Name _____________________________________________ Date _______________________

Effects of Lactase Enzyme on the Breakdown of Lactose

Introduction:
Lactose, the sugar found in milk, is a disaccharide composed of the monosaccharides, glucose and galactose Sucrose,
ordinary table sugar, is also a disaccharide composed of glucose and fructose.

Enzymes act as organic catalysts to regulate the rates of reactions to maintain normal homeostasis within the body of
living organisms. They have a specific molecular structure which binds to a specific structure of the substrate, or
molecule that they act upon. The enzyme and substrate will bind together at an area on the enzyme known as the active
site. It is like the teeth on a key fitting into a specific lock. If they don’t have complementary shapes, they cannot bind.
This is called the Lock and Key theory.

Enzyme-
Enzyme Substrate Enzyme
complex
https://biochem80p.files.wordpress.com/2014/03/enzyme-working-mechanism.jpg

Lactase is an enzyme that breaks lactose down into glucose and galactose. 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 lactose down (in skim milk) into galactose and glucose. You will also observe what
happens if the shape of lactase is changed due to heating. This process is called denaturing. The shape of the active site
gets altered much like melting the teeth on a key. It won’t work in the lock anymore. Denaturing of enzymes in our
bodies is very difficult due to our body’s attempt to maintain homeostasis. In this lab, we had to boil the enzyme (placing
a test tube with the enzyme in a beaker of boiling water) for thirty minutes.

Purpose: This lab will examine how enzymes are specific to a substrate. You will also observe what happens when the
enzyme is denatured by heat and thus, changing its active site.

Materials:
Enzyme (Lactase) solution Skim Milk
Water Sucrose solution
10 ml Graduated cylinder 5 Test Tubes
Test Tube Rack Marking Pencil
Labeling tape 6 Glucose Test Strips
Stirring Rod

Procedures:
1) Put on your goggles and gather your equipment
2) Set up 5 test tubes with the following labels:

A. Skim milk + enzyme

B. Skim milk + water.
C. Skim milk + denatured enzyme
D. Sucrose + enzyme
E. Sucrose + water.
3) In the first column in the Table 1, predict whether or not the disaccharides, (sugars), in each test tube would
be broken down.

a) 4) In test tube A, add 2 ml of skim milk and 1 ml of enzyme solution.

b) 5) In test tube B, add 2 ml of skim milk and 1 ml of water.
c) 6) In test tube C, add 2 ml of skim milk and 1 ml of denatured enzyme solution.
d) 7) In test tube D, add 2 ml of the sucrose solution and 1 ml of enzyme solution.
e) 8) In test tube E, add 2 ml of the sucrose solution and 1 ml of water.

4) With a glass stirrer, be sure to mix the solutions of each test tube…BE SURE TO WIPE DOWN THE
GLASS STIRRER BETWEEN EACH TEST TUBE SO YOU DO NOT CROSS CONTAMINATE THE
MIXTURES.
5) Set timer for 4 minutes after you have filled ALL the test tubes.
6) Test each test tube solution for glucose with the glucose test tape but dipping the glass stirring rod into the
solution and then putting 2 drops onto the tip of the glucose test tape.
NOTE: Glucose test strips are normally a teal color. They will change from teal to green or brown
with a higher concentration of glucose. Any change from this teal is a positive result.
17) If there was glucose present, mark a '+' in the table. If glucose was absent, mark a '-' in the table. Record
this data in table 1.

Table 1: Glucose presence in solutions

 Glucose test Strip
Prediction Test Tube Treatment results
( + or -)
CIRCLE ONE:

Lactose
Will Break Down/
1 Skim milk + Enzyme
Will NO T Break

CIRCLE ONE:

Lactose
2 Skim milk + water
Will Break Down/
Will NO T Break

CIRCLE ONE:

Lactose Skim milk + Boiled

Will Break Down/
3 enzyme
Will NO T Break

CIRCLE ONE:

Sucrose
4 Sucrose + Enzyme
Will Break Down/
Will NO T Break

CIRCLE ONE:

Sucrose
5 Sucrose + water
Will Break Down/
Will NO T Break

Conclusion/Questions Answer each question in a COMPLETE SENTENCE. Be sure to include

EXPLANATIONS / JUSTIFICATIONS.

1. What is an enzyme?

2. Label the diagram below with the following terms: Enzyme, Active Site, Substrate, Enzyme-
Substrate Complex and Product
3. What part of the enzyme will bind to the substrate (the molecule it acts upon)? ___________________

4. What was produced when lactase acted on the sugar lactose? _____________________________

5. How do you know the reaction was successful, turning lactose into glucose and galactose?

6. Glucose test strips will indicate if glucose is present in a solution. If sucrose is made up of the
monomers glucose and fructose, why did the glucose strips NOT detect glucose in the
‘sucrose + enzyme’ solution?

7. What part of the enzyme got altered when the enzyme lactase was boiled? ______________________

8. What two conditions can affect the effectiveness of an enzyme?

9. Lactase is supposed to work in the stomach which normally has an acidic pH level between 2-6.
a. Would effect would lowering the pH of the lactase’s environment to a pH level of 4 affect the
enzyme? Explain.
 b. Would lowering the pH of the enzyme’s environment to a pH level of 0 affect the enzyme?
Explain.

10. What type of reaction is this? Dehydration synthesis or Hydrolysis? Explain your answer using these
four terms monosaccharide, disaccharide, water, and bonds.

11. THINK ABOUT IT…If the enzyme lactase breaks down the sugar lactose, and maltase breaks down
maltose; what enzyme is necessary to break down sucrose?