Abstract

This lab’s intention was to test how catalase and hydrogen peroxide affect one another. I

assumed that the more hydrogen peroxide (or catalase , depending on which part of the lab) there

was, the faster the discs would float. My data for varying hydrogen peroxide concentrations

shows an increase in reaction rate (in inverse seconds) and then a slight decrease. For varying

catalase, the data shows a gradual increase in reaction rate (in inverse seconds). When there was

more hydrogen peroxide, the discs did float faster (ergo my hypothesis was supported), whereas

when there was more catalase the discs were being soaked in, the discs floated slower (my

hypothesis was not supported).

Introduction

Background

Enzymes are used to allow certain reactions with the use of a substrate. Enzymes are used

for everyday things, such as digesting food. “Most of the time enzymes ‘lock up’ with the

substrate they are supposed to break. However, sometimes non-substrate materials, such as

mercury and arsenic as water soluble compounds, can hook up with an enzyme, and the result

can be deadly.” (Science Learning Hub, 2011) The rest of the time, however, specific substrates

do connect with the enzyme it fits with. The region of where the substrate and enzyme connect is

called the active site. Once the enzyme and substrate are connected, the enzyme begins to break

apart the substrate to create the specific reaction needed. “Enzymes are also used in other

everyday ways. For example, acetaminophen, or Tylenol, has an enzyme used to target specific

substrates that create symptoms of fever or swelling. It is difficult to find the correct enzyme

substrate/enzyme combination and know how it will react on an actual person with the
symptoms.” (Rye, Wise, Desaix, Jurukovski, Choi, Avissar, 2016) However, most people do not

pay attention to enzymes in their everyday lives, because to them, there may seem like there is

no reason to. However, enzymes are what make us feel better when we feel sick and feel full

when we are hungry. Figure 1 shows the enzyme breaking the substrate into the products.

Figure 1

This figure shows all four steps of an enzyme

breaking apart a substrate. First, they find each

other, then they hook up, then the substrate starts

breaking, and lastly the substrate is broken.

Purpose/Objective

The purpose of this lab was to test the speed, or rate, at which enzymes and substrates

react upon one another and which component it affects more. The question being asked during

this experiment was: At what rate do enzymes and substrates act upon each other, and which is

affected more? My hypothesis for the varying hydrogen peroxide was the more hydrogen

peroxide there is, the quicker the discs will float. I concluded this because “bubbles float because

they have a lower density, and objects with a lower density and higher buoyancy rate. float on
more dense objects with a lower buoyancy rate.” (NCFL, 2014-2018)My hypothesis for the

varying calf liver extract was the more extract, the quicker the disc will float.

Variables

The independent variable was the concentrations, the first half of the experiment having

variations of hydrogen peroxide, and the second half of the experiment having variations of

catalase (calf liver extract). The dependent variable was the rate of reaction. The controlled

factor was the beaker that only had water because those discs will never float because the water

does not react with the catalase.

Controlled Variables Method of Control

Water beaker with catalase-soaked discs Choosing a material (water) that doesn’t react

with calf liver juice (catalase)

Water soaked discs in hydrogen peroxide Choosing a material (water) that doesn’t react

with hydrogen peroxide (anything that doesn’t

react with hydrogen peroxide is not a catalase)

Materials and Procedures

Materials

The materials needed are as follows*;

● Baby calf liver extract/juice

● Paper discs (this version was using the paper that falls out of a hole puncher after

punching holes

● Tweezers
 ● 3% hydrogen peroxide

● Water stopwatch/timer

● 1 50 milliliter beaker per group

● Pipette/eyedropper

● Graduated cylinder(s)

*No living materials are required.

Safety

When carrying out this lab, one should keep their hair tied back, wear safety glasses, and

wear close toed shoes because hydrogen peroxide is a bleaching agent. If already wearing

glasses, the participant may just wear those but if they are, for some reason, worried about their

glasses, may use some of the safety glasses provided.

Procedure

Varying H202 Concentrations

The person(s) doing this lab should first set up sixth beakers. All the following

measurements should be done in a graduated cylinder and then poured into the beaker because

beaker graduations can be anywhere from four to six milliliters off. The first beaker should have

40 milliliters of water. The second should have 10 milliliters of hydrogen peroxide and 30

milliliters of water. The third should have 20 milliliters of hydrogen peroxide and 20 milliliters

of water. The fourth should have 30 milliliters of hydrogen peroxide and 10 milliliters of water.

The fifth should have 40 milliliters of hydrogen peroxide. The sixth and final beaker should have

about 12 milliliters of the catalase. To extract this from its container, the people performing the
lab should use a pipette.Then the people should get 25 discs. The discs (they may be done

separately or in groups) should be put in the sixth beaker and pressed down with the tweezers, so

the top is also getting covered. The discs should not float. If they do you either have weird paper

or you did something wrong. Then, after 45 seconds, the discs should be removed and

immediately placed in a beaker, five in the ones that do not have catalase in them. The timer

should start. It is easier to do one beaker at a time for this part because it is not as easy to look at

25 discs as it is to look at five. If using a timer that can lap times, that is recommended because it

is easier to press a button than write while looking at the beaker. The times should then be

recorded, along with the average time (trial 1+ trial 2+ trial 3+ trial 4+ trial 5/5) for each beaker

and the rate (1/the average, or 1/[trial 1+ trial 2+ trial 3+ trial 4+ trial 5/5])for each beaker.

