ABSTRACT: Enzymes are catalysts, speeding up of chemical reactions, of biological systems by lowering the activation energy (Transitioned from the AP Biology Lab Manual). In addition, in order to determine the rate of an enzymatic reaction, one must measure a change in the amount of at least one specific substrate or product over time. We were curious about determining the effects of pH and heat on enzymatic activity because these are factors that usually affect the shape of an enzyme. We measured enzyme activity using an indicator for product at different pHs and temperatures. Upon mixing them and waiting, some of the tubes changed colors and helped us identify which ones were being affected more/less by pH. On the other hand, for the heat
…show more content…
We placed it in a test tube rack in order to observe the change in a prolonged period of time. We utilized the color palette (Figure 1-A) to see the change in color of the test tube. Since we used an indicator for oxygen, and guaiacol has a high likeness for oxygen, it will bind with oxygen creating a brownish color. Determining the Effect of pH on Enzymatic Activity:
This experiment dealt with the effect of pH on peroxidase. First, we made 6 pairs of test tubes. Half of them contained substrate solution and the other half contained enzyme solution. Unlike our previous experiment, this time we substituted different pH buffers for distilled water. Furthermore, for each substrate test tube we added 7mL of distilled water, 0.2 mL of guaiacol, and 0.3 mL of hydrogen peroxide for a total volume of 7.5 mL.
In the enzyme tubes, we added 6.0 mL of pH solution, beginning at pH 4 in the first tube and changing the pH to 5 on the second test tube, so on and so forth. We also added, 1.5 mL of peroxidase for a total volume of 7.5 mL. Afterwards, we mixed the solutions individually and then combined the enzyme solution to the solution of the substrate. Upon placing the tubes back on the rack, we began to time the reaction and observed the color change using the color palette (Figure
We placed it in a test tube rack in order to observe the change in a prolonged period of time. We utilized the color palette (Figure 1-A) to see the change in color of the test tube. Since we used an indicator for oxygen, and guaiacol has a high likeness for oxygen, it will bind with oxygen creating a brownish color. Determining the Effect of pH on Enzymatic Activity:
This experiment dealt with the effect of pH on peroxidase. First, we made 6 pairs of test tubes. Half of them contained substrate solution and the other half contained enzyme solution. Unlike our previous experiment, this time we substituted different pH buffers for distilled water. Furthermore, for each substrate test tube we added 7mL of distilled water, 0.2 mL of guaiacol, and 0.3 mL of hydrogen peroxide for a total volume of 7.5 mL.
In the enzyme tubes, we added 6.0 mL of pH solution, beginning at pH 4 in the first tube and changing the pH to 5 on the second test tube, so on and so forth. We also added, 1.5 mL of peroxidase for a total volume of 7.5 mL. Afterwards, we mixed the solutions individually and then combined the enzyme solution to the solution of the substrate. Upon placing the tubes back on the rack, we began to time the reaction and observed the color change using the color palette (Figure