Kera Pezzuti

Per. 4

In this lab, we observed the effect of varying times in a reaction involving an enzyme
called catalase. Before we could get to the actual lab, several values needed to be obtained that
would be important during the calculations of the reaction. First, we observed the reaction to be
created, H2O2  H2O + O2 with the assistance of catalase. In contrast, another experiment was
held to determine the rate of hydrogen peroxide decomposition spontaneously. “Spontaneous”
means that the solution will be left alone for twenty-four hours and the reaction will occur
without any enzymes. Also to be determined was the baseline—an index of the initial
concentration of hydrogen peroxide in solution. This value was utilized to establish the amount
of H2O2 used in the multiple timed reactions. Finally, the timed reactions were done by using the
same procedure in the test of catalase activity. These solutions were allowed to progress at
several times from one to six minutes. By comparing the timed reactions with the uncatalyzed
reaction, we could determine the effectiveness of catalase.
First, the reaction was observed by adding 1 mL of catalase to 10 mL of 1.5% H2O2 and
bubbles of oxygen were seen exiting the solution. From this we could determine the equation of
the reaction, H2O2  H2O + O2, and that the enzyme was catalase. To be certain the gas that was
created was water, titration similar to that done in the dissolved oxygen lab had to be performed.
A very similar procedure was followed to determine the base line, but catalase was left out and
replaced by H2O. 10 mL of sulfuric acid was also added to the solution. 5 mL were removed
from and then titrated with potassium permanganate. This showed us the initial amount of
hydrogen peroxide in the solution to be used throughout the course of the lab. In our group, we
came up with a baseline of 3.8 mL of potassium permanganate that was needed to change the
color of this solution.
We determined the rate of spontaneous conversion of hydrogen peroxide to dihydrogen
oxide and dioxide without catalase. In order to complete this, a small amount of 1.5% H2O2 was
put in a beaker. After being left for twenty-four hours, this solution was titrated just like the
baseline including adding the water, sulfuric acid, and potassium permanganate. By subtracting
this new titration value by the baseline a percent of spontaneously decomposed hydrogen
peroxide was found. Ours was 26.3%
In the main part of this lab, we measured how much substrate disappeared over various
increments of time. To find these values, the solutions went through the same procedure, but the
amount of time allowed for catalase to catalyze differed. In seven beakers, 10 mL of 1.5%
hydrogen peroxide were inserted and followed by 1 mL of catalase extract. Each beaker was then
swirled for a different amount of time—10, 30, 60, 90, 120, 180, and 360 seconds. After
whichever increment of time, 10 mL of H2SO4 was added to stop the reaction so measurements
could be made. For 10 seconds, .7 mL of hydrogen peroxide was used. For 30 seconds, .9 mL of
hydrogen peroxide was used. For 60 seconds, 1.8 mL of hydrogen peroxide was used. For 90
seconds, 2.1 mL of hydrogen peroxide was used. For 120 seconds, 2.2 mL of hydrogen peroxide
was used. For 180 seconds, 2.0 mL of hydrogen peroxide was used. For 360 seconds, 1.3 mL of
hydrogen peroxide was used.
Although some of our data breaks the mold, the general trend is that more substrate is
converted as the time goes on. By comparing the uncatalyzed reactions to the solutions that
included catalase, it was evident the enzyme had a great impact on the progress of the reactions.
Through all the different solutions created and titrated in this lab, one can conclude that time is a
great factor in the progress of a reaction and that enzymes make a remarkable difference.