Name: Date: Period: Amylase Lab Background Information: Enzymes are present in all living cells. Our bodies contain many different enzymes. One enzyme that is used everyday is amylase. Amylase is found saliva and it is responsible for breaking down carbohydrates. Starch is a large molecule commonly found in foods such as grains, potatoes, and pasta. Essentially, it is a very complex form of sugar. For our bodies to use starch, it must be broken down into smaller, simpler sugars. A common simple sugar is known as glucose. In our mouths, the amylase that is found in saliva actually begins the break down of starch into glucose. Have you ever eaten a piece of bread or a cracker slowly and allowed it to dissolve in your mouth? You may have noticed a faint sweet taste or sensation. This is actually because your spit is breaking down the starch from the grains into a sugar! Try it! Purpose: - To determine the effects of high temperatures on enzyme activity. - To determine that amylase breaks down starch into sugar. Materials: Safety goggles Starch solution Test tubes (6 per group) Test tube rack Benedict’s solution 400 ml beaker Iodine solution Bunsen burner Amylase solution (or your own spit – you choose!) Information about the solutions: Starch solution: This is going to symbolize the grain food that we eat. Think of the starch solution as a piece of bread or a cracker Iodine solution: This is used as a starch indicator. Iodine changes color to a dark blue/black when it comes into contact with starch. Therefore, it lets us know if starch is present in a solution Name: Date: Period: Benedict’s solution: This is used as a glucose indicator. Benedict’s solution changes color to a reddish/pink when it comes into contact with glucose. Therefore, it lets us know if glucose is present in a solution. In order to work properly though, Benedict’s solution must be warmed up using a warm water bath Amylase solution: This represents the enzyme found in your own spit that is responsible for breaking down starch into glucose. You may use this solution, or, if you would like to be more realistic, try using your own spit instead Procedures: Part 1: Testing the enzyme to see if it works (Complete the data table of observations as you go.) A: Test for Starch: 1. Obtain 4 test tubes and label them “Test Tube 1”, “Test Tube 2”, “Test Tube 3” and “Test Tube 4” 2. Pour approx 10 drops of starch solution to each of the test tubes 3. In “Test Tube 1” and “Test Tube 2”, add an additional 10 drops (approx) of amylase solution (or your own spit). Mix them well by flicking the test tube. Allow them to marinate while you proceed to step 4. 4. In “Test Tube 3”, test for starch by adding one drop of the iodine solution. 5. Repeat step 4 for “Test Tube 1”. You are now finished with “Test Tube 1” and “Test Tube 3” Sample Contents Test Tube 1 Test Tube 3 starch Starch and amylase Reaction with Iodine (indicate color change) No colour change Blue/black B: Test for Glucose: 6. In “Test Tube 2” and “Test Tube 4”, add approx 5 drops of Benedict’s solution. 7. One at a time, carefully place “Test Tube 2” and “Test Tube 4” into a warm water bath. Name: Date: Period: Sample Contents Test Tube 2 Starch and amylase Test Tube 4 Starch Reaction with Benedict’s solution (indicate color change) Yellow/orange/pink No change Part 2: Effect of Temperature 1. Obtain 2 test tubes and label them “Test Tube 5” an “Test Tube 6” 2. Pour approx 10 drops of starch solution into both test tubes. 3. In a separate test tube, pour a small amount of amylase solution (or your own spit). Boil this test tube using the hot water bath. 4. Add your boiled amylase/spit solution to “Test Tube 6”. 5. Test both test tubes for starch by adding a drop of iodine solution to each. Sample Contents Reaction with Iodine (indicate color change) Test Tube 5 starch Test Tube 6 Starch and boiled amylase Blue/black Blue/black Concluding Questions: 1. Biochemists write the relationship among substrate, enzyme and product with the equation: enzyme Substrate -----------> product Write this equation using the substrate, enzyme and product of this amylase lab. (3 marks) amylase Starch ------------- Glucose 2. List three things that will denature an enzyme. (3 marks) Heat, high acidity, high alkaline (basic) Name: Date: Period: 3. In part 2 of the experiment, what happened to the amylase enzyme when it was boiled? Did it still work? What is happening to the active site? (3 marks) The enzyme was denatured, no it did not work because starch was still detected. The active site is being changed so that the substrate can no longer bind. 4. The temperature inside the human body where enzymes can function is 36.6°C. What might happen if a person has a fever of 40°C or higher? (2 marks) The enzymes would denature and not work properly because the substrate would no longer bind to the active site. 5. What was the purpose of adding the iodine solution and Benedict’s solution to our test tubes? (2 marks) Iodine: detects starch Benedict’s detects glucose