Effect of Initial PH on Growth Characteristics

and Fermentation Properties of Saccharomyces

cerevisiae
Xingyan Liu, Bo Jia, Xiangyu Sun, Jingya Ai, Lihua Wang, Cheng Wang, Fang Zhao, Jicheng Zhan, and Weidong Huang

Abstract: As the core microorganism of wine making, Saccharomyces cerevisiae encounter low pH stress at the beginning
of fermentation. Effect of initial pH (4.50, 3.00, 2.75, 2.50) on growth and fermentation performance of 3 S. cerevisiae
strains Freddo, BH8, N˚.7303, different tolerance at low pH, chosen from 12 strains, was studied. The values of yeast
growth (OD600 , colony forming units, cell dry weight), fermentation efficiency (accumulated mass loss, change of total
sugar concentration), and fermentation products (ethanol, glycerol, acetic acid, and l-succinic acid) at different pH stress
were measured. The results showed that the initial pH of must was a vital factor influencing yeast growth and alcoholic
fermentation. Among the 3 strains, strain Freddo and BH8 were more tolerant than N˚.7303, so they were affected
slighter than the latter. Among the 4 pH values, all the 3 strains showed adaptation even at pH 2.50; pH 2.75 and 2.50
had more vital effect on yeast growth and fermentation products in contrast with pH 4.50 and 3.00. In general, low initial
pH showed the properties of prolonging yeast lag phase, affecting accumulated mass loss, changing the consumption rate
of total sugar, increasing final content of acetic acid and glycerol, and decreasing final content of ethanol and l- succinic
acid, except some special cases. Based on this study, the effect of low pH on wine products would be better understood
and the tolerance mechanism of low pH of S. cerevisiae could be better explored in future.

M: Food Microbiology
Keywords: Fermentation, growth, initial pH, Saccharomyces cerevisiae

Introduction
Saccharomyces cerevisiae is the core microorganism in fermenta-
tion, especially in wine making. Most S. cerevisiae strains grow at
pH values between 2.50 and 8.50, but they are acidophilic organ-
isms and grow better under acidic conditions (Carmelo and others
1996). The optimal pH range for yeast growth can vary from pH
4.00 to 6.00, depending on temperature, the presence of oxygen,
culture, and the strain of yeast (Narendranath and Power 2005).
During wine making, S. cerevisiae are always encountered with
adverse environmental conditions, for example, at the beginning
of fermentation, yeast cells are affected by osmotic stress because
of the high sugar concentration, as well as low pH (Cardona and
others 2007).
External environmental pH especially caused by weak acid can
affect the cell wall structure and alter the conformation of proteins
protruding from the plasma membrane; meanwhile, it also has an
impact on the lipid organization and function of cellular mem-
branes and the perturbation of the function of membrane embed-
ded proteins. The loss of plasma membrane integrity increases cell
permeability to ions and other small metabolites, which leads to
the stimulation of passive diffusion of protons from the exterior to
the cytosol. In this case, the reduction of internal pH in weak acid
challenged cells and the dissipation of the electrochemical poten-
tial maintained across this membrane (a driving force for secondary
transport; Mira and others 2010) are easily occurred. Eventually,
pH value affects the growth and fermentation rate of yeast and
influences the constitution of fermentation products (Ough 1966;
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Buzas and others 1989; Nielsen and Arneborg 2007; Yalcin and
Ozbas 2008; Arroyo-López and others 2009). Recently, a lot of re-
searches about low pH or weak acid stress on S. cerevisiae had been
reported, but most were involved in acetic acid (Zhao and others
2014), lactic acid (Abbott and others 2009), critic acid (Nielsen
and Arneborg 2007), or weak acid preservatives such as benzoic
acid (Hazan and others 2004) and sorbic acid (Papadimitriou and
others 2007). There are also some literatures about the effect of
grape initial pH on S. cerevisiae, mostly studied the pH value above
3.00 (Ough 1966; Yalcin and Ozbas 2008).
How about pH lower than 3.00 affecting S. cerevisiae? To the
best of our knowledge, there was little study on this issue. It is
worth to point out that pH value in grape musts normally ranges
from 2.75 to 4.20 (Belloch and others 2008); however, the pH
value of Chinese Vitis amurensis grape is always as low as to pH
2.50 to 2.90 (Zhao and Wang 1996; Jiang and others 2008). So it
is necessary to study the effect of low pH on S. cerevisiae.
In this study, our aim is to investigate the effect of low pH
on the growth and fermentation properties of S. cerevisiae during
alcoholic fermentation. However, natural musts show a variable
composition among different years that can influence the yeast
growth (Arroyo-López and others 2009). For this reason, a de-
fined synthetic must was chosen as the most appropriate growth
medium to overcome this variation in this study. This work would
give some help to wine making and lay a foundation for tol-
erance mechanism research of S. cerevisiae at low pH value in
future.

MS 20141730 Submitted 10/17/2014, Accepted 1/7/2015. Authors are with

Materials and Methods
College of Food Science & Nutritional Engineering, China Agricultural Univ., 100083
Beijing, China. Authors Liu is with College of Food Science, Sichuan Agricultural
Yeast strains and culture medium
Univ., 625000, Sichuan Ya’an, China. Direct inquiries to authors Huang and Zhan Twelve S. cerevisiae strains were used in this study, including
(E-mail: huanggwd@263.net and jczhan@263.net). eleven commercial strains and a wild strain named BH8 isolated
from the “Beihong” wine grape variety, cultivated by the Inst. of

C 2015 Institute of Food Technologists

 R

doi: 10.1111/1750-3841.12813 Vol. 00, Nr. 0, 2015 r Journal of Food Science M1

Further reproduction without permission is prohibited
 Effect of initial pH on yeast . . .

Table 1–List of yeast strains used in this study.

Strains Abbreviation in
designation Factories Purposes this study
AWRI R2 Marivin, Australia White wine A
Aroma white Enartis, Italia White wine AW
BH8 Wild B
CY3079 Lallemand, France White wine C
DV10 Lallemand, France Champagne, white wine D
EC1118 Lallemand, France White wine, ice wine E
Ezferm Enartis, Italia Prevent and cure sluggish and EZ
stuck fermentation
Freddo Erbslöh Geisenheim AG, Germany White wine F
LVCB DSM, the Netherlands White wine L
N˚.7303 DSM, the Netherlands Red wine N
Red Fruit Enartis, Italia Red wine RF
XR Lamothe-abiet, France Red wine XR

Botany, the Chinese Academy of Sciences, Beijing, China (Li and
others 2010).
YPD medium: 1% yeast extract, 2% peptone, 2% dextrose, add
2% agar when required.
Model synthetic medium (MSM): was described previously
(Marullo and others 2004; Masneuf-Pomarede and others 2006). It
could simulate the composition of grape must. It was a chemically
well-defined medium and in advantage allowed modifications in
M: Food Microbiology
trifugation (6000 r/min 5 min, 4 °C), washed twice with sodium
phosphate buffer (50 mmol/L, pH 6.0), and then resuspended in
different initial pH LMSM (1% inoculation proportion, final cell
density was [2.5–5.0] × 105 CFU/mL, and pH was adjusted by
6 mole/L NaOH and 3 mole/L H2 SO4 , pH range was 2.50 to
6.00,actual pH value = set value ± 0.05 when pH ࣘ 3.0,whereas
pH = set value ± 0.10 when pH > 3.0], then put them in a shaker
at 28 °C and 150 rpm with the ration of flask volume/medium

