Enzyme Activity Lab Report

In this lab we tested the effect of temperature has on the rate of enzyme activity. The way we figured this out was by taking four different temperatures and testing the difference absorbance levels they produced every 20 seconds for about 2 minutes straight using a spectrophotometer. The important part of this experiment was the temperature the enzyme concentration was made at. What we got from the experiment was at lower temperature we got very low numbers for the absorbance, which gave us a lower rate for the enzyme reaction to complete, and vice versa for higher.
Materials an d Methods The first thing that we did in the lab was to put our safety glasses on and then we proceed with the experiment. We took and fresh Horseradish
Test tube one 1was filled with 5.0mL of buffer, 2.0mL of H2O2, and 1.0 mL of guaiacol. Tube 1 will be to calibrate the spectrometer. Tube 2-3 will be filled with Dil 0.25, tube 2, 4, 6 & 8 will contains 2.0 of 10mM H2O2 and 1.0 of 25mM of guaiacol. Tube 3 will contain 4.75 of 0.1 Buffer in mL and 0.25 of enzyme extract in mL. Tube 4-5 will be filed with Dil 0.5, but tube 5 will contain 4.5 of 1.0 Buffer in mL and 0.5 of enzyme extract in mL. Tube 6-7 will be filled with Dil 1.0, for instant tube 7 will contain 4.0 of 0.1 Buffer in mL and 1.0 enzyme extract in mL. Tube 8-9 will be filled with Dil 2.0; in addition tube 9 will contain 3.0 of 0.1 Buffer in mL and 2.0 of enzyme extract in mL. After we were done, we turned the spectrophotometer one and let it warm for about 5 minutes, we set the wavelength to 500nm we set it to a point that we could only read the absorbance until 1.999. We took a cuvette rack and placed about 5 cuvette tube. We took a clean cuvette and transfer all the contents of the tube 1 into the cuvette and wipe the outside with a kimwipe and insert the cuvette into the sample holder. After doing the previous step we turn the 100% transmittance control knob and control it until the reading said
When starting this experiment, we started with 0.25 of the dilution and at 20 seconds, we had an absorbance of .357, this process when thru 120 seconds we had reached the maximum of 1.999. From the dil of 0.50, 1.0 and 2.0 we started and ended with 1.999, with the wavelength of 500nm for each of the dil that we did in this experiment. After collecting the absorbance with the time, we got the rate of reaction by taking the concentration and dividing it. Giving you the rate of reaction of each of the concentration. Now by knowing the rate of reaction of the concentration we can determine the rate of reaction of each of the temperature that was provided to us, which are in (figure 3). How we can see, the higher and lower temperature gave us the lowest absorbance in all 120 seconds. And the only one that gave us the maximum room temperature which was 25°C. After getting the absorbance for the temperature, we could find the rate of reaction of the absorbance in the temperature by doing the same step in the previous step for (figure 2). And as said, that the room temperature was the highest, in the (figure 4) it shows where the other temperatures were place in the graph and in the rate of