Cellular Respiration in Disaccharides with (S.

Cerevisiae) Yeast and the Amounts of Carbon

dioxide released
Jennifer Gorcak
Red Deer College

Abstract

This paper offers information in how carbon dioxide levels change depending on the disaccharide
going through the process of cellular respiration. Disaccharides such as the ones offered in our experi-
ment, (sucrose, lactose, maltose) suggest that because they are made from two monosaccharides, when
they are put through the process of cellular respiration they produce twice as much carbon dioxide as
monosaccharides, as well as twice as much ethanol and energy. By testing different types of disaccha-
rides, we are able to study how depending on the sugar given, how much carbon dioxide will be produced
or not. In the procedure, after placing our tubes in the 40 degree water bath with 4.5 mL of 0.5 M solu-
tion of the desired glucose, the tubes were taken out one at a time and mixed with 1.5mL of yeast solu-
tion, then pipetted into a Durham tube and inverted back into the test tube, leaving the durham tube in the
original test tube it came from. After being placed back into the 40 degree water bath for 20 minuets, the
level of carbon dioxide produced in the tube was measured. To ensure our loss of carbon dioxide was
minimal to the air, these last steps were performed quickly and efficiently, for less error and better results.
This experiment investigates how having different glucose concentrations could affects the ability of yeast
to metabolize glucose in the process of cellular respiration. The results forged shows a consistent release
of carbon dioxide in Sucrose and Maltose, as Lactose showed no release and distilled water was our con-
trol. In conclusion, carbon dioxide release is sufficient with Sucrose and Maltose, but not with Lactose.

Introduction

Yeast is a unicellular organism that produces energy from organic compounds. It need a supply of

energy (sugar) for it go live and grow. In a respiration process, yeast uses oxygen to make energy from the

sugar. The more sugar, the more active the yeast will be. (Hewitson, Hill, 2018) By either aerobic or

anaerobic respiration, a sequence of catabolic reactions under go that involve enzymes which produce

energy (ATP) and release waste such as carbon dioxide. The difference between these to types of respira-

tion is that anaerobic does not need oxygen in its process, and aerobic respiration does. Aerobic respira-

tion, which is practiced in our lab, is a much more efficient way to produce ATP quicker. This is known as

oxidative phosphorylation. In aerobic respiration, when the process of glycolysis makes pyruvate which is

transferred to the mitochondrial membrane from the cytoplasm; if oxygen is present. While the aim of this

lab was too find which sugar has the best effect of a yeast's metabolism, this can be measured when taking

into account the amount of foamy bubbles that appear after mixing the glucose solution with the yeast.
These foamy bubbles (carbon dioxide) are released for the walls of the cell as a waste product, due to the

osmotic pressure in the cell as it's reaction undergoes. (Angustia, Chan, Dinneen, Hortamani, & Muta-

baruka, 2014)

Sucrose, Maltose, and Lactose are different isomers of sugars. Their chemical formula is the

same, (C12H22O11). Depending on the different types of isomers and the different enzymes that they

have, they have different ways in how they breakdown the yeast and perform cellular respiration success-

fully. These disaccharides are formed when a dehydration reaction occurs between 2 monosaccharides.

This means an OH- and and H+ formed a molecule of water and was released, thus being replaced by a

covalent bond. This bond is referred to as a glycosidic bond (Koshkina). Sucrose is a molecule that has a

glycosidic bond between glucose and fructose, while maltose is a molecule having a bond between 2 glu-

cose molecules, and lactose having the bond between glucose and galactose. Within out study, we were

able to test these different disaccharides to obtain information on how they metabolize in yeast and pro-

duce different levels of carbon dioxide. Because of the different monosaccharides they are made up of,

once they are broken down into their simplest monosaccharides (some of them), they're metabolized dif-

ferently. In lactose, yeast does not have the enzymes to breakdown lactose into its monomer sugars. By

using the enzyme lactase, an enzyme which breaks down lactose in our stomachs, it breaks it into those

monomers we can use for energy (ATP). The difference in these glycosidic bonds is that the monomers

are joined at different locations around the monomer, thus changing the chemical structure and thus

changing the type of disaccharide. It also has do do with the type of carbon ring. For instance, fructose, a

monomer in sucrose, is a 5 carbon ring. In different, Glucose, both monomers in Maltose, is a 6 carbon

ring. The way all of these different monosaccharides metabolize defines how much carbon dioxide they

can produce. (Reece, Minorsky, Moyes, Urry, Jackson, Scott, Cain, Rawle, Walde, Wasserman,2018, pp.

