Aim: To observe how enzyme concentration can affect the initial rate of reaction.

Summary: The site of the reaction occurs on the surface of the protein called the
active site. At low enzyme concentration there is a great competition for the active sites and the rate of reaction is low. Though as the enzyme concentration increases, there are more active sites and the reaction can proceed at a faster rate.

Hypothesis: Investigating the effect of the reduction in enzyme concentration on the

rate of reaction, this will result in the protein to breakdown by the protease enzyme.

Apparatus: - Casein (Trypsin milkprotien) - Protease enzyme - Distilled water - Colorimeter - Cuvettes - Syringe - Beakers - Pipette (if required) - Stop clock Prediction: I predict that the higher the enzyme concentration the faster the protein
Trypsin (casein) will be broken down, therefore a faster rate of reaction.

Procedure:
(1) Add 1cm3 of protease enzyme in a beaker using a syringe to a beaker. (2) Add another 1cm3 of casein using a syringe to a beaker. (3) Set up the colorimeter and reset at 0.00. (4) Set the stopwatch. (5) Fill a cuvette with distilled water. (6) Place that cuvette in the colorimeter. (7) Remove the cuvette when remains 0.00 (this is known as the reference) (8) Add the protease enzyme to the casein. (9) Place it into the colorimeter.

(10) (11) (12) (13)

As soon as the cuvette is place, start the stopwatch and record values Record the results in a table. Remove the cuvette containing the enzyme and the casein. Add different amounts of water, in order to change the percentage of

every 30 seconds.

concentration.

Results:
0.5M Trypsin
Time (seconds) 30 60 90 120 150 180 210 240 270 300 330 Absorbance 0.43 0.39 0.34 0.30 0.25 0.22 0.19 0.18 0.17 0.17 0.17

Explanation: - From the results collected throughout the experiment, we notice that the further the time passes by, the less amount of light is absorbed by the cuvette, till it reached a constant value from 270 seconds to 330 seconds of an absorbance rate of 0.17. This shows that both variables are inversely proportional. - (Independent variable is the concentration of the Trypsin, Dependant variable is the absorbance rate).
50% 0.5M Trypsin and 50% distilled water Time (seconds) 30 60 90 120 Absorbance 0.52 0.49 0.46 0.43

150 180 210 240 270 300 330 -

0.41 0.39 0.37 0.35 0.33 0.32 0.32

Also, as the time increases the absorbance rate decreases. This shows that at low enzyme concentration there is a great competition for the active sites and the rate of reaction is low. However, as the enzyme concentration increases, there are more active site, and the reaction can proceed at a faster rate. Eventually, increasing the enzyme concentration beyond a certain point has no effect, because the substrate concentration becomes the limiting factor.

25% 0.5M Trypsin and 75% distilled water Time (seconds) 30 60 90 120 150 180 210 240 270 300 330 Absorbance 0.51 0.49 0.47 0.44 0.42 0.40 0.39 0.36 0.33 0.32 0.32

- The percentage of Trypsin is much less than the percentage of the distilled water. Therefore, the results show that as the time increases the absorbance rate also decreases. This also shows that both variables are inversely proportional. - Also, we notice at the beginning of the experiment, the reaction will proceed quickly. However, as the substrate is used up, there are fewer substrate molecules to bind with the enzyme and the reaction slows down and eventually stops.

Evaluation of results:

The initial rate of reaction is directly proportional to the enzyme concentration, because there are more enzymes present, therefore there are a greater number of active sites to form enzyme substrate complexes. Thus, when more substrates are added, more enzyme substrate complexes can be formed, as there are enzyme substrate collisions. Conclusion:
I conclude that the solution containing the protein became clearer as the concentration of the protease enzyme increased. Also, increasing the concentration of the substrate will not affect the rate of reaction as the same amount of enzyme will be available. But, if you increase the enzyme concentration, then the rate of reaction will increase as there are more enzymes to work on the substrates, but even the rate reaction will continue to increase as there is a limited amount of substrate, which is the limiting factor. By superageous