Enzyme Experimentation

INTRODUCTION:

In this experiment you will observe the action of the enzyme amylase on starch. Amylase changes starch into a simpler form: the sugar maltose, which is soluble in water. Amylase is present in our saliva, and begins to act on the starch in our food while still in the mouth.

Exposure to heat or extreme pH (acid or base) will denature proteins. Enzymes, including amylase, are proteins. If denatured, an enzyme can no longer act as a catalyst for the reaction.

Benedict’s solution is a test reagent that reacts positively with simple reducing sugars like maltose, but will not react with starch. A positive test is observed as the formation of a brownish-red cuprous oxide precipitate. A weaker positive test will be yellow to orange.

MATERIALS:

Cornstarch
Distilled water
Saliva
Vinegar
Benedict’s qualitative solution
3 graduated cylinders (10mL)
250-ml beaker
Stirring rod
3 test tubes (16 x 125mm)
Test tube rack
Wax pencil
Hot plate

PRE-LAB:

AID 1g of cornstarch to a beaker containing 100ml of cold distilled water. While stirring frequently, heat the mixture just until it begins to boil. Allow to cool.

PROCEDURE:

1. Fill the 250-mL beaker about ³/₄ full of water and place on the hot plate for a boiling water bath. Keep the water JUST AT BOILING.

2. Mark 3 test tubes A, B and C. “Spit” between 1 and 2 mL of saliva into each test tube.

3. Into tube A, aID 2 mL of vinegar. Into tubes B and C, aID 2 mL of distilled water. Thump the tubes to mix.

4. Place tube B into the boiling water bath for 5 minutes. After the five minutes, remove from the bath, and place back into the test tube rack.

5. AID 5 mL of the starch solution to each tube and thump to mix. Allow the tubes to sit for 10 minutes, occasionally thumping the tubes to mix.

6. AID 5 mL of Benedict’s solution to each tube and thump to mix. Place the tubes in the hot water bath. The reaction takes several minutes to begin.

OBSERVATIONS:

Tube A: Starch + saliva treated with vinegar (acid)

Was the test positive or negative?

What does this indicate?

Tube B: Starch + saliva and water, treated in a boiling water bath

Was the test positive or negative?

What does this indicate?

Tube C: Starch + saliva

Was the test positive or negative?

What does this indicate?

Please provide a lab report that explains your procedure, results and conclusions.  Remember to include pictures of the lab set-up and results.  Also, I want you to expand this initial experiment by proposing and testing (in the form of an aIDitional experiment) a testable hypothesis that involves the above general concept.  You will be graded on your use of the scientific method and the creativity/complexity of your experiment.  Both the above experimentation and your original experiment may be included together into one experiment