EFFECTS OF PEROXISOMES IN AN EXTRACT OF LEAF AND FLOWER
OF HIGHER PLANTS (PAWPAW)
AIM: An experiment to determine the rate of reaction of peroxisomes in the
leaves and flowers of pawpaw using the rate of oxygen gas evolution.
INTRODUCTION
Microbodies are a heterogenous group of small vesicle-like organelles
concerned with oxidation largely. They are found in the liver and kidneys of
vertebrates; in the leaves and seeds of plants as well as in protozoa, yeast
and other fungi. Microbodies consist of peroxisomes, glyoxysomes,
hydrogenosomes and glycosomes.
Peroxisomes are found mostly in photosynthetic cells in the leaves of higher
plants. They contain two enzymes associated with the process of
photosynthesis. The first is glycolate oxidase which controls a reaction in
which hydrogen peroxide, a toxic substance, is produced as a by-product.
Peroxisomes also contain catalase enzymes which break down hydrogen
peroxide (H2O2) to water and oxygen, both of which are harmless.
2H2O2 →2H2O + O2
The rate of this reaction can be determined by measuring the rate of evolution
of oxygen. One indirect but simple way of doing this is to soak discs of filter
paper in tissue extract and then drop them into a solution of hydrogen
peroxide. The paper sinks at first, but as bubbles of oxygen collect on its
surface, it will float up. The time taken from putting the paper into the solution
until the paper floats gives a relative velocity. The effect of enzyme
concentration upon the activity is investigated by this method.
MATERIALS NEEDED
Young pawpaw flower, pestle, mortar, 50ml measuring cylinder, six 50ml
beakers, forceps, 10ml size cork borer, filter paper, 3%hydrogen peroxide
solution, distilled water and stopwatch.
PROCEDURE
A leaf extract was prepared by grinding 30g of leaves in a mortar. 100ml of
distilled water was added and stirred well. The leaf fragments were allowed to
settle and the supernatant fluid decanted into a beaker. This stock solution
was assumed to be 1M. A measuring cylinder was used to prepare in suitable
labelled conical flasks, 50ml of 0.8M; 0.6M; 0.4M; and 0.2M. Two other flasks
of 50ml of the 1M solution and 50ml of distilled water were added respectively.
The flask was labelled and corked.
The procedure above was repeated using young flowers of pawpaw.
30 discs of filter paper of size 10mm in diameter or 30 squares of filter paper
(exactly 5mm* 5mm) were cut out using the cork borer. A filter disc is picked
with a forceps and then dropped into a test tube containing 20ml of 3%
hydrogen peroxide solution. As the filter paper was being dropped into the 3%
hydrogen peroxide solution, a stopwatch was started and stopped when the
paper floated back to the surface.
The time was noted and recorded and the steps were repeated 8-10 times
more. These measurements were repeated using the other concentrations of
leaf and flower extracts. Fresh 20ml of 3% hydrogen peroxide solution for each
concentration or treatment. A reciprocal of the times recorded were taken. A suitable table
was prepared for the results.
FLOWER EXTRACT
Concentration /M Average Time/s T-1/s
0.00 0.00 0.00
0.20 10.72 0.09
0.40 6.61 0.15
0.60 5.41 0.20
0.80 3.75 0.27
1.00 3.35 0.30
LEAVE EXTRACT
Concentration /M Average time/s T-1/s
0.00 0.00 0.00
0.40 6.18 0.16
0.60 4.96 0.20
0.20 7.60 0.13
0.80 3.19 0.26
1.00 3.41 0.32
PLOTTED VALUES
Concentratio 1.00 0.80 0.60 0.40 0.20 0.00
n/ M
Pawpaw leaf 0.32 0.26 0.20 0.16 0.13 0.00
T-1/s
Pawpaw 0.30 0.27 0.20 0.15 0.09 0.00
flower T-1/s
DISCUSSION
The higher the concentration of catalase, the faster oxygen is produced hence
lesser time.
It was observed that the concentration of enzymes in the leaf is more than in
the flowers because taking 1M of both, it takes less time for reaction to occur
in leaf extract as compared to flower extract.
The filter squares were all of the same size so they will all absorb an equal
amount of extract to allow the reactions on the different filter papers to be
equal.
The squares were immersed in the extract for the same length to allow all the
squares to absorb equal amounts of extracts. If the extracts are placed in
different lengths of time, some will absorb more extract than the others.
The reciprocal of the time was taken and used to plot the graph in order to
ensure that the rate of reaction increases with the increasing change in the
time depicted in the formula.
CONCLUSION
It was observed that the concentration of catalase enzyme was proportional to
the inverse of the time taken, hence a straight line graph attained.
Conclusively, the rate of oxygen evolution is dependent on concentration of
catalase enzymes present.
LABORATORY REPORT (CELL STRUCTURE)
NAME: ELLEN NSIAH ASARE
INDEX NUMBER: 7629321
PROGRAM: HUMAN BIOLOGY 1
12TH MAY, 2022
