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)
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