Enzymes and Temperature
Purpose (Objectives): The purpose of this experiment was for students to be able to understand the enzyme-facilitated reaction and explain how enzyme activity can be affected by environment conditions.
This experiment mainly delt with enzymes and the reaction that enzymes have with different solutions in various temperatures. Three different exercises were done. The first exercise had to do with enzymes and temperature. During this exercise three different test tubes were used for three different temperates. The first test tube which involved 0 ºC water bath had a total conversion of starch to sugar in the first 15 minutes, and the second test tube which involved 37ºC water bath had a total conversion of starch to sugar after the first five minutes. While the third test tube took a little longer. The second exercise had to do with enzymes and substrate concentration. This exercise took longer for the all of the starch to be removed from the test tubes, and it involved 37ºC water bath for all five test tubes. The last exercise had to do with enzymes and pH. This exercise involved four test tubes all put into a water bath of 37ºC. The fourth test tube had a total conversion of starch to sugar in the first five minutes while the other three took a little longer to convert.
Experiment & Observation:
First the water baths and the 1% Alpha-amylase was prepared. Three different water baths were prepared. One being set at 0ºC, the next being set at 37ºC and the last one being set at 100ºC. Next a 96-well microplate was marked with times and numbers based on the test tubes number. Then using a pipet one drop of iodine was placed into each well (1,2,3) up to total of 30 minutes. Then the test tubes were marked 1cm and 6cm from the bottom. Afterward, 1cm of alpha-amylase was pipetted into each test tube. Then, test tube 1 was incubated at 0ºC, test tube 2 was incubated at 37ºC, and test tube 3 was incubated at 100ºC for five minutes. After that 1% starch solution was added to the 6cm mark. Next it was put back into its assigned water bath for another 5 minutes. Then two drops of solution was put into its corresponding number on the microplate in the 5 minute row. The color was immediately recorded. The steps were repeated for a interval of 30 minutes or until each well had an amber color in it. Test tube 3 did not change within the 30 minute interval so it was placed into the 37ºC water bath for another 30 minutes. Then one drop of solution and one drop of iodine was added into the well. There was still no color change, so another 1cm of alpha-amylase was added to it and was incubated for another 30 minutes, the color became amber.
Table 1. Effect of Temperature on Amylase Enzyme Conversion of Starch to Sugar
|Time/Minutes |Test Tube 1 / 0ºC |Test Tube 2 / 37ºC |Test Tube 3 / 100ºC | |5 |+ |- |++ | |10 |+ | |++ | |15 |- | |++ | |20 | | |++ | |25 | | |++ | |30 | | |++ | | | | | | |60...