Affecting Enzyme Activity

Ian Chachana, Emilie Rose, and Allison Hocking

Purpose: To test the reactivity of peroxidase in extreme temperatures or pH.

Hypothesis: The peroxidase will react faster in higher temperatures and

slower in lower temperatures. It will not react in extreme pH.

Procedure:
- Cut potato into 5, 3.5g pieces and placed two in beakers of water, two in
empty test tubes, and one in a empty petri dish.
- Placed one of the beakers full of water on a hot plate and heated to 650 C,
the other beaker had ice cubes placed in it with the potato and was chilled to
50 C.
- Put 5 mL of 0.1 mol/L hydrochloric acid in a test tube with a piece of
potato, and put 5 mL of 0.1 mol/L sodium hydroxide in the other test tube.
- Waited for 5 minutes and removed the pieces of potato from their acid,
base, heated water, or ice water and put them in separate petri dishes.
- Poured 3 mL of 3% hydrogen peroxide on each potato and recorded the
results of the experiment.

Results:
Change to Potato Amount of Oxygen Bubbles Unreacted Peroxide
Hot Very little Two Most of it

Cold None One All of it

Acid None None All
Base None None All
Neutral Very little Many (50-75) 3/4 of it

The only one of the “modified” potatoes reacted with the hydrogen peroxide
and that was the heated potato. The potato produced much less bubbles and
oxygen then the “unmodified” potato but still reacted however slightly. The
hydrogen peroxide decomposed the potato however. The edges became
transparent and small pieces broke off. I believe this is because the potato
was in water that was too hot for too long and this caused the proteins
holding the potato to denature. When the hydrogen peroxide was poured
onto the potato it damaged the molecules inside the cells and caused small
parts to detach from it.
Analyze:
1. These laboratory procedures indicate that peroxidase does not react with
under extreme pH or low temperatures. Peroxidase will not react as strongly
in high temperatures because the heat destroys the cell functions and the
hydrogen peroxide destroys the cell before the reaction can take place.

2. I believe if the temperature was lower then the reaction would have
worked faster than the “unmodified” potato because the potato would not
have broken down and the reaction could have worked.

3. HCl reacted with the peroxidase because the peroxidase is made to keep
the cells safe and the HCl is dangerous to the cell in that concentration. The
peroxidase reacted with the HCl and could not react with the H2O2.

4. I believe that just like the HCl, the NaOH reacted with the peroxidase and
prevented the expected reaction from happening.

Conclude and Apply:

5. My data rejects part of my hypothesis and supports an other part of it.
I stated that the peroxidase would not react in extreme pH values or low
temperatures and it would react faster in high temperatures. The peroxidase
did not react in the extreme pH or cold like I expected but it did not react as
quickly in the high temperature as I hypothesized. If done over I would heat
to a lower temperature to achieve a faster reaction.

6. The peroxidase reacted best at room temperature in a neutral pH, although

it should have reacted faster in the high temperature but I heated it too high
and the reaction failed.

7. The purpose of control samples was to measure the different reactions

without having a variable from an external source. It lowered the margin of
error so that only one variable was being altered one each piece of potato so
the results could be measured accurately.

8. Hydrogen peroxide foams when it touches an open wound because our

bodies have peroxidase in them. When we are injured the blood flow to that
area is increased and it carries peroxidase in the blood to protect the tissues
that have been injured from infection. When hydrogen peroxide is applied it
reacts with the peroxidase.