the concentration of specific components to study the effect of of 5/2 for 48 h. Samples were taken at certain time interval to
only one factor. Its constituents are as follows (expressed in g/L): determine OD600 to draw growth curve. At the 1st 24 h, samples
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glucose (100.0), fructose (100.0), tartaric acid (3.0), citric acid were taken every 3 h, and at the next 24 h, taken every 6 h.
(0.3), l-malic acid (0.3), MgSO4 (0.2), and KH2 PO4 (2.0). Nitro-
gen sources were adjusted to 190 mg/L as (NH4 )2 SO4 (0.3 g/L) Selection of pH
and asparagine (0.6 g/L). Mineral salts (mg/L): MnSO4 ·H2 O (4.0), Use MSM as medium, yeast grew at 28 °C on liquid
ZnSO4 ·7H2 O (4.0), CuSO4 ·5H2 O (1.0), KI (1.0), CoCl2 ·6H2 O YPD medium until mid-log phase (OD600 = 1.30–1.50) be-
(0.4), and (NH4 )6 ·MO7 O24 ·4H2 O (1.0), H3 BO3 (1.0). Vitamins fore they were inoculated in ratio of 1% to 500 mL Erlen-
(mg/L): mesoinositol (300.0), biotin (0.04), thiamin (1.0), pyri- meyer flask containing 400 mL MSM (final cell density was
doxine (1.0), nicotinic acid (1.0), pantothenic acid (1.0), and p- [2.5–5.0] × 105 CFU/mL). Erlenmeyer flasks were sealed by rub-
amino benzoic acid (1.0). Fatty acids (mg/L): palmitic acid (1.0), ber plugs equipped with a fermentation bolt containing 5.0 mL
palmitoleic acid (0.2), stearic acid (3.0), oleic acid (0.5), linoleic H2 SO4 , thus gas in flasks can spread, but gas or microbe outside
acid (0.5), and linolenic acid (0.2). Before yeast inoculation, the cannot enter flasks. Then cells grew aerobically in a shaker at 28 °C
medium was filtered by 0.45 μm nitrate cellulose membrane. Low and 120 rpm. Flasks were weighted every day until fermentation
sugar MSM (LMSM): To avoid high osmotic pressure from sugar, finished (constant weight for consecutive 3 d). Total mass loss was
the concentration of reduced sugars in MSM was adjusted to 20 counted.
g/L according to YPD (Li and others 2010).
Determination of cell growth characteristics
Selection of strains Use the same method as pH selection, the only difference was
To find different tolerance to low pH, 12 S. cerevisiae strains Erlenmeyer flask had an opening on the side sealed by an anaerobic
(Table 1) were used to study. Yeast strains grew at 28 °C on liquid plug. Samples were taken one or more days until fermentation fin-
YPD medium until mid-log phase (OD600 = 1.30–1.50, deter- ished. Cell growth characteristics were determined by measuring
mined by using a spectrophotometer [UV-1800, SHIMADZU]) OD600 , CFU, and biomass. OD600 and CFU were determined by
before cells were harvested by centrifugation (6000 r/min 5 min, the same method as above, biomass was determined by use of the
4 °C), washed twice with sodium phosphate buffer (50 mmol/L, dry weight method. Accordingly, samples were centrifuged, then
pH 6.0), then resuspended in the same volume of YPD or pH 2.75 washed twice with distilled water and dried for constant weight at
LMSM (adjusted by 6 mole/L NaOH and 3 mole/L H2 SO4 ), then 80 °C.
cells grew aerobically in a shaker at 28 °C and 150 rpm with the
ration of flask volume/medium of 5/2. Samples were taken out Determination of fermentation rate and fermentation
at 3 h. Colony forming units (CFU) were determined by plate products
counting, appropriate dilutions were plated on YPD agar in trip- Use the same method as effect of initial pH on growth char-
licate. YPD agar plates were incubated at 28 °C for 2 to 3 d. acteristics of S. cerevisiae. Accumulated mass loss and change of
Relative viability was expressed by the ration of log10 CFU in pHtotal sugar concentration were used to reflect fermentation rate of
2.50 LMSM to log10 CFU of itself in YPD. S. cerevisiae. Fermentation products determined included ethanol,
glycerol, acetic acid, l-succinic acid, and l-lactic acid. Accumu-
Measurement of growth curve of S. cerevisiae lated mass loss was counted by weighting every day until fermen-
Yeast strains grew at 28 °C on liquid YPD medium until mid-log tation finished. Change of total sugar (d-glucose and d-fructose)
phase (OD600 = 1.30–1.50) before cells were harvested by cen- concentration and fermentation products were both counted by

