Abstract To figure out how temperature effects the change in the rate of an enzyme, test tubes were filled with solutions of buffer with a pH of 5, peroxidase, H2O2, and guaiacol and placed different water baths of the following temperatures: 4°C, room temperature (23°C), 32°C, and 60°C. To help determine the rate of reaction of peroxidase, a spectrophotometer was used to measure the absorbances of the solution. There was also a recovery portion to check the functionality of the enzyme after being tested under varying temperatures. We had discovered that there were increases in the rate activity of reaction of peroxidase around 4°C and 32°C and that the enzyme had ceased in activity once it reached 60°C. This indicated that the rate of reaction …show more content…
Two test tubes were then paired with each of the following temperatures: 4°C, room temperature (23°C), 32°C, and 60°C. 4.0 ml of buffer with a pH of 5 and 1.0 ml of peroxidase was filled into one test tube from each of the temperatures. The other test tubes were filled with 2.0 mL of H2O2 and 1.0 mL of Guaiacol dye. As the 4°C, 32°C, and 60°C test tubes set in its corresponding water bath for 15 minutes, the room temperature test tubes proceeded with the experiment. A blank test tube was used to calibrate the spectrophotometer to zero. After calibrating the spectrophotometer, the room temperature test tubes were mixed together by pouring its contents back and forth twice. Once mixed, the solution was poured into a cuvette and placed in the spectrophotometer. The absorbance of the solution was analyzed in 20-second intervals for two minutes. After 15 minutes elapsed, the other test tubes were taken out of their baths and the mixing and absorbance processes were repeated. To test if peroxidase can regain its functionality after it has been affected by the temperature, the procedure was repeated but the solutions were allowed to return to room temperature after being its respective temperature baths for another 15