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3.1: Enzyme Protocol

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  • Introduction to Sugars and Enzymes

    The food you eat contains many different types of molecules, including two types of sugar molecules: monosaccharides and disaccharides.

    Three types of monosaccharides are:


    Two types of disaccharides are:


    1. Each disaccharide is made up of two __________________________ .

    Monosaccharides from the food you eat are absorbed from your gut into your blood and carried to all the cells in your body where they are used for energy. Each disaccharide molecule must be broken down or digested into its monosaccharide components before it can be absorbed into the blood.


    2. When a sucrose molecule is digested, which monosaccharides are produced?






    The digestion of the disaccharide lactose to the monosaccharides glucose and galactose occurs very very slowly unless there is an enzyme to speed up the process. The enzyme that speeds up the digestion of lactose is called lactase. Lactase and most other enzymes are proteins.


    Each enzyme has an active site where a substrate molecule binds. For example, the substrate lactose binds to the active site of the enzyme lactase. Notice that the name of the enzyme lactase was created by adding the suffix "–ase" to part of the name of the substrate lactose.


    3. Circle the active site in the lactase enzyme in the figure.


    An enzyme speeds up a chemical reaction which converts a substrate molecule or molecules to a product molecule or molecules. The products are released from the enzyme and the enzyme returns to its original state, so the enzyme is ready to act on another substrate molecule. Thus, an enzyme molecule can be reused over and over. For example, a single molecule of the enzyme lactase can speed up the digestion of many many molecules of lactose.


    4. The following equation shows the digestion of lactose.


    Use E to indicate the enzyme, S to indicate the substrate, and P to indicate the products. Circle the molecule that is a protein, and use asterisks to indicate the molecules that are sugars.


    Experiment 1 - Can the Sugar Lactose be Digested without any Enzyme?

    To test whether the enzyme lactase is needed to digest the sugar lactose, you will test whether lactose breaks down to glucose and galactose in two different conditions:

    1. With no enzyme

    2. When the enzyme lactase is present.


    5. First, predict what you think will happen; for each column, circle the equation that describes what you think will happen.

    Prediction with No Enzyme Prediction with the Enzyme Lactase
    clipboard_eae253da736753d8bc1b52d2cfa5d372b.png clipboard_e7975baba703a4f4ca2a980fa95a2194f.png

    To test whether your predictions are correct, you will use glucose test strips to test whether glucose has been produced.



    • One member of your group (experimenter 1) should label test tube 1 and add 10 mL of lactose solution.
    • Another member of your group (experimenter 2) should label test tube 2 and add 10 mL of lactose solution and 1 mL of lactase solution. Put on a glove, put your thumb on the top of the tube and turn the tube upside down several times to mix the two solutions.
    • Wait 3 minutes to allow time for lactose to break down to glucose and galactose.
    • While you are waiting, experimenter 1 and 2 should each get a test strip. Record the original color of your test strip. ______________________________ In the next step, if your test strip changes color, this will indicate that glucose is present.
    • After the 3 minute wait, each experimenter should use a glucose test strip to test for glucose in the solution in his or her test tube. Your teacher will give you the instructions for using your glucose test strip.

    • Record your results in the table in question 6 below.

    • Clean your test tubes.


    6. Record your results in this table.

      Tube 1 - 10 mL of lactose solution

    Tube 2 - 10 mL of lactose solution

               + 1 mL of lactase solution

    Test strip color after a 3-minute wait


    Was there any change in the color of the test strip?


    ___ no glucose produced

    ___ some glucose produced

    ___ no glucose produced

    ___ some glucose produced


    7. Was lactose digested without any enzyme? ___ yes ___ no

    8. Was lactose digested when the enzyme lactase was present? ___ yes ___ no

    9. Do your results match your predictions? ___ yes ___ no

    If yes, what conclusion do your results support? If no, what do you think is the reason for the difference between your predictions and results?






    10a. In Tube 2, there were over 5000 lactose molecules for each lactase molecule. How can a single lactase molecule break down many many lactose molecules? (Hint: See question 3.)






    10b. A catalyst is defined as a substance that increases the rate of a chemical reaction without itself undergoing any permanent chemical change. Is the enzyme lactase a catalyst? Explain why or why not.






    Experiment 2 - Can the Same Enzyme Digest Lactose and Sucrose?

