Matthew Wright

Biology Lab

Materials and Methods

Potato extract was prepared by our instructor from 12 grams of potato that was
mixed with 50 mL of pH 7 buffer. This was blended together to give us our material for
testing the peroxidase enzyme. Peroxidase helps break down hydrogen peroxide into
water and oxygen and with a spectrometer we are able to measure the reaction rate of
the enzymes. The spectrometer is actually measuring a special die called guaiacol that
turns brown when it is oxidized. This oxidation shows us that the reaction is taking place
and we are able to quantify the results.

Our group tested the reaction rate at different levels of pH. We first tested the
absorption with a pH of 3. The pH buffer solutions were already prepared for us by the
instructor. After mixing the potato extract with the pH 3 buffer and then adding some
guaiacol die, we took the solution in a test tube to the spectrometer for testing. The rest
of the group did the similar method for pH 5 and pH 7. The other half of the class did the
temperature testing of the enzyme. Instead of the adding the pH buffer, they heated the
solution to test for changes in the reaction rate of the enzymes. The following section
shows the results of our tests.

Results

Temperature and pH are very important for enzymes to operate in there optimum
conditions. Table 1 clearly shows the relationship between temperate and the rate of
reaction for the enzyme peroxidase. The reaction rate peaked around 32 °C, and quickly
dropped as the temperature increased.

Table 1

Table 2 shows the more dramatic affect pH has on the reaction rates of the enzyme. A pH
of 5 is optimal for the enzyme to react with the hydrogen peroxide. The enzyme slows down
very quickly once the solution starts to get acidic. It seems to handle the base conditions much
better than increases in acidity.

Table 2

Peroxidase reacts in optimum conditions when the pH and temperature are around
pH 5 and 32°C respectively. It seems to be more sensitive to hotter temperatures and
higher acidity, with those to variables having the most dramatic affect on the reaction
rate of the enzyme. Peroxidase is a very important enzyme in our body that seems to be
well adjusted to the environments that it will encounter either with a potato or a human.