THE EFFECT OF NATURE OF SUBSTRATES ON THE RATE OF CELLULAR

RESPIRATION IN YEAST (Saccharomyces cerevisiae)

ABSTRACT

Cellular respiration is a metabolic process that happens in

all cells of living organisms providing and supplying
energy for their survival. The objective of the study is
focused on determining the effect of the nature of the
substrate on the rate of cellular respiration in yeast
(Saccharomyces cerevisiae.). The study used Smith
Fermentation Tube Method in determining the amount of
carbon dioxide produced by different substrates diluted
with the yeast solution. Height, volume was calculated to
determine the rate of cellular respiration of each substrate.
The data suggested that the simpler the substrate, the faster
the rate of cellular respiration.

INTRODUCTION

Cellular respiration is the breaking down of food into usable energy for cells through a

long series of chemical reactions (Schroeder, 2003). Glucose is synthesized in cellular respiration

through glycolysis and oxidative mitochondrial respiration (Takeda, 2015).Cellular respiration is

a metabolic process that happens in all cells of living organisms providing and supplying energy

for their survival. Food is broken down into carbon dioxide and water, and the energy is released

forming cell’s energy carrier known as ATP or adenosine triphosphate (Lea et. al, 2000).

Cellular respiration has two types: aerobic respiration (oxygen-requiring) and anaerobic (without

oxygen) respiration (Schroeder, 2003).

 Aerobic respiration is a type of respiration that occurs in the presence of oxygen. In this

process, oxygen is utilized to release all the available energy in the glucose molecule. The

product of the reactants, glucose and oxygen, yields the release of six molecules of carbon

dioxide (CO2), six molecules of water (H2O) and ATP (adenosine triphosphate). Anaerobic

respiration is the metabolic breakdown of glucose without using oxygen. It undergoes two

processes including glycolysis and fermentation. Glycolysis processes and breaks down glucose

into pyruvate and ATP (energy) having carbon dioxide as its by- product. The second step,

fermentation, uses pyruvate fermenting it to produce ethanol or lactate (lactic acid). These two

processes take place in the cytoplasm of the cell. In these two chemical processes, glucose is

converted yielding energy (Gillespie, 2018).

According to Lewin (2017), carbohydrates are classified into two basic groups: simple

and complex. Complex carbohydrates are composed of multiple simple sugars, joined together

by chemical bonds. The more chains and branches of simple sugars, the more complex a

carbohydrate is and in turn, the longer it takes to be broken down. Simple carbohydrates can be

categorized into monosaccharides and disaccharides. Monosaccharides are a one sugar molecule

like glucose and fructose. On the other hand, disaccharides are a two-sugar molecule such as

sucrose and lactose. Glucose is a simple sugar that is the chief main source of energy. Sucrose is a

non-reducing disaccharide composed of glucose and fructose linked via their anomeric carbons.

According on Libretexts (2018), proteins are hydrolyzed by variety of enzymes into amino acids

which undergoes glucose catabolism through pyruvate, acetyl CoA and components of citric acid

cycle. All amino acids possess at least one amino group, this nitrogen-containing functional

group must be removed prior to entry into respiration. On the other hand, fats are synthesized to

form Acetyl CoA molecules being primed from aerobic respiration.

 The general objective of the study aimed to determine the effect of the nature of the

substrate on the rate of cellular respiration of yeast. It can be hypothesized that substrates that

have simple structure have a faster rate of cellular respiration. On the contrary, the structure of

the substrates does not affect the rate of cellular respiration in yeast.

The study was conducted at Institute of Biological Sciences Laboratory, University of the

Philippines- Los Baños, Laguna, Philippines on October 9, 2018.

 MATERIALS AND METHODS

The study used Smith Fermentation Tube Method in determining the amount of carbon

dioxide produced by different substrates diluted with the yeast solution. Five Smith tube was

used for five different substrate solutions. The substrates were poured having 15mL amount in

each tube. The first tube contains albumin (protein). The second tube contains oil (lipid). The

third tube contains glucose (monosaccharide). The fourth tube contains sucrose (disaccharide)

and the fifth tube contains distilled water (H2O). After pouring of substrates, 15 mL of distilled

water (H2O) and 15 mL of 10% yeast suspension was added into each tube. Gently, it was mixed

to provide uniformity all throughout the tubes. To make sure that there were no bubbles trapped

in the vertical arms of the tubes; it was tilted slowly for the bubbles to move and pushed through

the body where air is present. Cotton was plugged into the opening of Smith tubes. The tubes

were tied together at their vertical arms to keep them from upright position.

