Enzyme Lab: Catalase Activity in S.

cerevisia

Introduction
This experiment presents an investigation on the effect of different factors on the rate of
reaction of the enzyme catalase in living cells. The reaction investigated in this
experiment is the breakdown of hydrogen peroxide (H 2O2) by the enzyme catalase from
S. cerevisiae cells. H2O2 is a by-product of cell respiration and it is toxic for cells.
Catalase is an enzyme present in almost all organisms that are exposed to oxygen.
Catalase binds to H2O2 (substrate), and breaks it down into H2O and O2 (products),
which are not toxic for cells. The rate of this reaction can be determined by the rate at
which O2 is produced.

2 (H2O2) + Catalase ----> 2 (H2O) + O2 + Catalase

Saccharomyces cerevisiae is a unicellular organism commonly known as baking yeast.

S. cerevisiae uses oxygen to generate ATP by aerobic respiration and like most cells
that use oxygen, it naturally produces the enzyme catalase. In this experiment, living S.
cerevisiae cells will be used as a source of catalase enzyme.
Experimental Design
This lab aims to measure the effect of an independent variable
(substrate concentration, enzyme amount (max 5ml), or
temperature) on the rate of reaction of catalase in S.
cerevisiae, measured by the volume of gas (oxygen) produced
in 30 seconds, using a water-displacement system. Each
student will choose an independent variable and will watch the
videos showing the experiment conducted in our lab at JIS.
In the experiment, 10 ml of H2O2 (substrate) and 5ml of S. cerevisiae (source of the
enzyme catalase) are combined into a sealed conical flask, which has two tubes
connected. One of the tubes goes to a syringe which holds the enzyme (S. cerevisiae)
or substrate (H2O2)solution and the other tube leads to a water displacement tray that
allows escaping gas to bubble up into a graduated cylinder filled with water. As the gas
bubbles into the graduated cylinder, the gas pushes the water out of the graduated
cylinder and into the tray: the water inside the measuring cylinder is displaced by gas
released from the enzymatic reaction, into the tray. The enzymatic reaction begins by
combining the yeast cells and the hydrogen peroxide and the volume of oxygen gas is
measured by the water displaced from the measuring cylinder.
 ● Dependent variable: Volume of oxygen produced during 30 seconds, measured
by water displacement in a 100 ml measuring cylinder. Experimental setup video.
● Independent variables:
○ Substrate concentration [H2O2] - Range {1-5%} - VIDEO
○ Enzyme concentration [S. cerevisiae] - Range {2-10%} - VIDEO
■ NOTE you can vary the amount of yeast/enzyme solution (up to
5ml max) but not concentration per se
○ Temperature of the enzymatic reaction - Range {10-60 °C} - VIDEO

Assessment Goals
In this experiment you will be assessed on:
● Exploration: (Formatively assessed)
● Analysis & Communication: (Summatively assessed)

Deadline for submission in Turnitin: End of Class on 7th December