OBJECTIVE

To observe the hydrolysis of amylose (starch) by enzyme amylase.

INTRODUCTION

Enzymes are biological catalysts that can significantly increase the rate of a chemical
reaction that take place within cells. Enzymes are very specific to the reactions they catalyze
and the substrates that are involved in the reaction.

Enzymes are specific to the substrates as they have a specific active site, which is a
small part of the enzyme where the catalyzed reaction take place, that only allow certain
substrates to bind to the active site. This is because the specific shape of the active site that
other substrates cannot fit and bind to it. Therefore, the lock and key hypothesis states that only
certain substrates fit an enzyme's active site, like a lock and a key would.

Enzyme amylase is an enzyme that catalyzes the hydrolysis of the polysaccharide

starch (amylose and amylopectin) into maltose, a disaccharide. In human body, enzyme
amylase is predominantly produced by the salivary glands and the pancreas.

Furthermore, enzyme amylase is widely distributed in plant tissues. However, it is most

abundant in seeds to initiate the breakdown of stored starch in the seed to disaccharides during
germination process.
 RESULT

Color of the area

Initial color of after flooding with Interpretation
Area of the plate
the area iodine solution
Soaked seed Starch is broken down into
Colorless Colorless
area maltose by enzyme amylase.
No starch is broken down into
Boiled seed area Colorless Blue-like color
maltose.
Starch is broken down into
Dry seed area Colorless Colorless
maltose by enzyme amylase.
Saliva control Starch is broken down into
Colorless Colorless
area maltose by enzyme amylase.

The result obtained in this study has shown the color changes in different area of the
plate before and after flooding the plate with iodin solution. It is known that the iodine solution
will react with amylase in starch and turn to blue-black color while it does not react with
disaccharide maltose. Hence, the interpretation for the area of the plate that has change from
colorless to blue-like color is that the starch in that area is broken down into maltose by enzyme
amylase. In contrast, the areas that do not change the color is said that no starch is broken
down into maltose by amylase.
 DISCUSSION

Based on the result, the boiled seed area is the only area of the plate that has changed
the color from colorless to blue-like color after flooding with iodine solution. While the soaked
seed area, dry seed area and saliva control area maintain colorless after flooding with iodine
solution.

The change of color of the boiled seed area is because the iodine solution has reacted
with the amylose in the starch and give a blue-like color. The presence of amylose in the boiled
seed area indicates that there is no amylase (especially α-amylase) presence in the boiled seed
area. This is because amylose of starch is the substrate of amylase enzyme. Starch is made up
of 20% amylose and 80% amylopectin (Hazard, B., Zhang, X., Colasuonno, P., Uauy, C.,
Beckles, D. M., & Dubcovsky, J., 2012). All reactions are faster at a higher temperature.
However, the enzyme-catalyzed reactions become slower or stop if the temperature becomes
too high. When the amylase is under a high temperature (higher than 55oc), the enzyme is
undergoing irreversible denaturation as protein can be denatured by heat. At a high
temperature, the active site of the enzyme will change its shape and cannot bind to the specific
substrate, thus losing its function (Daniel, R. M., Peterson, M. E., Danson & Lee, C. K., 2010).
Since there is no breakdown of starch into disaccharide maltose by amylase, the starch-agar
area changes from colorless to blue-like color when the triiodide ion (I3-) complex of the iodine
solution slips into the coil of the starch.

In the other hand, soaked seed area, dry seed area and saliva control area remain
colorless after flooding with iodine solution because the starch in the area is broken down into
maltose by amylase. When the corn seeds are soaked in the water for a period of time and are
incubated for 30 minutes, the embryo of the seed is activated to start the germination process.
The activated embryo will synthesize and releases gibberellins hormone which triggers the
aleurone layer of the seed to synthesize α-amylase and other digestive enzymes (Lee Ching, J.
Arunasalam, 2011). Although the dry seeds do not undergo germination process, the seeds still
contain α-amylase. Since the corn grain is cut longitudinally and the cut surface is facing down,
onto the agar surface, the α-amylase enzyme in both seeds can catalyst the hydrolysis of starch
into maltose. When the starch in the starch-agar area for soaked seed and dry seed is
breakdown into maltose, the starch-agar remains colorless as iodine solution do not react with
disaccharides.
 For saliva control area, the saliva that is applied on the starch-agar area contains
abundant amount of enzyme amylase. Therefore, the starch of that area is then broken down
into disaccharides, maltose by the amylase in saliva (Peyrot des Gachons, C., & Breslin, P. A.,
2016). Since iodine solution do not react with disaccharides, the starch-agar in saliva control
area maintain colorless.
 CONCLUSION

In a nutshell, the purpose of this experiment was to observe the hydrolysis of amylose
(starch) by enzyme amylase. The goal was accomplished by conducting the experiment with
“boiled seed”, “soaked seed”, “dry seed” and saliva control in a starch-agar plate. The result
shown that starch-agar will turn into blue-like color after flooding with iodine solution if the
amylase is absence at that area as the starch is not broken down (Rani, A., & Ali, U., 2021).
However, if the starch-agar remains colorless after flooding with iodine solution, it indicates the
presence of amylase at that area as the amylase will catalysts the hydrolysis of the starch in the
starch-agar into disaccharides maltose. Since iodine solution does not react with maltose, the
starch-agar remains colorless.

From this experiment, the hydrolysis of amylose (starch) by enzyme amylase can be
observed by observing the change of color after flooding with iodine solution. This observation is
useful for better understanding of hydrolysis of enzyme amylase to the substrate, amylose. The
α-amylase enzyme in seed is very important to a seed as it is used to breakdown the stored
starch in seed into disaccharides that is essential for a seed germination process. In the other
hand, salivary amylase is most associated with the breakdown of starch in carbohydrates,
making it a centerpiece of study for diabetes and obesity research. In addition, salivary amylase
also binds to bacteria in the mouth and on teeth, which has implications for dentistry, like excess
plaque and the development of cavities in the teeth (F.A. Scannapieco, G.I. Torres, M.J. Levine,
1995).
 REFERENCE

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F.A. Scannapieco, G.I. Torres, M.J. Levine. (1995). Salivary Amylase Promotes Adhesion of Oral
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Hazard, B., Zhang, X., Colasuonno, P., Uauy, C., Beckles, D. M., & Dubcovsky, J. (2012). Induced
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