Varying Calf Liver Juice

This experiment should have two beakers. Again, all measurements should be done with

a graduated cylinder. The first beaker will always be at 40 milliliters of hydrogen peroxide. The

second beaker should change. The second beaker that will only have water the participants do

need to make because, as shown in the first part of the experiment, catalase and water do not

react, so it is just wasting water to make this beaker. The second second beaker should have 2.5

milliliters of catalase and 7.5 milliliters of water. The third second beaker should have 5

milliliters of catalase and 5 milliliters of water. The fourth second beaker should have 7.5

milliliters of catalase and 2.5 milliliters of water. The fifth and final second beaker should have

10 milliliters of catalase. I referred to all these as second beakers because there is no reason to try

to make this experiment go faster. This is because the second beakers are what is soaking the

discs. Each second beaker should have five total discs soaked for 45 seconds, again pressing
down using the tweezers. After the discs are all soaked, they should be put (using tweezers) into

the 40-milliliter beaker with the hydrogen peroxide. The discs should be timed, again lapping

times if easier. When done timing all the discs and recording the answers, the average for each

beaker that the discs were soaked in should be recorded, along with the rate using the equations

noted in the varying H2O2 concentrations section of the procedure section.

Results

Qualitative Data

When conducting experiments, the first and foremost thing to do is recognize the basics.

In both parts of this experiment it should be noted that catalase is an enzyme that catalyzes, or

accelerates, reduction of hydrogen peroxide. In the case of this lab the catalase is the calf liver

juice/extract.
Quantitative Data

Varying H2O2 Concentrations

Beaker Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average Rate

([Trial 1 + Trial (1/Average)

+ Trial 3 + Trial

+ Trial 5]/5)

1- 0.00% Will never Will never Will never Will never Will never N/A N/A

H2O2 ffloat float float float float

2- 0.75% 0 1 3 3 4 2.20 0.45

H2O2

3- 1.50% 3 5 5.50 6 7 5.30 0.19

H2O2

4- 2.25% 5 9 9 9 10 8.40 0.12

H2O2

5- 3.00% 1 7 7 8 8 6.20 0.16

H2O2
Varying Calf Liver Juice

Beaker Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average Rate

([Trial 1 + Trial (1/Average)

2 + Trial 3 +

Trial 4 + Trial

5]/5)

1-0% Calf Will never Will never Will never Will never Will never N/A N/A

Liver Juice float float float float float

2- 25% 31 33 35 42 44 37 0.03

Calf Liver

Juice

3 -50% 13 14 14 15 16 14.4 0.07

Calf Liver

Juice

4 –75% 10 11 13 14 14 12.4 0.08

Calf Liver

Juice

5- 100% 9 10 11 11 11 10.4 0.10

Calf Liver

Juice

*These tables numbers are rounded to the nearest hundredth.

Uncontrolled Variables

In this lab while there were controlled variables such as the beakers with water, there

were also uncontrolled variables. For example, if the liver extract were contaminated by another

group with hydrogen peroxide or spit, our results may not be accurate. Below are graphs of the

averages and rates of the time for both parts of the experiments.
* The y- axis represents the rate of reaction in seconds, while the x-axis represents the beakers

with varying concentrations. Numbers are rounded to the nearest tenth in the graph. For the

numbers above the bars are the results rounded to the nearest hundredth.

In the graphs above, there are the rates of the trials averages. The colors are to

differentiate between bars. The two water only beakers, which are both beaker 1, had no

movement because water is not a catalase nor hydrogen peroxide, so there could not be a

reaction. There is definitely an outlier, which is beaker two’s rate of reaction (0.45 inverse

seconds) in the Varying Amounts of Hydrogen Peroxide bar graph.

Conclusion

Purpose and Hypothesis

The purpose of this lab was to determine the rate of reaction for paper discs soaked in

catalase and put into hydrogen peroxide. My hypothesis for varying hydrogen peroxide (the more

hydrogen peroxide there is, the quicker the discs will float) was supported by the results because

beaker five (0.16 inverse seconds) had a quicker reaction rate than beaker four (0.12 inverse

seconds). My hypothesis for varying catalase (the more extract, the quicker the disc will float)

was not supported by the results because beaker two had the quickest reaction time (0.03 inverse

seconds), followed by beakers three (0.07 inverse seconds), four (0.08 inverse seconds), and five

(0.10 inverse seconds).

Evaluation

This experiment would have had more accurate results if we had done more trials per

concentration, and if we had compared our results to the rest of the class because there would be

less room for error if there are more things being averaged together. “The catalase/hydrogen

peroxide reaction is similar to a lactase/lactose reaction because lactase (the enzyme) reacts on

lactose (the substrate)”. (Biology Dictionary, Editors) Another variable we could have tested was

having uniform hydrogen peroxide and catalase, with varying water percentages in the catalase

to dilute it and see how that would affect the paper discs.

Bibliography

Rye, C., Wise, R, Jurukovski, V., Desaix, J., Choi, J., Avissar, Y., 2016, Biology 6.5 “Enzymes”

National Center for Family Learning, 2014-2018, “Why do Bubbles Float?”

Science Learning Hub, 2011, “Digestive Enzymes”

Biology Dictionary, 2014. “Substrate”