Critical Reading

1. What is the material of the dress?

cotton

2. In this case narrative, Rob learned that the stain near the neckline of the dress contains starch. What specific

types of macromolecules are starch and cellulose?

 Polysaccharides

3. What monomers are starch and cellulose made of?

Glucose
4. Describe the overall structure of starch.

polysaccharide comprising glucose monomers joined in α 1,4 linkages

5. To remove the stain from the dress, Rob is treating the stain with a cleaner containing a hydrolytic enzyme

amylase. How did Rob know that the amylase enzyme would not damage the fabric of the dress?

The dress was made of cotton, which is mostly cellulose made from configurative links. The links aren’t
substances for amylase because cellulose isn’t made from a-configuration links.
6. What happens to the starch at the molecular level when it is digested by amylase?

When starch encounters amylase, it cuts off the bond between 2 glucose molecules. The enzyme breaks the

broken starches into simple sugars (maltose).

Analyze and Design an Experiment

To further investigate starch and its components, first you will analyze an experiment. Then you will design your own.
The experiment as outlined in the diagram below.

First they use iodine (Iodine-potassium-iodide, IKI) and Benedict’s solution as indicators to test the effect of amylase on
starch. Benedict’s is normally light blue. In the presence of simple sugars, Benedict’s solution gives color change to
green for a positive result. In the present of long-chain starch, iodine’s positive result is orange to dark blue/black.

Four test tubes were set up. To see the contents of each tube, see the table below.

Tube Reagent 1 Reagent 2 pH Boiled? Time Temp. (oC) Results Results

IKI Benedict’s
color color
1 Starch Water 7.0 No 60 37 Black Lt. blue
+ -
2 Starch Amylase 7.0 No 60 37 Orange Green
- +
3 Amylase Water 7.0 No 60 37 Orange Lt. blue
- -
4 Water Maltose 7.0 No 60 37 Orange Green
- +

The tubes were incubated at 37o C (body temperature) for 60 minutes. None of the tubes were exposed to extreme heat
or boiling temperatures. Half of the contents in tubes 1-4 were poured into tubes 1A-4A. The contents of tubes 1A-4A
were tested with IKI (iodine). The remaining contents in tubes 1-4 were tested with Benedict’s solution. The results are
below and the next set of questions will help you analyze the results of both tests.
Table 1.1 Test Tube Contents (check off the reagents/ingredients found in each test tube AFTER the reaction)

Reagent Test tube

1 2 3 4
Starch x
Amylase x
Buffer pH 7.0 x x x x
Maltose x x
Water x x

1. The test tubes (1A-4A) are shown at the top left of the figure. The color change shows the results. If starch is
present, the iodine’s positive reaction is the dark bluish/black color. The light shade (which is actually
yellow/orange). Which is the only test tube (1A-4A) positive for starch?

Test tube 1A

2. Why is the iodine test negative for test tube 2A?

While test tube 2A did contain starch, amylase was also added to the tube. Amylase broke down the starch during the
incubation period; therefore, when iodine is added to the tube there is no starch left for it to react with.

3. What is the purpose of adding iodine to the other test tubes (1A & 3A)?
Iodine is added to 1A to test the effectiveness of the iodine solution because the tube contains starch the contents
should turn a dark blue. Test tube 3A is a control to show that a combination of iodine and amylase does not lead to a
blue solution, it should remain light yellow.

4. Look at the results for the Benedict’s solution test (those are shown on the right). A dark shade indicates a
positive test for simple sugars. The light shade indicates a negative test. Does a positive Benedict’s test indicate
the presence of starch, amylase, or maltose?

Benedicts solution must be a test for maltose. The tests with starch alone and amylase alone produced a negative result.
The test with maltose alone and with starch and amylase produced positive results. The benedicts solution test reveals
that amylase does break down starch into maltose.

A positive result indicates maltose that result from starch being digested by amylase.

5. What is the relationship between starch, amylase, and maltose?

Amylases digest starch into smaller molecules, ultimately yielding maltose, which in turn is cleaved into two glucose
molecules by maltase.

Designing an Experiment

6. Consider either the iodine test or the Benedict’s test. Design an experiment to test for the effect of amylase on
starch under one of the following conditions: pH, incubation, temperature, incubation time, boiling, or freezing.

Which question are you considering? Does a change in pH level affect how well amylase breaks down starch?

Which test will you simulate using? Iodine and benedicts test `

Restate the question as a hypothesis and predict your outcomes.

If the pH is raised as high as possible, then the enzyme will fail to break down starch.

In Table 1.2 describe the contents of your test tubes. Check off the reagents/ingredients in the tubes then describe the
experimental conditions. Although it is likely that you might have more than one control tube, describe just one of your
controls for test tube 1. Describe your experimental treatment in test tube 2.

Table 1.2 Test Tube Contents & Conditions

Reagents Test Tube 1 (control) Test Tube 2 (experimental)

Starch x x
Amylase x x
Experimental Conditions
pH 7 14
Incubation Time 1 hour 1 hour
Incubation Temperature 37 degrees C 37 degrees C
Boiling
Freezing
 7. In Table 1.3, indicate the results you would expect if your hypothesis were supported (use + and – for positive
and negative for the test you chose).

Table 1.3 Predicted Results, Supporting Hypothesis

Tests Test Tube 1 (control) Test Tube 2 (experimental)

Benedict’s Blue - Blue -
IKI Black + Black +

8. In Table 1.4, indicate the results you would expect if your hypothesis were NOT supported (use + and – for
positive and negative for the test you chose).

Table 1.4 Predicted Results, NOT Supporting Hypothesis

Tests Test Tube 1 (control) Test Tube 2 (experimental)

Benedict’s Green + Green +
IKI Orange - Orange -

Off the Wall: Starch Degradation Investigation

Hildy planned to surprise her parents by remodeling their living room while they were away for the weekend. First she
had to remove the wallpaper so that she could paint. When she started scraping at the edge of the wallpaper, only a few
pieces came off. “What’s up with this wallpaper?” Hildy asked herself, “It’s just not coming off!”

Hildy got a spray bottle and filled it with warm water. She sprayed the walls to moisten the large areas. After several
minutes, she scraped at the wallpaper again. Larger pieces came off this time, but big patches of hardened paste
remained. Hildy couldn’t spend the whole day scraping! She rummaged around the house and found some alcohol and
some vinegar. Unsure of what these substances would do to the walls, she also went out and bought two different types
of commercial wallpaper remover. She wondered which would work the best.

To test which one would work best, she chose a section of the wall behind the couch and applied the five substances to
a small section of the wall. She labeled each patch to remember which substance had been applied to each area. After
30 minutes, she noted how much wallpaper she could remove with one scrape from each patch.

Hildy’s results.

9. Which substance appears to work the best? What does that tell us about the composition of the wallpaper
paste?

The substance with .5% amylase removed the most wallpaper. This tells us that the wallpaper past must contain starch.

10. How did the best remover work to remove the paste (what’s the chemical mechanism)?

The amylase in the solution broke down the bondages that held the starch monomers together.

11. Explain the results seen with the use of the acid & vinegar. What is happening to the molecules of the paste?

Acids work to hydrolyze starches, which results in a small area to be cleaned.

12. Why was it important that Hildy also test plain water?

The water was used as a control group to prove that water alone would not break down starch like the other substances
would.