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Effect of initial pH on yeast . . .

Table 2–Relative viability of yeast strains. of the 3 strains was significant affected, but the extent of influence
Strain Relative viability was different, the growth of N was most seriously affected. At pH
2.50, growth of the 3 strains was almost totally inhibited, which
F 92.22% ± 0.98%a was in accordance with the results of Carmelo and others (1996).
A 92.19% ± 0.30%a
RF 91.84% ± 0.22%ab
Based on these results, it can also be concluded that the low pH
D 91.17% ± 0.37%abc tolerance of strain N was the weakest among the 3 ones.
C 90.66% ± 1.29%abc
B 90.17% ± 0.61%bcd PH selection
EC 89.36% ± 1.52%cd
XR 88.68% ± 2.14%de
Effect of initial pH on total mass loss was shown in Table 3. As
L 88.44% ± 1.14%de shown in Table 3, total mass loss was ranging from 11.67 to 36.06 g.
EZ 88.38% ± 0.20%de Effect of initial pH on the 3 strains had some difference, but the
AW 87.24% ± 0.33%e trend was similar. To be specific, at pH 3.50, 4.50, and 5.50,
N 87.05% ± 0.57%e there was no significant difference in all the 3 strains, which was
a
Values were presented as means ± standard deviation from 3 independent experiments. because of the tight control of intracellular pH (Orij and others
b
Values in columns followed by different letters indicated significant differences at P <
0.05.

supernatants of fermentation broth determined by HPLC during

fermentation process.
HPLC analysis. A Waters Alliance 2695 HPLC system with
Waters 2414 RID Detector (Waters Corp., Milford, Mass., U.S.A.)
was used to simultaneously separate and analyze total sugar and
fermentation products. The system was run at 0.5 mL/min
using an Aminex HPX-87H ion exchange column (300 mm

M: Food Microbiology
× 7.8 mm) protected with a Bio-Rad guard cartridge (30 mm ×
4.6 mm). The column was thermostated at 55 °C. Mobile phase

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was 5 mmol/L H2 SO4 in purified water (Du 2011). The injection
volume was 10 μL. Waters Empower Software Chromatographic
data software was used for data acquisition, peak integration, and
standard calibration. Peaks were qualitatived with retention time
and quantified with external standard calibration based on areas.
External standard including d-glucose, d-fructose, ethanol, glyc-
erol, acetic acid, l-succinic acid, and l-lactic acid were all bought
from Sigma. H2 SO4 was analytically pure grade made in China.

Statistical analysis
All the experiments were performed in triplicate. Values were
presented as means ± standard deviation from 3 independent ex-
periments. Significant differences were determined by analysis of
variance (ANOVA), followed by the Duncan test at a level of P <
0.05. Data were analyzed by using IBM SPSS Statistics software,
version 17.0, and OriginPro software, version 8.5.

Results and Discussion

Strains selection
Relative viability of the 12 strains was shown at Table 2.
From Table 2 it can be seen that different strains had differ-
ent relative viability at low pH, which was between 87.05% and
92.22%. Among all commercial strains, strain F and strain A had
the highest relative viability, and strain N and AW had the lowest
relative viability. Although the only wild strain B had moderate
relative viability. So strain F, B, and N were selected to next study.