68-70)

Doing this experiment, we will be able to see how different combinations of monosaccharides can

affect how disaccharides are metabolized, and how much carbon dioxide their cellular respiration reac-

tions can produce. This is in turn determines the yeast's ability to break the glycosidic bonds. Our testable
hypothesis is that if we put disaccharides through cellular respiration, then we would see different

amounts of carbon dioxide developed from different disaccharides. We will see different levels of carbon

dioxide because the monomers that make up the disaccharides differentiate in shape, size, and where they

connect to form disaccharides. We should see carbon dioxide result from sucrose and maltose, because

distilled water is the control and lactose cannot be broken down by this yeast (needs lactase). When these

disaccharides break, these different monomers are separated which fluctuate the levels of carbon dioxide

and ATP produced.

Materials and Methods

To begin, get 12 test tubes and place them in a test tube rack. Label them by their disaccharide/

control (distilled water, sucrose, lactose, maltose) and their trial number (1, 2, 3) for a multiplied total of

12 tubes. Next, obtain approximately 20mL of 0.5M solution for each saccharide. To do this, obtain 4

40mL beakers. The first containing 20mL of distilled water, and the other containing 20mL of 0.5M of

your disaccharide solution. In our case, one of our disaccharides did not come in a 0.5M solution. To

achieve the right 0.5M solution for any concentration you may have, use the concentration/volume equa-

tion : C1V1=C2V2. Our solution was 2M, so we would need 5mL of our 2M solution and add 15mL of

distilled water to get 20mL of 0.5M solution. After making up your solutions, pipette 4.5mL of the re-

spected solution into its respected test tube. Make sure to use different pipettes for different solutions. Af-

ter all the tubes contain their respected 4.5mL liquid, place the rack into a 40 degree water bath for a min-

imum of 10 minuets before starting your experiment. Using a 50 mL beaker, achieve approximately 20mL

of yeast, and place it aside which will later be mixed into your glucose solution test tubes. After 10 min-

uets, grab your first test tube, pipette 1.5mL of yeast into the tube, invert it with the parafilm over it. After

inverting, transfer that solution with the pasteur pipette into the durham tube. After making sure the liquid

is filled to the top, invert the durham tube back into the test tube and place it in the 40 degree water bath

for a total of 20 minuets. When you get the tube back, measure the amount of carbon dioxide bubbles in

mL. Repeat this for all the tubes (Querengesser & Froggatt, 2016).
 In this procedure, we used different species of disaccharides to describe the release of carbon

dioxide going through the process of cellular respiration. The statistical procedure we performed on

Numbers. We created a table, showing us our mean average and standard deviation. These were then

compared and displayed on a graph, the (x) value being the disaccharide and the (y) was the average

amount of carbon dioxide . Having a sample size of 3 gives us a quick, more averaged reading on how

certain disaccharides affect carbon dioxide being produced as a product.

Results

From our data, we obtained mostly positive results from our disaccharides and yeast solutions.

They were consistent with our original hypothesis, which stated that lactose was no going to produce any

carbon dioxide because the enzymes in yeast are not able to metabolize lactose into its two monosaccha-

rides, glucose and galactose, shown in the figure below (Figure 1). The minimum and maximum CO2

release is 3.87 and 3.89, maltose and sucrose respectively (Table 1).
Table 1. Effectiveness of disaccharides on the rate of cellular respiration in S. Cerevisiae, n=3.

Figure 1. The effect of different disaccharides undergoing cellular respiration with yeast on release of car-
bon dioxide from the cell wall, n=3.
 Discussion

The key findings that we found with are experiment was that sucrose and maltose did produce

significant levels of carbon dioxide. Our control was to show us how much CO2 will be released in the

absence of sugar. On the other hand, the results we got from lactose were accurate as well, considering the

knowledge we know about lactose. Like in our bodies, this disaccharide cannot be processed by yeast be-

cause like the intestinal b-galactosidase, it lacks it's enzyme source to metabolize lactose. (Mahoney,

1998). Enzymes, such as Saccharomyces cerevisiae in yeast are able to metabolize the other disaccharides

into their monomers. (Amaya-Delgado, Hidalgo-Lara, & Montes-Horcasitas, 2006)

The experiment was performed in 2 hours or so, but I think we might have been able to fit in an-

other trial of each disaccharide, which would of given us a more detailed graph, since only 2 of 3 pro-

duced CO2. More trials would make our average of carbon dioxide more reliable and realistic. We might

be able to see a less deviation in our results if the water bath was closer to our work bench. If we assured

that when the tubes were taken out of the water bath for the same amount of time, we might see more sim-

ilar amounts of CO2.

In conclusion, we found that when multiple different disaccharides under go cellular respiration

through yeast, depending on the structure of the monosaccharides, and where the glycosidic bond is be-

tween the monosaccharides, disaccharides will produce significant amounts of CO2 in sucrose and mal-

tose, and none in lactose.

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