Jeffrey F.

Giannini
Introduction to Research (BIOL3600)
Professor Dr. Zwick
Annotated Bibliography
18 March 2014

How is fermentation by Saccharomyces cerevisiae affected by

pressure?

Fermenting various alcoholic beverages has been a passion of mine for the
past 5 years. Recently in my endeavors I have begun to ferment in closed containers
where pressures have been building higher as fermentation progresss. I have begun to
wonder what effect this pressure has on the resulting fermentation. During the course
of my search I started to understand just how much pressure like temperature can be
used to bring about certain flavors in beer by inhibiting certain esters and fusels more
so than others. I found out the pressures I deal with are far below where yeast start to
see sever changes for the worst. It was also quite interesting to read about
fermentation under a near vacuum and how yeast seemed to thrive in that sort of
environment as well. I think there is much left to be explored in this area not only in
bringing about desired flavors in beer but by telling us how to build better more efficient
bio reactors, and bio-fuels.

1. Classic Source
Ramalingham A, Finn RK. 1977. The Vacuferm Process: a New Approach to Fermentation
Alcohol. Biotechnol Bioeng; 19(4): 583-589.
In finding a classic source cited over 50 times I found google scholar to be my search
engine of choice. The search terms I found most useful in this query were yeast, ferment, and
pressure. In progressing through my research I learned to stray away from the term stress.
When the term stress was included the studies were mainly concerned with genetic
mechanisms in yeast, as opposed to quantitative analysis of fermentation.
The following paper stood out among other in that it was cited more than any other
paper I came across while being pertinent to my topic. In addition to being widely cited the
paper introduced me to the concept of fermentation under a vacuum which I found immensely
interesting. The paper for me specifically, was useful for understanding my topic better by

mentioning a minimal pressure which the yeast need in order to survive. Initially I believed as
pressure increased fermentation would slow so it was useful for me to know that an absolute
vacuum would kill the yeast. Upon seeing the words Vacuferm Process my attention
immediately peaked and I knew I had found some useful information.
PassageThe maximum cell density for the vacuum run was nearly one and one-half times
higher than that for the atmospheric run; the exponential growth phase for the vacuum run was
extended by nearly 2hr. Near the end of exponential growth, the residual sugar level fell faster in
the vacuum run than in the atmospheric run. However this higher rate of disappearance of sugar
is accounted for by the higher cell density at this stage in fermentation.
ParaphraseCarrying out fermentation under a near vacuum leads to an extended growth cycle producing
more yeast cells than otherwise found under normal atmospheric pressure (Ramalingham and
Finn 1977).

2. Source Older Than 2003:

As I stated previously I encountered problems being overwhelmed in my searches with
multiple copies of papers concerned with yeast genetics. The path I chose in finding this paper
was looking at the list of references in a paper that was concerned with pressure, however not
for the purposes that interest me. I found this to be a great way of moving around between
papers ounce I was in my niche I found most papers referenced one another. Seeing the name
of the journal alone was enough for me to give it further examination.
I chose this specific source because it goes into detail about how the flavor profile is
affected by pressure during fermentation. I found it interesting that pressure affects certain
esters and fusels differently allowing for more room in tweaking flavors.
PassageThe concentrations of esters and fusel alcohols formed during brewery fermentations were
significantly reduced by increased carbon dioxide pressure. However, the overall formation of
fusel alcohols was less affected than that of the esters.
ParaphraseIncreased Carbon Dioxide pressure can be used to lower ester levels while having less of an
effect on total fusel alcohols.(Renger et al. 2001).
3. Recent Source:

In searching for my recent source I again used google scholar as my search engine of
choice. I limited my searches to the articles being published from 2011-present while using the
key words yeast fermentation and pressure.
Selecting this article was easy due to the vast majority of papers of interest to me being
at least 5 years old. This article offered the most data points for various pressures which was
useful for seeing how the effects play out across a broad range not just extremes. I think this
article can be of particular use to me because it offers a clear level of pressure at which things
start to change drastically as well as warning me that toxic methanol production doubles after
3atm. This papers title almost mirrors my topic question so I felt it was a great fit, setting me off
on a good direction for my other sources.

Passageethanol productivities were not significantly different at pressures of 13 atm, but were
significantly lower for 5 and 7 atm (27.0 and 20.5 g/L d, respectively).The acetaldehyde
concentration was low to negligible for all the pressures tested (Table 3). Methanol showed a
more than 2-fold increase above 3 atm compared to 2 atm (7298 and 33 mg/L, respectively).
ParaphraseAs pressure increases above 3atm toxic methanol concentrations more than double while
desirable ethanol concentrations drop (Charis et al.2012).

