Name: Missy Arabella C.

Pame Date Performed: July, 2021

Course and Year: BSN1-C Name of Lab Instructor: Ma’am Guadalupe Oracion

Experiment No. 9
ENZYMES

OBJECTIVES

1. To be able to describe the effect of temperature to the action of enzymes towards

a substrate.
2. To demonstrate the catalytic action of enzymes.

RESULTS AND DISCUSSION

I. Oxidase from Fruits

- After leaving the fruits for a period of time, the fruits would then start to turn
its color to brown. The enzyme responsible for the browning is called
polyphenol oxidase. In the presence of oxygen, the PPO enzymes changes
substance known as phenolic compounds into different compounds called
quinones. The quinones then react with other compounds to form melanin.
Melanin is the same dark brown pigment that colors hair, skin and the irises
of our eyes. It also turns fruit and vegetables brown. This reaction, however,
usually does not happen within fresh fruits and vegetables because the
PPO and the phenolic compounds are separated in produce plant cells. The
enzymatic browning process is only triggered when PPO, phenolic
compounds and oxygen come in contact with one another. This is exactly
what happens when a fruit is cut, falls or is knocked around too much. When
fruit tissue is damaged because of heat, cold, age or mechanical stress, its
cells break open and the phenolic compounds and the enzyme are released
and mix with oxygen in the air. As a result the damaged tissue turns brown
almost immediately.
 - The chemical reaction:
Polyphenol Oxidase + O2 Melanin (color brown) with Oxygen.
Oxygen activates the compound polyphenol oxidase in the fruit to turn the
fruit brown.
II. Catalase
a. From Potato
- In procedure A, I have observed that the boiled solution has lesser bubbles
than the solution that is not boiled. This is because the heat degraded the
catalase enzyme, making it incapable of processing the hydrogen peroxide.
The room temperature potato produced has the most bubbles that is
produced because catalase works best at a room temperature. Catalase
acts as the catalyzing enzyme in the decomposition of hydrogen peroxide.
Nearly all living things possess catalase, including us! This enzyme, like
many others, aids in the decomposition of one substance into another.
Catalase decomposes, or breaks down, hydrogen peroxide into water and
oxygen.
b. From Liver
- In procedure b, I have observed a foam or bubble formation was evident
after the addition of hydrogen peroxide. The gas that is liberated is oxygen.
When the enzyme catalases comes in contact with its substrate, hydrogen
peroxide, it starts breaking it down into water and oxygen. Oxygen is a gas
and therefore wants to escape the liquid. However, the dish soap that you
added to all your solutions is able to trap the gas bubbles, which results in
the formation of a stable foam. As long as there is enzyme and hydrogen
peroxide present in the solution, the reaction continues and foam is
produced. Once one of both compounds is depleted, the product formation
stops. If you do not add dish soap to the reaction, you will see bubbles
generated but no stable foam formation. If there is no hydrogen peroxide
present, the catalase cannot function, which is why in cup one you shouldn't
have seen any bubble or foam production. Only when hydrogen peroxide is
 available, the catalase reaction can take place as you probably observed in
the other cups.
III. Amylase
- In this procedure, I have observed that when you add the Fehling’s reagent,
T1 A (boiled), it appeared as light blue, and meanwhile T2 A appeared as
yellow.
- In the next procedure where iodine solution was added, T1B (boiled)
appeared as almost black and T2B appeared as light brown and some
precipitate.
- This procedure indicates that T1A has a negative result, meaning that
amylase didn’t work well because of the high temperature when it was
boiled. Since the boiled test tube has no or little working amylase, this
indicates that starch was not broken down. This is the explanation ny blue
black color was the result of the iodine test.

CONCLUSION

Enzymes are essential for survival. The proteins made by our cells help transform
chemicals in our body, enzymes function as catalyst. A catalyst gets reactions started
and makes them happen faster, by increasing the rate of a reaction that otherwise might
not happen at all, or would take too long to sustain life. However, a catalyst does not take
part in the reaction itself—so how does this work? Each chemical reaction needs a
minimum amount of energy to make it happen. This energy is called the activation energy.
The lower the activation energy of a reaction, the faster it takes place. If the activation
energy is too high, the reaction does not occur.

Enzymes have the ability to lower the activation energy of a chemical reaction by
interacting with its reactants (the chemicals doing the reacting). Each enzyme has an
active site, which is where the reaction takes place. These sites are like special pockets
that are able to bind a chemical molecule. The compounds or molecules the enzyme
reacts with are called their substrates. The enzyme pocket has a special shape so that
only one specific substrate is able to bind to it, just like only one key fits into a specific
lock. Once the molecule is bound to the enzyme, the chemical reaction takes place. Then,
the reaction products are released from the pocket, and the enzyme is ready to start all
over again with another substrate molecule.

Catalase is a very common enzyme that is present in almost all organisms that are
exposed to oxygen. The purpose of catalase in living cells is to protect them from oxidative
damage, which can occur when cells or other molecules in the body come into contact
with oxidative compounds. This damage is a natural result of reactions happening inside
your cells. The reactions can include by-products such as hydrogen peroxide, which can
be harmful to the body, just as how a by-product of a nice bonfire can be unwanted smoke
that makes you cough or stings your eyes. To prevent such damage, the catalase enzyme
helps getting rid of these compounds by breaking up hydrogen peroxide (H 2O2) into
harmless water and oxygen. Do you want to see the catalyze enzyme in action? In this
activity you will disarm hydrogen peroxide with the help of catalase from yeast.

APPLICATION

1. What does the following enzymes do?

a. Catalase
- It is an enzyme in the liver that breaks down harmful hydrogen peroxide
into oxygen and water. When this reaction occurs, oxygen gas bubbles
escape and create foam.
b. Amylase
- It is an enzyme that catalyzes the hydrolysis of starch into smaller
carbohydrate molecules such as maltose (a molecule composed of two
glucose molecules). Three categories of amylases, denoted alpha, beta,
and gamma, differ in the way they attack the bonds of the starch molecules.
2. What is the optimum temperature for the enzyme activity? What would happen to
the enzymes above the optimum temperature?
- The optimum temperature for most enzymes is about 98.6 degrees
Fahrenheit (37 degrees Celsius). There are also enzymes that work well at
lower and higher temperatures. For example, Arctic animals have enzymes
adapted to lower optimal temperatures; animals in desert climates have
enzymes adapted to higher temperatures. However, enzymes are still
 proteins, and like all proteins, they begin to break down at temperatures
above 104 degrees Fahrenheit. Therefore, the range of enzyme activity is
determined by the temperature at which the enzyme begins to activate and
the temperature at which the protein begins to decompose.

REFERENCES

Lohner, S. (2019). Fruits Gone Bad? Discover Enzymatic Browning. Bring Home Science.
https://www.scientificamerican.com/article/fruits-gone-bad-discover-enzymatic-
browning/

Catalase and Hydrogen Peroxide Experiment. Science Project.

https://www.education.com/science-fair/article/activator/

Science Buddies. (2012). the Liver: Helping Enzymes Help You. Bring Science Home.
https://www.scientificamerican.com/article/bring-science-home-liver-helping-enzymes/

Britannica, The Editors of Encyclopedia. (2020). Amylase.

https://www.britannica.com/science/amylase

Resource. Effect of temperature on enzymatic reaction. Comprehensive Technology

Information. https://www.creative-enzymes.com/resource/effect-of-temperature-on-
enzymatic-reaction_50.html