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Biology

The effect of temperature on the growth of ​Saccharomyces cerevisiae ​populations.

Aryan Suri
Mrs. Henderson
August 31 2017
IB Biology HL-2
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Biology

Table of Contents

SEC 1. Aim 3

SEC 2. Engagement 3

SEC 3. Background 3

SEC 4. Variables 4

SEC 5. Materials 6

SEC 6. Procedure 6

SEC 7. Safety and Ethical Issues 7

SEC 8. Data and Calculations 8

Table 1.1 Growth of Yeast (by ml) after 26 hours at specific temperature
Table 1.2 Average Growth of Yeast (by ml) after 26 hours at specific temperature
Table 1.3 Standard Deviation of Growth of Yeast (by ml) after 26 hours at temperature
Table 1.4 Qualitative Data Marked
Graph 1.5 The Average change in Yeast after 26 hours at Temperature
Table 1.6 ANOVA Statistics Calculations

SEC 9. Conclusions 10

SEC 10. Improvements and Evaluation 11

SEC 11. Works Cited 12

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Biology

SEC 1. Aim

The aim of this Internal Assessment is to ​investigate​ how (and by how much) temperature
affects the growth, stability, and survivability of ​ Saccharomyces cerevisiae ​, commonly known
as Yeast. This Assessment will use the common statistical test known as ANOVA test (Gooch
2011), to determine if both factors listed actually affect the growth of Yeast and if they are
statistically significant.

SEC 2. Engagement

As a high school Senior, and the plethora of majors open to me in college, I decided to only
study what I loved the most and that I think could help the most people. All throughout my life,
science has been at the forefront, biology and chemistry being the most significant to me. During
my Junior year, while visiting Bales Laboratory at University of California, Davis, I understood
the Biotechnology was the study for me. This internal assessment will be focused on using
technology such as Carbon dioxide sensors, dynamic heat and cold baths, all to address the
biological question on what is the optimal environment for a fungus, like ​Saccharomyces
cerevisiae​, to grow? The research question, unlike papers that research affects independently like
by Rodney P. Jones ,“Effect of carbon dioxide on yeast growth and fermentation” (Jones, 1982).
These papers, however do not address confounding variables; This Internal Assessment will
work to answer how to temperature and concentration of Carbon Dioxide in an environment
work in conjunction or against each other to create an optimal environment for the yeast and
work to create a replicable procedure which takes guide from a University of Nebraska lab
method to test “What affects yeast growth?”(University of Nebraska)

SEC 3. Background
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All organisms on the plant grow effectively in an optimal environment, and the Yeast strain
Saccharomyces cerevisiae, ​with the major factors being pH of the surrounding
environment(Pampulha, 1998), the pressure of the dish the yeast is in(Hartman, 2003), the
temperature and the concentration of essential nutrients. Even though the pH and pressure won’t
be experimented with, It is important to show how they affect yeast growth . Pressure seems to
show a parabolic relationship to the growth of Yeast colonies with one study stating that
increased pressure deforms the yeast cell(Pampulha, 1998), while other sources state that a large
increase in pressure leads to death of Yeast cell by “anesthesia”(Stockar 8) effect. pH, which is a
measure of the concentration of Hydrogen ions in a solution, can readily affect Yeast growth
because it can denature, “The process of partial or total alteration of the native structure of a
macromolecule resulting from the loss of tertiary or tertiary and secondary structure”
(MacNaught, 1997), And once the Yeast protein denatures, it can’t grow. Temperature is one of
the largest factors that affect Yeast growth with many academics from (Heard, 1988) and (White,
1951) stating that as temperature increases, the population of yeast colonies increased, as well as
the byproducts of yeast. One sources quotes that, “ it becomes evident that at 30~ practically a
100% population of Saccharomyces yeast was established within 3-5 days.” (Scharf 1983). This
means that during this Internal Assessment, the experimented temperatures will have large
variability to see by how much increasing temperature will increase the population of yeast. As
the concentration of Carbon Dioxide in the environment increases, it is expected that the rate of
growth for the yeast cells will decrease, because yeast emits Carbon Dioxide; Homeostasis
dictates that yeast will have to grow at a lower rate to balance the large amount of Carbon
Dioxide. ON study conducted stated that. “Pco~ 1 atm only 0.03% of the total count survived”
(Jones, 1982). The study continues to state that at lower pressures, more of the Yeast survives,
proving that Carbon Dioxide pressure is an inhibitory factor. Overall while Carbon Dioxide
concentration is seen as inhibitory and increased Temperature leads to increased population, This
INternal Assessment will delve into how specifically temperature affects the overall population
of ​Saccharomyces cerevisiae​.

