Lab 5 Enzyme Activity Lab, Recommended to wear gloves and safety goggles, take precautions with hot

water in part 2

Objective: To identify reagents/substrates and enzymes, to identify the products of a specific enzyme/substrate
reaction, to determine the optimum or preferred environment/conditions for an enzyme to function.

Hypothesis: YOU WILL PROVIDE THIS

Materials needed:

1. Clear plastic, 4 ounce (3)

2. Clear plastic, 6.5 to 8 oz. minimum (1)
3. Measuring spoons: ¼, ½. 1 teaspoon
4. Measuring cups, ¼ and ½ cups
5. Yeast, bakers, 3 envelopes
6. Hydrogen peroxide, 3%, 100ml
7. Plastic test tubes, 15 ml capacity (6)
8. Transfer pipets (5)
9. Ruler
10. Marker, sharpie
Student provides:
11. Timer
12. Thermometer (optional)
13. Tap water or distilled
14. Paper towels
15. Goggles and gloves
16. Optional: masking tape

Procedure:
Watch the TeleLearning video part 1 Enzyme Concentration first to get an idea of what you are doing.
Then once your supplies are organized,
1. begin by labeling 3 cups A, B and C. You will create 3 concentrations of the test subject, yeast. Yeast is
an organism which produces the enzyme catalase similar to our cells.
2. In first cup measure ½ teaspoon of yeast.
3. Carefully mix ½ cup of lukewarm water with yeast. You will need to determine this. To be a careful
scientist you could check the temperature with a digital thermometer you use for yourselves or the kids.
These report in Fahrenheit so you can practice conversions! Hooray! Note the F = 1.8 (C) + 32 so if I
need to be about 30C what should I be in F? My microwave usually takes 30 seconds for 1 cup of cold
tap water. You want to prepare about 2 cups of lukewarm water.

For a comparable experience to the video and to be considerate to those possessing mostly English measures, I
will use both. Note that we do not have to use precisely the same volumes as the demonstrator but should be
close.

MIX THOROUGHLY and make sure that all the yeast is dissolved = solution A
Need to work QUICKLY to do dilutions and make sure your solutions are well mixed
Now remove ¼ cup (60 ml) of this yeast solution (A) and add to another clean glass
Add ¼ cup (60 ml) of warm water to this and mix THOROUGHLY = solution B
Now remove a ¼ cup (60ml) of this yeast solution (B) into another clean glass
Add ¼ cup (60 ml) of warm water to this and mix THOROUGHLY = solution C

Congratulations you may have just made your FIRST serial dilutions.
Now you are ready to note the effect of enzyme concentration on the amount of product formed.

1. Label 3 test tubes 1 , 2 and 3

2. Add 1 teaspoon or 2 transfer pipets (5 ml) of hydrogen peroxide (H2O2) to each test tube.
Do the following one cup at a time since this reaction goes VERY quickly = 20 seconds
2. Mix yeast solution A thoroughly and add 1 teaspoon or 2 transfer pipets (5 ml) to hydrogen peroxide in
test tube 1
3. Time about 30 seconds
4. Measure the height of the bubbles in centimeters or millimeters with a ruler. Do not include the volume
of liquid in the measure.
5. Do the same for solution B (1 teaspoon or 2 transfer pipets (5 ml)) to test tube 2 of H2O2
6. Do the same for solution C (1 teaspoon or 2 transfer pipets (5 ml)) to test tube 3 of H2O2

Analysis:
Record your data in the lab report.
What happened? Did you prove your hypothesis?
Dispose of the solutions down the drain and Wash up all the equipment so it can be reused.

Conclusions:
What affect does enzyme concentration have on the amount of product formed? How did you know?

Part 2: watch TeleLearning video: Effects of temperature on Enzymes before performing

1. Label 3 test tubes 1, 2, and 3
2. Add 1 teaspoon or 2 transfer pipets or 5 ml of H2O2 to each
3. Set tube #1 in freezer for 15 minutes
4. Set tube #3 in a large glass with hot water (this should be 85C at least) for 15 minutes.
Note before you begin this experiment, make sure your test tube will not melt in this!!! The test tube needs to fit
without floating and tipping over.
5. Leave #2 at room temperature. You can record this from the thermostat in house.
While waiting for #1 to cool and #3 to heat you can proceed with #2.
6. Add 1/4/teaspoon of yeast to #2
7. Mix yeast with H2O2 by swirling the test tube gently to make sure it dissolves
8. Time 30 seconds
9. Measure the bubble height as you did in part 1
After the 15 minutes have passed you can do steps 6, 7, and 8 for #1 and #3.

Analysis:
Record your data in the lab report.
What happened? Did you prove your hypothesis?
Clean up your equipment, wash hands and dispose of solutions.

Conclusions:
What affect does temperature have on enzyme activity? How did you know?