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.
we need to break down lactose, lactase breaks and decomposes galactose to glucose

2. What type of reaction is this? Dehydration synthesis or hydrolysis?

lactase reaction
hydrolysis breaks it apart
hydrolysis of lactose and dehydration of lactase

3. Which type(s) of sugar did the enzyme lactase break down? Explain your answer.
lactose is the type of sugar that it broke down and can be confirmed but its not breaking down
glucose because its already broken at its lowest. Milk and water did not have the glucose because
water doesn't affect it, but in the lactase was gonna have glucose regardless. For maltose it would
end positive even if it was added water or lactose solution because maltose had glucose and
would test positive.

4. What happened when the lactase enzyme was boiled?

It stayed negative because
if you change the form of lactase, with heat, then it breaks it down and the function won't be able
to do the same thing. Altho u had the enzyme it didn't do what it was supposed to do because the
form was changed.

5. Would lowering the pH of the lactase solution affect the enzyme? Explain.
if u change the ph it would affect the solution because if its lowered its making it break more
when u pore acid, the enzyme breaks down and ruins the shape and it wouldn't be working the
same. Ph does affect the enzyme negatively because the enzymes need a certain homeostasis and
a certain temperature otherwise they will break down and it changes the protein which would be
the enzyme.