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Starch Digestion by Amylase Lab Report

Mercline Okengo

Department, Institutional Affiliation

Human Anatomy & Phys II Lab

Ms. Walton

22nd March 2022

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Starch Digestion by Amylase Lab Report

Proteins' primary function is to act as enzymes or catalysts, which speed up practically all

cells' chemical reactions. Enzymes are biological catalysts that help chemical reactions move

faster (Şentürk, 2017). As a protein, the structure of an enzyme can be influenced by a variety of

factors. For example, a rise in temperature can enhance an enzyme's rate, but it can also denature

and become inactive. Starch has to be broken down for human beings to utilize energy. Enzyme

amylase catalyzes this process making it more efficient. Enzyme amylase is produced in the

salivary glands and pancreas to facilitate food digestion (Şentürk, 2017). Starch is a polymer that

consists of molecules of glucose linked through covalent bonds. In the presence of a disaccharide

like an amylase, starch is broken down into maltose. The IKI (Iodine-Potassium Iodide) test can

be used to monitor amylase's effect on starch; however, IKI does not stain maltose. Benedict's

solution is sometimes used to detect maltose by a solution turning red when heated.

Purpose

The experiment aims to see whether breaking down starch into maltose is faster in the

presence of enzyme amylase or in its absence and whether or not the enzyme performs better

when it has been boiled at 100oC for three minutes.

Hypothesis

The experiment's hypothesis is that there will be no reaction occurs when starch and

amylase are boiled at 100 degrees Celsius in distilled water. With room temperature amylase, the

reaction can be seen since amylase has not been denatured.

Materials and Methods

Materials
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To perform the experiment, the materials used were a rack of six test tubes, three spot

plates, a hot plate, a large beaker, disposable droppers, and a marker. Wet materials used were

amylase, starch solution, maltose solution, IKI (dropper), Benedict's solution, and distilled water.

Methods

2 mL of distilled water was placed in a clean test tube and named #1 water. 2 mL of

amylase was then poured into two test tubes and labeled #2 amylase and #3 boiled amylase. The

test tube labeled #3 boiled amylase was then placed into a boiling water bath for five minutes.

After the five minutes, it was removed, and cold water was run over its end to speed up the

process of it cooling to room temperature. The spot plates' three columns were then labeled as

follows: Column 1- #1 water, Column 2- #2 amylase, Column 3 - #3 Boiled amylase. The spot

trays were then arranged in a single line while ensuring all wells were aligned.

Testing for Starch with IKI Reagent

A drop of Iodine solution was placed in each well of each spot plate. The experiment was

begun by adding 2 mL of starch to each test tube. With this addition, tube #1 had 2mL starch and

2mL water, tube #2 had 2mL starch and 2mL amylase, and tube #3 had 2mL starch, and 2mL

boiled amylase. Each test tube was shaken to mix the components, and immediately two drops of

these solutions were added to each start well. After a minute, two more drops of these solutions

were added to the "1 minute well". The addition of two drops of the solutions process was done

after every minute for a total of 11 minutes. The starch reactions were then recorded as positive

(+)and negative (-) in the results table.

Testing for Maltose (Glucose + Glucose) with Benedict's Reagents

Each test tube was then tested for the presence of maltose (glucose + glucose) by adding

equal amounts of Benedict's reagent to the remaining solutions in each test tube. A millimeter
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ruler was used to measure the height of the remaining solution in each test tube. The exact height

in millimeters of Benedict's reagent was then added. The three test tubes were then placed in the

boiling water bath for four minutes. During the four minutes, time for each test tube to begin

changing color was recorded in seconds or minutes.

Results

The results of control identifications, which are color-changing reagents used to identify

positive controls, are shown in the two parts of the experiment. In the first part, testing for starch

with IKI reagent, the second well, which contains starch and amylase, showed a gradual color

change (brown-black to brown) with each testing minute. The first and third wells row exhibited

no color change since the first wells row retained its yellow-orange color and the third-row

solutions retained the blue-black color. Despite the first row retaining its yellow-orange color, its

contents were supposed to turn blue-black at first when starch and water were added to iodine.
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Fig 1; Spot plates aligned and labeled 0 to 11 minutes in each row and water, amylase, and boiled

amylase in the three columns to determine when starch digestion began and if there was the

presence of starch at time = zero

The second part, testing for maltose (glucose + glucose) with benedict's reagents, of the

control technique shows that the test tube containing maltose has changed color from blue to red.

