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Investigating Trypsin

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Investigating Trypsin

Aim- to investigate factors which affects the trypsin in different
temperatures.

Key factors- Trypsin is one of a group of proteases secreted by the
pancreas into the upper (small) intestine. Factors that affect the
activity of enzymes are the temperature. If you increase the
temperature you will increase the random movements of the molecules
and increase the chances of substrate and enzyme colliding. If the
temperature is raised too much the enzymes will be destroyed. The pH
level also affects enzymes. Certain enzymes work better in different
levels of pH. For example the digestive enzymes in the stomach work
best in acidic conditions. While those in the small intestines work
best in alkaline conditions.

I will investigate how the temperature affects the trypsin. I will see
how long it takes to break down the protein on a photographic film.
The protein is called gelatin. I will measure its affect on the rate
of reaction it takes to for trypsin to break down the gelatin in
minutes and seconds. I will do this at different temperatures to see
which temperature trypsin works best in. Trypsin specifically
catalyses the splitting of peptide bonds on the carboxyl side of
lysine and arginine residues. It works best at slightly alkaline pH
(around 7.6 to 8) and body temperature. It is classified in the serine
protease family because of the presence of a vital serine amino acid
residue in the active site.

Prediction- I predict if the temperature is too cold the enzyme
trypsin will not break down the gelatin very well. This is because of
a lack of energy to produce random movements. I say around 35ºc the
trypsin will work best. I this will happen because the trypsin will
have more energy to break down the gelatin. Also because this is the
temperature inside the Stomach and more random movements the trypsin
will make. Trypsin is found in the stomach and trypsin works best in
acidic conditions. Gelatin is a protein, which is found on the
photographic film. If the film is acidic the trypsin will break down
the gelatin. When trypsin reacts with gelatin, the gelatin will be
broking down to amino acids. But if the temperature is shape the
trypsin will lose its shape and will start to destroy itself. Also I
have evidence from my preliminary test that around 30ºC that the
trypsin will break down the gelatin on the photofilm.

Secondary sources

Biology for life.

Method

I will first get my equipment which is:

· Photographic film

· Trypsin

· Test tube holder

· Stopwatch

· Heated tub

· Water

· Test tubes

· Thermometer

· Measuring beaker

Then set it up to look like above. To measure which temperature
trypsin works best in. I will put the photographic film in a test tube
of trypsin and heat up the test tube in the heated tub. I will make
sure I have the right temperature by using a thermometer. As soon as
we start heating the test tube I will use the stopwatch to time how
long it takes for the trypsin to break down the gelatin. I will know
this when the photographic film is clear. The temperatures I will use
to do this are 20ºc, 25ºc, 30ºc, 35º, 40ºc, 45ºc, 50ºc and 55ºc. Also
I will do the test three times to make the test reliable and fair. I
will time how long it takes for trypsin to break down the gelatin in
seconds and minutes. At the end do an average. To make the test fair I
will make sure the temperature doesn't change from the amount I said
it would be. I will always use the same amount of trypsin in each test
tube. I will use one photographic film and a different one for each
test tube. Also 50ml of Trypsin. After we have done this I will put my
results onto a table.

Safety

For safety I have to make sure no water gets in the sockets where the
heated tub will be. Wear eye protection incase trypsin goes in my
eyes. Do not run in lab. If any trypsin goes on my hand wash it off
using cold water.

Preliminary work

From this I will do 20ºC to 55ºC going up in 5.

Time taken

For film to

Go clear secs

Temperature

Test 1

Test 2

Test 3

Average

20ºC

1170

1156

1190

1172

30ºC

350

364

378

364

40ºC

421

379

418

406

Fair Test

The experiment must be a fair test or I will be getting a lot of
anomalous results and there may not be a clear pattern in the results.
To make the experiment a fair test I am going to make sure that
volumes of trypsin and the size of the photographic film is the same.
I must also make sure that the temperature of the water is right and
the trypsin and to keep the experiment accurate. I will also need to
make sure that the photographic film is completely clear before I stop
the timer. When I think it is clear I will press the red button on the
timer and if the film goes no clearer I will take the time that I
paused the timer at. If it does go clearer I will continue the timer
and stop it when it goes clearer.

Analyst and conclusion

From the graph I see no anomalous. I think the best-fit line is
correct because it curves down, and then goes up and down. After the
time will keep increasing. The time is very high because trypsin has
very little energy to react with the gelatin at 20ºc. Also I notice
the time is starting to rise after 55ºc. This is due to the trypsin
making more random movements and will start to destroy itself, and
lose shape. My prediction was right trypsin reacted best with gelatin
around the 35ºc. Also I was right that if the temperature was too
cold. The results show that trypsin will digest the protein gelatin
best between 30ºc-40ºc. This is due to good energy levels and lots of
random movements. These temperatures will be the same in the human
body. I was also right with my prediction that I say around 35ºc the
trypsin will work best. I this will happen because the trypsin will
have more energy to break down the gelatin. Also because this is the
temperature inside the Stomach and more random movements the trypsin
will make. Trypsin is found in the stomach and trypsin works best in
acidic conditions. I believe that my results do support my prediction
because of what I have just explained.

Evaluation

I believe my results are accurate and reliable because this is the
temperature in the stomach. Also because I did the test very fairly.
With no anomalous in my results. I recorded all my data correctly and
also did the test three times to make I accurate and reliable. There
is no unusual result or anomalous in my results. Which means I did the
test right. There were no sources of error in my test. If I was to do
the test again. I would test out whether the pH level will make a
difference. Which I think it will. Then I could see which pH and
temperature worked best together. Also I believe that if some one were
to do the experiment again they would get the same results. I would
also use a measuring beaker to the nearest 1ml. This would help me
measure the trypsin more accurately, which would make my results
fairer.

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"Investigating Trypsin." 123HelpMe.com. 24 Apr 2014
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