4. Personal Choice:
For this article I used all of my previous terms while adding carbon dioxide to my search. I found
this article while searching through summon. I selected this article because it deals with wine
where my other selections generally dealt more with beer or just sugar. This article mentions in
the abstract something about yeast being conditioned to continuously ferment at high pressure
however I was unable to understand anything further in this paper at this time.
PassageTable 2 shows the effects on yeast growth of alcohol concentration [adjusted as given in
Materials and Methods to give 10, 11, and 12% (v/v)]. These samples were maintained at 5 atm
of pressure. All samples were inhibited at this pressure, as compared with multiplication at
atmospheric pressure (see Table 1). The extent of inhibition was dependent on alcohol
concentration and pH. The results in Tables 1 and 2 show that pressure is so inhibitory that
adjustments in pH and alcohol concentration do not allow yeast multiplication to be as rapid as
at atmospheric pressure. Even at the lowest alcohol and highest pH level, yeast multiplication
under pressure never reached 100-fold increase.
ParaphraseWhen presented with less than satisfactory conditions yeast seem least able to rebound
when subjected to pressure opposed to ph or high alcohol(Kunkee and Ough 1966).

5. Review Article:
I searched this article through Stockton summon search I used the terms yeast stress
and pressure. Again I selected this article due to the title nearly mirroring my topic question. I
selected this because it gives a pressure where yeast morphology are affected and another
where wild type yeast die. It was particularly interesting to continue to examine yeast respond to
pressure in similar ways they respond to heat. Also the paper mentions how yeast can develop
a barotolerance if pretreated for high pressure.
PassageYeast viability during hydrostatic pressure treatment decreases with increasing pressure and
this effect is more pronounced when cells are submitted to pressures above 100MPa, while at
220 MPa all wild-type cells are killed. A pressure of 50 MPa is not sufficient to kill or to alter
yeast cell morphology.
ParaphraseYeast cells remain unchanged when submitted to pressures under 50Mpa however as
pressures approach 220Mpa all wild types will perish(Fernandes 2005).

6. Popular New Article:

I found this article after sorting through many ones I found useless for the topic. I started my
search with the local newspapers website which offered plenty of articles about beer, however
none that delved into anything particular useful for my purposes. I used the key words
fermentation, yeast, and beer.
I selected the current article because I think it is really outstanding. It sort of brought my
topic full circle now knowing how pressure effects fermentation this article let me know how the
wine and beer industry are using the same knowledge I just gained to create flavors. I feel like it
addressed many other aspects of how our increasing knowledge of science is changing
traditions that have remained somewhat unchanged for thousands of years. I have always
known that yeast were responsible for the production of these flavors however after completing
this project I now better understand another tool the brewing industry, and now myself has to
bring about flavors in the products they create.
Passage-

Adjustments are also permitted in the level of carbon dioxide in fermenting wine, which affects
a wine's acidity and fruitiness. Adding unfermented grape juice sweetens the wine. Enzymes
lock in color. Yeasts control the level of fermentation. Tannins, naturally occurring chemical
compounds in grape skins and wood, are used in powdered form to further enhance a wine's
taste and feel in the mouth.

Advances in yeast cultivation have now made it an ingredient for taste as well. Chardonnay
producers looking for a toasty, buttery taste use a special yeast that enhances those qualities.
Another example is a yeast that gives a banana flavor and aroma, originally introduced 10 years
ago in Beaujolais.
Marty Bannister, the founder of Vinquiry, a wine analysis and consulting firm in Sonoma, Calif.,
said yeast was ''the essential fermentation tool.'' But now, she added, ''people also look toward it
for flavor.''
ParaphraseYeast are increasingly being looked at today not only to transform sugar into alcohol but to
impart a variety of specialized aromas and flavors.(Feiring 2001).

Bibliography
1. Classic Source:
Ramalingham A, Finn RK. 1977. The vacuferm process: a new approach to fermentation
alcohol. Biotechnol Bioeng; 19(4): 583-589.

2. Pre 2003 source

Renger R, van Hateren S, Luyben K. 1992. The formation of esters and higher alcohols during
brewery fermentation; the effect of carbon dioxide pressure. Jnl Institute Brewing, 98(6): 509513.

3. Recent source 2011-present:

Charis M, Galanakis A, Kordulis C, Kanellaki M, Koutinas A, Bekatoroub A, Lycourghiotis A.
2012. Effect of pressure and temperature on alcoholic fermentation by Saccharomyces
cerevisiae immobilized on c-alumina pellets. Bioresour Technol. (114)0:494-498.

4. Personal Choice:

Kunkee R, Ough C.1966. Multiplication and fermentation of saccharomyces cerevisiae under

carbon dioxide pressure in wine. Appl Microbiol. (14)4: 643-648

5. Review paper:
Fernandes P. 2005. How does yeast respond to pressure? Braz J Med Biol Res, (38)8: 12391245.

6. Popular New Article:

Feiring A.Business; For Better or Worse, Winemakers Go High Tech.26 August 2001 NY
Times[Internet][Cited 2014 March 18];[about 3 screens]. Available from:
http://www.nytimes.com/2001/08/26/business/business-for-better-or-worse-winemakers-go-hightech.html