Investigating Yeast Metabolism

Dilbag Dosanjh

Saturday, April 20, 2024

Monday, April 29, 2024

Ms. Englehart

SBI4U0-B
Background Information
Baker's yeast (Saccharomyces cerevisiae) is a type of fungus which is commercially important
for the production of bread; it is the CO2 released by the fermentation of S. cerevisae that causes
the bread to rise. Other, similar yeasts are needed for the production of wine, beer and industrial
chemicals.

Purpose
To investigate the metabolism of sucrose by yeast, and to determine what products are produced
from the anaerobic metabolism (fermentation process) of yeast.

Materials:
• Goggles • Scoopula
• 250-mL Erlenmeyer Flask • Tap water
• 1 test tube • Hot plate
• Yeast • Electronic Balance
• Tubing • Thermometer
• 600-mL beaker • Wooden Splint with Lighter
• Thermometer • Sucrose
• Parafilm

Procedure:
1. Prepare a 35◦C water bath solution (tap water!!!) in a 600-mL beaker. Ensure that throughout
your experiment it does not go above 37◦C or below 33◦C

2. Use the electronic balance to measure 2g of yeast. Add the yeast to a 250mL Erlenmeyer
Flask.

3. Use the electronic balance to measure 10g of sucrose. Add the sucrose to the same 250mL
Erlenmeyer flask (from step 2).

4. Add 100mL of tap water to your yeast and sucrose mixture. Swirl the Erlenmeyer flask until
the sucrose has dissolved.

5. Place your Erlenmeyer flask (yeast solution) into the water bath that you prepared in step 1.

5. Add some parafilm to the top of the flask. Poke a hole in the parafilm to allow a piece of
tubing to be able to fit into the top of the flask. Connect the tubing to a large test tube. Be sure to
seal the test tube using the parafilm. You will need to complete this step in order to collect the
gas that is produced. Run this reaction for at least 20 minutes. If your reaction is overflowing,
then stop the reaction and determine your products.

6. To determine the products you have collected, perform a splint test. Light a flaming splint and
insert the splint into the test tube. Observe the reaction and record the result. Remove the
parafilm from the flask and waft the products. Indicate the smell produced. Use table 1 (next
page) to help you with the splint test results:
Table 1: Splint Test Results
Splint Test Result Gas Produced
Glowing/Flaming Oxygen
Pop Noise Hydrogen
Flame Extinguished Carbon Dioxide

Observations:
● What happened to the flaming splint when it was inserted into the test tube? What is the
identity of the gas that was produced?
○ Once Inserted into the test tube a popping sound was identified, indicating that
hydrogen gas was present
○ The splint began flaming and glowing which indicated the presence of oxygen
allowing it to burn and glow further
○ Once the flame was extinguished it resulted in the release of carbon dioxide.
● What smell did you notice when you wafted the yeast solution? What product was
produced?
○ The smell most noticeable when wafting the yeast solution was the smell of
alcohol that was somewhat sweet.
■ The Smell most likely resulted from the fermentation.
○ The product that was produced from the yeast fermentation of sucrose was
Ethanol

Discussion Questions:

1. In this experiment, you fed sucrose to your yeast. Do you think glucose would be a better
food source for the yeast? Which sugar (glucose or sucrose) do you think is the best food
source for yeast? Explain your thinking in terms of cellular respiration.

In my opinion, I believe that glucose would be a better food source and essentially the
best sugar food source for yeast. This is due to the fact that in the glycolysis cycle, glucose does
not require any additional energy in order to convert it into alcohol. On the other hand as Sucrose
is a disaccharide, meaning it is made up of two monosaccharides (simple sugars) which in this
case are glucose and fructose. Both glucose and sucrose are carbohydrates sources that can be
metabolized by the yeast to create energy through cellular respiration. The issue comes with the
amount of sugar molecules, breaking down sucrose would require more time and energy since it
is made of two sugar molecules compared to the single glucose. Therefore glucose would be a
better food source as it can be broken down and metabolized more efficiently .
 2. Why was it important to ensure that the water was 35℃ and did not go too high? Why
was it important that it was not too cold?

It was important to maintain and ensure that the water remained at 35℃ and not higher
or lower as it could affect the fermentation process. Yeast is an organism meaning that its
metabolic activity can be changed by outside factors such as temperature. A higher temperature
may lead to the death of cells slowing the fermentation rate, although with a lower temperature
the metabolic reactions can slow, such as the fermentation leading to the same result as a much
higher temperature.

3. Refer to your observations AND your knowledge of the fermentation process, to identify
what gas was collected in your balloon. What simple test could you perform to confirm
that this gas was collected. Hint: think of the common gas tests. Show your teacher
your hypothesis for what gas was produced and perform the test at the front of the
room to confirm your hypothesis.

If the reaction between yeast and sucrose occurs under anaerobic conditions, the
production of carbon dioxide would occur as the reaction of fermentation would happen due to
the absence of oxygen. The test conducted to identify what was collected was the splint test.
When using the splint test and addition to the observations. In the observations it was stated that
a glowing splint is attributed to oxygen being present, a pop noise being hydrogen and what
occurred in this citation being Carbon Dioxide. Therefore when taking advantage of the splint
test to verify our hypothesis it proves it to be true.

4. What other factors might affect fermentation rates? Would these factors affect rates of
other biological processes? If so, how? What does this imply about all organisms?

Factors that could affect the rate of fermentation could be concentration, pressure and
temperature. A higher concentration creates more reactants for the product or particles to react
with which leads to more collision and quicker reactions. These sped up reactions lead to a faster
fermentation process, this also applies for pressure as increasing pressure can create close
quarters where particles are in closer proximity to each other creating more collisions.
Temperature also has a large impact on the fermentation rate whether it is through an increase or
decrease in the temperature. An optimal temperature would usually be a higher temperature as it
allows for faster reactions. Due to how the collision theory functions a higher temperature would
create more energy leading to quicker reactions, it also works in the reverse as a slow
temperature would create slower reactions as the molecules do not have as much energy/speed.
Extremes in either direction can inhibit and damage the speed/quality of fermentation rates. A
high temperature could lead to the death of the yeast and this also goes for a low temperature as
yeast is a living organism that needs certain qualities to survive.

These factors can also impact the rates of other biological processes due to their ability to
affect metabolic processes such as protein synthesis or cellular respiration. All organisms have
methods of controlling their enzyme activity and metabolic process so they can continue to keep
homeostasis, creating changinges that can affect how things function can mess with it.
 5. State sources of error (experimental, not human) that might have occurred during the
experiment and suggest improvements for the experimental design.

Sources of error that could have occurred during the experiment may have been the
leakage of gas as well as fluctuation in the temperature. The leakage of gas may have occurred
through the connections between the flask and test tube, leading to changes in or loss of gas that
was produced during fermentation. The leakage leads to inconsistencies and errors within the
results as the fermentation process has been altered/messed with. The leakage occurs in the
process where the parafilm is applied onto the flask and test tube, when creating the connection
and applying the parafilm small holes are inevitably made. In order to minimize and deal with
this issue, regular checks of the parafilm to ensure there are no leaks can be effective as well as
creating multiple layers of parafilm. Another source of error that may have occurred during the
experiment may have been the fluctuation and changes with the temperature. This error occurs as
there is no proper method of properly monitoring and maintenance of the temperature of the
water bath. This leads to fluctuations in temperature leading to changes in the range of yeast
fermentation. The optimal range of temperature was 33℃ - 37℃, was a challenge to keep
constant as the temperature measuring apparatus was not able to make up for the possible heat
that continues to remain on it. This can lead to small temperature increases over time leading it
out of the optimal range. In order to minimize this error, the only viable option is constant
temperature changing and monitoring, as well as taking the flask off of the heating apparatus.
These two methods of temperature regulating can ensure that the temperature remains within the
optimal range but require lots of focus. In order to improve these issues in general an automated
form of temperature regulation could assist with the fluctuations in temperature as well as a
premade apparatus that seals and contains the gasses for the fermentation process.

6. Fermentation can also produce vinegar. Research this biochemical process. (Include
in-text citations and a reference page)
(a) Identify the organism responsible for producing vinegar from ethanol.
(b) Is vinegar production aerobic or anaerobic? Explain.
(c) Describe how the relationship between the yeast and this organism is a form of
symbiosis.

(a) The organism that is responsible for producing vinegar from ethanol is a genus of
bacteria called Acetobacter. Acetobacter is acetic acid bacteria that has the ability to
convert ethanol into acetic acids when in the presence of oxygen (Gomes et al., 2018).

(b) The process of producing vinegar is mainly an aerobic process, as the conversion of
ethanol to acetic acid through the means of acetic acid bacteria is key in the reaction of
the production of vinegar, however this reaction only occurs in the presence of oxygen.
Therefore, the production of vinegar is aerobic (Gullo et al., 2014).

(c) The relationship between yeast and the bacteria of Acetobacter showcases a form of
symbiosis known as mutualism. Mutualism is a form of symbiosis where both organisms
benefit from the association; in this scenario the mutualistic symbiosis between yeast and
the acetobacter involves reciprocal metabolic activities that help support the growth of
each other as well as their metabolic process (Furukawa et al., 2013).
 References

Furukawa, S., Watanabe, T., Toyama, H., & Morinaga, Y. (2013). Significance of microbial

symbiotic coexistence in traditional fermentation. Journal of Bioscience and

Bioengineering, 116(5), 533–539. https://doi.org/10.1016/j.jbiosc.2013.05.017

Gomes, R. J., Borges, M. de F., Rosa, M. de F., Castro-Gómez, R. J. H., & Spinosa, W. A.

(2018). Acetic Acid Bacteria in the Food Industry: Systematics, Characteristics and

Applications. Food Technology and Biotechnology, 56(2).

https://doi.org/10.17113/ftb.56.02.18.5593

Gullo, M., Verzelloni, E., & Canonico, M. (2014). Aerobic submerged fermentation by acetic

acid bacteria for vinegar production: Process and biotechnological aspects. Process

Biochemistry, 49(10), 1571–1579. https://doi.org/10.1016/j.procbio.2014.07.003