SEC 4. Variables
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Biology
Independent Variable How is it controlled / tested
Temperature After placing the yeast within
the sample plates, ten plates
will receive an ice bath, five
plates will stay room
temperature, while ten more
plates will sit on a hot plate. It
will be tested for accuracy
with a digital thermometer
Dependent Variable How is it controlled / tested
Growth of ​ ​Saccharomyces The growth of the Yeast will
cerevisiae be recorded by change in
millilitres of yeast from the
original .1 ml of Dry yeast
Controlled Variable How is it controlled / tested
Pressure Pressure will be controlled by
keeping all samples within
the exact same type of sample
plates as well inside the same
room. Pressure, because of
this, will not change
Amount of Yeast Each sample plate will have
the same amount (in
microlitre​) of yeast, and will
be placed by an advanced
pipette
Uncontrolled Variables How it is monitored
Surrounding Light Sources During the night the lights of
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Possible Obstacles
There could possible some
issue with the yeast dying in
excessive heat or cold. Also
the sample plate might not be
equipped to handle large
changes in temperature
Possible Obstacles
Not applicable
Importance
Yeast requires an optimal
(​atm​) pressure to thrive and
grow in a natural
environment, therefore steps
must be taken to keep the
atm. Pressure constant.
It is important to have the
exact same amount of Yeast
so once to experiment is
concluded so there can be an
accurate count of Yeast
Importance
While light can be a factor for
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the room the samples will be yeast growth, since it is not a

stored in, while light doesn’t
affect Yeast growth largely, it
could have some affect.
Therefore the Yeast will go
through a light dark cycle
premier variable, it is not too
important to create a constant
light source for.

SEC 5. Materials

1. [ 25 gram] sample of Dry Saccharomyces cerevisiae Yeast

2. [ 30] micro test tube
3. [ 6 x 10 milliliter] Micropipette
4. [ 1.00 litre] Deionised water
5. [ 100 gram] Ice
6. [ 6 x 1.00 litre] Beaker
7. [ 1] Water Bath
8. [ 1] Sharpie pen
9. [ 30 centimetre] Scotch tape
10. [1] Styrofoam Rack
11. [ 1] Rag
13. [ 30] White Labels

SEC 6. Procedure

1. Remove Saccharomyces cerevisiae sample from freezer or packaging

2. Remove 30 micro test tube from packaging
3. Label each set of 5 of the micro test tube:
3.1. First 5: Control (18 C)
3.2. Second 5: Over 1 (25 C)
3.3. Third 5: Over 2 (30 C)
3.4. Fourth 5: Over 3 (35 C)
3.5. Fifth 5: Over 4 (40 C)
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3.6 Sixth 5: Over 5 (45 C)

4. Using a micropipette, pull 1 milliliters of the Yeast sample
5. Take one micro test tube and expel the 1 microlitre of Saccharomyces cerevisiae 6. Then close
the micro test tube with the top and tape over
6. Add 1 millilitre of water into the micro test tube.
7. Repeat Steps 4 through 6 for all micro test tubes
8. Create an water bath by placing water into the water bath
10. Each set of five receives a different temperature controls:
10.1. No special condition applied
10.2. Place the five micro test tube into water bath until temperature reaches 18
10.3. Place the five micro test tube into water bath until temperature reaches 25
10.4. Place the five micro test tube onto water bath until temperature reaches 30
10.5. Place the five micro test tube into water bath until temperature reaches 35
10.6. Place the five micro test tube into water bath until temperature reaches 40
10.7. Place the five micro test tube into water bath until temperature reaches 45
11. Each treatments should be placed in the water bath for FIVE MINUTES
12. Place each batch in a separate beaker
13. After a temperature control is applied to a set, wait 26 hours then pipette the remaining
Yeast
14. Record the milliliters of Yeast after the time period, then record the change from the original
1 milliliters
15. Clean up all apparatuses

