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CatalaseEnzymeLab ADA

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17 views

CatalaseEnzymeLab ADA

Uploaded by

SeungJang Park
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Biology 10, Foothill College

Enzymes and Reaction Rates


Objectives
At the end of this lab you should be able to:
1. Define the terms enzyme, substrate, reactants, products.
2. Identify the enzyme we are using in lab today, and the substrate on which it acts.
3. Explain how temperature, pH, concentration of substrate, and concentration of enzyme
each influence the rate of enzymatic reactions.

Introduction
Most chemical reactions would occur at extremely slow rates if it were not for enzymes. Enzymes
are proteins that speed up reaction rates by lowering the required activation energy. Enzymes
are very specific, meaning that they only interact with other molecules that fit into their active site,
just as a key must fit into a lock. The molecule that an enzyme binds to is called a substrate. In
the picture below, the enzyme helps to break bonds in the substrate molecule, resulting in two
products. Other enzymes help to form bonds between reactant substrate molecules, thus
building larger products. Because enzymes are not altered in chemical reactions, they are
reused in many more reactions with additional substrate molecules.

Humans, and all other living things, have a wide variety of different enzymes that are responsible
for speeding up all of the chemical reactions that occur in cells all the time. Enzymes are affected
by many environmental factors, including temperature and pH. Increasing temperatures typically
increase reaction rates, but if temperatures are too high, they may denature proteins (including
enzymes), changing their 3-dimensioanl shape, therefore destroying their function.

Image from http://www.corescience.co.uk/


One way to write a general enzyme-catalyzed reaction is this:

Enzyme
Substrate(s) (reactant) Product(s)

The enzyme name is written over the arrow since the enzyme remains unchanged in the reaction.
As soon as it converts one substrate to product, it does that same thing again to a new substrate
molecule, and so on.

Page 1
Biology 10, Foothill College

In this lab activity we will examine one specific chemical reaction that occurs in almost every
living organism (plants, animals, bacteria) that is exposed to oxygen. The enzyme that catalyzes
this reaction is called catalase; it acts on the substrate (also known as the reactant) hydrogen
peroxide to form (produce) both water and 02 molecules, known as the products. Using the
general reaction above as a reference, write the specific chemical reaction we are exploring
today:

Hydrogen peroxide is formed in cells as a byproduct of metabolism, but is extremely toxic to cells;
it can damage proteins, membranes and DNA. Therefore, catalase is crucial as it degrades this
damaging compound into non-toxic water and oxygen.

Two approaches to determining the rate of an enzymatic reaction are to keep track of rate of
disappearance of the substrate, or to keep track of the rate of appearance of the product (Morgan
and Carter 2011). In our experiment, we will keep track of the rate at which the oxygen (product)
appears. Depending on your table’s assignment, you will test the effect of one of these on the
rate of reaction:
 Temperature
 Substrate (H2O2) concentration
 Enzyme (Catalase) concentration

Materials per lab table


 5 small beakers
o 1 to hold potato puree (source of catalase)
o 4 for different experimental conditions
 forceps
 15 filter paper discs
 stopwatch
 3% hydrogen peroxide

Procedure
The enzyme: make as a class
1. Peel and then cut a large potato into chunks and place into a blender
2. Add 400mL of water
3. Blend until the potato is ground fairly smooth
4. Pour the blended mix into cheesecloth that is placed over a large beaker
5. Allow the liquid to drain into the beaker while you label your other beakers, etc.

Preparing your experiment


1)Each group will receive more detailed instructions for their specific experiment on the day of
lab. Check your tray to see which of the 3 different independent variables you’ve been
assigned to study:
a)Temperature
b)Substrate Concentration
c) Enzyme Concentration

2)Obtain about 40mL of the potato puree (catalase solution) in a small beaker.
3)Label the 4 other small beakers using Sharpie with the various conditions being carried out in
your experiment. These will each contain 30mL of substrate (H2O2) solution.

Page 2
Biology 10, Foothill College

Carrying out your experiment


1. Holding a disc with the forceps, soak it in the catalase (potato) solution for 10 seconds.
2. Hold it above the solution for a few seconds to allow excess solution to drip off
3. One team member should have the stopwatch ready.
4. With the forceps, place the disc at the bottom of the H2O2 solution and as soon as you let it
go, start the stopwatch. Stop it when the disc reaches the top of the solution. Do you
see the O2 bubbles, carrying the disc to the surface?
5. Repeat steps 1-4 2 more times for each condition. (You’ll perform the same experiment 3
times for each of your 4 conditions.)
6. Record the times in the data table.
7. Once you carry out the experiment 3 times in each of your 4 conditions, clean up as
described by your instructor before you go discuss results with other groups.

Data Collection and Graphing


Data Table: Use this table to record your raw data. Then, calculate the average (Add up the 3
times and divide by 3) of the time for the 3 trials in each condition and put that in the data table.
Then, take 1/average time to get the reaction rate. This is what you will use to graph.

Average Rate
Independent
Time for Disk to Average Time of Reaction
Variable Trial
Rise (sec) for Disk to Rise (1 disc/ avg
____________
time)
1
2
3
1
2
3
1
2
3
1
2
3

Graphing
Make a graph of your data using the format we’ve discussed (independent on x axis and
dependent (reaction rate) on y axis). Be sure everyone in the group participates in the graphing
and be prepared to present your graph to the class as a whole.

Then, you will take your graph and go discuss it with the other groups who carried out the same
experiment as you and answer the questions in your instructions. Everyone in the group should
be prepared to present the graph to the class.

All students are responsible for knowing how each of the independent variables affect the
reaction rate. This means you need to pay attention as other groups present and take notes as
needed.

The idea for this lab came from several sources, listed here.
1) Melia, M. 2015. Biology 40series lab materials. Foothill College.
2) Morgan, J.G. and M.E.B. Carter. 2011. Investigating Biology Laboratory Manual, 7th edition. Benjamin Cummings,
San Francisco, CA.

Page 3

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