Name: Brooke Jordan

Title of lab: catalase lab- a biology enzyme activity lab

Aim:
1. To investigate the effect of concentration on enzyme activity.
Apparatus:
Ice bath, potato extract, four test tubes, test tube rack, test tube holder,
tweezers, filter paper, hydrogen peroxide, forceps, vials and stopwatch

Method:
One vial of catalase was obtained at 100 units/ml. each dilution of 3.0%
h2o2: 10ml 3%h2o2, 1.75% h2o2: 10ml 3%h2o2 +10ml distilled water,
0.75% h2o2: 10ml 1.5%h2o2 +10ml distilled water, 0.38% h2o2: 5ml
0.75%h2o2 +5ml distilled water, 0.0% h2o2: 10ml distilled water was made.
The filter paper was dipped in the catalase, drained on a paper towel and
dropped in the 3% h2o2. The time it took for the filter paper to rise to the top
was recorded. The procedure was repeated for each of the substrate
dilutions. The results were recorded in an appropriate table.

Results:
Hydrogen
peroxide
concentration/%
0

Hydrogen
peroxide
depth/cm
7.0

Data time/s

Data rate

0.38
0.75
1.5
3.0

6.8
6.9
7.1
6.9

70
40
24
16

0 (filter paper
didnt rise to the
surface)
0.097
0.17
0.296
0.431

Rate= distance time

Analysis: Graph showing concentration vs rate

Chart Title
3.5
3
2.5
2
1.5
1
0.5
0

Column3

Precautions:
1. Potato extract was shaken well before pouring into test tube to ensure
that there is an even distribution throughout the solution.
2. Test tubes and measuring cylinders were washed thoroughly to prevent
cross contamination

Sources of error:
1. All filter paper were not cut the same size and thus different
amounts of hydrogen peroxide was taken up and to ensure that
the same amount of oxygen gas is required to lift it to the
surface.

Limitation:
1. Room temperature at 37 c may change.

Discussion:
Enzymes are biological molecule (proteins) that act as catalyst that helps
speeds up chemical reactions that take place in the body. A catalyst is a
substance that lowers the activation energy for a chemical reaction and
therefore increases the rate of reaction without being used up in the process.

Enzymes have a special feature in which possess an active site. In this

reaction the substrate binds to the active site of the enzyme and form
enzyme-substrate complex with the enzyme. The enzyme breaks the bonds
of this substrate but however it remains unchanged at the end of the
reaction. Hydrogen peroxide is a poisonous by product of metabolism that
can damage cells if it is not removed which is used in this lab as the
substrate. When broken down oxygen and water is released.

2H2O2

2H2O + O2 (g)

There are many factors which affect the rate of reaction of an enzyme
however in this lab the effect of concentration is being examined. Catalase
the enzyme used was extracted from potato and the substrate hydrogen
peroxide. This solution (enzyme) was left on ice to ensure that the enzymes
were inactive. Pieces of filter paper are dipped in the catalase solution at
different concentrations before they are submerged in the hydrogen peroxide
solution (other factors such as pH and temperature are kept constant). The
oxygen produced in this reaction will cause the filter paper to rise and the
time it took was recorded.
When the filter paper was dipped in the catalase and then submerged in 3%
hydrogen peroxide solution (10 ml h2o2) the filter paper took 16 seconds to
rise to the surface of the solution. This trial possessed the fastest time and
thus had the fastest rate of reaction. This solution containing the hydrogen
peroxide only the enzyme had more substrate to react with. When the
concentration of a substrate in excess, all the enzyme molecules are
saturated with substrate molecules and therefore the enzymes active site
will bind to a substrate thus resulting in a fast rate of reaction.
When the catalyze filter was submerged in the 1.75% hydrogen peroxide
solution it took 24s for the filter paper to rise at the surface of the solution.
Like the previous solution this substrate was made with 10ml hydrogen
peroxide but this time 10ml distilled water was added. This resulted in a
lower concentration of hydrogen peroxide and therefor the enzyme had less
substrate to react with and as a result, the rate of reaction is decreased.
In the next experiment the concentration was further decreased to 0.75%
hydrogen peroxide and then 0.38%. When the filter paper was dipped in the
potato extract solution (enzyme) and then submerged in the hydrogen
peroxide alternatively a time of 40s and 70s were obtained. Both having a
decrease of rate of reaction, 0.75% possessed a higher rate of reaction than
0.38 solution. This occurred because the less substrate molecules there are
around the less often the enzymes active site can bind. Thus there is a slow
rate of reaction.

When the filter paper was dipped into the catalase and submerged in the 0%
hydrogen peroxide solution which contained distilled water only the filter
paper didnt float to the top of the solution. This happened because there
was no substrate molecules to bind to the active site of the enzyme causing
no reaction to take place. Also because there was no hydrogen peroxide to
decompose, oxygen was not produced which initially caused the filter paper
to rise to the top of the solution.

Conclusion:
In this lab the effect of substrate concentration on enzymes activity was
investigated. It was seen that at the lowest concentration of 0.38% hydrogen
peroxide, the rate of reaction was very slow but as the concentration was
increased up to 3.0% hydrogen peroxide the rate of reaction also increases.
Where there was no substrate present no reaction occurred. Thus it was
concluded that the effect of substrate concentration on enzyme activity is
that as substrate concentration increases (less diluted solution) the rate of
reaction also increases.