
terms
of tar-
taric
acid
(g/L)
volatile
acids in
terms
to
acetic
acid
(g/L)
Tartaric
acid
(g/L)
Malic
acid
(g/L)
The
ratio of
tartaric
acid to
malic
acid
Citric
acid(g/
L)
Lactic
acid
(g/L)
Acetic
acid
(g/L)
Commo
n sugar
glu-
cose/
fruc-
tose
(g/L)
The
volume
frac-
tion of
the
ethanol
(% v/v)
Concen
tration
of total
pheno-
lic com-
pounds,
(mg/L)
Aroma -
tic
230106
5.67±
0.35
**0.53
±0.02
2.9±
0.24
**0.98
±0.02
*2.95±
0.1
**0.09
±0.02
**1.21
±0.03
0.39±
0.08
**1.14
±0.04
13.79±
1.08
270.0±
30.41
Odessa`s
Muscat
230107
*4.80±
0.1
*1.15±
0.1
**1.41
±0.02
**1.29
±0.04
*1.09±
0.1
**0.10
±0.01
*0.85±
0.1
1.20±
0.2
1.07±
0.08
15.55±
0.8
279.0±
22.4
Opalovy
(Opaline)
230103
5.19±
0.1
0.69±
0.05
**1.11
±0.03
1.70±
0.2
0.65±
0.07
**0.23
±0.02
**0.67
±0.03
**0.25
±0.03
*1.61±
0.1
14.94±
1.6
248.8±
16.43
Ovidio -
polskij
230104
6.67±
0.2
0.43±
0.09
5.04±
0.3
1.94±
0.06
*2.59±
0.1
**0.09
±0.02
1.45±
0.06
**0.23
±0.02
2.3±
0.2
*7.02±
0.1
147.0±
13.2
Selena
230101
*6.21±
0.1
*0.85±
0.1
3.27±
0.2
1.55±
0.2
2.10±
0.2
***0.0
8±0.01
**0.69
±0.04
**0.42
±0.02
*1.53±
0.1
*14.15
±0.1
841.8±
11.0
Sukholi -
mansky
230102
5.8±
0.5
*0.68±
0.1
*3.01±
0.1
*1.84±
0.1
*1.63±
0.1
***0.07
±0.01
**0.76
±0.02
***0.1
2±0.01
1.83±
0.07
13.4±
0.4
384.6±
11.0
Zagrey
230105
6.47±
0.2
**0.90
±0.05
*4.2±
0.1
*2.24±
0.1
1.87±
0.09
***0.06
±0.007
**0.41
±0.04
**0.54
±003
**1.05
±0.05
9.92±
0.4
282.1±
13.0
Fig. 2. Morphology of Saccharomyces cerevisiae
yeast culture isolated from white grape variety
«Opalovy (Opaline)» MAFF-230103:
the volume fraction of alcohol (ethanol) produc-
tion — 14.94 v/v%; stained by Gram method;
magnification —
×720
Fig. 3. Morphology of Saccharomyces cerevisiae
yeast culture isolated from white grape variety
«Selena» MAFF-230101:
the volume fraction of alcohol (ethanol) produc-
tion — 14.15 v/v%; stained by Gram method;
magnification —
×720
Note: P–value * P ≤ 0.1; ** P ≤ 0.05; *** P ≤0.01.