    To test whether the same enzyme can digest lactose and sucrose, you will test whether the enzyme lactase can digest the disaccharide sucrose.

    11a. Write the procedures for your experiment to test whether lactase can digest sucrose. You will have available the same supplies as you used in Experiment 1 plus a sucrose solution.






    11b. Create a data table to record the results of your experiment.






    After your teacher has checked your procedures and data table, carry out your experiment and record your results.


    12. Did lactase break down sucrose? How do you know?






    13. Does the same enzyme digest lactose and sucrose? ___ yes ___ no

    Your results illustrate a general principle called enzyme specificity. Each enzyme acts only on a single substrate or several chemically similar substrates.

    14. Which part of an enzyme do you think is responsible for this enzyme specificity? Explain your reasoning. (Hint: See the bottom of page 1.)

    Because of enzyme specificity, our bodies need lots of different enzymes to digest different types of food molecules. For example, our small intestine has the enzyme lactase to digest lactose and a different enzyme to digest sucrose.






    15. What do you think is the name of the enzyme that digests sucrose? (Hint: See the figure in Question 2.)






    16. Complete the following equation to show the digestion of sucrose. Include the enzyme and the products.


    Experiment 3 – Does the Enzyme Lactase Digest the Sugar in Milk?

    17. Suppose you have mixed 10 mL of milk with 1 mL of lactase solution, waited 3 minutes, then tested for glucose and found that your mixture of milk and lactase contained glucose. What are two possible interpretations of this result? (Hint: Think about the possibility that, before digestion, milk contains either a disaccharide or a monosaccharide.)






    18. Design an experiment to test whether lactase is needed to digest the sugar in milk. For this experiment, you will have available the same supplies you used in Experiment 1 and milk. Write your procedures and create a data table.









    • After your teacher has checked your procedures and data table, carry out your experiment and record your results.


    19. Is lactase needed to digest the sugar in milk? ___ yes ___ no 

          How do you know?






    20. Which sugar does milk contain? ___ glucose ___ lactose ___sucrose?

          How do you know?






    The Digestive System and Lactose Intolerance

    Molecules like sugars and protein enzymes are much much smaller than a whole person. For example, a person is roughly a billion times longer than a protein molecule like an enzyme. So where are these tiny molecules located in our bodies?

    This diagram shows a simplified outline of the human digestive system, including the small intestine where enzymes like lactase and sucrase break down food molecules into smaller molecules. Most digestion occurs in the small intestine.


    21. In this diagram, use D with arrows to show how disaccharides like lactose and sucrose get from food to the small intestine.

    - Use E to indicate the location of the enzymes, lactase, and sucrase.

    - Use M to indicate where monosaccharides like glucose are produced from disaccharides.

    22. Suppose that the cells in a person's digestive system do not make lactase. What do you think would happen to the lactose molecules in the milk that person drinks?






    Human babies and the babies of all other mammals depend on milk for their nutrition. Almost all babies produce the enzyme lactase to digest lactose, which is the main sugar in milk.

    In contrast, many adults produce very little lactase, so they can only digest a little bit of lactose at a time. When a person who produces very little lactase consumes large amounts of lactose in a short time period, most of the lactose is not digested in the small intestine and lactose reaches the large intestine where it is digested by bacteria. This can result in symptoms such as diarrhea, flatulence, and discomfort. This condition is called lactose intolerance.

    23a. A person who is lactose intolerant can drink lactose-free milk to gain the benefits of the protein, calcium and vitamin D that milk provides. Explain how a manufacturer can use lactase to make lactose-free milk.






    23b. Do you think lactose-free milk contains glucose? ___ yes ___ no

    If yes, where did the glucose come from?






    What Happens to the Digested Food Molecules?clipboard_e9b14160d23aa6beb3e882856cc84ee66.png

    The digestive system and the circulatory system work together to provide nutrients to cells in all parts of your body. Digested food molecules are absorbed from the small intestine into the blood. The heart pumps the blood through the blood vessels to carry the digested food molecules to all the cells in your body.


    Your cells use digested food molecules for two basic purposes:

    • To provide the building blocks for synthesizing needed molecules
    • To provide the energy for cellular activities such as muscle contraction and synthesizing needed molecules.




    24. Describe the steps by which disaccharides in your food end up as monosaccharides in cells throughout your body. Write several sentences and/or add arrows and labels to these diagrams. Include the words blood, enzymes, heart, and small intestine in your explanation.