MONTESSORI SCHOOL

SCIENCE
DEPARTMENT
HIGH SCHOOL BIOLOGY MANUAL
Enzymes laboratory

Goal: The student recognizes the effect of the enzymes over a substate and would be able of
recognize the variables that can affect their correct functioning

Materials:

 5 beakers or transparent cups of 250mL

 Fresh Pineapple (Alternatives are: Kiwi and Papaya)
 Boiling Water
 2 Unflavor Gelatin Packages
 Vinegar or bleach
 Spoon or Spatula
 Bunsen burner/Oven
 Goggles, gloves and lab coat

Introduction

Enzymes are biological catalysts. A catalyst is a substance which speeds the rate of reaction but
remains unchanged in the reaction. Catalysts reduce the activation energy needed for a reaction.
Enzymes are proteins and occur naturally in living biological systems, acting in many metabolic
pathways.

The activity of enzymes are affected by pH, temperature, enzyme concentration and substrate
concentration. Enzymes have optimum pH and optimum temperatures, at which they experience
maximal activity. Enzymes are essential for life and an understanding of how they operate is
important for medicine and industrial biochemistry.

How enzymes operate – the active site theory of enzyme action.

1. The active site of the enzyme has a shape closely complementary to the substrate.
2. The substrate locks into the enzyme’s active site.
3. The shape of the active site changes slightly gripping the substrate more tightly and straining
it; an enzyme-substrate complex is formed.
4. The substrate undergoes a chemical change the new substance (the product) is released from
the active site. The active site springs back to its original shape and is ready to receive fresh
substrate.
Note: For enzymes to operate they must be able to make contact with the substrate; the enzyme
or substrate or both must be in solution, i.e. in rapid random motion in water. Our cells are about
90% water and water is the medium in which most of our biochemistry takes place.

In this laboratory we will see the action of the enzyme “Bromelain” from the pineapple or “papain”
from the papaya, participating in the degradation of the collagen present in the gelatin, also we
are going to observe the effect of temperature and pH in the enzymes activity.

What do you think is going to happen? Make your hypothesis

Hypothesis:

Procedure:

1. Dissolve the content of the 2 unflavored gelatin packages in hot water, make sure to put one
cup more of hot water that the indicated in the instructions, the idea is obtain a semisolid
gelatin.
2. Distribute the dissolved gelatin in 5 beakers or transparents cups and put it on the fridge.
3. Take the fruit and cut it in medium pieces (The size depends of the width of the cup)
4. Distribute the pieces as follow:
 One portion for boil in hot water for 5 minutes
 One portion for freeze
 One portion for put it in vinegar or bleach for 5 minutes
 The last portion keep it fresh

5. Take out all the cups from the fridge, make sure the gelatin is semisolid and cover the surface
of the gelatin with various pieces of pretreated fruit as follow:

Control

6. Observe and compare.

Lab report

1. What is meant by an enzyme?

Enzymes are biological catalysts; catalysts accelerate chemical reactions. Also, enzymes are
used in many biological functions within the body.

2. What kind of enzyme is present in the fruit of the experiment?

Pineapple-Bromelain.

3. Explain the phenomenon that you observe in the experiment (your results: include please your
pictures). Also, give your conclusions.

The Phenom is the changes in the enzymes properties with heat. I can conclude that the
enzymes work only when they are in normal conditions, if they are exposed to heat, we can
see that the reaction would be totally different. We can see this evidenced in the experiment,
the difference between both glasses.

4. Do you consider the time you left the experiment running enough? Explain.

Yes, the results expected of the experiment happened in the time established for the
experiment.

5. Have you identified, and controlled, all relevant independent variables? enumerate some of
these variables.

In this experiment we can identify many independent variables, some of them are: application
of vinegar to pineapple, boiling of pineapple, freezing the pineapple, the time the gelatin was
left in the fridge, and the amount of pineapple used. These are some of the independent
variables identified in this experiment.

6. What is the main goal of using a negative control condition (no enzyme)?

The negative control condition is the gelatin that we didn’t put anything on, the main goal of
the negative control condition is to be able to compare it with the other two results.

7. Can you include a positive control condition to confirm the enzyme activity? Explain.

No, because as we saw in the experiment, which each cup, something different happened, so if
we used one of those as the control or the normal one, the results won’t be accurate or right
according to the experiment that you are performing.

8. Indicate three safety precautions that you used.

I use safety implements such as: Goggles, gloves and lab coat.
I was very careful with the boiling water to avoid getting burn with it.
I used a pan that have a plastic handle, for avoiding burnings.
I off the gas immediately after the water boiled, and I put cold water to lower down the heat of
the pan.

9. State one source of error and suggest an improvement to minimize the error.

One source of error can be to have the gelatin too cold, too hot, or in the incorrect temperature
for the experiment. The improvement is to control constantly the temperature of all objects, to
have a precise result of the experiment.