ENZYME PRACTICAL

Aim: To investigate how temperature factor affects the activity of a

named enzyme.

Hypothesis: Our prediction is based on, that protease enzyme is

going to destroy through the increase of temperature, and when it
reaches certain amount of time, in this case 3 minutes approximately.
Also, temperature would affect enzyme in a negative way and would
cause enzyme decrease or increase its activity.

Variables:
Independent: temperature (we can manipulate it by changing
temperatures (60, 50, 40, and 30)
Dependent: time (we can measure time with a chronometer
(Every 10 or 20 sec)
Controlled: amount of substrate, time, and amount enzyme

Materials:
- Pipette
- Spotting tile
- Glass rod
- Measuring cylinder
- Water bath at: 60, 50, 40, 30 degrees
- 1% protease solution
- Biuret solution

Method:
1. Add one drop of solution to each well of a spotting tile.
2. Setup the water bath at 30, 40, 50, or 60 degrees.
3. Take water at different temperatures.
4. Pour water in the test tubes more than the half.
5. Then put in the test tube some of the enzyme and then introduce
it into the water bath at different temperatures.
6. Past 5 minutes, take the test tube out from the water bath.
7. Then, add the protease to the enzyme and start the stopwatch.
8. Every 10 or 20 seconds add one drop of the mixture

DATA TABLE
GRAPH

Time taken for colour to change Obsevations Negative / Positive

Temperature
There was no change,
Stayed the same, no noticeable
the protease dident Negative
change
60 c break up the protein.
The protease took a lot
of time to break up
50 c 2 minutes 20 sec (140 sec) Positive
compared with oter
times.
Took few time to break
40 c 1 minute 40 sec (100 sec) because temperture Positive
was kind of controlled.
It break up the protease
30 c 1 minute 30 sec (90 sec) Positive
with good time.

ENZYME ACTIVITY (TEMP VS TIME)

70

60 60

50 50
TEMPERATURE (oC)

40 40

30 30

20

10

0
N/A 140 100 88
TIME (SECONDS)

Conclusion

In my opinion, the hypothesis we had was correct, as we were right

about our predictions, and we could reinforce it with the data we got.
Meanwhile the practical, what happened with the mixture was that
when we added a drop of it into the spotting tile every certain amount
of time, it changed to another color step by step, sometimes it wasnt a
very notorious change, but other times it was. We tried to be as
organized as possible, but we were kind of slow when following
instructions and that could have affected our results and the
recollection of data. We did expect to get similar results to do ones we
got, as we mentioned it in the hypothesis. Another improve we could
make, is to add more trials into the experiment with the same enzyme
in order to get much more reliable results. In addition, it would be
interesting trying the experiment with three different enzymes and
solutions in order to get in deeper research about enzymes activity and
its affectance by temperature. Despite all of this, we were lucky to
enjoy and get the results we wanted, so we could learn throughout an
example how enzymes really work and until what point they are able to
change.

Evaluation

During the practical, the results we got were good because even
thought they could have been taken in an organizer way and could be
improved by trying more than one enzyme, they were precise, reliable,
and the results we expected. I think our method was right, because
while doing the experiment we got at least 2 steps for each of the trials
which create more reliable and precise results.

Weakness: Time taking not correct

How does affect the result: This could affect the result as you wouldnt
know in what exact moment you should drop the mixture, and as time
passes the enzyme change notices the more, so it wouldnt be such a
fair test.
Improvements: We can improve it, by being careful and measuring
precisely every 10 or 20 second (depends on the test).

Weakness: Not having the same measurements

How does affect the result: This could affect the results as the amount
of liquid in each test depends on the test trial. This could affect the
results got on each test.
Improvement: This can be improve by having constant and equal
amounts of measurement in each test and making sure to mix it with a
measuring cylinder to have more precise results.

Weakness: Affectance in room temperature

How does affect the result: Factors such as fans, air conditioner, or a
lot of sun through the window, can affect the test by making the
temperature in the surroundings of the solutions decrease or increase
affecting the enzyme.
Improvements: It could be improve by turning off the fans, air
conditioner, or the majority of things that could affect the room
temperature.