Growth curve of S. cerevisiae

Growth curve of strain F, B, and N were shown at Figure 1.
As it was shown, initial pH of medium had vital influence on
the growth of strains. pH 4.00 to 5.00 was the optimum range to
yeast growth, which was in good agreement with the results of
Buzas and others (1989).When pH ࣘ 3.0, the lag period of yeast Figure 1–Growth curve of strain F, B, and N in LMSM of different initial pH
increased and the maximal OD600 decreased. At pH 2.75, growth value.

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 Effect of initial pH on yeast . . .

Table 3–Effect of initial pH on total weight loss. facilitated the growth of wine yeast. However, too low pH value
pH F B N could inhibit the growing of wine yeast (Vine and others 1997).
Effect of initial pH on growth characteristics of strain F, B, and
2.50 11.67 ± 0.94c 22.44 ± 0.18d 20.02 ± 0.95d N was shown in Figure 2.
2.75 29.96 ± 1.20b 29.59 ± 1.43c 26.35 ± 0.60c
As shown in Figure 2, initial pH affected growth of the strains.
3.00 33.13 ± 0.58a 31.65 ± 0.36b 31.75 ± 0.40b
3.50 35.03 ± 0.28a 35.63 ± 0.21a 35.04 ± 0.52a Among the 4 pH values, pH 2.50 severely affected the growth
4.50 34.91 ± 0.69a 36.06 ± 0.74a 35.39 ± 0.09a of yeast with its lag phase much longer and the maximal OD600
5.50 34.04 ± 0.21a 34.36 ± 0.02a 35.34 ± 0.14a value much lower than other pH values. With the increase of pH,
a
Values are presented as means ± standard deviation from 3 independent experiments. the values of maximal OD600 became higher. Although, when the
For each treatment, values in columns followed by different letters indicate significant highest OD600 value was reached, it declined. In the 3 strains,
differences at P < 0.05.
strain N had the longest lag period at pH 2.75 and 2.50, so was
the whole fermentation time, which was also reflected its weakest
2011). Although at pH 3.00, 2.75, and 2.50, total mass loss had low pH tolerance.
significant difference with each other, and also with that of at pH OD600 value included all cells both live and dead, whereas CFU
3.50, 4.50, and 5.50 (P < 0.05). reflected only live cells. As shown in Figure 2(B), initial pH value
In consideration of the growth curve and effect of initial pH also affected CFU of strain F, B, and N. With the increase of pH,
on total mass loss, pH 4.50, 3.00, 2.75, and 2.50 were selected to the maximal log10 CFU became higher, and pH 2.50 induced log10
next study. CFU of strain N and F decreased after inoculated, and increased a
few days later, but the maximal log10 CFU was lower than pH 3.00
and 4.50. From this, it can be seen pH 2.50 was a very severe stress
Effect of initial pH on growth of S. cerevisiae to S. cerevisiae, and cells made use of its stress response to survive.
Decrease in initial pH value of must inhibited metabolic activ- As shown in Figure 2(C), initial pH affected cell dry weight
ities of contaminated bacteria during wine fermentation, which as well. In general, cell dry weight firstly increased, and then
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Figure 2–Effect of different initial pH value on OD600 (A), CFU (B), and dry weight (C) of strain F, B, and N during fermentation in MSM.

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Effect of initial pH on yeast . . .