SEC 7. Safety and Ethical Issues

This lab is not dealing with any animals not plants, therefore no specific ethical danger persists.
This lab deals with Yeast cells, and they are living, so the uptomst care will be taken while
holding them.
This lab does not utilise any sharp or glass equipment, however a hot water bath is used ( That
could reach up to 45 degrees therefore precaution such as gloves must be taken. Also no plastic
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(the micro test tube) will be placed on a small styrofoam rack to prevent any melting of the
plastic.
SEC 8. Data and Calculations

Table 1.1 Growth of Yeast (by ml) after 26 hours at specific temperature
Temperature in
Celsius 18 C 25 C 30 C 35 C 40 C 45 C
Set #1 0.10 ml ± .01 0.30 ml ± .01 0.1 ml ± .01 0.20 ml ± .01 1.00 ml ± .01 1.00 ml ± .01
Set #2 0.10 ml ± .01 0.70 ml ± .01 0.75 ml ± .01 0.20 ml ± .01 1.00 ml ± .01 1.00 ml ± .01
Set # 3 0.40 ml ± .01 1.00 ml ± .01 1.00 ml ± .01 0.60 ml ± .01 0.60 ml ± .01 0.00 ml ± .01
Set # 4 1.00 ml ± .01 0.52 ml ± .01 0.50 ml ± .01 0.70 ml ± .01 0.50 ml ± .01 1.00 ml ± .01
Set #5 0.60 ml ± .01 0.10 ml ± .01 0.50 ml ± .01 1.00 ml ± .01 0.25 ml ± .01 0.80 ml ± .01

n
1
Average = μ = n * ∑ x
i=1
Sample Calculation:
Propagation of Error: Only .01 from the test tube vial
n 3
1 1
μ = n * ∑ x = 3 * ∑ 1.000 = ​2.2/ 5 =​ .44± .01
i=1 i=1

Table 1.2 Average Growth of Yeast (by ml) after 26 hours at specific temperature
Temperature in
Celsius 18 C 25 C 30 C 35 C 40 C 45 C
Average 0.44 ml ± .01 0.524 ml ± .01 0.57 ml ± .01 0.54 ml ± .01 0.67 ml ± .01 0.76 ml ± .01

√
N
Standard deviation = σ = 1/N ( ∑ (x − u)2
i =1

Propagation of Error: Only .01 from the test tube vial

Table 1.3 Standard Deviation of Growth of Yeast (by ml) after 26 hours at temperature
Temperature in Celsius 18 C 25 C 30 C
Std. Deviation 0.378 ml ± .01 0.349 ml ± .01 0.334 ml ± .01
Temperature in Celsius 35 C 40 C 45 C
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Std. Deviation 0.343 ml ± .01 0.327 ml ± .01 0.433 ml ± .01

Table 1.4 Qualitative Data Marked

The Yeast came from dry dormant package, so it must be awakened with water
Once the Yeast was placed into the water, it began to froth and expand.
All treatments had some effect on the Yeast, but the higher temperatures create a solid slab of
yeast in the test tube.

Graph 1.5 The Average change in Yeast after 26 hours at Temperature

Table 1.6 ANOVA Statistics Calculations

The ANOVA Statistics Calculation will be used to determine if the averages presented in the
means of the treatments are ​significantly​ different, all a significance value of .05 will be used.
The ANOVA Statistics Calculation will be calculated externally on this website (Jeremy
Stangroom. 2018.) and the data is presented as:
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ANOVA Statistics Calculations

Source SS df MS

Between-trea 61.5849 4 15.3962 F = 2.96813

tments

Within-treat 103.7435 20 5.1872

ments

Total 165.3284 24

The ​p​-value is .044724, therefore​ the result is significant​ at ​p​ < .05. (Jeremy Stangroom,
2018).

