Introduction

Biologists are very interested in enzymes organic catalysts that control virtually all of the reactions that occur in living organisms. Enzymes are used in all metabolic
reactions to control the rate of reactions and decrease the amount of activation energy necessary for the reaction to take place. Enzymes are specific for each reaction
and are reusable. Enzymes have an area called the active site to which a specific substrate will bond temporarily while the reaction is taking place. The maximum rate at
which an enzyme can catalyze a reaction (in this case the breakdown of a substrate) is known as Vmax. In this activity, you will simulate the reaction of an enzyme with
its substrate. You will also determine Vmax.

Materials (per team of three people)

toothpicks
stopwatch
calculator
paper clips
gloves
ice
In this activity, the toothpicks represent a substrate and your thumbs and index fingers represent the enzyme, toothpickase. The
function of toothpickase is to catalyze the reaction of a whole toothpick breaking into two pieces. When you break a toothpick, the
place where the toothpick fits between the fingers of both your hand represents the active site of the enzyme. Place the toothpick
between the thumb and the first finger of each hand. You must use both hands to break the toothpicks. You can only break one
toothpick at a time.

Draw the reaction (reactant and products):

Draw the enzyme/substrate complex (label the

enzyme subunits, substrate, and active site)

Part 1: Rate of Enzyme Activity

1. Count out 50 toothpicks and place them in a shallow bowl.
2. In your group of three, one person will act as the enzyme toothpickase, the second will be the timer, and the third person will be the recorder.
3. The enzyme is to break the toothpicks without looking. Close your eyes! All of the product (broken toothpicks) must remain in the bowl.
4. The experiment is conducted in 20-second intervals. The timer calls out start and then marks each 20-second interval. The recorder tallies the cumulative number
of toothpicks broken as the timer announces each interval. You do not stop the timer, but record the total number of broken toothpicks at each interval. The
Toothpickase should break toothpicks during the entire time. Record the results in the data table below.
5. Graph the number of toothpicks broken as a function of time (20, 40, 60, 80, & 100 seconds.) Be sure to title your graph and to label the x and y-axis.

Table 1:
Time (seconds)
20
40
60
80
100
Graph 1:

Total # of toothpicks broken

6
7
8
8
7

Questions/Analysis:
1.
2.
3.
4.
5.
6.
7.

How many subunits does the toothpickase enzyme have? 3

Where is your active site? Fingers were used as the active site
What happens to the reaction rate as the supply of toothpicks runs out?
What would happen to the reaction rate if the toothpicks were spread out across the table so that the breaker has to
reach for them? There would be a lower initial substrate concentration and slower reaction rate
What would happen to the reaction rate if more toothpicks (substrate) were added? Increase initial substrate
concentration--> Vmax Same but longer
What would happen to the reaction rate if there were more breakers (more enzymes)? It would be slower.
For each time interval, calculate the rate of reaction of the toothpickase (divide the number of toothpicks broken for
each interval by the amount of time in that interval). Record your data in Table 2:

Time interval (seconds)

0 to 20
20 to 40
40 to 60
60 to 80
80 to 100

Rate of reaction of toothpickase (toothpicks broken/second)

0.3
0.35
0.4
0.4
0.35

8. Based on the table above, what is the V max of toothpickase?

9. What would happen to the reaction rate if the toothpickase wears bulky gloves? Count out 50 unbroken toothpicks and
repeat the data collection to find out! Record your data in table 3

Table 3:
Time (seconds)
20
40
60
80
100

Total number of toothpicks broken (wearing gloves)

2
0
1
4
1

10. Graph this data using a different color pen/pencil on Graph 1.

11. Describe the effect of the gloves on the reaction rate. Compare this to real enzymes. They slow it, like noncompetitive
inhibitors
12. Summarize what you have done in the activity so far. Acted as an enzyme and an enzyme with an inhibitor

PART 2: Reaction Rate vs. Substrate Concentration

1. Remove the broken toothpicks from the shallow bowl. Place 100 paper clips in the bowl. The paper clips represent a solvent in which the toothpicks are
dissolved. Mixing different numbers of toothpicks in with the paper clips simulates different concentrations.
2. For the first trial, place 10 toothpicks in the bowl with the paper clips. The enzyme has 20 seconds to react (break as many toothpicks as possible). Remember that
the enzyme is to break the toothpicks without looking. Record the number broken at a concentration of 10.

3. Remove the broken toothpicks and repeat with concentrations of 20, 30, 40, etc. up to 100 toothpicks. The enzyme only reacts for 20 seconds each time. Record
your data in the table below.

Table 4:
Substrate concentration
10
20
30
40
50
60
70
80
90
100
Questions/Analysis:

Rate of reaction of toothpickase (toothpicks broken/second)

0.2
0.3
0.5
0.5
0.5
0.45
0.45
0.55
0.7
0.75

1. When was the reaction rate the slowest? The fastest? Slowest= Smallest concentration /10/ Fastest= Largest
concentration /100/
2. Explain the relationship between substrate concentration and reaction rate.
Higher concentration=faster reaction rate
3. Summarize what you did in this part of the activity.

Part 3: Reaction Rate vs. Temperature

1. Select 10 toothpicks. Time how long it takes to break the 10 toothpicks as fast as you can. Then, place your hands in a bucket of ice
water for 30 seconds. Time how long it takes to break 10 toothpicks as fast as you can. 23 seconds

Questions/Analysis
1. Calculate the rate of enzyme action in toothpicks per second for each temperature. Show all of your work.
2. Compare the 2 rates. Explain what happened and why it happened. Relate this to real enzymes.
3. Summarize what you did in this part of the activity

FINAL DISCUSSION Based on your results in parts 1, 2 and 3, discuss the rate of enzyme activity under various conditions: