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Rapid Beer Fermentation: The Effect of Vacuum Pressure on a Pilot Scale Lager
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DOI: 10.1080/03610470.2019.1669416

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 Journal of the American Society of Brewing Chemists
The Science of Beer

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Rapid Beer Fermentation: The Effect of Vacuum

Pressure on a Pilot Scale Lager Fermentation

Mario Guadalupe-Daqui & Andrew J. MacIntosh

To cite this article: Mario Guadalupe-Daqui & Andrew J. MacIntosh (2019): Rapid Beer
Fermentation: The Effect of Vacuum Pressure on a Pilot Scale Lager Fermentation, Journal of the
American Society of Brewing Chemists, DOI: 10.1080/03610470.2019.1669416

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JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS
https://doi.org/10.1080/03610470.2019.1669416

Rapid Beer Fermentation: The Effect of Vacuum Pressure on a Pilot Scale Lager
Fermentation
Mario Guadalupe-Daqui and Andrew J. MacIntosh
Food Science and Human Nutrition Department, University of Florida, Gainesville, Florida, U.S.A.

ABSTRACT KEYWORDS
Changes to industrial fermentation processes are rare, largely due to the impact on viability for Carbon dioxide; high
Saccharomyces spp., related to environmental stresses during the process such as: osmotic pres- gravity; low pressure; rapid
sure, ethanol concentration, dissolved CO2. This research studied yeast behavior during a High fermentation; vacuum
Gravity (HG) fermentation under partial vacuum pressure during standard brewing operations. This
was accomplished by controlling the temperature and pressure of 30 L pilot fermentations. An
experimental lager wort recipe with “Diamond” lager yeast (Saccharomyces pastorianus) was used
in this study. The modified process conducted at 24.1 kPa of pressure in the headspace was com-
pared to a control process at standard atmospheric pressure (101.3 kPa). This research showed
that at 15
C, lager fermentations (14–15
P) fermented up to 30% faster under vacuum, with an
increase in the number of yeast cells in suspension of up to 100%. The rate of sugar consumption
under vacuum was greater compared to the control, thus increasing the rate of ethanol produc-
tion. The removal of volatile CO2 is one hypothesized mechanism for these results. The total etha-
nol production, pH and yeast viability did not differ significantly between the control and
modified process. The application of vacuum during brewing resulted in an increased fermenta-
tion rate and number of yeast cells in suspension during fermentation.

Introduction 19th century, the nature of the fermentation process was

Historically, the industrial fermentation process has been
slow to change due to the potential impact upon the viabil-
ity of typical brewer’s yeast. The impact of environmental
conditions on brewing species (Saccharomyces cerevisiae and
Saccharomyces pastorianus) is related closely to specific
stresses present during the process. Throughout fermenta-
tion, levels of osmotic pressure,[1] ethanol concentration,[2]
and carbon dioxide[3,4] all affect yeast viability.[5] If too
severe, the effect of the stress upon the yeast may result in
non-viable cells, forcing the fermentation either to slow
down or cease without reaching the desired extract concen-
tration (also termed attenuation). Yeast cells undergo adap-
tation to various stresses during the brewing process.[6] One
such mechanism of yeast to resist diverse stresses is the for-
mation of different protective compounds, such as trehal-
ose[7] and glycerol.[6,8,9] One method to reduce carbon
dioxide concentration is through fermentation under a par-
tial vacuum; the reduced total pressure results in a low CO2
partial pressure, thus a lower amount of CO2 dissolved in
the wort.
Fermentation is a critical process to the brewing industry,
in which the main goal is to use yeast to convert sugars into
ethanol and carbon dioxide.[10] Around 1854, Louis Pasteur
began researching on fermentation,[11] and by the end of the
clearly established. While many other industrial fermenta-
tions have changed from batch to continuous, semi-continu-
ous or controlled atmosphere conditions,[12] the brewing
industry continues to utilize batch fermentations under one
atmosphere (101.3 kPa), carbon dioxide headspace.
Previous work by Cysewski and Wilke[13] explored two
different methods of controlled atmosphere yeast fermenta-
tion: a semi-continuous and a continuous process, where a
constant feed of 10% glucose media was fed to the fermen-
tor. Due to the accumulation of compounds not metabolized
by the yeast, especially under a semi-continuous process, the
viability decreased to 60% of viable cells after 55 h. The vac-
uum continuous fermentation studied included a constant
volume of media fed to the fermentor, as well as a specific
volume of fermented broth withdrawn to avoid the accumu-
lation of toxic metabolites. The cell mass was kept constant
at 50 g/L for 13 days. Ethanol production rate increased
from 7 g/Lh1 at atmospheric continuous conditions to 40 g/
Lh under vacuum pressure. Continuous vacuum fermenta-
tion was able to completely ferment a 33.4% glucose feed
media, whereas an atmospheric fermentation was not able to
do this.
Ramalingham and Finn[14] compared a batch fermenta-
tion at atmospheric and using vacuum conditions. Similar to
previous research, this research included the removal of

