Introduction

Living cells require energy in order to progress with cellular processes that occur.

Research question
What are the effects of sugar concentration vs yeast in regards to measuring the volume of
carbon dioxide produced

Background research
Yeast (Saccharomyces cerevisiae) is a single celled organism that performs cellular
respiration to survive. This investigation will investigate the effects of different sucrose
concentrations on cell respiration in yeast to measure the volume of carbon dioxide
produced over a twenty minute period. During the fermentation process, ethyl alcohol is a
byproduct of anaerobic respiration during glycolysis where it occurs when oxygen is absent
and yeast is present and glucose is being broken down. The chemical reaction that takes
place transfers energy from glucose to the cells.

C6H12O6 → 2CH3CH2OH + 3CO2

Sucrose is a disaccharide which is composed of the monosaccharides, glucose and

fructose. Yeast allows the fermentation of monosaccharides more efficiently than other
carbohydrates, therefore sucrose must be digested prior to being fermented. Conveniently,
the enzymes responsible for the pre- fermentation process are found in the yeast cells.

Figure 1: The Structure of Common Monosaccharides

In order to simplify the understanding of the process, the fermentation reaction can be split
into three parts. The sugar must be digested into the simplest and most basic form. Since
yeast is fermenting, it needs to be digested into the original monomer component. Vitally,
one should take notice that yeast can’t metabolise lactase, which is not produced by yeast
cells. Therefore, through the use of genetic recombination, scientists were able to genetically
modify yeast cells to be able to produce and even secrete lactose. The reaction taking place
would be considered as an invertase digestion which can be seen in figure 2.

Figure 2: Invertase Digesting Sucrose into Glucose and Lactose

Once sucrose has been digested, the second step would be to break down the glucose into
two pyruvate molecules. Pyruvate, also known as pyruvic acid, is an amino acid which
happens to also be a key factor in forming ethanol. This occurs under glycolysis, lysis
meaning breakdown. Lastly, the most important step of fermentation is production of ethanol.
Pyruvate is transformed into an intermediary form called acetaldehyde, from then it will
undergo a series of redox reactions.

Figure 3: Generalised Equation for Ethanol Fermentation

Carbon dioxide that is released in the fermentation would be the indicator that will evidently
measure the rate of reaction.

Hypothesis
As the sucrose concentration is increased, the rate of which respiration takes place will also
increase hence the volume of CO2 will also increase. This is due to having higher sucrose
concentration which indicates that more reactant will be available for the yeast to ferment,
which means there is a greater percent change in carbon dioxide produced. Additionally,
there is a slight possibility that a plateau would be seen in the data. This can indicate that the
yeast cells are working at an optimal rate.

Variables
Independent variable
Concentration of the sucrose solutions.
- The concentrations would be 0.0, 0.2M, 0.4M, 0.6M, 0.8M, 1.0M.
Dependent variable
-
- Production of Carbon dioxide

Table 1: control Variables

Variable value/unis / How it was why it was

uncertainties controlled controlled

temperature of 41
water bath C
(+- .1 )

mass of yeast 1g
(.1)

same type of yeast

- Time period )
over 20
minutes

pH

Surface area No needed

Volume 100ml per rial

Material list and Apparatus

- Water bath
- 5 boiling tubes
- 5 gas syringes
- Balance
- 5 lids
- 5 orange thingies with the thingy coming out
- Test tube rack
Experiment setup

Method
1. Prepare the water bath and ensure the temperature of the water is 40-41°C
2. Place in the test rack with the test tubes in them
3. Prepare the yeast by measuring 1.00 grams of yeast in the lids then pouring it in the
test tubes, 5x for each concentration
4. Pour the sucrose solution in the measuring cylinder then dilute the remaining with
water, 5x for each concentration
5. Pour solution in test tubes that contain the 1.00 gram of yeast.
6. Swiftly, cover the test tubes by putting in the orange hinges with the tube connecting
the gas syringe.
7. Measure for twenty minutes, shake the test tube so they would mix every few
minutes or when it starts forming excessive bubbles and foam.
8. Rinse used apparatus including the test tubes with the bunges and tubing.
9. Repeat steps 3-9 for each concentration

Risk assessment
Experiment is harmless and can go down the drain. No need to wear protective gear

Procedure for data collection

Preliminary experiment

Preparing the sugar concentration and amount of yeast

Setting up the experiment to capture the volume of CO2 produced

Data processing

Table 2: amount of CO2 produced during 20 minutes amongst 5 trials

C
a
l
c
u
l
a
tions

Table 3: the mean and SD of amount of volume of CO2 produced

Qualitative Data

● In the 0.00 sucrose concentration, there were a few bubbles and it wasn’t producing
CO2 rapidly. After twenty minutes most of the five trials had produced a volume of
carbon dioxide ranging from 0-7. Interestingly enough, trail four had not produced
any carbon dioxide while trail two had produced a total volume of 7dm3.

