How Temperature Affects the Rate of Enzyme Activity

Research Question: What is the effect of temperature on the rate of

reaction between catalase enzymes and hydrogen peroxide to produce
oxygen gas, as measured by a gas pressure sensor?
Aim: This experiment aims to observe and record the effect of temperature
on the release of oxygen gas as one of the products of the reaction
between hydrogen peroxide and catalase enzymes. Catalase is an enzyme,
a globular protein which functions as a biological catalyst which catalyses
hydrogen peroxide (H2O2), a poisonous by-product of respiration, into the
non toxic products of oxygen (O2) and water (H2O). By measuring the speed
at which these products are formed, the rate of reaction under different
variables can be concluded and used to support the presented hypothesis.
Hypothesis: As the temperature of the solution increases, as will the rate of
reaction; up to the point at which the catalase enzyme denatures and no
further enzyme substrate complexes can be formed, resulting in a sharp
drop in rate of reaction. This is supported by the collision theory, which
concludes that increasing kinetic energy in molecules, as would be
achieved through increased temperature, increases their movement and
therefor the likelihood that they collide and react with another molecule.
However, due to the catalase being a human enzyme, it has an optimum
operating temperature of approximately 37.5C (human body
temperature), if the enzymes are heated too far past this point, the amino
acids within the proteins will vibrate so much as to break the
intermolecular bonds in the enzyme, irreversibly changing its structure so
that its active site is no longer suitable to its complimenting substrate,
rendering it useless for its purposes. This hypothesis would predict that the
rate of reaction (as measured by oxygen production) would increase and
peak among the points nearest to human body temperature (35C and
40C) and fall rapidly after that point, resulting in a potential for no reaction
at all to occur among the higher temperatures due to denaturation.
Independent Variable: The temperature of the catalase and hydrogen
peroxide solutions (5C intervals from 10C to 60C). Temperatures will be
achieved through the use of a water bath to maintain the various
temperatures required for each test.
Dependent Variable: The oxygen produced by the reaction will be
measured, as an indication of the rate of reaction, by a gas pressure
sensor, sealed to to the test tube that the reaction will occur in by a rubber
bung. The rate of reaction will be measured in kPa/min and recorded for 2
minutes by the gas pressure sensor.
Control Variables:
Time of trial will be kept at 2 minutes throughout the experiment;
the gas pressure sensor will do this automatically. The duration is
kept constant to ensure that it is only the IV that is affecting the

How Temperature Affects the Rate of Enzyme Activity

result, as a longer trial would potentially show a quicker/slower rate

of reaction compared to other trials.
The volume of Hydrogen Peroxide (substrate) will be kept at 1cm3
for each separate trial and it will all originate from the same sample
in order to assure equal concentration/potency as to not allow
increased or decreased rates of reaction due to the
volume/concentration of substrate.
An equal volume (2cm3) of catalase enzyme solution will be used for
each trial to ensure that it does not affect the rate of reaction.
The same sized test tubes will be used for each trial in order to
maintain a comparable pressure output to me measured by the gas
pressure sensor.
The temperature for each trial will be maintained at a constant level
throughout the trial (2 minutes) to ensure that the results arise
directly from the recorded temperature. This will be done through the
use of a water bath to maintain temperature and measured with a
temperature probe inside the bath.

Monitored Variables:
Room temperature will be monitored to record any potential
significant increase or decrease in temperature there is in the
substrate or enzyme solutions before the test begins as this could
affect rate of reactions within the trials.
Method:
Apparatus:
Risk Assessment: