The Factors that Affect the Rate of Breakdown of the Protein Gelatine by Trypsin

The Factors that Affect the Rate of Breakdown of the Protein Gelatine by Trypsin

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The Factors that Affect the Rate of Breakdown of the Protein Gelatine by Trypsin

Aim

To investigate factors that affects the rate of breakdown of the
protein gelatine by trypsin.

Key Factors:

Possible factors that I could change-

pH- Different types of enzyme work best at different pH level. The
best pH level for an enzyme to be effective depends on its site of
action. An example of this is; enzymes in the stomach have an optimum
pH value of about 2. This is because the stomach is acidic, where as
enzymes in intestines, have an optimum pH of about 7.5.

Temperature- as the temperature increases, so does the reaction rate.
This is due to heat energy that causes the enzyme and the substrate to
collide. But however, very high temperatures damage enzymes by
denaturing them.

The amount and concentration of trypsin- The more trypsin there is
would increase the rate of digestion, as the gelatine will be broken
down quicker.

The size of the gelatine strips-More gelatine would take longer to
digest, as there is a greater ratio of gelatine to trypsin.

Type of factor

Quantity

Unit

Method of Measurement

Independent.

Temperature.

°C.

Thermometer

Dependent.

Time.

mins /secs.

Stop watch

Controlled.

Amount of Trypsin.

ml.

Measuring Cylinder

Controlled.

Amount of Gelatine.

cm.

Ruler

Prediction:

I am going to change the temperature that the trypsin works at. The
time taken for the trypsin to digest the gelatine will change because
of this.

I predict that the temperature that the trypsin works at best (and so
the time that the gelatine is digested quicker at) will be about 37°C,
because this is body temperature.

Hypothesis:

Trypsin is one of the three principle digestive enzymes which are
called Proteases. the other two are Pepsin and Peptidases. During the
digestion process, Trypsin acts with the other Protease enzymes to

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break down protein molecules to their component Peptides and amino
acids.

Trypsin is produced in the pancreas but works in the small intestine.

Gelatine is a protein, and will be digested by the Trypsin.

I predicted that the trypsin will work best at 37°C. This is because
enzymes break down the substrate by a method known as the "lock and
key" method. The enzyme traps the molecules that they react with (the
substrate.)

The shape of the enzyme is vital for this. To be effective, it must
fit exactly with the substrate. It is often described as a key (the
substrate) fitting into the lock (the enzyme)

If the temperature is too cold, the shape of the enzyme is altered
slightly and the reaction cannot happen because there is less kinetic
energy and fewer collisions. This is because the substrate can no
longer fit the enzyme. But however, if the enzyme is warmed up again,
its shape is restored.

If the temperature is too hot (above about 38°C.) then the shape of
the enzyme is altered again, this means that the reactions are
affected (slowed down or stopped)

But in this case however, the higher temperatures can permanently
destroy the enzyme.

The best temperature, therefore is when the shape of the enzyme best
fits the substrate molecule. For humans, our body temperature is 37°C
as this is what our enzymes work best at.

Apparatus list

15 test tubes

measuring cylinder

pipette

5 water baths set up at 15°C, 25°C, 35°C, 45°C and 55°C.

Trypsin enzyme

Gelatine strip

Thermometer

5 stop watches

2 test tube racks

Tweezers

Method

Set up 5 different water baths at 15°C, 25°C, 35°C, 45°C and 55°C.
These temperatures were decided with the help of my preliminary
experiment and these are grouped around my predicted value for the
best temperature which is 37°C.

Safety

- Wear safety glasses at all times

- Keep all test tubes, in the test tube racks.

- Keep the tables and floor area clear with stools under the desks.

Fair test:

Factors that will make the experiment fair.

- Same size of gelatine.

- Same amount of trypsin.

- Same pH level of trypsin.

- Keep temperatures the same in all water baths.

- If the temperature in the water baths goes over 60°C the gelatine
may melt, which would lead to unfair results.

- Pickup the gelatine using tweezers as fingertips may damage the
film.

Fill 5 test tubes with 8 ml of trypsin, using a pipette, then put 1 of
theses into each of the different temperature water baths.

Use a thermometer to check to see if the trypsin has reached the
temperature of the water bath.

Then put a strip of gelatine into each of the 5 test tubes. Start a
stopwatch for each one at the same time as u do this.

When the gelatine strip is no longer black and goes completely clear
in each test tube of trypsin, it means that the gelatine has been
digested. Stop the correct stopwatch and record the time in a table.

Repeat this experiment, three times to ensure that any odd results are
fixed so that the results are reliable.

Results

Time Taken (minutes)

Temperature °C

1

2

3

15

23.02

24.04

24.13

25

13.06

11.53

11.25

35

11.18

7.28

8.34

45

9.14

7.24

8.36

55

7.51

7.26

7.16

Averages:

25°C - 23.07mins.

30°C - 12mins.

35°C - 9mins.

40°C - 8.25mins.

45°C - 7.31min

Evaluation

Overall, I felt that the experiment went reasonably well. Although, it
was not a perfect experiment I felt we got the results we expected. A
result that doesn't seem to fit can be seen in the result's table
where 55 seems to be the lowest. This is because when we obtained the
data we saw that the film was clear but the solution was still murky,
we only found out later that this meant that the solution hadn't
digested completely. We assume that this result is wrong as it does
not fit any pattern. I felt the method that we used was a good method
and helped us get most things right. The results have shown us that
the enzymes work best at 35 , this may be because 35 is around the
same temperature as body temperature, so enzymes would work better.




Analysis
The pattern I was expecting to see was something with a U shape,
instead I got more of a W. This may have been because there was a
wrong result which leads to me getting the W. As the temperature began
to rise the speed of reaction increases until it gets to 35 ,then the
speed starts to go down because the enzymes are then denatured. At 15
there is not enough heat to get the enzymes to work. At 35 this is
almost the right temperature for the enzymes to work as it is almost
body temperature. At 55 the temperature is too hot and some of the
enzymes are denatured and unable to work.
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