CONTACT Andrew J. MacIntosh andrewmacintosh@ufl.edu

Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/ujbc.
Presented at the 2018 ASBC Annual Meeting, San Diego, CA.
ß 2019 American Society of Brewing Chemists, Inc.
2 M. GUADALUPE-DAQUI AND A. J. MACINTOSH
alcohol from the fermentor, maintaining a constant level of
alcohol below 6% by volume. Their study concluded that the
yeast growth rate during the exponential phase did not differ
when fermenting under either condition. However, vacuum
fermentation resulted in a higher cell density than atmos-
pheric fermentation. Under vacuum pressure, the cell density
was one to one-half times higher than at one atmosphere.
Also, the extract fell faster, reaching a higher cell density
under vacuum pressure than atmospheric conditions. This
study also mentioned that the removal of alcohol led to better
sugar utilization. It is noteworthy that this study did not con-
trol the acclimation of yeast with oxygen.
Literature has emphasized the importance of oxygen dur-
ing a fermentation process, related to the formation of ster-
ols and fatty acids.[14] Cysewski and Wilke[13] concluded
that dissolved oxygen promoted the rate of fermentation.
Although it was not clearly defined as the mechanism, this
is very likely due to the growth of new yeast cells, as well as
the formation of protective compounds. Their study
included the use of air instead of oxygen, resulting in a
reduced rate of ethanol production. However, the literature
discussed did not represent a conventional brewing operation
system, since it was more focused on a semi-continuous or
continuous fermentation. Therefore, it is possible that the
process of manipulating pressure can be used as a practical
modification for industrial brewing fermentations (thus
increasing the rate of fermentation), provided the oxygen
requirements for the yeast are optimized, and a simple pro-
cess for the removal of CO2 from the headspace is developed.
The objective of this research was to evaluate and study
the behavior of yeast within the high gravity lager wort fer-
mentation under vacuum pressure (24.1 kPa) compared to
normal fermentations (101.3 kPa). Specifically, to determine
the effect upon ethanol production, number of yeast cells in
suspension, and viability of cells, in order to assess the feasi-
bility of using this technology in the brewing and industrial
fermentation industry. To accomplish the objective, this
study controlled the temperature and pressure of a 30 L pilot
fermentation at several extract concentrations, while moni-
toring attributes such as: extract, pH, number of yeast cells
in suspension, viability, and ethanol concentration.
Experimental
In order to assess the effect of vacuum pressure on typical
brewing fermentations, conditions were mimicked as closely
as possible over a series of pilot scale fermentations, within
a vacuum rated 130 L fermentor. The experiment was
designed to run three duplicates of vacuum and control fer-
mentations, six runs in total at 14.5
P ± 0.5
P (each run
took 1.5 weeks including propagation and cleaning).
Sufficient wort was created for a control and vacuum fer-
mentation from each mash. After pitching, these worts came
to 14
P, 14.5
P and 15
P. Therefore, the six experiments