Fig. 4. Morphology of Saccharomyces cerevisiae
yeast culture isolated from red grape variety
«Charivniy (Magic)» MAFF-230100:
the volume fraction of alcohol (ethanol) produc-
tion — 12.27 v/v%; stained by Gram method;
magnification —
×720
Fig. 5. Morphology of Saccharomyces cerevisiae
yeast culture isolated from red grape variety
«Odessa`s Black» MAFF-230109:
the volume fraction of alcohol (ethanol) produc-
tion — 11.22 v/v%; stained by Gram method;
magnification —
×720
Експериментальні статті
101
Table 2. Organic acid content (g/L) after fermentation and formation of wine stocks from grape varieties
of Tairov`s selection. (red wine grape varieties)
Wine
stocks
from
white
grape
vari-
eties
MAFF
Collec -
tion
number
in
Genbank
of Japan
Titrata -
ble
acidity
in terms
of tar-
taric
acid
(g/L)
volatile
acids in
terms
to
acetic
acid
(g/L)
Tartaric
acid
(g/L)
Malic
acid
(g/L)
The
ratio of
tartaric
acid to
malic
acid
Citric
acid
(g/L)
Lactic
acid
(g/L)
Acetic
acid
(g/L)
Commo
n sugar
glu-
cose/
fruc-
tose
(g/L)
The
volume
frac-
tion of
the
ethanol
(% v/v)
Concen
tration
of total
pheno-
lic com-
pounds,
(mg/L)
Chariv -
ny
(Magic)
230100
6.0±
0.2
*0.69±
0.1
2.74±
0.2
*1.45±
0.1
*1.88±
0.1
0.09±
0.03
0.72±
0.06
0.24±
0.02
*1.55±
0.1
12.27±
0.5
675.9±
13.8
Illychev -
sky
Early
230110
*10.71
±0.1
*1.49±
0.1
2.27±
0.2
0 –
0.09±
0.04
6.71±
0.2
3.60±
0.2
5.38±
0.2
*10.54
±0.1
1004.2
±19.5
Odessa`s
Black
230109
7.28±
0.2
***0.1
8±0.01
5.81±
0.2
**1.36
±0.05
4.27±
0.04
0.19±
0.04
**0.49
±0.05
***0.02
±0.005
*1.62±
0.1
11.22±
0.3
543.6±
11.0
Odessa
Pearl
230108
11.89±
0.2
1.6±
0.2
2.76±
0.2
0 –
***0.1
7±0.01
*7.51±
0.1
3.1±
0.3
8.25±
0.2
8.11±
0.3
1123.4
±43.7
Ruby
Jubilee
230099
7.49±
0.2
0.44±
0.04
4.21±
0.2
*1.09±
0.1
3.86±
0.02
0.07±
0.02
0.45±
0.04
**0.23
± 0.05
4.15±
0.3
7.2±
0.3
654.3±
18.7
Tairov`s
Ruby
230098
*6.05±
0.1
**0.76
±0.05
2.84±
0.2
*1.75±
0.1
*1.62±
0.1
0.11±
0.02
0.59±
0.04
0.45±
0.03
6.23±
0.2
12.27±
0.5
645.4±
15.3
Note: P–value * P ≤ 0.1; ** P ≤ 0.05; *** P ≤ 0.01.
Table 3. Normal range of organic acid content (g/L) after grape must fermentation and formation
of wine stocks from grapes
Group
of wine
stock
Titratable
acidity in
terms of
tartaric
acid (g/L)
volatile
acids in
terms to
acetic
acid
(g/L)
Tartaric
acid
(g/L)
Malic
acid
(g/L)
The
ratio of
tartaric
acid to
malic
acid
Citric
acid
(g/L)
Lactic
acid
(g/L)
Acetic
acid
(g/L)
Commo
n sugar
glu-
cose/
fructose
(g/L)
The
volu me
fraction
of the
ethanol
(% v/v)
Concentr
ation of
total phe-
nolic com-
pounds,
(mg/L)
pH
For
white
dry
wines
not less
than 3.5
1.20 1.5–5
From
traces
up to
5
3 and
more
no
more
than
0.8
from
traces
up to
0.5–5
0.4–
1.5
less
than
4.0
8.5–15
1200–
1500
2.8–
3.8
For red
dry
wines
5.0-7.0 1.50 1.5-5 0-5
3 and
more
no
more
than
1.0
1–5
0.4–
1.5
less
than
4.0
10–15
1800–
3700
3.0–
4.6
Note: There are no strict standards of content for the concentration of organic acids in dry grape wines. However
research Laboratories established their own standards based on grape growing regions.
