Joseph Latif

Sports, Exercise and

Health Science: Digestion
of Protein Practice
Internal Assessment

Joseph Latif

Results
Pepsin Activity
Test
Tube #

Contents of Tube

pH Level

Water + Hydrochloric
Acid
Pepsin
Pepsin + Sodium
Hydroxide
Pepsin + Water
Pepsin + Hydrochloric
Acid

2
3
4
5

5.5

Relative
Amount of
Coating
4

Relative
Pepsin
Activity
2

7
10

5
2

1
4

7.5
5.5

3
1

3
5

Table 1 Observed Results

Graph 1 Relationship between Relative amount of coating and Relative pepsin activity
6
5

R = 1

4
Relative Amount of Coating

3
2
1
0
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5
Relative Pepsin Activity

Joseph Latif

Conclusion and Evaluation

Concluding:
This investigation aimed to illustrate the effects of pH on the activity of an enzyme.
The investigation specifically focused on the effects that external factors, including
pH level and temperature, had on the proteolytic enzyme pepsin.
The activity of pepsin was measured using two factors that were visually measured
according to a rank from 1 to 5, where 1 is the least and 5 is the most.
These two factors were the Relative amount of coating and Relative pepsin
activity. It was hypothesised that tube 5 would provide an environment where pepsin
would be most active; therefore the film would have the least amount of coating in
tube 5. These factors shared a perfect negative correlation ( r 2 = 1) this is
illustrated by the r 2 value given in Graph 1. Although the r 2 purely represents
the correlation between the two variables, the slope of the trend line indicates that the
variables share a negative relationship. This means that as one variable increases, the
other decreases.
As hypothesised, pepsin was most active in tube 5 in the presence of hydrochloric
acid (HCl), having a relative pepsin activity of the maximum score of 5. Thus, due to
the converse relationship between variables, tube 5 also held the minimum score of
relative amount of coating at a score of 1. Pepsins activity is optimal in the range of
pH 1.5 to 2.21. The pH level of the environment in tube 5 may explain pepsins
strong activity, as it was the most acidic of all the tubes (pH level of 5). Therefore,
pepsins strong activity in tube 5 may be explained by the fact that it was in an
environment closest to the optimal range of all the tubes.
Conversely, the tube that held the least ideal environment for pepsin to operate was
tube 2, ranked with a relative pepsin activity score of 1 and relative amount of coating
score of 5. Tube 2 solely contained pepsin, however was heated using a boiling water
bath. The conclusion can be made that because the pepsin was placed in a boiling
environment, it was exposed to temperatures >100 . The optimum temperature
interval for free pepsin [is] 30 - 40 2. Therefore, it can be said that the
temperature of the environment denatured the pepsin molecules, thus inhibiting the
function of the enzyme.
Test tube 1, which contained only water and hydrochloric acid, had more of an effect
on breaking down the proteins on the film than tube 2, which contained pepsin. Test
tube 1 ranked 4th and 2nd in relative amount of coating and relative pepsin activity
respectively. This may be explained by the role of hydrochloric acid in denaturing and
subsequently catabolising proteins. Although hydrochloric acid does not completely
catabolise proteins, it has some effect on the proteins on the film. Therefore, because
hydrochloric acid has some catabolic effect on the proteins, it had a slightly stronger
impact than the pepsin in tube 1, which was completely denatured by the extreme
temperature.
1 Schlamowitz,Max,andLinnU.Peterson.StudiesOnTheOptimumPhForTheActionOfPepsin.
1sted.NewYork:JournalofBiologicalChemistry,1959.Web.6July2015
2 Altun,GamzeDurgun,andSenayAkkusCetinus."Immobilizationofpepsinonchitosanbeads."
FoodChemistry100.3(2007):964971.Web.6July2015

Joseph Latif
Furthermore, tube 3 contained the highly alkaline chemical, sodium hydroxide
(NaOH), as well as the enzyme pepsin. However, although pepsin was present in the
tube, it is highly unlikely the pepsin was able to have a large impact on the proteins on
the film, given that it resided in an environment with a highly alkaline pH of 11
which is very far from pepsins optimal range. If this is true, NaOH must have been
responsible for the breaking down of proteins on the film. This may be explained
through a process called Alkaline Hydrolysis3, where the peptide bonds in proteins are
broken by water.
Test tube 4, contained pepsin and water, and ranked 3rd in both variables. The reason
that the solution in tube 4 did not succeed tube 3 and 5 in terms of relative pepsin
activity may be explained firstly by the fact that the water diluted the concentration of
the pepsin. Secondly, the pH level of the tube was 7.5, which is relatively far from the
optimal pH range that pepsin operates in.
Interestingly, test tube 2 and 1, which both ranked 1st and 2nd in relative pepsin activity
and 5th and 4th in relative amount of coating respectively, both had no pepsin that acted
upon the proteins on the film. This may suggest that without pepsin, there would be
great difficulty in breaking down proteins, especially in the human body.
Ultimately, these conclusions could be drawn as a result of the investigation being
executed with precision. These conclusions are reliable as the results had been
compared by all groups that took place in this investigation, and all found similar, if
not exactly the same results. Furthermore, a similar experiment named The Effect of
Various Acids on the Digestion of Proteins by Pepsin4 showed that pepsin operated
optimally in the presence of hydrochloric acid. This affirms part of this experiment,
especially the results produced by tube 5.

3 GordonI.Kaye,PH.D,PeterB.Weber,PH.DandWilliamM.Wetzel.TheAlkalineHydrolysis
Process.Articles2004.Web.6July2015.
4 Northrop,J.H."Theeffectofvariousacidsonthedigestionofproteinsbypepsin."TheJournalof
generalphysiology1.61919:pg607612.Web.6July2015

Joseph Latif
Evaluating Procedures/Improving the Investigation:
As a general rule, the reliability of any investigation can be improved through using
more time, more subjects, more trials and more precise equipment. However, in this
investigation, there were specific limitations that could have been amended if some
extra processes were executed.
Given that this experiment was conducted in a school environment, there were many
safety concerns that may have impaired the validity of the results. For example, when
dealing with chemicals such as sodium hydroxide, which is alkaline enough to burn
skin, small concentrations of the chemical were used to prevent this from occurring.
Therefore, this may have limited the validity of the results, as it does not truly
replicate the biological environments where protein digestion takes place.
There were also some prominent weaknesses in the experiment:
Identified Weakness
The inability to keep the
temperature of tube 2 at a
constant boiling temperature
the equipment used in the
investigation did not allow
for a constant temperature
to be kept so that tube 2
could remain boiling.

Significance of Weakness
This was of rather low
significance as the
temperatures were extreme
enough the denature the
pepsin so it could not
function and therefore it did
not matter if the temperature
varied by 10

Suggested Improvement
Precise equipment such as a
laboratory oven could have
been equipped in order to
control the temperature of
the boiling water. The
experiment could possibly
recommend executing the
investigation in a scientific
environment where such
equipment is readily
available

The method was not

completely followed the
method instructed to allow
10 to 15 minutes for some
procedures, however these
were cut short due to the
lack of time.
Accurate recording methods
were not used qualitative
data that was observed was
not recorded, but rather was
remembered.

This is of mild significance

as the results may have been
more valid if the method was
completely followed

Delegated segments of time

should be dedicated to
performing the experiment
to allow for the most valid
results

This may have effected

results as there may have
been a lack of consistency in
recalling the data. This is of
mild significance as for the
purpose and accuracy of the
experiment, the data was
adequate in providing sound
results. However, if this
experiment were to be done
in a much more formal
environment, this would be of
high significance.

Accurate data recording

methods should take place,
such as electronically
recording the qualitative
data and/or taking
photographs of the data