were performed in alternating pairs (initial run at 14
P, first
control then vacuum; second run at 14.5
P, first vacuum
then control; final run at 15
P, first control then vacuum.).
For these fermentations, dried yeast was rehydrated and
propagated in a 5 L bioreactor prior to pitching into the
130 L primary fermentor. Fifteen to twenty samples were
taken throughout the fermentations and extract, pH, etha-
nol, number of yeast cells in suspension, and viability were
assessed. Each attribute was modeled and statistically com-
pared to determine if any difference between fermentation
pressures occurred. The overall experimental preparation is
outlined in Figure 1.
Media (wort)
The wort used for this research was based upon an experi-
mental lager wort recipe as described in Table 1. The time
and temperatures used during the mash are presented in
Table 2. Hops were added to the boil in two parts: Hallertau
Hersbruckera was added 20 min after boiling was initiated.
Additional Hallertau Hersbruckerb was added at the end of
the boil (70 min after the first addition). UK Pale Ale
(Propino) 2 row, and Rahr standard 6 row malt were uti-
lized in this research and were provided by Muntons plc.,
and BSG Inc., while Hallertau Hersbrucker hops were
from Yakima Chief Hops (YCH) and flaked rice was from
Briess Malt & Ingredients Co., and were purchased from
Hoggetowne Ale Works in Gainesville, FL. The water used
for this research was filtered and sterilized using Impact LC
Series commercial reverse osmosis equipment (IWP water
systems, Austin, TX) with an UV lamp attached.
Yeast rehydration and propagation
A dried Saccharomyces pastorianus strain, commercially des-
ignated “Diamond Lager” Active Dry Yeast (ADY), was used
in this study. Multiple packages containing the same pro-
duction lot were provided directly by Lallemand Inc. These
were stored under refrigeration at 4
C, in their supplied
packages, prior to use. Survival of yeast during rehydration
and storage is determined by many factors such as: osmotic
pressure, temperature, and the medium.[15] Therefore, the
rehydration and propagation procedures are vital to reduce
variability between fermentations and these were carefully
controlled to ensure a consistent inoculum for the fermenta-
tion trials. The ADY was rehydrated at 25
C for 1 h as per
the method described by Jenkins et a1.[16] After rehydration,
approximately 15  106 yeast cells per mL were pitched into
a 5 L BioFloV R 310 Benchtop Bioreactor for 48 h of propaga-
tion at 20
C. Air was supplied to the bioreactor with a con-
stant flowrate and 100% relative humidity, while a constant
agitation of 100 rpm was maintained. The main objective of
this process was to promote yeast growth under aerobic
conditions, the number of yeast cells, yeast viability, pH
drop and extract reduction rate were monitored. Figure 2
shows the yeast cells in suspension and viability data during
propagation, prior to each fermentation run.
Fermentation
The primary fermentation was completed in an Applikon
BioPlot 130 L cylindroconical fermentor. A condenser was
 JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 3

Figure 1. Process description for creation of experimental wort for control and vacuum fermentations.