declined, which was in accordance with Figure 2(A) and (B), and F, B, and N during fermentation in MSM medium of different
also in accordance with the character of yeast growth. initial pH were shown in Figure 3(B).
As shown in Figure 3(B), 3 strains showed both some similar and
different characters. The similarity was that at low pH especially
Effect of initial pH on fermentation rate of S. cerevisiae pH 2.50, total sugar concentration decreased very slowly and the
Alcoholic fermentation is the anaerobic transformation of sugars final sugar concentration was high, whereas at pH 4.50 and 3.00,
(mainly glucose and fructose) into ethanol and carbon dioxide. total sugar concentration decreased sooner, the change trend at
The release of carbon dioxide would lead to mass loss, therefore pH 4.50 and 3.00 was similar and their final concentration was 0.
mass loss can always be used to an index of fermentation rate. The difference was that the change was closely related to strain.
Effect of initial pH on fermentation rate of S. cerevisiae was In regard to strain F, the trend at pH 2.75 or 2.50 differed severely
shown in Figure 3. with that at pH 4.50 and 3.00; in regard to strain B, the trend at
As shown in Figure 3(A), initial pH affected accumulated mass pH 2.75 was almost the same with that at pH 4.50 and 3.00, but
loss of strains. Accumulated mass loss of all the 3 strains at pH 4.50 was severely different to that at pH 2.50, whereas to strain N, the
was the maximal among the 4 pH value, which was in accordance trend at pH 2.75 or at pH 2.50 was almost the same.
with Lin and others (2012). In early days, accumulated mass loss As shown in Figure 3, it can also be seen total fermentation
of strains at pH 3.00 and 4.50 was similar, but at the later days it time was different related to initial pH and yeast strain. In regard
was slower than pH 4.50, and the maximal accumulated mass loss to strain F, its fermentation time at pH 4.50, 3.00, 2.75, and 2.50
was lower. Although accumulated mass loss at pH 2.75 and 2.50 was, respectively, 13, 15, 17, and 13 d; in regard to strain B, its fer-
obviously much lower than that at pH 4.50 and 3.00, and strain mentation time at pH 4.50, 3.00, 2.75, and 2.50 was, respectively,
N at pH 2.50 had the longest lag period. 13,17, 17, and 15 d; whereas in regard to strain N, its fermentation
Sugar was substrate of alcoholic fermentation, yeast tend to time at pH 4.50, 3.00, 2.75, and 2.50 was, respectively, 24,26, 50,
utilize glucose firstly in all sugars, which could also be observed and 48 d. From this point of view, the much longer fermentation
in this study (data not shown). Total sugar concentration of strains time of N was also reflected its lower pH tolerance.

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Figure 3–Effect of different initial pH
value on accumulated mass loss (A) and
total sugar content (B) of strain F, B, and N
during fermentation in MSM.

