Wildred Arroyo

Enzymes 2 Lab Report

Experiment #1: Enzyme-Substrate Specificity (Catalase)

Purpose I will be testing the enzyme catalase, which defends cells by destroying toxic hydrogen peroxide, to see
if it is specific to its substrate or if it can affect other substrates.

Procedures We’ll add ½ tbsp. of potato extract, 1 tbsp. of water, and ½ tbsp. of hydrogen peroxide… then, we’ll
stir the substances together to observe the reaction it makes. This process will repeated again by using another
substrate and a non-carbonated drink; although, the amount of the potato extract and water will remain the
same.

I.V.: Testing which substrate (hydrogen peroxide, tea, and sparkling water) will cause a reaction that makes the
bubbles increase their quantity and height.

D.V.: Recording if the catalase enzyme specific to one substrate.

C.V.: The amounts of the substances stay the same (1/2 tbsp. of potato extract, 1 tbsp. pf water, and ½ tbsp. of
the tested substrate) throughout the entire experiment; also, the time for the stirring process remains equal (1
min.).

Hypothesis Hydrogen dioxide will be the substrate that reacts to the enzyme catalase by increasing the amount
of bubbles that will form on top of the mixed substance. This will be positive control experiment.

Results

Image 1.1 shows what happens Image 1.2 shows what Image 1.3 shows what
once the potato extract, water, and happens once the potato happens once the potato
carbon dioxide are mixed together. extract, water, and tea are extract, water, and sparkling
mixed together. water are mixed together.

PS: My last name is on the left

side of this picture! I was focused
on photographing the enzyme
reaction that I forgot to take a
better picture with my name on it,
but my last name is showed on
this small notebook.
 Substrates Observations Measurement Experiment
Hydrogen Once the H.P mixed with the other two 16 millimeters tall Positive
Peroxide substances (potato juice and water), bubbles Control
started to form at the bottom of the pill vial and +
rose up to the surface creating a thick bubble
layer.
Tea Tea didn’t make a huge reaction when it was 5 millimeters tall Negative
mixed with the water and the potato extract. Control
Clear bubbles formed in the surface of the -
substance.

Sparkling Water This substrate didn’t make any thin layer, 4 millimeters tall Negative
instead all the bubbles formed on the edges of Control
the surface. Please refer to Image 1.3. -

Conclusion Finally, my hypothesis was correct! Hydrogen peroxide is the substrate that reacts to the enzyme
catalase by increasing the amount of bubbles that formed on top of the mixed substances (potato extract and
water). This lab is considered a positive control experiment because a reaction was recorded. In the attached
images from above, you will see the three different experiments that took place on this lab for us to identify
which catalase enzyme reacted to the specific substrate.

Experiment #2: Enzyme Concentration (Catalase)

Purpose I will be testing the enzyme catalase to see if increasing the amount of enzyme speeds up the reaction.

Procedures For this experiment, I will add more potato extract (1 tbsp.) and less water (1/2 tbsp.), I will repeat
this action twice to record their reaction. Then, I will do this experiment by only using 1 ½ tbsp. of potato
extract and no water, this part will also be done twice to record both results. Once this is done, I will average
my results for each concentration.

I.V.: Testing which enzyme concentration will react to the alteration of the amounts that will be poured into the
pill vial.

D.V.: Averaging the results we recorded for both enzyme concentration experiments (water and no water).

C.V.: Stirring the substances for one minute and recording the measurement of the bubbles that form at the top
of the pill vial.

Hypothesis Adding more potato extract (1 tbsp.) and ½ tbsp. of water will increase the amounts of the bubbles
that will form at the top of the enzyme, this concentration will be react to when the stirring process is over.

Results
Image 2.1 shows the reaction when potato extract and Image 2.2 shows the reaction when potato extract is
water is present in the experiment. only present in the experiment.

Enzyme Observation Measurements

Concentration
1 tbsp. of potato Once the stirring process ended, some clear bubbles were visible 8 millimeters
extract on the surface, creating a layer around the edges of the tube. tall
½ tbsp. of water
1 ½ tbsp. of potato A thin layer was visible on top of the enzyme concentration but 4 millimeters
extract the height of the bubbles was not as higher as it was for the tall
No water previous experiment (where water was added).

Conclusion To conclude, my hypothesis was correct! There was an effect when adding more enzyme to our
experiment, adding more potato extract (1 tbsp.) and ½ tbsp. of water did make a reaction. When the stirring
process was over for both experiments (water and no water), the pill vial that presented the highest
measurement of bubbles was the once that had potato juice and water added. This proves that both enzymes
(potato extract and water) are important to the creation of bubbles forming the top of the substances.