After the positive controls, the actual process begins. Tube #1, which contained distilled

water and starch, showed no color change in its corresponding well in Lugol's test. Color

changes were seen in tube #2 well (starch and amylase at room temperature) during 2-3 minutes,

changing from brown/black to yellow. There was no color change in the wells of tube #3 that had

boiled amylase and starch. The Benedict solution test is conducted at the end of the procedure. A

color shift from blue to red was seen in test tube #2, which contained starch and room

temperature amylase after 4 minutes boiling water bath. Test tubes #1 and #3 exhibited no color

change after the 4-minute boiling water bath as they retained the blue color.
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Discussion

Enzymes are proteins that speed up the rate of a reaction. The IKI (Lugol's) test was used

to show a positive control. Starch's presence was shown by the black color in wells in row two

labeled starch and amylase. When Benedict's solution is used, the maltose in the test tube turns

red, indicating a positive test. The hypothesis was found to be correct since the results supported

it. The starch was not hydrolyzed in boiled amylase in the first portion of the experiment since

amylase was denatured in tube #3. High heat either speeds up the reaction or induces

denaturation, which proved irreversible in this experiment. However, by observing the color

shift, hydrolysis of starch was noticed in the well containing room temperature amylase labeled

starch and amylase. The color shift from blue-black to brown in tube #2 can be observed since

amylase breaks down the starch present to simpler sugars (Şentürk, 2017). Benedict's solution

was positive for test tube #2 in the final stage of the experiment, as shown by a shift in color

from blue to red. There was no breakdown of starch and thus no color change since the amylase

enzyme was denatured in the boiling amylase test tube labeled #3. This was also the case in tube

#1 since there was no color change.

Possible Sources of Error

Despite the results supporting the hypothesis, there was an error in the first part of the

experiment when testing for the presence of starch using an IKI reagent. In the 1st row labeled

starch and water, the solutions in the wells were supposed to turn blue-black. However, this is not

the case since they retained a yellow-orange color showing that there was an error. Possible

sources of the error could be improper cleaning of test tube #1. In the future, the error can be

avoided by thoroughly cleaning test tubes and being keen when placing solutions into every test

tube.
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Practical Use of Information from the Experiment

The vital information picked from this experiment demonstrates a better grasp of how

the enzyme amylase aids in the breakdown of starch. Salivary amylase is linked to the

breakdown of starch in carbohydrates. Therefore, this information can be used in obesity and

diabetes studies. Chemical digestion of a piece of bread and other carbohydrates occurs in the

mouth when enzyme amylase coats them. In the process, the enzyme amylase binds to germs in

the mouth and on teeth, which has direct dental repercussions (Scannapieco et al., 2016). Excess

plaque and the formation of cavities are linked in dentistry.

Clinical Implications

Some health problems require the intake of enzymes to mitigate them. Exocrine

pancreatic insufficiency is a condition in which the pancreas is unable to produce enough

digestive enzymes to aid in the digestion of foods in the intestine. Malabsorption, diarrhea, high

susceptibility to infections, and other major health problems are all symptoms of pancreatic

insufficiency. In such cases, the standard treatment is for the patient to take enzymes like

amylase (Brennan & Saif, 2019). Such use of amylase to mitigate pancreatic insufficiency

improves human health.

Further Experiments

Further experiments can be done to test the stated hypothesis further. Reducing the

boiling time for tube #3 allows more experiments to explore alternative possibilities. By

changing the distilled water with frozen amylase, a new experiment can be carried out also.

Amylase in frozen form for later use in a room temperature experiment can also be used in a new

experiment. In the new hypothesis, a change in color of the reagents should be seen in the

previously frozen amylase.

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Conclusion

It's critical to understand the effect of amylase and the contents of each tube when doing these

tests. Since amylase belongs to the enzymes group, it is vital to know how each experiment

determines the capacity of amylase in the starch break down. If a scenario where starch and

amylase are present, the starch will be broken down by the end product, resulting in no starch

being detected and a negative Iodine test. Both tests should be negative if amylase is present, but

no substrate is available. When an enzyme lacks to facilitate the break down of the substrate, and

amylase is absent, but starch is present, the Iodine test will produce a change in color. The ability

of amylase to break down the starch in the tests was only observed at room temperature, showing

that enzymes can be denatured at very high temperatures.

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References

Brennan, G. T., & Saif, M. W. (2019). Pancreatic Enzyme Replacement Therapy: A Concise

Review. JOP : Journal of the pancreas, 20(5), 121–125.

Scannapieco, F., Torres, G., & Levine, M. (2016). Salivary α-Amylase: Role in Dental Plaque

and Caries Formation: . Critical Reviews in Oral Biology & Medicine, 4(3), 301-307.

https://www.deepdyve.com/lp/sage/salivary-amylase-role-in-dental-plaque-and-caries-

formation-tJh20IzsZn?key=sage

Şentürk, M. (2017). Enzyme Inhibitors and Activators. IntechOpen.

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