Investigating a factor affecting the activity of the enzyme peroxidase

The compound hydrogen peroxide (H

2
O
2
) is a by-product of metabolic reactions in most living organisms.
However hydrogen peroxide is a damaging molecule inside cells. !s a result nearly all organisms produce the
en"yme peroxidase which brea#s down hydrogen peroxide as it forms. $elery is one source of peroxidase.
%eroxidase facilitates the brea#down of hydrogen peroxide into water and oxygen. This reaction can be detected
by observing the oxygen bubbles generated.
&n this lab you design a procedure and carry out an experiment to test the effect of temperature OR substrate
concentration on peroxidase activity.
There are 2 parts to this lab. &n part ! you will use celery and observe the peroxidase reaction. &n part ' you
design a procedure and with your teachers approval carry out an experiment to test the effect of temperature
OR substrate concentration on peroxidase activity. Only part B requires a formal lab write up which will be
submitted for evaluation.
Part ! Observing the peroxidase reaction
(. %ut on your safety goggles. )se forceps to place a small amount of celery into a test tube.
2. )se a dropper to add 2 ml of hydrogen peroxide to the test tube.
*. Observe and record any evidence of a chemical reaction. +easure the maximum height of the bubbles.
,. Test for the production of oxygen by performing a glowing splint test on the test tube. To do this light a
wooden splint in the 'unsen burner flame and blow it out so that only a glowing tip remains. Then lower it
into the mouth of the test tube. &f bubbles are forming in the test tube brea# them with the splint to test the
gas inside them. -ecord your results.
.. The reaction in the test tube eventually stops. /oes it stop because all of the hydrogen peroxide has run out
or because the peroxidase has run out0
To test this ta#e the li1uid (hydrogen peroxide) from the test tube and pour it into a new test tube. !dd a
fresh piece of celery to the old hydrogen peroxide. -ecord your observations.
2. !dd new hydrogen peroxide to the old celery. -ecord your observations
3. &s peroxidase reusable0 )se your data to support your answer.
4. %our the celery from the test tubes into a waste bea#er clean each of the test tubes out and put all materials
away.
Part B! "esign an experiment
)sing the techni1ues you learned in %art ! design an experiment to test the effects of temperature OR substrate
concentration on celery peroxidase. $reate a research 1uestion for this experiment. 5nsure that you have
designed a method that controls the variables and allows the collection of sufficient relevant data. %resent this
lab as a typed report using sentence and paragraph structure. 6our lab should not exceed * typed pages. Include
only the sections mentioned below and be sure to meet the expectations of the rubric.
Introduction
&nclude brief bac#ground information about en"ymes generally and the importance of peroxidase.
6ou will need to develop a research question
#ypothesis!
)se your observations from %art ! and ma#e a prediction about how temperature OR substrate
concentration will affect the rate at which peroxidase brea#s down hydrogen peroxide. $reate a
hypothesis to explain your prediction.
$ariables!
&dentify the independent dependent and controlled variables for your lab.
Independent variable: the variable that you are studying (changing) in an experiment. &t is the only
variable you change.
Dependent variable: the factor which is measured in response to the change in the independent
variable
Controlled variables: all the factors that are held constant in an experiment. +inimum of .
controlled variables. 7uantify each if possible.
The best way to organi"e this information is to ma#e a table with columns for the different variables.
%aterials!
8ist all the re1uired material for the lab.
Procedure!
9rite the procedure as a numbered se1uence.
$onsider these 1uestions:
o How will you change and control your independent variable0
o 9hat variables;factors need to be controlled0
o 9hat data will you collect and in what form will you record it0
o 9hat method will you use to confirm the reaction has happened0
'e sure to include enough details in your procedure for a classmate to replicate the experiment.
Results!
Organi"e your results in a table. &nclude 1ualitative observations.
+a#e an appropriate graph of your results.
&onclusion and 'valuation!
<tate a conclusion based on an interpretation of the data. &t should be =ustified and relate bac# to your
hypothesis.
5valuate at least two biological wea#nesses and;or limitations of this lab and suggest realistic
improvements of the identified wea#nesses and;or limitations.
Due Date: _______________________________________
Design
Levels/Marks
Defining the question and selecting
variables
Controlling Variables Developing a method for
collection of data
Complete/4
Formulates a focused
problem/research question and
identifies the relevant variables with a
high degree of effectiveness.
Designs a method for the
effective control of the variables
with a high degree of
effectiveness
Develops a method that
allows for the collection
of sufficient relevant data with a
high degree of effectiveness
Mostl
Complete/!
Formulates a focused
problem/research question and
identifies the relevant variables with
considerable effectiveness
Designs a method for the
effective control of the variables
with considerable effectiveness
Develops a method that
allows for the collection
of sufficient relevant data with
considerable effectiveness
"artial/#
Formulates a focused
problem/research question and
identifies the relevant variables with
some effectiveness
Designs a method for the
effective control of the variables
with some effectiveness
Develops a method that
allows for the collection
of sufficient relevant data with
some effectiveness
$ot at all/%&'
Formulates a focused
problem/research question and
identifies the relevant variables with
limited to no effectiveness
Designs a method for the
effective control of the variables
with limited to no effectiveness
Develops a method that
allows for the collection
of sufficient relevant data with
limited to no effectiveness
(hinking and
)nvestigation /%'
Data collection and processing
Levels

*ecording ra+ data

"rocessing ra+ data

"resenting processed data
Complete/4
Records appropriate qualitative and
quantitative raw data, including units,
with a high degree of effectiveness
Processes quantitative raw data
with a high degree of
effectiveness
Presents processed data with a
high degree of effectiveness
Mostl
Complete/!
Records appropriate qualitative and
quantitative raw data, with
considerable effectiveness
Processes quantitative raw data,
with considerable effectiveness.
Presents processed data
appropriately, with considerable
effectiveness
"artial/#
Records qualitative and quantitative
raw data, with some effectiveness
Processes quantitative raw data
with some effectiveness
Presents processed data with some
effectiveness
$ot at all/%&'
Records qualitative and quantitative
raw data, with limited to no
effectiveness
Processes quantitative raw data
with limited to no effectiveness
Presents processed data with
limited to no effectiveness
Communication
/%'
Conclusion and ,valuation
Levels
Conclusion

,valuating procedure-s.

)mprovements
Complete/4
States a conclusion, with ustification,
based on a reasonable interpretation
of the data with a high degree of
effectiveness
!valuates wea"nesses and
limitations with a high
degree of effectiveness
Suggests realistic improvements in
respect of identified wea"nesses and
limitations with a high degree of
effectiveness
Mostl
Complete/!
States a conclusion based on a
reasonable interpretation of the data
with considerable effectiveness
#dentifies wea"nesses and
limitations with considerable
effectiveness
Suggests realistic improvements in
respect of identified wea"nesses and
limitations with considerable effectiveness
"artial/#
States a conclusion based on a
reasonable interpretation of the data
with some effectiveness
#dentifies wea"nesses and
limitations with some
effectiveness
Suggests realistic improvements in
respect of identified wea"nesses and
limitations with some effectiveness
$ot at all/%&'
States a conclusion based on a
reasonable interpretation of the data
with limited to no effectiveness
#dentifies wea"nesses and
limitations with limited to no
effectiveness
Suggests realistic improvements in
respect of identified wea"nesses and
limitations with limited to no effectiveness
(hinking and
)nvestigation /%'