Height of the area that carbon dioxide (CO2) occupied was measured in cm every five

minutes for thirty minutes using a ruler. The volume of carbon dioxide produced was calculated

using the following formula.

Volume of CO2 Produced (V: cm3) = πr2

Where: r = radius of the vertical arm
h = height of the space occupied by CO2
The average rate of carbon dioxide CO2 production was measured and used the formula:

Average Rate of CO2 production (r ; cm3/ min)

= final volume of CO2 produced / total time

 RESULTS AND DISCUSSION

Set-up
Time Tube 1 Tube 2 Tube 3 Tube 4 Tube 5
(s) (w/ Albumin) (w/ Oil) (w/ Glucose) (w/ Sucrose) (w/ dH2O)
Height Volume Height Volume Height Volume Height Volume Height Volume
(cm) (cm3) (cm) (cm3) (cm) (cm3) (cm) (cm3) (cm) (cm3)
0 0 0 0 0 0 0 0 0 0 0
5 0.30 0.75 0.30 0.75 1.00 2.51 1.05 2.64 0.40 1.00
10 0.50 1.26 0.70 1.76 3.00 7.54 3.30 8.29 0.60 1.51
15 0.50 1.26 0.70 1.76 6.30 15.83 5.80 14.58 0.80 2.01
20 0.50 1.26 0.70 1.76 9.10 22.87 7.80 19.60 0.80 2.01
25 0.50 1.26 0.70 1.76 11.00 27.65 9.80 24.63 0.80 2.01
30 0.50 1.26 0.70 1.76 12.20 30.66 11.20 28.15 0.80 2.01
Rate
(cm3/min) 0.24 0.32 3.57 3.26 0.35
Table 1.1. Height, Volume and Rate of CO2 Produced

Table 1.1 shows the change in height (cm) and volume (cm3) occupied by the carbon

dioxide (CO2) produced in the different substrate of each tube and the rate of cellular respiration

in each tube. For the first tube, it can be observed that it reached 0.3 cm and occupied a volume

of 1.76 cm3. This shows that carbon dioxide is produced since there is a change in height

indicating that cellular respiration happened. On the second tube, the substrate contained is oil. A

value of .7 cm was observed on the 30th minute interval. Additionally, a volume measuring 1.76

cm3 was calculated. For the third tube, which contains glucose as substrate, it gained 12.0 cm

after reaching 30th minute interval occupying a volume of 30.66 cm3. Since it is glucose, a

reactant needed for cellular respiration to occur, it is expected for generation and production of

carbon dioxide. The fourth tube contains sucrose as substrate. It reached a height of 11.2 cm and

a volume measuring 28.15 cm3. It is also expected to have yielded carbon dioxide since it is a

glucose-containing monosaccharide. Lastly, for the fifth tube which contains distilled water

(H2O), a change in height was also observed. A height of 0.8 at the end of 30th minute interval
was computed with an occupied volume of 2.01 cm3. It was highly unexpected to not produce

carbon dioxide since it does not contain any glucose or material needed for the metabolic process

of cellular respiration to occur.

13
12
11
10
9
8
Height (cm)

Albumin
7
Oil
6
5 Glucose
4 Sucrose
3 dH2O
2
1
0
0 5 10 15 20 25 30
Time (s)

Figure 1.1 Height of CO2 Produced every 5 minutes for 30 minutes

In the figure 1.1, it shows the different height that carbon dioxide has occupied in every 5

minutes for 30 minutes. The substrate that has the least height occupied is albumin amounting

only to 0.5 cm. It was followed by the substrate oil which is limited to 0.7 cm. Distilled water

occupied a height of 0.8 cm. On the other hand, sucrose was second to the highest in terms of

height with 11.2 cm. Amongst the substrates, the substrate glucose has the highest height

reaching a value of 12.2 cm after the 30th minute interval. This trend is somehow inconsistent

since it was only expected for only the substrates glucose, sucrose, albumin and oil to progress to

cellular respiration.
 4
Average Rate of CO2 Production (cm3 /
3.5

2.5
min)

1.5

0.5

0
Albumin Oil Glucose Sucrose dH2O
Substrates

Figure 1.2. Average Rate of CO2 Production of Different Substrates

After getting the height, volume of each substrate was calculated to get the rate of cellular

respiration of the substrates. Figure 1.2. shows the difference in rate of cellular respiration of the

substrates. It is predetermined that the substrate having a shorter height will have the slower rate

of cellular respiration for having lower volume. On the contrary, the substrate that has the longer

height will undergo a faster rate of cellular respiration for obtaining a higher amount of volume.