Fig. 6. Morphology of Saccharomyces cerevisiae yeast culture isolated
from red grape variety «Tairov`s Ruby» MAFF-230098:
the volume fraction of alcohol (ethanol) production — 12.27 v/v%: stained by Gram method;
magnification —
×720
BIOTECHNOLOGIA ACTA, V. 6, No 2, 2013
102
Table 4. Dispersion analysis (ANOVA) represents parameters of organic acid concentration
and other components in the wine stocks made from white and red grape varieties of the Tairov`s selection
Tested parameters
White dry wine stocks Red dry wine stocks
F P r F P r
Titratable acidity/ volatile acids 349.09 <0.0001 -0.47 50.47 <0.0001 0.82
Titratable acidity/tartaric acid 23.27 0.0004 0.95 17.44 0.001 -0.32
Volatile acids/ acetic acid 3.3 0.09 0.82 0.35 0.56 0.94
Tartaric acid/ acetic acid 21.6 0.0005 -0.38 6.38 0.03 -0.60
Tartaric acid/ malic acid 5.9 0.031 0.51 15.88 0.002 0.39
Tartaric acid/ citric acid 29.69 0.0001 -0.65 37.1 0.0001 0.48
Tartaric acid/ phenolic compounds 16.33 0.0016 -0.02 66.38 < 0.0001 -0.64
Tartaric acid/ sugar 7.11 0.02 0.41 2.22 0.16 -0.60
Tartaric acid/ ethanol 57.41 < 0.0001 -0.90 43.79 < 0.0001 -0.10
Tartaric acid/ lactic acid 15.18 0.002 0.32 0.22 0.64 -0.54
Acetic acid/ sugar 22.76 0.0004 -0.65 4.04 0.07 0.67
Acetic acid/ ethanol 108.93 < 0.0001 0.35 67.01 < 0.0001 -0.18
Citric acid/ lactic acid 31.58 0.0001 -0.07 3.59 0.08 0.19
Citric acid/ acetic acid 6.39 0.026 -0.13 3.03 0.11 0.06
Citric acid/ sugar 63.52 < 0.0001 0.1 5.38 0.04 0.07
Citric acid/ ethanol 116.73 < 0.0001 0.37 153.88 < 0.0001 0.04
Lactic acid/ acetic acid 4.71 0.05 -0.16 0.92 0.36 0.98
Lactic acid/ sugar 8.53 0.01 0.50 2.51 0.14 0.68
Lactic acid/ ethanol 101.75 < 0.0001 -0.38 21.07 0.0009 -0.34
Malic acid/ citric acid 92.98 < 0.0001 -0.11 7.0 0.02 -0.11
Malic acid/ total Phenolics 16.46 0.001 -0.11 66.82 < 0.0001 -0.92
Malic acid/ sugar 0.37 0.55 0.36 4.46 0.06 -0.56
Malic acid/ ethanol 88.19 < 0.0001 -0.63 100.5 < 0.0001 0.53
Malic acid/ lactic acid 14.35 0.002 -0.37 1.62 0.23 -0.95
Malic acid/ acetic acid 32.94 < 0.0001 -0.34 0.21 0.65 -0.93
Sugar/ ethanol 90.17 < 0.0001 -0.51 0.03 0.86 -0.42
Note: F — Fisher`s criterion. Dispersion analysis (ANOVA) (unifactorial model);
P — value of differences between the investigated groups;
r — Spearman`s rank correlation coefficient.
Експериментальні статті
103
samp les fermented using yeast cultures:
Charivny (Magic) MAFF-230100 more than
85% and Tairov`s Ruby MAFF-230098 more
than 85%. Comparative assessment shows
that some white wine grape varieties produce
much more ethanol compared to red wine
grape varieties. This indicates that fermenta-
tion activity and ethanol forming capacity of
tested wine stocks increases when using yeast
strains MAFF-230100 and MAFF-230098. In
both white and red grape varieties there exist
2–3 yeast strains which produce maximal
amount of ethanol over other yeast strains.
Those strains with high enzymatic activity
could be used in the alcohol industry to pro-
duce ethanol for the pharmaceutical and food
industries. Maximal concentration of tartaric
acid in wines received from white grape vari-
eties in Ovidiopolskij MAFF-230104 was more
than 480%. For red wine grape varieties max-
imal concentration in Odessa`s Black MAFF-
230109 was more than 548%. The maximal
level of malic acid concentration in white wine
grape varieties for Ovidiopolskij MAFF-
230104 was more than 120% and Zagrey
MAFF-230105 was more than 260%.
Red wine grape varieties showed maximal
concentration of malic acid using yeast culture
Charivny (Magic) MAFF-230100 and was more
than 45%, Tairov`s Ruby MAFF-230098 was
more than 75%. Such red wine grape varieties as
Illychevsky Early MAFF-230110 and Odessa`s
Pearl MAFF-230108 did not containe malic acid.
This means that spontaneous malolactic fermen-
tation was completed in these wines.