Figure 3: Picture of 0.M sucrose solution

● In the 0.2M sucrose concentration, the volume of carbon dioxide per trial had
increased over the period twenty minutes. The lowest value is 25dm3 and highest
was 65dm3. Despite having the same amount of yeast in each test tube , same
amount of sucrose concentration, values were very different.

● By 0.4M there was also a range

of values that weren’t nearly as
accurate as other trials. There
were bubbles and a massive
fluctuation in volumes when
comparing all trials.

● When moving on to 0.6M

sucrose solution, the
range of numbers was
relatively seen. As usual
there was an excessive
amount of bubbles and foam.
 ● The 0.8M sucrose solution had data with some differences in regards to the range of
the numbers. Bubbles were observed and foam

● In the last trial, the

volume of carbon dioxide
produced per trial was
relatively low compared to
the ones in between. It did
produce bubbles and slight foam but not excessively.

Conclusion
Carbon dioxide is a by-product of Yeast fermentation, which is responsible for digesting
glucose in ethanol. This process is catalysed by enzymes found in yeast cells. Glucose
molecules are digested into two molecules of pyruvic acid, which are then broken down into
ethanol and carbon dioxide molecules. This process is affected by various environmental
conditions, such as temperature, concentration. By using carbon dioxide as the indicator for
the rate of the yeast fermentation, the effect of sucrose concern can be meausred
quantitatively.

By going through with this concept, this investigation concluded that the rate of yeast
fermentation and the concentration of sucrose available for reaction are positively correlated
to a certain extent. For sucrose concentration between 0.2 to 0.8M, the rate at which carbon
dioxide was released had continued to increase throughout the twenty minutes. 0 sucrose
concentration solution and 1.00M concentration seems to stagnate the production of carbon
dioxide. This plateau indicates that the complete mass available reactant has been
fermented.

Interestingly, as the concentrations with more sucrose available had reached plateaus
before with less reactant available. In this case, sucrose was used as a reactant. As sucrose
is a disaccharide and therefore must be partially digested before undergoing fermentation,
there is an extra reaction occurring prior to fermentation. This further means that perhaps an
abundance of reactant is nevertheless limited by the digestion reaction of sucrose. Further
investigation suggests that sucrose is digested into Glucose and fructose by invertase.
Hence in order for fructose to become glucose, it must undergo isomerisation which is
fermentable. Both reactions which has to occur prior to the actual process of fermentation,
reaction is unaffected by the digestion of sucrose and the isomerisation of fructose.

The 0 concentration solution had a surprising increase in the production of carbon dioxide
considering there was no reactant provided for the glucose to react with. Referring back to
prior knowledge, in regards to cell structure, it must be inferred that the yeast cells
used stored sugars to undergo fermentation and/ or full cellular fermentation to sustain basic
cell functions.

Overall, this experiment concluded in finding that elevated sucrose solution concentrations
correlate to an increase in the rate of fermentation to a certain extent.

Evaluation
Precise and accurate results were essential to this experiment and the investigation upon it.
Even the slightest variation in data can completely throw off the results of the investigation.
Very reliable equipment choices were made to ensure the best accuracy. For instance, the
electronic scale used to measure the weight of the yeast was up to a ± 0.01g, the water bath
was left till reached 40°C. Regardless of the efforts taken to prevent error, random and
experimental forms still occurred and could have interfered with the results.

For starters, the water bath was difficult to maintain. Even though the setting on the hot plate
had corresponded twitch the approximate temperature of 40.4°C, the slight variations in the
room and exterior temperature could have affected it. The change in temperature could have
changed the rate at which glucose was being fermented to allow the production of carbon
dioxide, which would skew with the results.

Furthermore, after applying the yeast and the sucrose solution then covering it with the bung
in order to collect the carbon dioxide, it fired to be as quick as possible to ensure the process
of collecting carbon dioxide is done accurately. However, through the second trial, whilst
comparing the data with each other, one of the trials was producing low carbon dioxide so
assuming there was a leak in the bung itself thst was causing the low values. The bung was
changed but there were still low numbers in comparison to what the others were producing.
Bibliography