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 Effect of initial pH on yeast . . .
Effect of initial pH on fermentation products of S. cerevisiae
Alcoholic fermentation is the anaerobic transformation pro-
cessing of sugars into ethanol and carbon dioxide. Besides ethanol,
several other compounds are produced throughout alcoholic fer-
mentation such as glycerol, acetic acid, succinic acid, lactic acid,
and so on (Zamora 2009). Improved ethanol fermentation activity
can be achieved by controlling various parameters. In addition to
temperature and substrate concentration, pH is also a key factor
that affects ethanol fermentation (Gunasekaran and Raj 1999; Lin
and others 2012). Effect of initial pH on fermentation products
including ethanol, glycerol, acetic acid, and l-succinic acid were
shown in Figure 4 to 7. l-Lactic acid was also determined, but it
failed to check out, because of no malo-lactic fermentation in this
study.
Effect of initial pH on ethanol concentration was shown in
Figure 4. As shown in Figure 4, the maximum ethanol concen-
tration of 3 strains was at pH 4.50, which was in accordance with
the study of Lin and others (2012) and Reddy and Reddy (2011).
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Figure 4–Effect of different initial pH value on ethanol concentration of Figure 5–Effect of different initial pH value on glycerol concentration of
strain F, B, and N during fermentation in MSM.
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For strain F, the maximum ethanol concentration at pH 4.50, 3.00,
and 2.75 was almost the same, but the change trend at pH 2.75 was
different to that at pH 4.50 and 3.00; for strain B, the maximum
ethanol concentration at pH 4.50, 3.00, and 2.75 was almost the
same too, but was vitally different to that at pH 2.50; for strain
N, the maximum ethanol concentration at pH 4.50 and 3.00 was
much higher than that at pH 2.75 and 2.50.
Glycerol, contributing to the smoothness of wine, is an im-
portant constituent of wine formed as a by-product during the
fermentation process and is important for high osmotic stress
response. Except for carbon dioxide and ethanol, glycerol is the
most abundant constituent with the typical levels found in wine
varying from 1 to 15 g/L (Scanes and others 1998). According to
the report, the glycerol production was significantly affected by
the composition of the must, such as pH, nature of the osmotic
agent, and carbon source (Perez-Torrado and others 2002).
Effect of initial pH on glycerol concentration was shown in
Figure 5. As shown in Figure 5, the maximum glycerol concen-
tration of strain F, B, and N at the 4 pH value was 6.50 to 8.74,
strain F, B, and N during fermentation in MSM.
Effect of initial pH on yeast . . .
5.78 to 7.91, and 5.60 to 7.39 g/L, respectively. It is obvious that
the maximum glycerol concentration of different pH was closely
related to strain. For strain F, the maximum glycerol concentration
appeared at pH 2.50; for strain B, the maximum glycerol con-
centration was also at pH 2.50, and then the maximum glycerol
concentration was significant increased (P < 0.05) along with the
decrease of pH; but for strain N, the maximum glycerol concen-
tration appeared at pH 4.50, perhaps this difference was because
of the weak tolerance of strain N. As shown in Figure 5, ini-
tial pH had an vital influence on glycerol content of yeast strain,
but high content of glycerol at low pH perhaps because low pH
inhibited cell growth and alcoholic fermentation, which lead to
sugar hardly been consumed, so the osmotic stress caused by high
sugar concentration was sustained, and the production of glyc-
erol as an osmolyte was required to counteract hyperosmotic stress
(Zuzuarregui and others 2005). Whether single pH affects glycerol
production? Perhaps further study can use LMSM to avoid high
osmotic pressure caused by sugar.
Figure 6–Effect of different initial pH value on acetic acid concentration Figure 7–Effect of different initial pH value on L-succinic acid concentration
of strain F, B, and N during fermentation in MSM.
Acetic acid, a byproduct formed during yeast alcoholic fermen-
tation, is the main component of volatile acidity. When present
in high concentrations in wine, acetic acid imparts an undesirable
‘vinegary’ character that results in a significant reduction in quality
and sales Cordente and others 2013. The OIV (2012) states that
the maximum acceptable limit for volatile acidity in most wines is
1.20 g/L of acetic acid.
Studies on the production of volatile acidity by S. cerevisiae under
winemaking conditions showed that this acid is mainly formed at
the beginning of alcoholic fermentation (Vilela-Moura and others
2011) and its production is affected by different factors, including
the yeast strain, anaerobiosis, very low pH (<3.1), or very high
pH (>4; Pascal Ribereau-Gayon and others 2006; Vilela-Moura
and others 2006).
Effect of initial pH on acetic acid concentration was shown in
Figure 6. As shown in Figure 6, the change trend and maximum
acetic acid concentration of strain F, B, and N was closely related
to initial pH value. Acetic acid was generated by strain F, B, and
N after inoculated 3, 2, and 2 d, respectively, the maximum value
at 4 pH was soon reached with the values of 0.43 to 1.25, 0.30 to
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of strain F, B, and N during fermentation in MSM.
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 Effect of initial pH on yeast . . .

0.91, and 0.42 to 1.00 g/L, respectively. The maximum acetic acid Bo Jia participated in writing,; Cheng Wang and Fang Zhao par-
concentration at pH 2.50 was huge higher than that at the other ticipated in statistical analysis.
3 pH values. For strain B, the lower the initial pH, the higher
the maximum acetic acid concentration, and the maximum acetic
acid concentration at 4 pH was significantly different to each References
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