Name: Florencia & Oneal

Pak Andrea Okem

Chemistry – 9.1 - Title

4th October 2022

1. INTRODUCTION

I like to eat at Chinese Restaurant. Pickle is one of the fermented foods. Pickle is also a
common side dish in Chinese restaurant. The taste is sour and refreshing. Pickle was a cucumber
that has been through fermentation process.

Fermentation is a metabolic process in which an organism converts a carbohydrate, such as

starch or sugar, into an alcohol or an acid. For example, yeast performs fermentation to obtain
energy by converting sugar into alcohol. Yeast is essential for the treatment of wastewater and
the production of biofuels. Fermentation occurs when pyruvate produced by the metabolism of
glucose is broken down into ethanol and carbon dioxide by yeasts (and some bacteria). One of
the examples using yeast for fermentation is brewing, so beer is traditionally made from four key
ingredients: malted cereals, water, hops, and yeast. During fermentation, yeast cells convert
cereal-derived sugars into ethanol and CO2. Variation in these metabolites across different yeast
strains is what allows yeast to so uniquely influence beer flavor.
Besides fermentation, in this experiment we get carbon dioxide or CO 2 as our final product.
Basically, carbon dioxide is a gas that includes one part of carbon and two parts of oxygen in
their product. It is colorless, odorless, non-flammable gas, and can turn into liquid or solid
(“Carbon Dioxide”). It is one of the most important gases on the earth. We also use it in our daily
life, such as for fire extinguishers, inflating life rafts and life jackets, blasting coal, foaming
rubber, and plastics (“Carbon Dioxide | Definition, Formula, Uses, and Facts”). Carbon dioxide
also most known of their process of photosynthesis for the plants on the earth. In this experiment,
through fermentation, we can also produce carbon dioxide using yeast. In the process, we need
warm water and sugar to combine with the yeast. Later on, the yeast will start to produce carbon
dioxide as their product.
The chemical equation for the fermentation that produces carbon dioxide is:
The process is through one glucose molecule converted into two ethanol molecules and two
carbon dioxide molecules, based on that statement expressed as
C6H12O6 (glucose)⟶2C2H5OH (ethanol) + CO2 (carbon dioxide)
Before fermentation takes place, one glucose molecule is broken down into two pyruvate
molecules. This is known as glycolysis.
There are physical and chemical factors that affect the fermentation process. First, the physical
factors are temperature, the effect of temperature on microorganisms that involves in the
fermentation process is numerous, temperature is the most important factor that determines the
rate of growth, survival, multiplication, and death of all living organisms. High temperatures
may reduce fermentation process because of damage microbes by transport carries, proteins, and
denaturing enzymes. At very low temperatures fermentation process may also be reduced
because membranes of bacteria may also solidify, and enzymes also do not function properly, so
in this case the right temperature of water that we should use is around 100 F (Mill), the reason is
because in that number, the yeast can run the fermentation better than higher or lower
temperature. Second, fermentation process may also be affected by pH because microbial growth
is strongly affected by the pH of the medium, so the right rate of pH that we need to use is
around 4.0 to 6.0. This means that the cells of the yeast require a slightly acidic environment to
run the fermentation (“E-unit: pH and Fermentation”). During the fermentation process, with the
utilization of nutrients and the accumulation of metabolites by the bacteria, the pH of the
fermentation broth will inevitably change. pH can be controlled by adding ammonia, urea,
ammonium carbonate, or calcium carbonate in the middle. There will be 25 trials, with 5
different amount of yeast, each amount of yeast in 5 trials. The amount of sugar will be 6
grams/trial and the yeast will be 0, 2, 4, 6, and 8, the fermentation will take 20 minutes.
There are multiple chemical factors. First is carbon; simple to complex carbohydrates can
be added to media in fermentation as a source of carbon. Second is nitrogen; source is required
for most organelles to synthesize protein, nucleic acid and other cellular compounds. The third
factor is oxygen;the availability of oxygen is a crucial element in aerobic fermentation.
Depending on how much oxygen is needed by the organisms, it may be provided as pure oxygen
infrequently or in the form of air, which contains roughly 21% volume by volume oxygen. The
oxygen needs of the organism might vary significantly depending on the carbon source. The
bioreactor receives filtered, sterilized air for most of the fermentation process. Last is water; all
fermentation processes, apart from solid state fermentation, require a lot of water. In addition to
being an essential element of all media, water is also necessary for ancillary functions including
heating, cooling, cleaning, and rinsing. Therefore, it is crucial to have a steady source of huge
amounts of clean water.
In the end, the objective or the purpose of this investigation is to observe the effect of the
amount of yeast to the CO2 production in fermentation. We can measure the amount CO2
production in fermentation by measuring the diameter of the balloon in its final product. One
example of the use and results of a mixture of sugar and yeast in our lives is when we make cake
dough, and need yeast and sugar to make the cake, yeast to make the dough rise and soft, then
sugar to add sweetness to the cake. If we use too much yeast, the cake dough will failed to rise,
the dough gets overly gassy so kneading time is reduced which lowers the quality of the bread,
and If we let the dough rise too long, it will start having a yeast or beer smell and taste and
ultimately deflate or rise poorly in the oven and have a light crust. Then if we use too much sugar
the taste will be too sweet, and of course it is not good for health, for example it can cause
diabetes.
1.1 Research Question (Florencia)

How does the amount of yeast affect the CO 2 production by measuring the diameter of the
balloon during fermentation?

