Enzyme Lab

Background:
Human saliva contains the enzymes amylase and lipase. Amylases break down carbohydrates (mostly starches) and
lipases break down lipids. Example reactions for these enzymes are shown below.

There are many other enzymes throughout the various digestive organs, including additional ones to further break down
carbohydrates and lipids. For the purpose of this lab, however, we will focus in on human saliva and substrates
dissolving or ‘getting mushy’ in your mouth (our identifier of breaking down into products). Keep in mind the enzymatic
schematic discussed in the powerpoint:

Purpose:
Students will experimentally design a test to determine the effectiveness of the enzymes amylase and lipase in human
saliva in various conditions.

Structure:
Ø In your assigned group, determine what feature you will be testing about amylase that would impact its
effectiveness
o Choose either an environmental factor (changing the pH by eating something slightly acidic or basic
with it, changing the temp of the food or holding an ice cube in your mouth prior to eating) or a
substate factor (simple vs complex carbs, different types of simple carbs)
Ø In your assigned group, determine what feature you will be testing about lipase that would impact its
effectiveness
o Choose either an environmental factor (changing the pH by eating something slightly acidic or basic
with it, changing the temp of the food or eating the lipid with other macromolecules at the same time)
or a substate factor (saturated vs non-saturated, different types of lipids)
Ø Look around your kitchen/pantry.
o What do you have that would qualify as a lipid and a carbohydrate that you are willing (and allowed) to
eat? Do you have substances that would meet what you chose as your feature above?
o How does what you have compare to what your lab mates have?
 Ø As a group design your two experiments for amylase and lipase. Fill in the template below before you get
started.
o You’ll need to define “gets mushy/dissolves” so that everyone uses the same stopping point for equally
comparable results.
Ø Perform the experiment. Remember to do multiple trials. I would recommend between 3 and 5 per person.

Experimental Design Templates

Experiment 1: Amylase
1. Hypothesis:

If the pH level in the mouth is lowered with lemon juice, then it will take less time to break down the substance,
because acid helps to break down substances.

2. Variable Identification (DV/IV/control):

The dependent variable is how long it takes for the food to break down, the independent variable is the lowering of the
pH, and the control group is doing the experiment without lemon juice.

3. Materials:
- Mini rice cakes
- Lemon Juice
- Measuring Spoon
- Stopwatch
4. Method: (remember someone should able to read these by themselves and repeat exactly what you do)
a. The general gist here is you should be putting it in your mouth and timing how long it takes to
digest/breakdown/get mushy.
b. Remember to do multiple trials here!
1. Place one half of mini rice cake in mouth
2. Start stopwatch and time until substance turns into a paste-like consistency
3. Repeat steps 1-2 for 3-5 trials, record results
4. Measure ¼ teaspoon of lemon juice, and swish in mouth for 10 seconds
5. Place one half of mini rice cake in mouth
6. Start stopwatch and time until substance turns into a paste-like consistency
7. Repeat steps 4-6 for 3-5 trials, record results

5. Results:

Time (min:seconds) Trial 1 Trial 2 Trial 3 Average

6. Conclusion:

This portion of the lab was used to test the effectiveness of amylases in saliva. Mini rice cakes served as the
carbohydrate, were tested in a normal pH, then tested in a more acidic pH. When tested in a normal pH of saliva, around
6.2 to 7.6, the saliva broke down the carbohydrate on an average of 26.23 seconds. After swishing around lemon juice in
the mouth, the pH was lowered to a more acidic level for the second set of trials which decreased the average time to
13.65 seconds. After nearly cutting average in half, it is clear to see that amylases work faster in a setting with a lower
pH, or more acidic environments, confirming the hypothesis.
Experiment 2: Lipase
1. Hypothesis:

If the pH level in the mouth is lowered with lemon juice, then it will take less time to break down the substance,
because acid helps to break down substances.

2. Variable Identification (DV/IV/control):

The dependent variable is the time it takes for the cashew to break down, the independent variable is adding lemon
juice, and the control group is breaking down the cashew without lemon juice.

3. Materials:
- Cashews
- Lemon Juice
- Measuring Spoon
- Stopwatch
4. Method: (remember someone should able to read these by themselves and repeat exactly what you do)
a. The general gist here is you should be putting it in your mouth and timing how long it takes to
digest/breakdown/get mushy.
b. Remember to do multiple trials here!
1. Place one half of cashew in mouth
2. Start stopwatch and time until substance turns into a paste-like consistency
3. Repeat steps 1-2 for 3-5 trials, record results
4. Measure ¼ teaspoon of lemon juice, and swish in mouth for 10 seconds
5. Place one half of cashew in mouth
6. Start stopwatch and time until substance turns into a paste-like consistency
7. Repeat steps 4-6 for 3-5 trials, record results

5. Results:

Time (min:seconds) Trial 1 Trial 2 Trial 3 Average

6. Conclusion:

This portion of the lab was to test the effectiveness of Lipases at breaking down lipids in saliva. Cashews were
representative of lipids, tested in a normal pH of saliva, then again in a lower pH. The control group was tested in
saliva with a normal pH and took an average of 1:24.67 to break down into a paste. Afterwards, lemon juice was
used to lower the pH of saliva, creating a more acidic environment for the cashews and increasing time to 2:21.33.
The group reported similar findings, with most trials returning with longer times after adding acid. It can be
concluded that Lipases are most effective at a neutral to basic pH rather than acidic, contradicting the hypothesis.