Peroxidase Lab
Abstract
The enzyme peroxidase has been shown to break down H2O2. Enzymes are known to increase the rate at which a chemical reaction occurs. We looked at factors that affected the breakdown of hydrogen peroxide. These effects are the different temperatures and pH levels the enzymes were placed in. We found that the optimum, or best condition, temperature for the enzymes tested was about 22 degrees Celsius. The optimum pH level for the enzyme was 7.
Introduction
Enzymes are biochemical that catalyze, or increase the rate, at which a chemical reaction occurs. All enzymes are proteins that have a specific shape that is vastly determined by their unique amino acid sequence (Vodopich and Moore 2011). Enzymes run on a method similar to that …show more content…
The materials needed for this experiment were four medium sized tubes, a spectrophotometer, a buffer with a pH of 5, H2O2, Peroxidase, and Guaiacol Dye. We as a group had four tubes labeled two, three, four, and five. In the tube labeled two we had a solution of 2.0 mL of H2O2 and 1.0 mL of Guaiacol Dye for a total solution of 3.0 mL. In the tube labeled three we had a solution of 4.0 mL buffer and 1.0 mL Peroxidase for a total of 5.0 mL solution. In the tube labeled four we had a solution of 2.0 mL H2O2 and 1.0 mL Guaiacol Dye for a total of 3.0 mL solution. Last, in the tube labeled five we had a solution of 4.0 mL buffer and 1.0 mL Peroxidase for a total solution of 5.0 mL. We then set the spectrophotometer to a value of zero, using a blank containing buffer, H2O2, and Guaiacol Dye. After that, tubes two and three were mixed together and tubes four and five were mixed together for a large solution of with a total of 8.0 mL. Once the two solutions are mixed clean the tube with a Kim Wipe, so all fingerprints are removed. Place the tube with solutions of four and five in the ice bath for twenty minutes. While placing the tube in the spectrophotometer make sure the white strip that is on the tube is facing the direction of the person who is handling placing the tube I the spectrophotometer. Furthermore, just let the spectrophotometer act on the solution for the desired length of time while recording the data at each specific …show more content…
Also, a given slope for pH 3, 5, 7, and 9.
Figure 1.1: Graphical representation of pH on peroxidase activity in 20 second increments for two minutes.
The next set of data was used in relation of different temperature levels in degrees Celsius instead of pH. In all four groups there was a change in value.
Table 1.2 Temperature data condensed
Time
4
22
32
48
Temp
Slope
20
0.438
0.444
0.207
0.255
4
0.0008
40
0.460
0.585
0.223
0.257
22
0.0056
60
0.474
0.700
0.242
0.260
32
0.0009
80
0.490
0.815
0.263
0.260
48
6E-0.5
100
0.509
0.920
0.282
0.260
120
0.514
1.010
0.300
0.261
Table 1.2: Temperature effects on Peroxidase at temperatures of 4, 22, 32, and 48 degrees
The enzyme peroxidase has been shown to break down H2O2. Enzymes are known to increase the rate at which a chemical reaction occurs. We looked at factors that affected the breakdown of hydrogen peroxide. These effects are the different temperatures and pH levels the enzymes were placed in. We found that the optimum, or best condition, temperature for the enzymes tested was about 22 degrees Celsius. The optimum pH level for the enzyme was 7.
Introduction
Enzymes are biochemical that catalyze, or increase the rate, at which a chemical reaction occurs. All enzymes are proteins that have a specific shape that is vastly determined by their unique amino acid sequence (Vodopich and Moore 2011). Enzymes run on a method similar to that …show more content…
The materials needed for this experiment were four medium sized tubes, a spectrophotometer, a buffer with a pH of 5, H2O2, Peroxidase, and Guaiacol Dye. We as a group had four tubes labeled two, three, four, and five. In the tube labeled two we had a solution of 2.0 mL of H2O2 and 1.0 mL of Guaiacol Dye for a total solution of 3.0 mL. In the tube labeled three we had a solution of 4.0 mL buffer and 1.0 mL Peroxidase for a total of 5.0 mL solution. In the tube labeled four we had a solution of 2.0 mL H2O2 and 1.0 mL Guaiacol Dye for a total of 3.0 mL solution. Last, in the tube labeled five we had a solution of 4.0 mL buffer and 1.0 mL Peroxidase for a total solution of 5.0 mL. We then set the spectrophotometer to a value of zero, using a blank containing buffer, H2O2, and Guaiacol Dye. After that, tubes two and three were mixed together and tubes four and five were mixed together for a large solution of with a total of 8.0 mL. Once the two solutions are mixed clean the tube with a Kim Wipe, so all fingerprints are removed. Place the tube with solutions of four and five in the ice bath for twenty minutes. While placing the tube in the spectrophotometer make sure the white strip that is on the tube is facing the direction of the person who is handling placing the tube I the spectrophotometer. Furthermore, just let the spectrophotometer act on the solution for the desired length of time while recording the data at each specific …show more content…
Also, a given slope for pH 3, 5, 7, and 9.
Figure 1.1: Graphical representation of pH on peroxidase activity in 20 second increments for two minutes.
The next set of data was used in relation of different temperature levels in degrees Celsius instead of pH. In all four groups there was a change in value.
Table 1.2 Temperature data condensed
Time
4
22
32
48
Temp
Slope
20
0.438
0.444
0.207
0.255
4
0.0008
40
0.460
0.585
0.223
0.257
22
0.0056
60
0.474
0.700
0.242
0.260
32
0.0009
80
0.490
0.815
0.263
0.260
48
6E-0.5
100
0.509
0.920
0.282
0.260
120
0.514
1.010
0.300
0.261
Table 1.2: Temperature effects on Peroxidase at temperatures of 4, 22, 32, and 48 degrees