The trend in the values of cellular respiration would be similar to the trend in difference of

height. As the data suggests, albumin would rank fifth among them. Oil to be ranked as fourth

while distilled water (H2O) to be placed at the third place. Sucrose, on the other hand, would get

the second position. Lastly, glucose was expected to have the fastest rate of cellular respiration to

get the rank being first.

Since it was expected only to have cellular respiration to proceed on the four substrates,

glucose, sucrose, albumin and oil, a comparison in their rate of cellular respiration could be

done. As seen on the table 1.1 and figure 1.2, glucose have the faster rate of cellular respiration

than sucrose. This phenomenon is due to the condition that glucose is the one needed for cellular
respiration. Sucrose, on the other hand, needs to be hydrolysed or broken down to glucose with

the presence of the enzyme invertase or sucrase (de Almeida et.al, 2005). Thus, the simpler

sugar, the faster rate of cellular respiration is (Estioko, 2012). Comparing albumin which

contains proteins and oil which contains fatty acids, oil have much faster rate of cellular

respiration. It is due to the fact that oils are synthesized to lipids directly and undergo cellular

respiration faster. On the other hand, proteins are first hydrolyzed to form amino acids.

Moreover, according on Libretexts (2018), these amino acids contain one amino group which is

needed to be removed in order to proceed to cellular respiration. With this, it needed to undergo

longer process before undergoing the process of cellular respiration. For water, a height was

observed but since it is highly unlikely for it to undergo cellular process due to lack of materials

needed to proceed to cellular respiration, contamination or other errors could have happened

when doing the experiment.

The possible sources of error could be due to contamination of some tubes to glucose-

containing material. Improper washing of tubes could also be the factor for the contamination

leading to the said inconsistency. The errors could be caused by the inconsistent sizes of the

Smith tubes used in which could have resulted to discrepancies of data gathered.

SUMMARY AND CONCLUSION

This study aimed to determine the effect of the nature of substrate through carbon dioxide

production in yeast was determined through the use of five Smith tubes containing 15mL of five

different substrates including albumin, oil, glucose, sucrose and distilled water (H2O) diluted

with 15mL distilled water and 15 mL of 10% yeast suspension in each tube. The height and

volume were calculated to compute for the rate of cellular respiration. The substrate that yielded

the slowest rate of cellular respiration was albumin while the substrate that yielded the fastest
was glucose. The hypothesis was accepted since substrates that contain the simplest structure

yielded faster cellular respiration. Because of that fact, it is concluded that the simpler the

substrate, the faster the rate of cellular respiration.

 LITERATURE CITED

De Almeida, A., De Araújo, L., Costa, A., De Abreu, C., De Andrade Lima, M., & Fernandez
Palha, M. (2005). Sucrose hydrolysis catalyzed by auto-immobilized invertase into intact
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http://www.academia.edu/4532179/Effect_of_the_Nature_of_Substrate_on_the_Rate_of_
Respiration_of_Yeast
Gillespie, C. (2018). Importance of Aerobic Cellular Respiration. Retrieved Ocyober 18, 2018
from https://www.thoughtco.com/cellular-respiration-process-373396
Lea C., Lowrie P., McGuican S. (2000). AS Biology for AQA. Jordan Hill, Oxford. Heinemann
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Lewin J. (2017, July 28). Sugar Explained [Blog post]. Retrieved from
https://www.bbcgoodfood.com/howto/guide/sugar-explained
Libretexts. (2018, June 06). 7.6: Connections of Carbohydrate, Protein, and Lipid Metabolic
Pathways. Retrieved from
https://bio.libretexts.org/TextMaps/Introductory_and_General_Biology/Book:_General_
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hydrate,_Protein,_and_Lipid_Metabolic_Pathways
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Takeda, K., Starzynski, C., Mori, A., & Yanagida, M. (17 april 2015). The critical glucose
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 THE EFFECT OF NATURE OF SUBSTRATES ON THE RATE OF CELLULAR

RESPIRATION IN YEAST (Saccharomyces cerevisiae)

Gabriel P. Banta

Bio 11.1 8L

October 19, 2018

i
A scientific paper submitted in partial fulfilment of the requirement in GeneralBiology I

Laboratory under Ms. Aimee Cagalawan, 1st sem., 2018-2019.