Table 1. Experimental lager wort recipe. Table 2. Process applied to make lager wort.
Ingredients Quantity Process Temperature Time
Malted barley Mashing 65
C 90 min
2-row (Muntons - Propino) 14.37 kg 79
C 10 min
6-row (Rahr - standard 6 row) 14.37 kg Lautering 79
C
Adjunct Boiling Phase 1 Boiling point 20 min
American flaked rice (Briess) 9.61 kg Phase 2 Boiling point 70 min
Hops
Hallertau Hersbruckera 147.91 g
Hallertau Hersbruckerb 295.83 g 101.3 kPa and 24.3 kPa respectively; both at 15
C, until the
Water
Mashing water 160.00 L
lowest extract content was reached. Temperature and pres-
Lautering water 40.00 L sure were controlled to ±0.5
C and ±3.5 kPa, while pH, dis-
a
First addition of hops to the boiling process (Phase 1). solved oxygen, extract, ethanol concentration, yeast cells in
b
Second addition of hops to the boiling process (Phase 2). suspension and yeast viability were monitored via intermit-
tent sampling.
attached to the top of the tank and connected to the vacuum
line during vacuum fermentations. The condenser was main- Wort sampling procedure
tained at 4
C during fermentations to retain evaporated
volatile ethanol to the fermentation during vacuum fermen- Control and vacuum fermentations were sampled (100 mL)
tation. A 30 L portion of 16.5
P wort was poured into the at appropriate intervals to assess the monitored fermentation
fermentor and diluted to three different extract concentra- attributes. An initial sample of fermenting wort was taken
tions (14.0
P, 14.5
P and 15
P). Diluted wort was oxygen- 30 min after pitching, to guarantee a good distribution and
ated until fully saturated using an inner bubbler (sparge) adaptation of the inoculum. Vacuum fermentation samples
and agitation at 100 rpm. The pitching rate was controlled at were taken by applying a vacuum pressure to the sample
15x106 viable cells per mL and mixed at 100 rpm for 5 min port, lower than the internal pressure of the tank, thus
to homogenize. The timing of the vacuum application was ensuring that there was no increase of pressure and air con-
an important attribute due to the oxygen requirements of tamination to the tank. A 5 mL aliquot of the fermenting
yeast during fermentation. Thus, the vacuum was not media was separated for cell count and viability determin-
applied until the oxygen had been fully depleted. The dis- ation of the yeast. The rest of the sample was degassed and
solved oxygen within the wort was monitored, and the vac- filtered immediately after the pH measurement. The samples
uum applied 12 h after pitching for each fermentation. were partially degassed by shaking in a tightly sealed tube,
Control and vacuum fermentations were then carried out at unscrewing the lid to relieve the pressure until no more gas
4 M. GUADALUPE-DAQUI AND A. J. MACINTOSH
Figure 2. Yeast propagation under aerobic conditions and at 20
C, prior to each fermentation. (A) for fermentation at 14
P, (B) for fermentation at 14.5
P, (C) for
fermentation at 15
P. (-) Cells in suspension and (- -) viability.
was released (approximately 3 times). Finally, the degassed
sample was vacuum filtered, using 1 g of diatomaceous earth
(DE) per 100 mL of sample. The filtered degassed sample
was then assessed using the Anton-Paar ALEX 500 to deter-
mine the ethanol and extract concentration.
Measured attributes: yeast cell in suspension, viability,
extract, ethanol, and pH determination
Yeast cells were counted after rehydration, and throughout
both propagation and fermentation, based upon the method
described by the American Society of Brewing Chemists
(ASBC) Yeast-4. A hemocytometer with a cover slip was
used for cell collection and the cells were observed through
a microscope with at 400 magnification (combining a 40
objective lens and 10 eyepiece). Yeast viability was assessed
using Painting and Kirsop’s[17] methylene blue technique.
The methylene blue technique is commonly accepted in the
industry to assess viability. However, it should be noted that
the accuracy of this technique is significantly reduced when
viability is less than 90%.[18] Methylene blue was used in
this research due to its simplicity, prevalence in industry,
and as the viability throughout the fermentation was
expected to be higher than 90%.
An AccumetV R AB15 pH meter was used to determine the
pH measurements. Thirty mL of degassed and filtered sam-
ple was used per test to determine extract, measuring the
density based on the oscillating U-tube technology using an
Anton-Paar DMA35. Ethanol concentration was calculated
based upon the ASBC Beer-4: Alcohol part C, then con-
firmed using an Anton-Paar ALEX 500 through an absorp-
tion measurement via Near Infrared Reflectance (NIR)
spectroscopy.
Models
Two models were used for the data analysis, a Step model
(Equation 1) and the Sigmoidal five-parameter (5P) model
(Equation 2). Due to the high sampling frequency, the Step
model described the number of yeast cells in suspension
during the fermentation more accurately than previously
discussed methods such as those described in the American
Society of Brewing Chemists (ASBC) Yeast-14 method
(Equation 1). The amplitude of this model described the
maximum quantity of yeasts in suspension during the fer-
mentation. It also incorporated the Heaviside function to
account for the final yeast population[19], when the yeast
was not removed via cold crash. The amplitude of the Step
model was selected for the statistical analysis of significant
differences with respect the maximum number of suspended
yeast cells. The Step model is described as follows:

