NAME : VIVIANA ANAK VERY MATRIC NO : 2018671504

OBJECTIVE

1. To investigate the rate at which the enzyme catalase converts substrate into product.
2. Determine how much substrate has been broken down by catalase at varying time.

HYPOTHESIS

The rate of enzyme will increases as the concentration of the catalase increases.

INTRODUCTION

Enzyme catalyzes a reaction by lowering the activation energy necessary for the
reaction to occur. The molecule that an enzyme acts on is called the substrate. In an enzyme
medicated reaction, substrate molecules are change and a product is formed.

In this experiment you will investigate the rate at which the enzymes catalase converts
substrate to product. You will allow catalase to react with hydrogen peroxide for varying
amounts of time and then stop the reaction by adding H2SO4. To determine the amount of
hydrogen peroxide that remains after the reaction, you will do a titration wih KMnO4.

Materials

 Catalase
 Hydrogen peroxide (H2O2)
 Hydrogen sulphide (H2SO4)
 KMnO4
 Pipette 10 ml
 Seven beaker 50 ml
 Burette
 Conical flask 250 ml
 Measuring cylinder 10 ml
PROCEDURE

1. 10 ml of H2O2 is added to each seven beakers.

2. 1 ml of catalase is added into the first beaker at 0 second.
3. The reaction occurred for the time shown on the label.
4. After the time up, the reaction stopped when 10 ml of H2SO4 added.

5. The procedure was repeated for 10 sec, 30 sec, 60 sec. 120 sec, 180 sec, 360 sec.
To determine how much hydrogen peroxide ( substrate) has been broken down by the
catalase at varying times, measures the amount of peroxide remaining in each flask.
6. 5 ml was removed from above sample for titration and transfer the sample to a clean
conical flask.
7. The initial burette reading of KMnO4 was recorded.
8. KMnO4 was added until a faint brown colours persists. This is the end point of the
titration ( the more KMnO4 you use, the more peroxide appears in the flask).
9. The data was recorded in table 2.1

ANALYSIS OF RESULT

Plot the graph, moles of product vs time (sec) .Calculate the rate of a reaction by
measuring over time, either the disappearance of substrate (as in our catalase example ) in
moles/second.

The rate of enzyme reaction = mole of product

Time

MOLES OF PRODUCT

Beaker 1

10  0.5 1
x = 4.75
1 2

Beaker 2

10  0.3 1
x = 4.85
1 2

Beaker 3

10  1.1 1
x = 4.45
1 2
Beaker 4

10  0.1 1
x = 4.95
1 2

Beaker 5

10  1.0 1
x = 4.5
1 2

Beaker 6

10  0.1 1
x = 4.95
1 2

Beaker 7

10  0.5 1
x = 4.75
1 2
GRAPH
 RESULT

Test Time Titration The rate enzyme

tube (second) (the amount of KMno4 used) reaction (moles/second)
1 0 0.5 ml 0
2 10 0.3 ml 0.485
3 30 1.1 ml 0.445
4 60 0.1 ml 0.495
5 120 1.0 ml 0.450
6 180 0.1 ml 0.495
7 360 0.5 ml 0.475

Table 2.1 : Titration of hydrogen peroxide

QUESTION

1. At what time interval is the enzyme working at its maximum velocity?

The best time interval for the enzyme working at its maximum velocity is at the early
phase. This is because at the early stage, the solution contain the highest
concentration. The more concentrated the solution, the more substrate molecule is
available. Hence, the more frequently they access the active site of the site of the
enzyme. However, the rate of reaction continuing to decreases when the concentration
of the solution decreases.

2. Suggest what should be done in order to keep the rate constant over the entire time
course.

Added more enzyme which will causes the concentration of the solution increases.
Hence, the rate of reaction will increase until the enzyme reach it maximum point.
When it reach the maximum point, the reaction will stay constant even though we
increase the concentration.

3. What is the role of sulphuric acid (H2SO4) in this experiment?

As we know, in order for enzyme to work efficiently, the pH value of the solution
needs to be optimum. The role of sulphuric acid (H2SO4) is to stop the reaction of the
enzyme with hydrogen peroxide because pH value of the sulphuric acid is not
optimum to the enzyme to work which is below than pH 7. It can stop the reaction
immediately.
DISCUSSION

An enzyme is a macromolecule that acts as a catalyst, a chemical agent that speeds up

a reaction without being consumed by the reaction. In this experiment, we use potato starch
as the catalyse. We are using hydrogen peroxide (H2O2) because this solution have very
optimum pH or the enzyme to work efficiently. Besides that, we want to find the rate of
reaction by changing the concentration of the substrate. Hence, we have 7 beakers with
different concentration of the substrate.

When we added 10 ml of Hydrogen peroxide (H2O2) and ml of catalase for an

interval of time for 7 beakers separately, we find out when we added 10 ml sulphuric acid
(H2SO4) to each beaker it show bubbles which is from the hydrogen peroxide when it reacted
it will produce water and oxygen gas.

Next, we take 5 ml of the solution and doing a titration with KMnO4 . However, when
we are doing the titration, we mostly got dark brown which is not the result that we
expecting. This shows that the KMnO4 was overshoot while doing the titration. It is
supposedly changing to light brown which indicate it reaching the end point. Furthermore,
the solution that more concentrated does not need a lot of KMnO4 and it will change to light
brown which the result that we fail to obtain. The rate of enzyme was determined by the
volume of titration that we used. There is error happened during the experiment which is the
correct technique to do titration.

Moreover, when we do the calculation, the rate o enzyme was all positive, but the
values that we got was almost similar which was not realistic. Because in different period of
times, the volume of titration of KMnO4 should be different because of the different
concentration of the solution. As I said, there are errors which is we might read the incorrect
reading on the pipette. We suppose to read it as our eyes are perpendicular to the scale of the
pipette in order to prevent parallax error.

CONCLUSION

The rate of reaction is depend with the period of time. The longer the time given for the
enzyme to react with the substrate, the faster the rate of reaction.

REFERENCE

 Neil A .Campbell, Lisa A.Urry, Michael L.Cain, Steven A. Wasserman, Peter

V.Minorsky, Jane B. Reece (2018), Biology A Global Approach Eleventh Edision,
Pearson Educational Limited