How is the amount of yeast (0gr, 2gr, 4gr, 6gr, 8gr) affect the production of CO2?

Hypothesis (Oneal)
The hypothesis of this experiment is the more amount of yeast used in the fermentation, the
higher the amount of CO2 produced. According to research, the rate of fermentation is dependent
on the concentration of yeast but independent of the concentration of sugar (…) This means that
the amount of CO2 produced can be different due to the different rate of fermentation caused by
the concentration of yeast.

The first theory to support this is according to research, when yeasts eat sugar and turn it into
energy, they also produce carbon dioxide. This type of reaction is known as alcohol
fermentation. During the fermentation, yeast will transform sugars to ethanol and carbon dioxide.
Therefore, the more yeast used, the more carbon dioxide will be produced as there is more yeast
available to transform sugars. The second theory is related to Saccharomyces cerevisiae, a well-
known species of yeast that has a significant role in alcohol fermentation. Due to its ability to
grow in aerobic and anaerobic conditions in the presence of glucose, this species enables yeast to
use sugars to undergo anaerobic respiration in the absence of oxygen, such as fermentation, in
order to produce ethanol and CO2 gas. Therefore, in the bottle that contain yeasts and sugar, the
yeasts should have made abundant carbon dioxide, which will inflate the balloon (…) the third
theory is according to research, the rate of CO2 released during fermentation depends on the
concentration of yeast. As previously mentioned, the rate of fermentation is dependent on the
concentration of yeast. This theory has been proven by an experiment done by Charles Pepin and
Charles Marzzaco, where their graph shows that the slope of the sample with 7 gr of yeast is
about twice as large as the one with 3.5 g of yeast. Meaning, that the sample with 7 gr of yeast
has a faster rate than the one with 3.5 g of yeast.
 Therefore, the higher amount of yeast used, the more CO2 will be produced. The supporting
theories behind this is because there will be more yeast available to transform the sugars, there
will be more of Saccharomyces cerevisiae to produce ethanol and carbon dioxide through
fermentation, and the rate of fermentation will be faster, which will increase the rate of CO
produced. Hence, resulting in a higher amount of CO2 produced.

https://wcww.scientificamerican.com/article/single-celled-science-yeasty-beasties/

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466055/#:~:text=During%20fermentation%2C
%20yeast%20cells%20convert,taste%20of%20beer%20are%20produced

https://uwaterloo.ca/chem13-news-magazine/april-2015/activities/fermentation-sugars-using-yeast-
discovery-experiment

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC539022/

file:///Users/leonarossanairah/Downloads/mayui,+sacch+Thurs+pm+final+2015+for+publication.pdf

Variables (Oneal)
1.1.1 Independent variables

The independent variable of this experiment will be the different amount of yeast used in the
process of sugar fermentation. This variable will be manipulated as it will be divided into 5
different values including 0 gr, 2 gr, 4 gr, 6 gr, and 8 gr, where each value will be repeated for
five different times in order to get an accurate result. For example, for the first IDV (0 gr), there
will be 5 trials of sugar fermentation containing 0 gr of yeast. For the second IDV (2 gr), there
will also be 5 trials of sugar fermentation containing 2 gr of yeast, and so on. These values were
made 2 gr distant away from each other in order to give more difference to the final result of the
fermentation. If they are only 1 gr or 0.5 gr distant away from each other, it’s possible that the
results will not show much difference, which will make it harder to make a conclusion out of the
results (McGrath). Another reason why the chosen values are small is because all the
fermentation of every IDV will be also made with a small amount of sugar, which is 6 gr. Hence,
a big amount of sugar might not be needed in order to make a difference in the amount of CO2
produced. Furthermore, they will be divided using a digital balance with the uncertainty of 0.005
gr, which comes from the smallest scale of the tool divided by 2. The reason why this experiment
uses a digital balance is because it has a small uncertainty, meaning the measurement can be
more reliable.

1.1.2 Dependent variables

The dependent variable of this experiment will be the amount of CO2 produced during sugar
fermentation. This variable will be measured by using a balloon's volume which indicates the
total amount of CO2 produced. For example, for the first IDV (0 gr), a balloon will be attached
to the bottle containing 0 gr yeast, sugar, and warm water. The balloon will be blown due to the
sugar fermentation which occurs in the bottle. After that, the diameter of the balloon will be
measured by a ruler with the uncertainty of 0.5 cm in order to find the volume of the balloon,
which will be using this formula:

4 3
𝑉= 𝜋𝑟
3

The bigger the volume of the balloon, the higher the amount of CO2 produced. This applies to
all other IDVs. The reason why this experiment uses a ruler instead is because it is one of the
tools to measure length and its small uncertainty can make sure the result is more accurate.

1.1.3 Controlled variables

1.1.4 Discontinuation and Sufficient Data

Independen The amount of the yeast combined with the sugar made CO2 production.
t Variety amounts of yeast that used in the fermentation of producing the
carbon dioxide, as in this experiment we use 20 gram yeast for each 5 trials
or 20 (0 gr, 2gr, 4gr, 6gr, 8gr) × 5 ¿100 grams for the whole experiment.
This data varies from 0 to 8 gram because we need to do some trials for
this experiment to make sure about the result from each of the amount of
yeast that will be used.

The amount of yeast that will be used surely will be measured using digital
 balance to make sure if using the right amount of the yeast.
Dependent The amount of the CO2 produces from the reaction. The amount of the
cabon dioxide from this experiment will be different base on the amount of
sugar that will be used in these trials. Since carbon dioxide is the final
product of this experiment, we will count the amount of the carbon dioxide
by measuring the diameter of the CO2 gas in the balloon by using the
4 3
formula of volume of sphere V = 𝜋𝑟
3
. In that way, it will show the volume of the balloon as the representative of
the amount of the carbon dioxide gas they produces.
Controlled Method of Control Reason
Variables
Amount of water Water should be measured Amount of water should matter in these
until the amount is exactly experiments. Using the water, the yeast
100 mL for each of the will be combined with the sugar easily
trials. Overall, it has to be in the whole trials. Water is not only a
100 × 25 trials for the major component of the experiments,
whole experiment. but it is also important fir ancillary
processes such as heating, cooling,
cleaning, and rinsing.
Temperature of Measure the temperature Since this experiment is about
water using thermometer and fermentation that used yeast as their
make sure it is 100 F. subject. The best number of
temperatures that can run the
experiment is around 100 until 110 F
since it uses yeast. The reason is
because yeast can run the fermentation
properly in that temperature.
pH of the water Using the pH meter, the pH Since the experiments use big amount
of the water should be of water, pH has a big role during the
measured perfectlyin the fermentation. Not just because its
 number of 4.00 to 6.00 microbial growth effect, it is important
to use a specific pH because if its vary,
it may disrupt the plasma membrane or
inhibit the activity of enzymes and
membrane transport proteins (Factors
Affecting Fermentation).
Amount of Sugar The amount of sugar should Since this experiment is about yeast
be measured using the fermentation, yeast should need food to
digital balance until 6 gr for be transferred into carbon dioxide. In
each of the experiments. here we are using 6 gr of sugar. Since
we use the multiple of 2 as the amount
of yeast (0, 2, 4, 6, 8) we use the
middle amount (6) as the balanced ratio
with the sugar. It has to be balance
because if it is not balance, there
should be chance for the experiment to
be failed.
Shape of Balloon Use balloons with similar To make sure the volume of the
shape. balloons is uniformed.
Time of The time should be counted Since the amount of yeast will be vary
fermentation until 20 minutes. and the sugar will be the same (1
amount) according to my hypothesis,
any number of yeast will not matter on
how much carbon dioxide they
produce. In other words, the carbon
dioxide is based on the amount of
sugar. But the thing is, the difference in
the experiment is not about CO2 they
produce, but the time they take to have
the final amount of CO2/faster and
slower.
2. PROCEDURE (Florencia)
1.1 Equipment

Equipment Amount
• 250 mL plastic bottles - 25 pieces
• 100 mL graduated cylinder - 1 piece
• pH meter - 1 piece
• Digital balance - 1 piece
• Thermometer - 1 piece
• Heating plate - 1 piece
• 30 cm ruler - 1 piece
• Timer - 1 piece

1.2 Materials

Materials Amounts
• Sugar - 25x6 grams
• Latex balloons - 5x5 pieces
• Yeast - 100 grams
• Water - 100x25 mL
1.3 Labeled Diagram

1.3 Methods
1) Prepare the bottle or the beaker that will be used during the experiment.
2) Measure 100mL of water with 100 mL graduated cylinder.
3) Heat the water using a heating plate and to make sure the temperature of the water hit 100
F at minimum.
4) Measure the pH of the water using pH meter that is already provided.
5) Put the warm water into the bottles
6) Measure the sugar
7) Mix the measured sugar into the water from each of the bottles
8) Combine the mixture with the variety amount of yeast in each of the bottles.
 Result:

- bottle 1 was filled by 6 grams of sugar, 0 gram of yeast, and 100 ml of warm water

- bottle 2 was filled by 6 grams of sugar, 2 grams of yeast, and 100 ml of warm water

- bottle 3 was filled by 6 grams of sugar, 4 grams of yeast, and 100 ml of warm water

- bottle 4 was filled by 6 grams of sugar, 6 grams of yeast, and 100 ml of warm water

- bottle 5 was filled by 6 grams of sugar, 8 grams of yeast, and 100 ml of warm water

- Method must consider all relevant variables, allow collection of sufficient, trials etc.

- Suitable quantitative data collection method (Planning table/ Template)

1) Put the balloon

2) Turn on the timer for 20 minutes
3) Measure the diameter of each balloon

1.4 Safety Concerned

Hazard (Safety for ourselves)

Skin contact if exposed directly to the mixture, for some

people who have sensitive skin, it may
cause irritation to the skin like redness. For
our safety we must wear protective
equipment (lab coat and chemical
protection gloves) can help you avoid direct
contact.
Balloon explosion If there is too much CO2 in the balloon, the
balloon will expand and cause an explosion,
so that it can shock people, cause heart
attack and traumatize them. ( for example:
pak okem is afraid of balloons)
 Hazards

Ingestion Respiring yeast should generate carbon

dioxide gas in the end. Over exposure to
carbon dioxide can cause asphyxiation.
Another is from ingestion: Excessive
ingestion of liquid substance of yeast could
lead into intestinal discomfort such as
diarrhea, bloating, cramping, and many
more.
Microorganism Microorganism is an organism that can
contaminate food and cause illness,
bacteria, viruses, even parasites in it. For
example, a kind of microorganism that also
needs to be careful is the yeast. Yeast can
have chance to cause food to spoil but not
causing foodborne illness.
Hazard (Ethical)

Excessive use of sugar and yeast After doing the experiment, the materials
that have been used will be thrown away.
Of course, we need a lot of ingredients for
some trials, but there are a lot of people out
there who need sugar and yeast.
We use a lot of water for the experiment, it
Excessive use of water means that we also waste a lot of water, we
need to save water, because every person on
earth need water to survive, and water is
needed by plants to grow.
1.5 Designed Table

1.5.1 Raw Data Tables

Table 1. Final Volume of NaOH Recorded from 5 trials after Titration with Milk Samples

IDV DV
(unit, (unit,
± ___)± ___) ± ___________± ___________
)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
0
2
 4
6
8

DV
(unit,
IDV
± ___________± ___________
(unit, )
± ___)± ___)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5

I F D I F D I F D I F D I F D
0
2
4
6
8

1.2 Processed Data Tables

Table 2. Titre (Volume Difference) of NaOH from 5 trials after Titration with Milk Samples

Day of Storage Difference of Final and Initial Volume of NaOH recorded in burette in
(unit,
ml
± ___)± ___)
(
±0.05 𝑚𝑙±0.05 ml
)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
0
2
4
6
 8

Table 3. Effect of Storage Duration of NaOH on Titratable Acidity of Raw Milk Samples from 5
Trials

Day of Titratable Acidity of Raw Milk Samplex

Storage
(
(unit,
% 𝐿𝑎𝑐𝑡𝑖𝑐 𝐴𝑐𝑖𝑑)% Lactic Acid)
± ___)± ___)

Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average

0
2
4
6
8

2 REFERENCES

REFERENCES (MLA)
Maicas, S. (2020) The role of yeasts in fermentation processes, Microorganisms. MDPI. Available at:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466055/ (Accessed: October 6, 2022).

“Carbon Dioxide | Definition, Formula, Uses, and Facts.” Encyclopedia Britannica, 6 Sept. 2022,

www.britannica.com/science/carbon-dioxide.

“Carbon Dioxide.” Wisconsin Department of Health Services, 15 June 2022,

www.dhs.wisconsin.gov/chemical/carbondioxide.htm#:%7E:text=Exposure%20to

%20CO2%20can%20produce,coma%2C%20asphyxia%2C%20and%20convulsions.
Mill, Bob’S Red. “What Temperature Kills Yeast.” Bob’s Red Mill Blog,

www.bobsredmill.com/blog/baking-101/what-temperature-kills-yeast. Accessed 7 Oct.

2022.

“E-unit: pH and Fermentation.” AgEdLibrary.com,

agriculturewithmrsskien.weebly.com/uploads/2/1/3/2/21329554/ph_and_fermentation.pd

f. Accessed 7 Oct. 2022.