Lactase Enzyme Lab

Name:______________________

Introduction:
Lactose, the sugar found in milk, is a disaccharide composed of glucose and galactose (both six
sided sugars). Sucrose, ordinary table sugar, is also a disaccharide composed of fructose and
glucose. Glucose is a six sided sugar and fructose is a five sided sugar.
Lactase is an enzyme that breaks down lactose into galactose and glucose. Lactase can be
purchased in pill form by people who are lactose intolerant. These people lack the enzyme
lactase and cannot break down the sugar lactose into its component parts.
Although lactose is similar to sucrose, lactase will break down only lactose- due to the shape of
the sugar.
In this lab, you will see lactase break down lactose into galactose and glucose. We will also
observe what happens if the shape of lactase is changed due to heating.

Materials:
(Note: All enzymes end in ase)

Per Group
Lactase Solution
10-15 mL Skim Milkthe source of lactose
6 Test Tubes & test
tube rack
Marking pencil
Timer
6 Glucose Test Strips
Water

Shared Materials (you will not use all of these materials)

sucrose solution- a disaccharide composed of fructose and glucose
Amylase- breaks down starch to maltose (a disaccharide)
catalase solution- an enzyme found in the liver that breaks down
hydrogen peroxide, a natural byproduct of metabolism
HCl- a strong acid **
NaOH- a strong base **
maltose- a disaccharide composed of 2 glucose monomers

** must use safety goggles

Lab Procedures:
1) Take your lab bin containing materials. There will also be some shared materials on the
lab table in the front of class.
2) Label 6 test tubes from A through F. The test tubes will contain the following:
A - skim milk (source of lactose) and lactase enzyme solution.
B - skim milk and water.
C - skim milk and denatured lactase enzyme solution.
D you will decide
E you will decide
F time permitting, you will decide
3) In test tube A add 2 ml of skim milk and 1 ml of lactase enzyme solution.
4) Time for 2 minutes to allow the enzyme to work, and then test for glucose with the glucose
test strip. Record this data in Table 1. If there was glucose present mark a plus '+' in the
table. If glucose was absent, mark a minus '-' in the table.
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5) In test tube B add 2 ml of skim milk and 1 ml of water.

6) Repeat step 4.
7) In test tube C add 2 ml of skim milk and 1 ml of denatured lactase enzyme solution.
8) Repeat step 4.
9) For test tube D , write procedural step that will prove that lactase will not work on other
sugars. (Hint: are other sugars available to you?)
Procedure for this test tube:
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Now, prove it using test tube D.
10) Repeat step 4.
11) For test tube E, write a procedural step below that will prove that another enzyme will not
work on lactose. (Hint: are other enzymes available to you?)
Procedure for this test tube:

Now, prove it using test tube E.

12) Repeat steps 4.
13) Time permitting, find an appropriate experimental use for test tube F that relates to this
lab. Consult with your lab partner. If your procedure involves either HCl, or NaOH, then
consult with Mrs. Plassio prior to performing this step.
Procedure for this test tube:
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What do you hope to prove in this test tube?
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Now, prove it using test tube F.
Results:
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Table 1: Glucose presence in the following solutions.

Test Tube
Contents (write in contents if blank)
A

Milk and enzyme solution

Milk and water alone

Milk and denatured enzyme

+/- for glucose

D
E
F

Analysis Questions (Answer the following quesitons using complete sentences):

1) Describe how this lab demonstrates why our bodies need enzymes.
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2) What happened when the enzyme was boiled?

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3) Another way to affect the enzyme is by lowering (or raising) the pH of the solution. However,
lactase is supposed to be able to work in the stomach. Would lowering the pH of the enzyme
solution affect the enzyme? Why or why not?
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4) Draw a picture to represent the chemical reaction, labeling the enzyme, substrate and products.

5) How is the enzyme lactase specific in its function?

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6) As your summary for this lesson, write a paragraph describing how an enzyme works. You
must use the terms enzyme, catalyst, substrate, active site, and activation energy in your response.
The terms must be used in the correct context.
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Teacher Notes/Procedure for solutions:

Notes:
students may perhaps work in groups of 4 to save resources also, it helps when
doing procedures D &E where they have to THINK! Surprisingly, many have
trouble doing this, theyll put two sugars together or 2 enzymes.
some of the solutions that will not cause a reaction can be dummy solutions made
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up of corn starch or salt or baking soda, for example (just make sure they dont
have glucose)
some lactase products already have glucose in them and will not work with this lab.
All solutions can be prepared the day before
Solution Preparation:
1) Enzyme Solution: Add 1 lactase tablet to 200 ml of water. Stir until the tablet has dissolved.
2) Sucrose Solution: Add 5 grams of sugar to 100 ml of water. Stir until the sugar has
dissolved. Repeat for maltose- if you dont have maltose, you can use sucrose- it will act the
same!
3) Denatured Enzyme Solution (completed prior to lab):
if the kids do it
a) Place 20 ml of Enzyme Solution into a pyrex test tube.
b) Add 200 ml of water to a 400 ml pyrex beaker.
c) Place the test tube in the beaker
d) Place the beaker and test tube on the hot plate.
e) Boil the water in the beaker for at least 45 minutes.
f) Let the solution cool to room temperature.
If you prepare it, simply boil it on the stovetop for 45 minutes, make sure you test it because it
may still work!
4) For amylase and catalase enzyme solutions, the Flinn Scientific catalog has recipes on how
to reconstitute these enzymes. If you dont have them in stock, or choose not to use them, they
can be dummy solutions since we dont expect a reaction.

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