tl 2
12 r !
ð1þHðtÞ SÞ
e A
PðtÞ ¼ A þ A (1)
1 þ HðtÞ S ð1 þ SÞHðtÞ
where P(t) is the cells in suspension at a specific time (t),
A is the amplitude, m is the midpoint, r is the width factor.
H(t) is the Heaviside step function and S modifies the height
of the step in this function.
The Sigmodal 5P model was used to describe the extract,
ethanol content and pH drop throughout the fermentation.
The slope of the Sigmoidal 5P model was selected for the
analysis of the extract and ethanol production rates during
control and vacuum fermentations. Propagation data was
analyzed using this model. The sigmoidal 5P model is
described as follows:
Pi  Pe
PðtÞ ¼ Pe þ (2)
ð1 þ s eBðtMÞ Þ s
1
where P(t) is a function in term of a specific time (t), Pi
is the upper asymptote of the curve, Pe is the lower asymp-
tote of the curve, M is the time of the inflection point of the
curve, B is the slope, which can also be considered as the
rate factor of the curve, and s modifies the shape of the
curve around the inflection point.
Statistical analysis
Experimental data was subjected to statistical analysis using
GraphPad Prism version 8.00 for Windows, GraphPad
Software, La Jolla, CA, www.graphpad.com. The level of sig-
nificance used to determine differences was established at
5%. The models adjusted to the data collected during the

control and vacuum fermentations were fitted with a 95%
confidence interval.
Results and discussion
Rehydration and propagation of yeast prior to
fermentation
After initial rehydration, yeast viability was measured to be
43.52 ± 12.84%. This low percentage of viable yeast was likely
due to the stress applied to the ADY during drying and
given the low accuracy of methylene blue at low viabilities,
this number is likely not accurate. Regardless, this initially
low number showed a need to improve the viability of the
yeast prior to the fermentation. Therefore, a 48 h propaga-
tion was applied and yeast viability increased to
92.83 ± 2.72%. Although 93% viability was not ideal, the ini-
tial viability was consistent among the trials, reducing any
potential variability that could originate from this attribute.
Figure 3. Yeast cells in suspension throughout a 14
P, 14.5
P and 15
P fermentation at 15
C. Vacuum conditions at 24.1 kPa (v) and atmospheric conditions at
101.3 kPa (c).
Figure 4. Extract and ethanol concentration throughout a 14
P, 14.5
P and 15
P fermentation at 15
C. Vacuum conditions at 24.1 kPa (v) and atmospheric
conditions at 101.3 kPa (c).
JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 5
The final count of the yeast cells in suspension over the
propagation was 155.25 ± 16.98  106 cells per mL; this rep-
resented a 10-fold increase. No significant differences at P
value  0.05 were found between the propagations per-
formed prior each fermentation with respect to the previ-
ously mentioned attributes. The data collected during the
propagations are shown in Figure 2. Thus, through consist-
ent propagation techniques, there was no significant differ-
ence in the inoculum used for the vacuum fermentations
and atmospheric controls.
Vacuum and control fermentation
Yeast cells in suspension
The effect of partial vacuum pressure applied to yeast cells
in suspension during fermentation compared to the control
process is shown in Figures 3–5. Under partial vacuum pres-
sure, higher numbers of yeast cells in suspension were
6 M. GUADALUPE-DAQUI AND A. J. MACINTOSH

Figure 5. Direct comparison of control (A) versus vacuum (B) fermentation at 15
P, with respect to five fermentation attributes (extract, pH, ethanol concentration,
viability and yeast cells in suspension).

Table 3. Summary of statistical results from fermentations.

Original extract Model Treatment Model fit R2 P value Null hypothesis Conclusion
Cells in suspension
14
P Step model Control 0.8856 0.0266 Yeast cells in suspension are the same Reject null hypothesis
Vacuum 0.9942 for control and vacuum fermentation
14.5
P Step model Control 0.8856 0.0001 Reject null hypothesis
Vacuum 0.9294
15
P Step model Control 0.9351 <0.0001 Reject null hypothesis
Vacuum 0.8988
Attenuation rate
14
P Sigmoidal Control 0.9992 0.0086 Attenuation rate is the same Reject null hypothesis
Vacuum 0.9967 for control and vacuum fermentation


14.5 P Sigmoidal Control 0.9978 <0.0001 Reject null hypothesis
Vacuum 0.9947
15
P Sigmoidal Control 0.9990 <0.0001 Reject null hypothesis
Vacuum 0.9991
Ethanol production rate
14
P Sigmoidal Control 0.9993 0.0106 Ethanol production rate is the same Reject null hypothesis
Vacuum 0.9967 for control and vacuum fermentation
14.5
P Sigmoidal Control 0.9978 <0.0001 Reject null hypothesis
Vacuum 0.9949
15
P Sigmoidal Control 0.9990 <0.0001 Reject null hypothesis
Vacuum 0.9991

measured as compared to atmospheric conditions; a 15%
increase for the 14
P wort, 25% for the 14.5
P wort, and
100% for the 15
P wort. Table 3 shows how the Step model
(Equation 2) fitted the yeast data collected based upon the
coefficient of determination (R2), and the P value for the
comparison of the models between the vacuum experiment
and atmospheric control. This result is similar to that found
by Ramalingham and Finn,[14] where an increase of
100–150% of the cell density was observed under vacuum
pressure compared to atmospheric in a continuous (non-
brewing) fermentation. It was also found that the increase in
yield of yeast cells in suspension under vacuum pressure
throughout the fermentation process was proportional to the
original extract level (initial fermentable sugar concentra-
tion). There were significant differences between each con-
trol and vacuum fermentation at the three original extract
levels measured, and none between duplicate experiments.
Yeast viability and pH drop
Yeast viability was measured during vacuum and control
fermentations and increased over each process. A rising
trend was observed in both vacuum and control fermenta-
tions. The viability of the yeast in each fermentation started
at 91.22 ± 3.75% and rose to 97.54 ± 1.02% at the end of the
fermentation. This yeast viability trend was greater com-
pared to the vacuum viability trend reported by
Ramalingham and Finn.[14] They found a decreasing viabil-
ity trend and theorized that this was due to the accumula-
tion of nonvolatile metabolites, a common problem with
continuous and semi-continuous fermentations, regardless of
pressure. Normally, pH reduces during fermentation due to
the yeast’s production of carbon dioxide and the yeast’s
secretion of organic acids.[6,20] The pH throughout both
control and vacuum fermentations was measured as
described at each sampling interval. It decreased as expected
and its slope showed no significant difference between the
vacuum and the control fermentations. This was likely due
to the fact that the carbonic acid (at a higher concentration
in the atmospheric fermentation) did not significantly con-
tribute to the pH of the wort, as compared to the produc-
tion of organic acids.
Extract and ethanol concentration
The extract and ethanol concentrations throughout the fer-
mentations under vacuum and atmospheric conditions are
shown in Figure 4. The rate of extract reduction under the