Rating quality of wines by the ratio of tar-
taric to malic acid was maximal and perfect for
white wine grape varieties Aromatic MAFF-
230106, the ratio was 2.95 for Ovidiopolskij
MAFF-230104. We found that quality of wine
depends upon the ratio between tartaric and
malic acids. We found that due to a high ratio,
of dry wine received using yeast culture
Odessa`s Black MAFF-230109 in perfect qua -
lity, where ratio was 4.27. Quality of dry wine
received using yeast culture Ruby Jubilee
MAFF-230099 is excellent quality, where
ratio was 3.86. Parameters of titratable
organic acids provide an information of the
balance between assimilated and produced
organic acids for separate Saccharomyces cere-
visiae yeast culture. For acidity, an important
role is played by malolactic fermentation.
Wines from red grape varieties were bright
and lively ruby-red color, with a clear aroma
and notes of wild berries, ripe and dried plums
with accents of dry vegetable mass. Taste for
red wine grape varieties was well structured,
dense, soft and rounded, with delicate acids
and almost without bitterness. Such wine
stock samples were received with Saccharo my -
ces cerevisiae yeast strains and produced high
level of ethanol: white wine grape varieties
Ovidiopolskij MAFF-230104 and Zagrey
MAFF-230105. Samples of wine stock received
using Saccharomyces cerevisiae yeast strains,
Odessa`s Black MAFF-230109 and Ruby
Jubilee MAFF-230099 obtained from red
grape varieties contained a high volume frac-
tion of ethanol production. The strains which
produced high amounts of alcohol (ethanol)
are going to be proposed for use in wine
biotechnology, pharmaceutical industry, and
food industry.
Information given in Table 4 shows posi-
tive, moderate and negative correlations, P-
value, Fisher-Snedecor test (unifactorial
model) frequency in the null distribution in
the analysis of variance between tested para-
meters in both groups of white and red wine
stocks (F). Existing high correlation between
tested parameters of titratable acidity and
volatile acids for wine stocks of red grape vari-
eties (r = 0.82; P = 0.0001). It is natural
because between titratable acidity and volatile
acids there exists normal correlation and
interdependence.
There is very high correlation between
titratable acidity and malic acid in white grape
varieties (r = 0.95; P = 0.0004). There was
high correlation between volatile acids and
acetic acid in wine stocks received from white
grape varieties (r = 0.82; P = 0.09). We found
very high correlations in wine stocks received
from red grape varieties (r = 0.94; P = 0.56).
Such indices are natural because between
volatile acids and acetic acid in wine stocks
there exists normal correlation and interde-
pendence. We found moderate correlation
between tartaric acid and malic acid in wine
stocks received from white grape varieties
(r = 0.51; P = 0.03) and there was specified
low, but statistically reliable correlation in
wine stocks received from red grape varieties
(r = 0.39; P = 0.002). We found moderate cor-
relation between lactic acid and sugar in wine
stocks received from white grape varieties
(r = 0.50; P = 0.01), also moderate correlation
was found for wine stocks received from red
grape varieties (r = 0.68; P = 0.1). We found
very high correlation between lactic and acetic
acids in wine stocks received from red grape
cultivars (r = 0.98; P = 0.3). We found low
correlation between malic acid and sugar in
wine stocks received from white grape
varieties (r = 0.36; P = 0.5). We noticed
BIOTECHNOLOGIA ACTA, V. 6, No 2, 2013
104
moderat e correlation between malic acid and
ethanol in wine stocks received from red grape
varieties (r = 0.53; P = < .0001). We found
low correlations between citric acid and
ethanol in wine stocks received from white
grape varieties (r = 0.37; P = < 0.0001).
We found moderate correlation between
acetic acid and sugar in wine stocks received
from red grape varieties (r = 0.67; P = 0.07).
We noticed low correlations between tartaric
and lactic acids in wine stocks received from
white grape varieties (r = 0.32; P = 0.02). We
found low correlations between tartaric acid
and sugar in wine stocks received from white
grape varieties (r = 0.41; P = 0.02) and also
low correlation between tartaric and citric
acids in wine stocks received from red grape
varieties (r = 0.48; P = 0.0001).
