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AP Biology
(Peroxidase + Hydrogen Peroxide Complex Peroxidase + Water + Oxygen)
2H2O2 2H2O + O2 (gas)
Learning Objectives
The student is able to design a plan for
collecting data to show that all biological systems
are affected by complex biotic and abiotic
interactions (2D1 & SP 4.2, SP 7.2).
The student is able to use models to predict and
justify that changes in the subcomponents of a
biological polymer affect the functionality of the
molecule (4A1
& SP 6.1, SP 6.4).
The student is able to analyze data to identify
how molecular interactions affect structure and
function (4B1 & SP 5.1).
Label carefully
Gather all materials
Plan your procedure and know what you are
doing before you start. Do a mock run
There is timing involved, so be prepared to
start the timer as soon as you mix the
materials.
Make data charts ahead of time so you have
a place to put your results.
guaiacol
We can see the reaction because we are
adding guaiacol, an indicator of oxygen due to a
color change that occurs in its presence. The
more oxygen the deeper the brown the color
becomes. The compound guaiacol has a high
affinity for oxygen, and in solution,it binds
instantly with oxygen to form tetraguaiacol,
which is brownish in color. The greater the
amount of oxygen produced, the darker brown
the solution will become.
Color palette
Time
minutes
0 1
Scale/
number
B ENB
3ml NB neutral Buffer pH 7
1ml E= Enzyme turnip peroxidase
A SPNB
1ml NB neutral Buffer pH 7
1ml P = guaiacol
2ml Substrate hydrogen
peroxide
Time
minutes
0 1
Scale/
number
B ENB
3ml NB neutral Buffer pH 7
1ml E Enzyme turnip peroxidase
A SPNB
1ml NB neutral Buffer pH 7
1ml P = guaiacol
2ml Substrate hydrogen
peroxide
Immediately observe by comparing to the color palette and begin timing!
Observe every minute for 5 minutes.
Calculate the rate for the baseline. Color change/% oxygen over time
Part 2
This part will test how different pH levels will
alter the reaction rates as compared to the
baseline.
Gather your materials. Plan your procedure.
Put in the correct amounts. Be careful with the
buffers! Acids and bases can harm your skin
and eyes!
Label the test tubes,Set up your test tubes in
pairs.
Tubes 1,2,4,9,11,12
1 ml NB neutral buffer
1 ml P=product indicator
guaiacol
2 ml S= hydrogen peroxide
Tubes 3,5,6,7,8,10
Make a data
chart!
Tubes 1,2,4,9,11,12
1 ml NB neutral buffer
1 ml P=product indicator
guaiacol
2 ml S= hydrogen peroxide
Be ready!
Tubes 3,5,6,7,8,10 Mix tube 1 with tube 3.
Observe at time zero and
every minute for 5 minutes!
1 ml E=turnip peroxidase solution
3 ml buffer of the correct pH for the tube. For
example tube 3 use pH 3, tube 5 pH of 5 etc!
rate
Calculate the rate for each tube.
How much oxygen produced in 5 minutes?
Based on the readings over the 5 minute
time period compared to the color change
or % of oxygen.
% change over time= rate
graph
Rate for each pH
Assessment Questions
#3.
If you omitted the enzyme?
If you omitted the substrate?
If you omitted the indicator?
Based on your answer to #4 develop a
specific question to test for part 3 of the lab.
Your group must submit an experimental
plan for approval.
Part 3