BIO30S

Enzyme Lab – Effect of Pepsin on Protein (Protein Farts!)

In this lab we’ll investigate several biochemistry topics that you’ll need to understand for this course as they pop
up in later units. These are Enzymes, pH, and acidity. You’ll investigate how pepsin (an enzyme in your stomach)
digests a protein (egg white or albumin)

Intro to Enzymes

Enzymes are proteins that act as a catalyst in living cells. As catalysts, they increase the rate of chemical reactions,
allowing the reactions to proceed rapidly when they otherwise occur only very slowly.

Enzymes are highly specific in their catalytic activity. The specificity of enzyme action is the result of “lock and key”
arrangement in which enzyme and the substance it reacts with (the substrate) join together to form an enzyme-
substrate complex. In this case the place of reaction, called the active site, is fixed. The “induced-fit” hypothesis
suggests that the active site is not a fixed arrangement but is flexible, allowing for a better fit.

When a reaction is completed, the enzyme and the newly formed reaction products separate, leaving the enzyme
unchanged. Enzymes are highly efficient catalysts. Only small quantities are needed to catalyze the reaction of
relatively large amounts of materials. Each enzyme has an optimum range of temperature and pH at which it
operates most efficiently.

Pre-Lab Questions

Discussion (Use your prior knowledge, notes, and textbook to answer these questions)

1. Explain what a metabolic pathway is and what enzymes have to do with them.
2. Define enzyme, substrate, and induced fit model of enzyme action.
3. How are enzymes usually named?
4. What are 5 factors that determine how fast an enzymes work?
5. Explain what the pH scale is and what a buffer is.
6. The substance with which and enzyme reacts is its _____________
7. Is an enzyme “used up” by the reaction it catalyzes? Explain.
8. In what way does an enzyme affect the reaction it catalyzes? How does the enzyme produce this effect?
9. What is the active site of an enzyme?
10. What is it meant by enzyme specificity?

Problem

What does pepsin do to proteins? Is hydrochloric acid needed?

Materials

5% Pepsin Graduated Cylinder Test tubes (4)

0.2% Hydrochloric acid Marking pen Incubator
Hard Boiled Egg 400 mL beaker Stoppers (4)
BIO30S

Procedure

a. Label the test tubes A, B, C, and D

b. Put a piece of hard boiled egg in each test tube. Try to make the pieces the same size and shape. For
example, a cube with 1 cm for each side.
c. Add the following substances to the test tubes:
- Tube A – 10 ml of water
- Tube B – 10 ml of 0.2% HCl
- Tube C – 10 ml of 5% Pepsin
- Tube D – 10 ml of 5% pepsin plus 2 drops of 0.2% HC;
d. Put stoppers in the test tubes. Stand the test tubes in a beaker like the picture shows.
e. Put the set-up in an incubator at 37 degrees Celsius. Leave it there for at least 24h. If you don’t have an
incubator, let the set up stay for 48h.
f. Look at the egg whites closely. Note any evidence of digestion (breaking down)

OBSERVATIONS
A WATER

B HCl

C PEPSIN

D PEPSIN + HCl

Post Lab Questions

1. What are the independent variable, dependent variable, and control for this lab?
2. In which tube did digestion occur?
3. What substances are needed for the digestion of protein?
a. Are these substances present your stomach juices?
b. Your stomach is made of protein. Why is it not digested by its own juices?
4. Which tube showed the most digestion? Does this make sense? Explain
5. Why were the test tubes incubated at 37 degrees Celsius?
6. Could life as we know exist without enzymes? Explain

Remember, you need to create a final conclusion as part of your lab report. Post-lab questions and
conclusions are two different things, but both are included in your lab report.