We found very high correlations between
titratable acidity and tartaric acid in wine
stocks received from white grape varieties (r=
0.95; P = 0.0004). We found high correlations
between titratable acidity and volatile acids in
wine stocks received from red grape varieties
(r = 0.82; P = < 0.0001).
We noted negative correlation between
tartaric and acetic acids for wine stocks
received from white grape varieties (r = 0.38;
P = 0.0005) and for wine stocks received from
red grape varieties (r = –0.60; P = 0.03). In
both groups there were negative, but pretty
reliable connections.
Negative correlations between tartaric
acid and phenolic compounds in wine stocks
received from white grape varieties (r =
–0.02; P =0.001) and in wine stocks received
from red grape varieties (r = –0.64; P = <
0.0001) were observed. In both investigated
groups there were negative, but pretty reliable
connections. However, between the investiga -
ted groups of wine stocks received from white
and red grape varieties there was appreciable
difference. Negative correlations also existed
between the content of tartaric acid and
ethanol for wine stocks received from white
grape varieties (r = –0.90; P = < 0.0001) and
for wine stocks received from red grape
varieties (r = –0.10; P = < 0.0001).
There is a high correlation between titra -
table acidity and volatile acids in wine stocks
received from red grape varieties (r = 0.82;
P = <0.0001). However, the same parameters
in white wine stocks received from white grape
varieties are absolutely different, because cor-
relation is negative (r = –0.47; P = <0.0001).
Such great differences results in opposite cor-
relations between white and red stocks. It
could be explained by the fact that red wine
stocks contained different ingredients inclu -
ding pigments, anthocyanins, and high pheno-
lic and tannine contents. Therefore, red wine
stocks have a high correlation compared with
the same in white wine stocks. Very high cor-
relation was indicated between titratable aci -
dity and tartaric acid in white wine stocks that
were received from white grape varieties
(r = 0.95; P = 0.0004). However, the same
parameters in red wine stocks received from
red grape varieties are absolutely different,
and correlation showed a negative result
(r = –0.32; P = 0.001).
According to our investigation there were
determined the following: a high and very
high correlation between volatile acids and
acetic acid in white wine stocks received from
white grape varieties (r = 0.82; P = 0.09) and
in red wine stocks received from red grape
varieties (r = 0.94; P = 0.56); a high correla-
tion between lactic and acetic acid in red wine
stocks received from red grape varieties
(r = 0.98; P = 0.36). However, the same para-
meters in white wine stocks received from
white grape varieties were absolutely diffe -
rent, where correlation showed a negative
result (r = –0.16; P = 0.05); a moderate corre-
lation was found between lactic acid and resid-
ual sugar for white wine stocks received from
white grape varieties (r = 0.50; P = 0.01) and
for red wine stocks received from red grape
varieties (r = 0.68; P = 0.14); a moderate cor-
relation between malic acid and ethanol. For
wine stocks received from red grape varieties
(r = 0.53; P = <0.0001) and for white wine
stocks received from white grape varieties cor-
relation was negative (r = –0.63; P =
<0.0001); prospective yeast cultures of
Saccharomyces cerevisiae for wine biotechnol-
ogy produced a high volume fraction of
ethanol isolated from white grape varieties:
Odessa`s Muscat MAFF-230107, Opalovy
(Opaline) MAFF-230103, Selena MAFF-
230101 and yeast culture Saccharomyces cere-
visiae isolated from red grape varieties:
Charivny (Magic) MAFF-230100, Odessa`s
Black MAFF-230109, Tairov`s Ruby MAFF —
230098.
The author wishes to thank: Svetlana Dre -
vo va, Head of the Department of Wine ma -
king. Tarasova Vita, Head of the Laboratory
Physical and Chemical studies of wine stocks.
Evgeniya Polukarova, Department of wine-
making at the Ukrainian Tairov`s Research
Institute of Viticulture and Oenology, for
their real help with the grape must and wine
stock samples, providing liquid chromatogra-
phy, spectrometry in studied samples during
Експериментальні статті
105
research. Irina Dudenko, Head of the
Laboratory of winery for her help with the
grape must and wine stock samples. Special
thank to Ludmila Gerus, the Department of
Genetics and Selection the Ukrainian Tairov`s
Research Institute of Viticulture and
Oenology, for her help with grape samples
from different grape varieties the Tairov`s
selection.
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BIOTECHNOLOGIA ACTA, V. 6, No 2, 2013
106
КОНЦЕНТРАЦІЯ ОРГАНІЧНИХ КИСЛОТ
У ВИНОМАТЕРІАЛАХ
ПІСЛЯ ФЕРМЕНТАЦІЇ ДРІЖДЖІВ
Saccharomyces cerevisiae
В. М. Байрактар
Одеський національний університет
ім. І. І. Мечникова, Україна
E-mail: vogadro2007@rambler.ru
Досліджували біохімічні показники вино-
матеріалів, що впливають на смак і якість
вина: об’ємну частку етанолу, концентрації
залишкових цукрів, фенольних сполук, вин-
ної, яблучної, лимонної, молочної, оцтової
кислот, титрувальну та летючу кислотність.
Використовували виноматеріали, отримані з
білих і червоних сортів винограду Таїровсь ко ї
селекції. Відзначено кореляцію в обох групах
між показниками титрувальної та летючої
кислотності у виноматеріалах із білих та чер-
воних сортів винограду. Високу кореляцію
спостерігали між концентраціями молочної та
оцтової кислот, летючою кислотністю і кон -
цент раціями оцтової кислоти та загальних
цукрів. Встановлено, що виноматеріали
з високою концентрацією етанолу отримано
з використанням штамів дріжджів Saccharo -
myces cerevisiae, що ферментували виноградне
сусло з високою швидкістю й активністю бро-
діння. Смак виноматеріалів залежить від
показників відношення винної кислоти до
яблучної кислоти. Показано значні відміннос-
ті між виноматеріалами, що їх одержано з
білих і червоних сортів винограду: у концен-
траціях органічних кислот, фенольних спо-
лук, залишкових цукрів, а також об’ємної час-
тки етанолу. Для обох досліджуваних груп
відзначено позитивну кореляцію між показни-
ками летючої кислотності та концентраціями
оцтової, винної, яблучної, молочної кислот
і загальних цукрів. Відібрано найбільш пер-
спективні культури дріжджів з високою
продуктивністю синтезу етанолу, які можуть
бути використані у біотехнології вино роб ст ва.
Ключові слова: органічні кислоти, виноградні
сорти, Saccharomyces cerevisiae.
КОНЦЕНТРАЦИЯ ОРГАНИЧЕСКИХ
КИСЛОТ В ВИНОМАТЕРИАЛАХ
ПОСЛЕ ФЕРМЕНТАЦИИ ДРОЖЖЕЙ
Saccharomyces cerevisiae
В. Н. Байрактар
Одесский национальный университет
им. И. И. Мечникова, Украина
E-mail: vogadro2007@rambler.ru
Исследовали биохимические показатели
виноматериалов, влияющие на вкус и качество
вина: объемную долю этанола, концентрации
остаточных сахаров, фенольных соединений,
винной, яблочной, лимонной, молочной,
уксусной кислот, титруемую и летучую кис-
лотность. Использовали виноматериалы,
полученные из белых и красных сортов вино-
града Таировской селекции. Отмечена корре-
ляция между показателями титруемой и лету-
чей кислотности в виноматериалах из белых и
красных сортов винограда. Высокая корреля-
ция наблюдалась между концентрациями
молочной и уксусной кислот, летучей кислот-
ностью и концентрациями уксусной кислоты и
общих сахаров. Установлено, что виномате-
риалы с высокой концентрацией этанола полу-
чены с использованием штаммов дрожжей
Saccharomyces cerevisiae, которые ферменти-
ровали виноградное сусло с высокой скоро-
стью и высокой активностью брожения. Вкус
виноматериалов зависит от показателей отно-
шения винной кислоты к яблочной кислоте.
Показаны значительные различия между
виноматериалами из белых и красных сортов
винограда: в концентрациях органических
кислот, фенольных соединений, остаточных
сахаров, а также объемной доли этанола. Для
обеих исследуемых групп отмечена положи-
тельная корреляция между показателями лету-
чей кислотнос ти и концентрациями уксусной,
винной, яблочной, молочной кислот и общих
сахаров. Отобраны наиболее перспективные
культуры дрожжей с высокой продуктив-
ностью синтеза этанола, которые могут быть
использованы в биотехнологии виноделия.
Ключевые слова: органические кислоты,
виноградные сорта, Saccharomyces cerevisiae.
