Enzymes

By: Jocelyn Vazquez

1. Introduction
a. According to Reece (2012), enzymes are macromolecules that act as a
biological catalyst. An enzyme speeds up a reaction by decreasing the
activation energy. Activation energy is when the energy barrier comes
down before a chemical reaction can begin. The energy needs to be
absorbed to weaken the bonds in order to form new bonds.
b. The main purpose of this experiment was to test the effects of temperature,
time, enzyme concentration and pH on enzyme activity. The independent
variables were temperature concentration, and pH. The dependent variable
was the enzyme activity.
c. Before starting the Renin Experiment, it was hypothesized that test tube #1
would have the most enzyme activity. Before starting the Urease
Experiment, it was hypothesized that test tube #3 would have the most
enzyme activity because it contained more of the solution than test tube
#4. Before starting the Catalase Experiment, it was predicted that test tube
#2 would have the most enzyme activity because it contained hydrogen
peroxide. According to Reece (2012), catalase speeds the breakdown of
hydrogen peroxide. Lastly, for the Effect of pH on Catalase Activity it was
predicted that test tube #2 to have most enzyme activity due to the fact that
it contained hydrochloric acid. Hydrochloric acid is a known to be a
strong, caustic acid (Reece, 2012).

2. Method
Rennin Experiment
We started this experiment by place marking three clean test tubes at 2 cm. Test tube
number one was filled with warmed 2% milk up to the 2 cm line. Next, we added three
drops of warmed renin (found in stomach lining), and placed the test tube in an incubator
(water bath) at 37 degrees Celsius. After 15 minutes, we examined the test tube and
observed that the substance had turned into a solid. For the second test tube, we filled it
to the 2cm line with refrigerated milk. Next, we added three drops of refrigerated renin
and placed the tube in the refrigerator. We waited fifteen minutes and recorded our
results. Then we placed the tube in an incubator (water bath). After conducting the
experiment for the second test tube we saw that there was not much enzyme activity.
Lastly, tube #3 was filled with warmed 2% milk up to the 2cm mark. Next, we added
three drops of boiled renin and placed the tube in an incubator at 37 degrees Celsius. We
waited 15 minutes. After the fifteen minutes was up, we added three drops of warmed
renin and placed the test tube back into the incubator. After the 15 minutes were up, we
noticed that the product had denatured because it was too hot.
Urease Experiment
This experiment was consisted of four different test tubes, each of them were marked
with a 2cm line. For the first test tube, we filled the test tube with Urea solution up to the
2cm line. We then recorded the color of the solution (yellowish-green). The second test
tube contained urea solution up to the 2cm line, but we added 30 more drops of the
solution until it became a violet- pink color. Test tube #3 contained the urea solution up
to the 2cm mark, this time we added 15 more drops, shook the mixture, and recorded our

results. Test tube #4 contained 2cm of the urea solution. This time we added 7 more
drops of the solution, shook the mixture, and recorded our results. We believe test tube #3
and #4 mustve been contaminated because we didnt observe much enzyme activity or
color change in the solution.
Catalase Experiment
We marked three different test tubes with a 3cm mark. The first test tube was filled with
hydrogen peroxide up to the 3 cm mark. Next, we added sand to the same test tube. We
noticed that no bubbling occurred. Next, test tube #2 contained hydrogen peroxide filled
up to the 3 cm mark. We added a cube of a fresh potato (known to be an enzyme catalase)
and saw that moderate bubbling occurred. For test tube #3, we filled it with hydrogen
peroxide up to the 3 cm mark. Then, we added a cube of macerated potato. We mashed
the potato by using a mortar and pestle. After adding the potato we saw that a very good
amount of bubbling occurred.
Effect of pH on Catalase Activity
The first test tube had a 2cm mark and a 6cm mark. We added distilled water up to the
2cm mark. Next, we added a cube of macerated potato. After three minutes were up,
added hydrogen peroxide to the 6 cm mark. We observed that very good bubbling
occurred. The second test tube contained a 2cm and 6cm mark. We filled the test tube
with hydrochloric (HCl) acid up to the 2 cm mark. Next, we added a cube of macerated
potato. After waiting for three minutes, we added hydrogen peroxide to the 6 cm mark,
and noticed that nothing happened. The third test tube had a 2cm mark and a 6cm. We
filled it with sodium hydroxide up to the 2cm mark. Next, we added a cube of macerated
potato and waited three minutes. After the three minutes were up, we added hydrogen

peroxide to the 6cm mark. Lastly, we observed that the there was not much enzyme
activity because it was too basic.
3. Results
Rennin Experiment
Tube

Explanation

1. Refrigerated Renin

Turned solid

2. Warmed Renin

Remained as a liquid

3. Boiled Renin

Denatured

*The following chart lists the results that occurred after waiting fifteen minutes
for each trial.
Urease Experiment
Tube

Number of

Start

Final

Total Time

Urease

Time

Time

for Change

8:03

8:13

10 minutes

Results

Drops
#3

15 drops

Not much
enzyme
activity
occurred.

#4

7 drops

8:08

8:13

5 minutes

Not much
enzyme
activity
occurred.

*The chart above lists our results for test tubes #3 and #4. The results may be
incorrect as a result of contamination.
Catalase Experiment
Tube

Contents

Bubbling

#1

Sand

No Bubbling

#2

Fresh Potato

Moderate
Bubbling

#3

Macerated Potato

Very Good
Bubbling

*The chart above lists the results we observed for each trial.
Effect of pH on Catalase Activity
Tube

Contents

Bubbling

#1

Water

Very Good
Bubbling

#2

HCl

No Bubbling

#3

NaOH

Moderate
Bubbling

* The chart lists the results we observed for each trial.

4. Conclusion
For the renin experiment, we predicted the test tube with the warmed renin
would have the most enzyme activity because heat can speed up a chemical
reaction, but too much heat can denature it. We believe we may have incorrectly
measured the results because not much happened to any of the test tubes.
Before starting the Urease experiment, we predicted test tube #3 would
have the most enzyme activity because it contained more Urease solution than test
tube #4. For both of the test tubes, we did not see much of a change. Once again, I
believe an error occurred such as contamination.
Our hypothesis for the Catalase experiment was correct. We predicted that
the test tube containing the macerated potato would contain the most bubbling.
Since a potato is a catalase, by adding HCl it caused the bubbling to increase.
However, when the potato was mashed, it increased the spread of the bubbling.
Our hypothesis for the Effect of pH on Catalase Activity was incorrect.
We predicted that the test tube containing HCl was going to have the most
bubbling. The only substance that bubbled was test tube #1, which contained
water.
I think some of our answers may be incorrect. We may have made
mistakes with monitoring the amount of time needed for each step. I also think the
issue of contamination may have caused our results to be biased. If I were to do
the experiment over, I would make sure all of the equipment is properly cleaned
to avoid any altering results. I also would make sure that I follow each and every
step carefully to avoid making mistakes, especially with the time.

References:
Reece, J., Taylor, M., Simon, E., Dickey, J., & Hogan, K. (2012). How Enzymes
Function. In Campbell biology: Concepts & connections (8th ed.). Upper Saddle River:
Pearson.