CATALASE LAB TEMPLATE 2019

An investigation on the effect of enzyme concentration on enzymatic decomposition of

hydrogen peroxide by yeast (Saccharomyces cerevisiae) catalase

Research question: How does yeast catalase concentration affect the rate of decomposition of
hydrogen peroxide?

Hypothesis: As catalase concentration increases from 1- 100%, the gas pressure measured over a
defined time (15 s) will increase, indicating:

- increased production of the product (oxygen gas + water)

- a direct relationship between enzyme concentration and reaction rate

Environmental and safety precautions

The hydrogen peroxide can cause skin and eye irritation upon contact. Wear chemical splash
goggles, chemical-resistant gloves, and a chemical-resistant apron. Wash hands thoroughly with
soap and water Hydrogen peroxide is a caustic substance but is considered safe to flush down the
sink.1.2

MATERIALS LIST

- 3X 50 ml beakers, each containing 20 ml of 1%, 50% and 100% fresh bakers yeast (COOP
Switzerland)
- 50 mL of 3% hydrogen peroxide (in 100 mL beaker)
- 6X 5 mL Syringes (‡ 0.1ml)
- 3X Vernier pressure sensors, connected to Vernier device (kPA(‡0.01)
- Test tube holders
- indelible marker pen
- 4 human assistants
- Laptop

METHODOLOGY

1. Prepare catalase solution in syringe

2. Label 3 syringes on top with 1%, 50%, 100%
3. Draw 2 ml of appropriate catalase solution into each syringe

1
Preparation of Vernier sensors

● Connect Logger pro equipment to laptop and switch on application

● Connect 4 gas pressure sensors to Logger pro, and confirm that all four sensors are visible on
computer
● screen
● Label each sensor BOTH on the screen AND on the sensor itself: 100%; 50%; 25%; 0%
● Under Experiment Set up' - set up sensor to take 1 reading every 2 seconds for 6o seconds
● Confirm that Green 'start' button is lit so that equipment is ready for use

Preparation of hydrogen peroxide solution

● Label 3 test tubes: 100%; 50%: 1%

● Using 2 ml syringe, add 2 ml of hydrogen peroxide to each test tube

Data collection

● Designate 1 student to each concentration of catalase. Each student will hold the
appropriately labelled
● test tube and syringe
● Students will simultaneously add catalase to peroxide in test tube, briefly swirl the solution
and then
● simultaneously place the rubber bung securely on the test tube. Thereafter tubes must be
held still.
● One student will press 'start' as soon as all tubes are sealed. Data collection will continue for
60 seconds.
● Data export
● Data is saved using Vernier and then UPLOADED to Google drive.

TABLE OF VARIABLES

Variables (Independent, Likely impact upon the investigation How the variable will be changed/
dependent, control, OR measured OR controlled
uncontrolled)

Independent variable    Indicate the SPECIFIC values been

Clearly define the IV, including units used for its measurement chosen for the IV (provide UNITS AND
(when appropriate) uncertainty(accuracy)] (usually 4 – 5
enzyme concentration by measures of specific values)
oxygen produced over period of time

2
 [Explain briefly why these values were
chosen]

Dependent variable Gas pressure Briefly outline how, and how often
[Clearly define the measurements be taken? (provide
dependent variable UNITS AND uncertainty(accuracy)]]
including the units used
for its measurement

Controlled variables Using bullet points, for each controlled Using bullet points, for each
Using bullet points, variable, explain how and why the controlled variable outline how each
identify all variables that variable could impact the data collected of these variables will be kept
must be kept constant to constant and if necessary how they
ensure a fair test?] will be monitored and/or measured]

Uncontrolled variables Using bullet points , for each [Suggest how the effect of the
Using bullet points, uncontrolled variable, explain how and uncontrolled variable could be
identify all variables that why the variable could impact the data minimized and/or monitored, if
might affect the collected possible]
investigation, but are
impractical to control]

3
MATERIALS LIST

● 3 X 50 ml beakers, each containing 20 ml of 1%, 50% and 100% fresh bakers yeast (COOP Switzerland)
● 50 mL of 3% hydrogen peroxide (in 100 mL beaker)
● 6 X 5 mL Syringes (± 0.1ml)
● 3 X Vernier pressure sensors, connected to Vernier device ( kPA(± 0.01)
● Test tube holders
● indelible marker pen
● 4 human assistants
● Laptop

METHODOLOGY
Prepare catalase solution in syringe
● Label 3 syringes on top with 1%, 50%, 100%
● Draw 2 ml of appropriate catalase solution into each syringe

Preparation of Vernier sensors

● Connect Logger pro equipment to laptop and switch on application
● Connect 4 gas pressure sensors to Logger pro, and confirm that all four sensors are visible on computer
screen
● Label each sensor BOTH on the screen AND on the sensor itself: 100%; 50%; 25%; 0%
● Under Experiment Set up’ - set up sensor to take 1 reading every 2 seconds for 60 seconds
● Confirm that Green ‘start’ button is lit so that equipment is ready for use

Preparation of hydrogen peroxide solution

● Label 3 test tubes: 100%; 50%: 1%
● Using 2 ml syringe, add 2 ml of hydrogen peroxide to each test tube

Data collection
● Designate 1 student to each concentration of catalase. Each student will hold the appropriately labelled
test tube and syringe
● Students will simultaneously add catalase to peroxide in test tube, briefly swirl the solution and then
simultaneously place the rubber bung securely on the test tube. Thereafter tubes must be held still.
● One student will press ‘start’ as soon as all tubes are sealed. Data collection will continue for 60 seconds.

Data export
Data is saved using Vernier and then UPLOADED to Google drive.