vacuum fermentation was greater in every trial than the
controls (at atmospheric pressure), thus increasing the rate
of ethanol production when vacuum was applied. The total
reduction in the fermentation time, when fermenting
under vacuum, was found to be 15%, 20% and 30% lower
than that of the control process, using extracts of 14
P,
14.5
P and 15
P, respectively. The removal of CO2 and the
higher number of yeast cells in suspension under vacuum,
is the hypothesized mechanism for the higher observed
rates of fermentation. Vacuum fermentation showed a
higher extract reduction and ethanol production rates
compared to atmospheric pressure. Similar to the max-
imum number of yeast cells in suspension discussed previ-
ously, these rates appeared to be directly proportional to
the original extract (initial fermentable sugar concentra-
tion). There were significant differences between the con-
trol and vacuum fermentation with respect to the extract
reduction rate and with respect the ethanol production
rate at the three original extracts performed. Table 3
shows how the Sigmoidal 5 P model (Equation 2) fitted the
data collected based upon the coefficient of determination
(R2) and the P value obtained in each trial. A direct com-
parison of vacuum versus control fermentation at 15
P is
shown in Figure 5.
Previous research conducted on the application of vac-
uum during yeast fermentation has been more focused on
semi-continuous or continuous processes. These processes
are less applicable to brewing industries, as they normally
operate on daily schedules. However, under vacuum pres-
sure, Ramalingham and Finn[14] observed a production of
ethanol 5-times higher than at atmospheric pressure. This
study shows that similar results can be attained by the brew-
ing industry during batch processes, and through manipula-
tion of pressure, brewers can increase the rate of production
and cell density. These results open multiple new avenues
for research into modified atmosphere fermentations with
direct applications to the brewing industry.
Conclusions
Vacuum fermentation increased the number of yeast cells in
suspension, resulting in a higher ethanol production rate
and a decreased overall fermentation time (up to a 30%
reduction in total fermentation time, when pressure was
reduced to 24.1 ± 3.5 kPa at the 12 h mark and then held
constant). This is potentially due to the reduction in total
CO2 concentration, a known stress factor for yeast. Yeast
viability under both vacuum and atmospheric conditions,
increased throughout all fermentations (from 91.22 ± 3.75%
to 97.55 ± 1.02%). The pH of the vacuum fermentations did
not differ statistically from the control fermentations.
The results obtained from the early research opens the
possibilities for novel research related to the brewing indus-
try. Further studies are needed on how carbon dioxide, spe-
cifically its reactive species such as carbonic acid,
bicarbonate ion or carbonate ion, can inhibit yeast fermenta-
tion, and these studies could help explain the increased rates
of fermentation observed during this work. It is possible
JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 7
that the carbon dioxide could be impacting the yeast as dis-
solved molecular CO2, carbonic acid, HCO3, CO32, or as
physical bubbles influencing the yeast cell, and further cap-
acity to release intracellular CO2. Due to the increased num-
ber of cells in suspension, it is hypothesized that this
research could be applied to very high extract fermenta-
tions (>20
P).
In summary, fermentation under vacuum pressure was
completed on a pilot scale, using an experimental wort. The
results of this study are immediately applicable for distilla-
tion and bioethanol production, where the profiles of aro-
mas and flavors are not of major importance compared to
the speed and efficiency of the process. To be utilized within
the brewing industry, additional research on volatile and
sensory changes are necessary, as well as modifications to
trap escaping volatiles from the vacuum apparatus. This
technology has the potential to significantly increase fermen-
tation efficiency, if the sensory characteristics and quality of
the product are not negatively affected by the process.
Future work will assess fermentations under high pressure,
as well as examine the impact upon the sensory and volatiles
present during vacuum fermentation.
Note
1. g/Lh: grams per liter per hour (g/liter-h).
Acknowledgments
The authors would like to acknowledge Lallemand Inc. and Muntons
plc. for their kind donations of yeast and malt, respectively, used
throughout this research.
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