Experiment #3: Enzyme Denaturation by pH (Catalase)

Purpose I will be using potato extract as a source of catalase and measuring the amount of bubbles produced
when I mix it with hydrogen peroxide.

Procedures For this part, I will add ½ tbsp. of potato extract, ½ tbsp. of hydrogen peroxide, and 1 tbsp. of base
(ammonia). I will stir these substances for 1 minute and record its reaction (This experiment twice to average
their results). Then, I will repeat this process again but I will replace the base with an acid (white vinegar), then
stir for one minute (this lab will also be done twice). After we gather our results, we’ll average the effect of
changing the pH on the reaction caused.

I.V.: Testing which pH (base and acid) will cause to the enzyme (potato extract and hydrogen peroxide) will
cause.

D.V.: Recording the effect of changing the pH on the reaction will do to the enzyme after the stirring process
ends.

C.V.: Maintaining the same amounts of potato extract and hydrogen peroxide throughout both experiments and
recording both parts after 1 minute of stirring the substances together.

Hypothesis In this experiment, mixing ½ tbsp. of potato extract, ½ tbsp. of hydrogen peroxide, and 1 tbsp. of
base (ammonia)… will show a reaction to the enzyme once the stirring process is done. The acid (white
vinegar) won’t show much reaction like the base will.
Results

Image 3.1 shows the effect that base (ammonia) had when it was mixed with the potato extract and hydrogen
peroxide.

pH Reaction Measurement
Base Once the ammonia was poured into the potato juice, the extract 5 millimeters
(ammonia) changed its color to clear and when the stirring process was over, there tall
were small clear bubbles at the top of the enzyme… forming a thin
layer.
Acid (white When the substance were mixed after a minute the color of the potato 3 millimeters
vinegar) extract changed to clear and no bubbles were seen. However, there was tall
a thin layer on the surface of the pill vial.
Conclusion As predicted, my hypothesis was correct! The effect of the base (ammonia) with the potato extract
and hydrogen peroxide did have an effect in the enzyme. Small bubbles were visible and they formed at the top
of the enzyme creating a thin layer. While, the acid (white vinegar) didn’t show much effect on the enzyme.
Please refer to Image 3.1, to see the effect the base had with the other two substances (potato extract and
hydrogen peroxide.

Enzyme #4: Enzyme Denaturation by Temperature (Catalase)

Purpose I will test this enzyme with different temperatures and pH values to reinforce the idea of denaturing.

Procedures Using the chemicals I placed in the refrigerator, I will add ½ tbsp. of cold potato extract, 1 tbsp. of
cold water, and ½ tbsp. of cold hydrogen peroxide to the pill vial and stir for a minute. I will repeat this step
twice and average both results. Then, I will add ½ tbsp. of room temperature potato extract, 1 tbsp. of water,
and ½ tbsp. of room temperature hydrogen peroxide to the tube and stir for a minute. I will also repeat this step
twice. Once both experiments (cold temperature and hot temperature) are done, I will average my results.

I.V.: Testing which reaction (cold and hot) will cause an effect to the enzyme.

D.V.: Recording what the effect of changing the temperature will cause the enzyme to react.

C.V.: Maintaining the same temperatures for the cold and hot test to prove a point and keeping the same
amounts of substances equal for both experiments (hot and cold).

Hypothesis Adding hot water to the potato extract and hydrogen peroxide, will cause an effect in changing the
temperature of the room temperature substances. While, the cold experiment won’t show much reaction to the
enzyme.

Results
Image 4.1 shows the effect the cold temperature had Image 4.2 shows the effect the hot temperature had
towards the enzyme. towards the enzyme.

Temperature Reaction Measurement

Cold When the cold substances where mixed together, there was not much 6 millimeters
effect on the enzyme. A thin layer of bubbles, where formed on the tall
surface but not a super reaction happened.
Hot When the hot water was included to the enzyme, the bubbles started to 11 millimeters
rise up and they form a thick layer of boiling bubbles. Also, after a few tall
minutes, I saw some potato extract at the bottom of the pill vial (kind of
like sand particles).
Conclusion In conclusion, there was an effect in the enzyme when the hot temperature was tested, meaning my
hypothesis was correct! The hot temperature experiment made the bubbles rise up to the surface of the enzyme
and the bubbles seemed to be boiling due to the high temperature. After a few minutes, I noticed, that there was
some residue at the bottom of the pill vial and I believe this was part of the potato extract. Please refer to Image
4.2. However, the cold temperature didn’t show much reaction, it just made some bubbles on top of the enzyme,
creating a thin layer of bubbles.