Lactase Enzyme Lab

Purpose:
This lab will examine the specificity of an enzyme (lactase) to a specific substrate (lactose).
Students will observe the actions of the enzyme and how shape is important to enzyme reactions.
Students will also observe what will happen when the enzyme is denatured.

Background Information:
Lactose​, the sugar found in milk, is a disaccharide composed of glucose and galactose (both
6-carbon sugars). ​Sucrose​, ordinary table sugar, is also a disaccharide composed of fructose and
glucose. Glucose is a six-carbon sugar and fructose is a five-carbon sugar. ​Maltose​ is a sugar
found in germinating grains (like barley) and is used to brew beer and sweeten food products.
Maltose is a disaccharide composed of two glucose molecules.

Lactase​ is an enzyme that breaks lactose down 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.

Materials:
● Test tube rack
● 7 test tubes
● Test tube marking pencil
● Distilled water
 ● Solutions of milk, lactase, sucrose, and maltose
● Glucose test strips

Procedure:
1. Label 7 test tubes with the letters A through G.
2. To each test tube, add the following:
● Test Tube A: 2 ml milk + 1 ml water
● Test Tube B: 2 ml milk + 1 ml lactase solution
● Test Tube C: 2 ml milk + 1 ml boiled lactase solution
● Test Tube D: 2 ml sucrose solution + 1 ml water
● Test Tube E: 2 ml sucrose solution + 1 ml lactase solution
● Test Tube F: 2 ml maltose solution + 1 ml water
● Test Tube G: 2 ml maltose solution + 1 ml lactase solution
3. In the Data Table, record your predictions: Which test tubes will you expect to find glucose?
4. For each test tube, use the glucose test strips to test for the presence of glucose. Record your
findings in the Data Table.

Data Table
Prediction:
Will it test positive Glucose Test
Test Tube Contents
for glucose? (+ or -)
(+ or -)
A milk + water - -
B milk + lactase solution + +
C milk + boiled lactase solution + -
D sucrose solution + water - -
E sucrose solution + lactase solution + -
F maltose solution + water - +
G maltose solution + lactase solution + +

Analysis Questions:
1. Diagram and describe the lactose and lactase reaction.

In the image above the enzyme is represented by the circle labeled lactase. Lactose and lactase
would connect and then break down into galactose and glucose. This works because the enzyme
and Lactose are made to be shaped to fit with each other.
2. What type of reaction is this? Dehydration synthesis or hydrolysis?
The reaction in the lab is hydrolysis because in the ones that tested that has glucose, hydrolysis
means a reaction of breaking something down.

3. Which type(s) of sugar did the enzyme lactase break down? Explain your answer.
The two types of sugar that lactase can break down is galactose and glucose. The reason
for this is that the enzymes and sugars fit together like a puzzle piece, which causes them to
break down.

4. What happened when the lactase enzyme was boiled?

The lactase enzyme becomes deformed/loses it shape which can prevent it from breaking
down the sugars.

5. Would lowering the pH of the lactase solution affect the enzyme? Explain.
Having a lower pH would affect the enzyme because it slows down the process of
digestion.