The experiment conducted was to observe the enzyme catalysis of Horseradish Peroxidase with different determining factors. In this experiment the horseradish peroxidase enzyme should have an optimal temperature of 35°C, should have an optimal pH of 6, and should catalyze rapidly with the substrate and enzyme concentration. We were able to observe that the enzyme reacts best with its optimal temperature of 35° C and optimal pH of 6. Moreover, when there’s a high the substrate concentration and a high enzyme concentration we can observe a rapid enzyme catalysis. The hypothesis was partially falsified for the optimal pH and the optimal temperature were both different than that stated in the hypothesis. For the Horseradish Peroxidase reacts best
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Consequently, five test tubes were prepared at different temperatures to observe which test tube's temperature is the optimal temperature of the enzyme. These are the test tube and their temperature; Tube 1- was blank, Tube 2- room temperature, Tube 3- 35° C, Tube 4- 45° C and Tube 5- 55° C. These tubes had 1 mL of the substrate mixture and 1 mL of pH 6 buffer. These tubes were placed in a spectrophotometer to measure their reaction. This data was collected by placing a timer and acquiring the reaction every five seconds for a minute. (see pg. 35 lab manual)
During the second experiment to observe the optimal pH of the enzyme. Additionally, five test tubes were prepared with different levels of pH. These tubes had 1 mL of the substrate mixture and every test tube had its own type of pH. Consequently, Tube 1- pH 3, Tube 2- pH4, Tube 3- pH5, Tube 4- pH 6, Tube 5- pH8. These tubes were placed in the Spectrophotometer, for 1 minute at a time and the data was collected every five seconds. (see page 35 lab …show more content…
The tubes contained 1 mL of the substrate mixture, 1 mL of peroxidase and .1 mL of enzymes. The test tubes had the following content; Tube 1- 2.5 x 10^-8 M peroxidase, Tube 2- 5 x 10 ^-8 M peroxidase, Tube 3- 1 x 10 ^ -7 M peroxidase, Tube 4- 2 x ^-7 M peroxidase, and Tube 5- 4 x 10^-7 M peroxidase. After the test tubes were inverted and placed in the spectrophotometer to measure the enzymes catalysis. A timer was placed for a minute and the data was collected every five seconds. (see pg. 36 lab manual)
Results:
Figure 1-The Rate of Reaction for optimal Temperature
In this graph we can clearly see that Horseradish peroxidase reacted the most with the enzyme at 25 ℃ . Therefore, the optimal temperature of the enzyme was 25 ℃, for it had the fastets reaction. Figure 2- The Rate of Reaction for Optimal pH
In this experiment, we were able to conclude that the Horseradish reacts the best with pH 3. The optimal pH for the enzyme that is added into the horseradish is pH 3.
Figure 3- The rate of Reaction for The substrate Concentration
This graph demonstrates that the test tube with the most the substrate had the fastest enzyme catalysis with 20mM of the substrate concentration. Figure 4- The rate of Reaction for Enzyme
Consequently, five test tubes were prepared at different temperatures to observe which test tube's temperature is the optimal temperature of the enzyme. These are the test tube and their temperature; Tube 1- was blank, Tube 2- room temperature, Tube 3- 35° C, Tube 4- 45° C and Tube 5- 55° C. These tubes had 1 mL of the substrate mixture and 1 mL of pH 6 buffer. These tubes were placed in a spectrophotometer to measure their reaction. This data was collected by placing a timer and acquiring the reaction every five seconds for a minute. (see pg. 35 lab manual)
During the second experiment to observe the optimal pH of the enzyme. Additionally, five test tubes were prepared with different levels of pH. These tubes had 1 mL of the substrate mixture and every test tube had its own type of pH. Consequently, Tube 1- pH 3, Tube 2- pH4, Tube 3- pH5, Tube 4- pH 6, Tube 5- pH8. These tubes were placed in the Spectrophotometer, for 1 minute at a time and the data was collected every five seconds. (see page 35 lab …show more content…
The tubes contained 1 mL of the substrate mixture, 1 mL of peroxidase and .1 mL of enzymes. The test tubes had the following content; Tube 1- 2.5 x 10^-8 M peroxidase, Tube 2- 5 x 10 ^-8 M peroxidase, Tube 3- 1 x 10 ^ -7 M peroxidase, Tube 4- 2 x ^-7 M peroxidase, and Tube 5- 4 x 10^-7 M peroxidase. After the test tubes were inverted and placed in the spectrophotometer to measure the enzymes catalysis. A timer was placed for a minute and the data was collected every five seconds. (see pg. 36 lab manual)
Results:
Figure 1-The Rate of Reaction for optimal Temperature
In this graph we can clearly see that Horseradish peroxidase reacted the most with the enzyme at 25 ℃ . Therefore, the optimal temperature of the enzyme was 25 ℃, for it had the fastets reaction. Figure 2- The Rate of Reaction for Optimal pH
In this experiment, we were able to conclude that the Horseradish reacts the best with pH 3. The optimal pH for the enzyme that is added into the horseradish is pH 3.
Figure 3- The rate of Reaction for The substrate Concentration
This graph demonstrates that the test tube with the most the substrate had the fastest enzyme catalysis with 20mM of the substrate concentration. Figure 4- The rate of Reaction for Enzyme