Diffusion/Osmosis and Enzymes

Name: ___________________________________ (1pt)

TA: __________________________________ (1pt)

Section: _________________ (1pt)

Diffusion/Osmosis

Diffusion Experiment A. Kinetic Energy of Molecules

Review the information in the manual and observe the video: https://youtu.be/s1mn-ud2Cx0

1. Is the movement of a single carmine particle - random or directional? (1 pt)

2. Does the movement ever stop? (1 pt)

3. Do smaller particles move more rapidly than larger particles? (1 pt)

4. Are you observing molecular movement? Explain. (1 pt)

5. How can molecular movement bring about diffusion? (1 pt)

1
Diffusion Experiment B. Diffusion of Molecules Through a Selectively Permeable Membrane

After reading the manual and observing the experimental setup:

1. Propose an idea that has measurable and controllable elements
2. Hypothesize about the selective permeability of dialysis tubing to the substances being
tested
3. Predict the results of the I2KI and Benedict’s tests based on your hypothesis (if/then).

Question (1pt):

Hypothesis (1pt):

Prediction (1pt):

Results (10 pts)

Complete the table as you observe the results.

Solution Contents Original Original WAIT 45 Final Color Final Test

Color Test for MINUTES for
Glucose Glucose
30% X
Bag glucose +
starch
H2O+I2KI X
Beaker

Discussion
1. What is the significance of the results for the movement of the iodine, starch and
glucose? Did the results support your hypothesis? Explain, giving evidence from the
results of your tests. (3 pts)
Iodine =

Starch =

Glucose =

2
 2. The results can be explained by concluding that the dialysis tubing is permeable to
_____________and ________________ but not ___________ (3 pts).

3. From your results, predict the size of the I2KI molecules relative to starch. Predict the size
of the glucose molecules relative to starch. (2 pt)

4. What colors would you expect if the experiment started with glucose and I2KI inside of
the bag and starch in the beaker? Explain. (4 pt)
Beaker final color =

Beaker glucose test =

Bag final color =

Bag glucose test =

Osmosis Experiment A. Osmotic Behavior of Animal Cells

Read the manual and observe the test tubes with blood and the demonstration slides.

Results
Macroscopic observations of solutions A, B, and C (6 pts)
Appearance of the Solution Can you read the Print?
(opaque/transparent)
A

Microscopic observations of solutions A, B, and C (6 pts)

Draw Appearance/Condition of Cell
A

3
Based on the demonstration and your microscopic investigation, which of the three solutions is
(3 pts)
Hypotonic to red blood cells?

Hypertonic?

Isotonic?

Osmosis Experiment B. Osmotic Behavior in Cells with a Cell Wall

Read the manual and observe the demonstration slides.

Results (4 pts)

Draw Appearance/Condition of Cells

Based on your predictions and observations, which solution is (2 pts)

Hypertonic? A
Hypotonic? B

4
 Enzymes

We will be examining the following reaction:

2 H2O2 (aq) -> 2 H2O (l) + O2 (g)
The enzyme catalase speeds up this reaction and the rate of reaction can be observed by bubbles
in the product.
Experiment 1: Identification of Controls
1. Label tubes.
2. Add the catalase and water to tubes
3. Add hydrogen peroxide and replace cap.
4. Invert tube 1 time.
5. Wait 20 seconds.
6. Measure bubbles using ruler.
7. Record results (6 pts)

Tube Catalase Hydrogen Water Height of

Peroxide Bubbles (mm)

1 1 ml 3 ml -

2 1 ml - 3 ml

3 - 3 ml 1 ml

Which is the substrate? Which is the enzyme? (2 pts)

Substrate =
Enzyme =
Are we measuring the rate of disappearance of the substrate or the appearance of a product? (1
pt)

Which is/are the positive control tubes? Which is/are the negative control tubes? (2 pts)
Positive =
Negative =

5
Experiment 2: Identification of Inhibitor
Note: The inhibitor must be added to the tube first otherwise the reaction occurs regardless
of the presence of the inhibitor. Sodium azide must be added carefully in the hood!
1. Label tubes.
2. Add the catalase and water to tubes.
3. Add sodium azide to tube and replace cap. Invert tube 1 time. Wait 60 seconds.
4. Add hydrogen peroxide and replace cap.
5. Invert tube 1 time.
6. Wait 20 seconds.
7. Measure bubbles using ruler.
8. Record results (6 pts)

Catalase Hydrogen Sodium Water Height of

Peroxide Azide bubbles (mm)

1 1 ml 3 ml 5 drops -

2 1 ml 6 ml 5 drops -

3 1 ml 3 ml - 5 drops

Competitive inhibition can be reversed (meaning the rate of reaction is the same as uninhibited
reaction) if the concentration of substrate is raised to sufficiently high levels while the
concentration of the inhibitor is constant.
In contrast, noncompetitive inhibition can not be reversed by increasing substrate concentration
(meaning the rate of reaction is less than the uninhibited reaction).
Is there a difference in the height of bubbles between Tube 1 and 2? (1 pt)

Therefore, is sodium azide a competitive or non-competitive inhibitor of catalase? (2 pt)

6
Experiment 3: Effect of Temperature, pH and Enzyme Concentration on Rate of Enzymatic
Reaction
Note: Each group of 4 students will do 1 experiment. Data will be shared by the class. If
you run short on time, the TA will supply data on CANVAS.
Temperature:
1. Place 1 ml of catalase at the noted temperature for 15 minutes.
2. Add hydrogen peroxide and replace cap.
3. Invert tube 1 time.
4. Wait 20 seconds.
5. Measure bubbles using ruler.
6. Record results (8 pts)

Catalase Hydrogen Peroxide Height of bubbles

(mm)

4C (refrigerator) 1 ml 3 ml

25C (room temp) 1 ml 3 ml

37C (body temp) 1 ml 3 ml

60C 1 ml 3 ml

pH:
1. Place 1 ml of catalase and 2 ml of buffer in tube.
2. Add hydrogen peroxide and replace cap.
3. Invert tube 1 time.
4. Wait 20 seconds.
5. Measure bubbles using ruler.
6. Record results (8 pts)

Catalase Buffer Hydrogen Height of

Peroxide bubbles (mm)

pH 3 1 ml 2 ml 3 ml

pH 5 1 ml 2 ml 3 ml

pH 7 1 ml 2 ml 3 ml

pH 9 1 ml 2 ml 3 ml

7
Enzyme Concentration:
1. Label Tubes
2. Place catalase and water in tube.
3. Add hydrogen peroxide and replace cap.
4. Invert tube 1 time.
5. Wait 20 seconds.
6. Measure bubbles using ruler.
7. Record results (6 pts)

Catalase Hydrogen Peroxide Water Height of bubbles

0 ml 3ml 2 ml

1 ml 3 ml 1 ml

2 ml 3 ml -

Construct three separate graphs to illustrate your results from these three experiments. See
manual for assistance in graph construction. (10 pts per graph, 30 points total)

The independent variable will be placed on the X-axis.

The dependent variable will be placed on the Y-axis.
Label the axes and use and appropriate scale. Note units.
Title each graph. Example:

Rate of Enzymatic Reaction vs. Amount of

Substrate
8
Height of Bubbles (mm)

7
6
5
4
3
2
1
0
1 2 3 4
Amount of Substrate (ml)