Required Practical 1-

The effect of temperature on the rate of the reaction catalysed by

trypsin

INDEPENDENT VARIABLE:
The temperature of the water bath
DEPENDANT VARIABLE:
The amount of time for the milk to be catalysed by the trypsin/for it to be clear enough to see
the cross.
RANGE:
The range would be 555-55=500 for our results.
The range would be 511.2-77.2=434 for the classes mean results.
DEGREE OF UNCERTAINTY:
The degree of uncertainty would be around 10 seconds, that would be the time taken for us to
figure out If we can see the cross or not.
CONTROL VARIABLES:
 The volume of milk
 The volume of trypsin
 The volume of pH7 buffer solution
 The amount of time in the water bath before mixing, to reach the correct temperature.
 The same timer
 The same person measuring the amount of trypsin, buffer solution and milk powder.
 Milk power from the same container- same concentration
 Same size crosses + water washing off the cross
 Measure the temperature of the enzyme and milk before mixing
 Look from the same angle
 Standardise the method

Trypsin
Trypsin is an enzyme in the first part of the small Intestine that starts the digestion of protein
molecules by cutting the long chains of amino acids into smaller pieces. It is a form of protease.
Trypsin is formed in the small intestine when the pro enzyme form, the trypsinogen produced in
the pancreas, is activated.
Trypsin catalyses the hydrolysis reaction of peptide bonds, breaking down proteins into smaller
peptides. It is a necessary step in protein absorption as proteins are generally too large to be
absorbed through the lining of the small intestine.
en.Wikipedia.org/wiki/Trypsin
Optimum temperature: 65 degrees Celsius
Optimum pH: 9.0pH
https://www.ncib.nlm.nih.gov > articles > PMC4691143
 Graph to show the rate of trypsin activity in digesting casein in milk
16.0

14.0

12.0
Rate of Trypsin Activity

10.0

8.0

6.0

4.0

2.0

0.0
15 20 25 30 35 40 45 50 55 60 65

Temperture (degrees Celcius)

Our experiment shows a trend that the optimum temperature would be around 40 degrees, even
though the enzyme should have been moist active at around 65 degrees. For our experiment, the rate
of Trypsin activity was actually the lowest at the temperature of 60 degrees.
Our graph shows the trend that they had been denatured at the higher temperature of 60 degrees, and
did not work as well at the lower temps because it was colder.

Limitations:
The limitations in this method are that we didn’t have experiments to show the effect of temperature
between 30-40-50 degrees. This would have shown a closer average to what the optimum temperature
would have been.
We also had different people doing the experiment, which means that the average is not as accurate,
as people may have made mistakes. Also, different people would have measured the time differently
as when we could no see the cross, which would have been different for each of the people.

Adapting the experiment:

To show the effect of pH or substrate concentration on trypsin, instead of changing the temperature in
the water baths, you would change the concentration of milk to trypsin, or the pH of the solution the
Trypsin is in, and leave them in the same water bath, at the same temperature.