SEC 9. Conclusion

The experiment yielded results of a positive linear correlation, stating that as the temperature of
the yeast during treatment increased, then the average growth of yeast over 26 hours also
increased. The R correlation value for the linear trend was .888 showing that while the trend
was moderate, it wasn’t strong not exact. An ANOVA statistical test, refer to Table 1.6, was
performed on the data average, and with an F value of 2.96 and p-value of .0447 the difference
between the points ​were significant. ​This means that this experiments , within the data set,
showed a significant difference between the outcome of the treatments. This trend is proven
within outside literature, where many biologist states that the optimal temperature is set on a
bell curve- where there is a strong range of temperatures that help the growth of yeast, while
extreme temperatures will inhibit it. The data presented has in increase in growth at ‘40 and
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Biology
45” Celsius, which is near the middle of the bell curve of optimal temperature. Even with these
correlations, there were few errors within the lab that affected the end values.
SEC 10. Evaluation and Improvements
Evaluation
Misreading of Temperature of
Yeast treatment - ​Random
Error ​(The end values were
randomly affected due to this
error)
Lack of temperature
sustainability during the 26
hour period ​- Systematic
Error​ (The end values were
systematically lowered due to
this error)
Inconsistency in lighting
during 26 hour period that
may have lead to inconsistent
heating ​- Random Error
Significance
The lab is testing the effect of
temperature on Yeast growth,
therefore it is imperative to
control the temperature true
the water bath as well as a
secondary thermometer to
test the water. The water,
while heating, might not heat
the micro test tubes to the
specific temperature because
only five minutes was given
to acclimate it
After the five minute
treatment period the beakers
of micro test tubes were left
in the external environment in
the classroom, which was
significantly was lower than
all tested temperatures. This
means that for the 26 hours,
all the yeast treatments were
operating on the same
temperature, effectively
making this IA testing “heat
shock” not temperature.
As the earlier evaluation
states, the beakers of tubes
were left on the classroom
counter, where some beakers
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Improvement
Suggested improvement
would be to increase the time
of treatment to fifteen
minutes, rather than five
minutes, because the extra ten
minutes would allow time for
the five micro test tubes to
better acclimate to the
temperature, as well as
privode in stability in stability
in temperature of the actual
water bath, so incossintesty in
temperature is not occuring
there
The improvement this lab
would have been to write in
the use of an “incubator” and
speratre the trials over five
days- each day perform one
treatment and place the micro
test tubes within the incubator
at the specific temperature, so
the 26 hour period would be
done at the same treatment
temperature, which would
have removed this error and
allowed the data to be testing
“temperature” not “heat
shock”
The improvement to this error
would be to either place the
beakers in an environment of:
The incubator as discussed in
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(The end values were were closer to a thermal lamp the previous evaluation. Or to
randomly affected due to this than others, possibly affect place all beakers in a
error) their temperature and yeast cupboard to remove ALL
growth. Light as a factor.

SEC 11. Works Cited

Alan D. MacNaught; Andrew R. Wilkinson, eds. (1997).​ Compendium of Chemical

Terminology: IUPAC Recommendations (the "Gold Book").​ Blackwell Science.

Gooch, J. W. (2011).​ Chi-square Test of Independence​. Encyclopedic Dictionary of Polymers,

973-974.

Heard, G.M (1988). ​“The effects of temperature and pH on the growth of yeast species during
the fermentation of grape juice”​ .Department of Food Science and Technology, The
University of New South Wales

Hartmann Christoph (2003). ​Numerical simulation of the mechanics of a yeast cell under high
hydrostatic pressure.​ Chair of Fluid Mechanics and Process Control, Technische Universität
München

Jones, Rodney P (1982) .​“Effect of carbon dioxide on yeast growth and fermentation”
Department of Chemical Engineering, Enzyme Microbiology Journal. Vol 4. July

M. E. Pampulha (1998). ​Combined effect of acetic acid, pH and ethanol on intracellular pH of

fermenting yeast ​.Laboratory of Microbiology, Gulbenkian Institute of Science.

Munns, D. J(1951)​“INFLUENCE OF TEMPERATURE ON YEAST GROWTH AND

FERMENTATION”​ White and Munns

Jeremy Stangroom (2018). “​One-Way ANOVA Calculato​r”. Social Science Statistics.

Stockar, von U (1994). ​The influence of pressure and temperature of compressed CO2 on the
survival of yeast cells​. Swiss Federal Institute of Technology

Scharf, R. (1983) ​The Effect of Temperature on Spontaneous Wine Fermentation ​Department of

Food Engineering and Biotechnology Technion, Israel Institute of Technology

What affects yeast growth. ​University of Nebraska, Lincoln. IFT Experiments