“The Effect of Temperature on Amylase Activity”
The objective of this experiment was to determine the effect of temperatures (0 °C, 25 °C, 55 °C, and 85 °C) on fungal (Aspergillus oryzae,) and bacterial amylase’s ability to break down starch. Our team monitored starch catalysis using the Iodine test. Iodine turns the most catalyzed starch yellow, and the least catalyzed blue. When the reaction turns blue-black or dark brown, it means that starch is present and, therefore, not catalyzed.
Each member of team four had a temperature assigned to work with. Each individual monitored the activity of amylase (fungal and bacterial.) Time also varied respectively ( 0 mins, 2 mins, 4 mins, 6 mins, 8 mins, and 10 mins.) The group marked the spot plates with time (on the side) and temperatures (across the top.) The team labeled 4 test tubes with different temperatures, the enzyme source (B or F) and the group’s number (4.) The group also marked the pipette for each temperature. Test tubes were placed into the respective temperatures. Finally, the team carefully carried out the two trials.
Without removing the test tubes from the water bath at different temperatures, the team added 2 drops of starch from each temperature to the first row of the spot plate (at 0 minutes) containing iodine. The team poured the same amount of iodine before each trial. The team proceeded, transferring the starch into the amylase (fungal and bacterial) at their respective water bath and mix these two without removing the test tubes from the water bath. Then, the team added two drops of this new mixture (starch-amylase) at their respective temperatures to the second row of the spot plate. The other rows were filled with the same mixture at the temperatures assigned with two minutes interval between each trial.
The following tables and pictures show the results group four (first) and all groups (last) obtained.
Group 4 Result
Data Collected for all Groups
Fungal Amylase vs. starch
Bacterial Amylase vs. starch
After conducting the whole experiment, the team collected the data for the results. The group observed that the least amount of starch was present at 55 °C at any time, for bacterial amylase and 25°C to 55 °C for fungal amylase (for most students.) The reasoning behind this statement might be that the mixtures at 55 °C turned light yellow in the presence of iodine (at all times except time 0) for bacterial amylase. The mixture became dark yellow (at all times except time 0) at temperatures 25 °C to 55 °C for fungal amylase, which could indicate that amylase activity was not affected. The team also noticed that the enzyme amylase denatures at higher temperatures (85 °C) for both types of amylase. The group made that assumption because the solution turned dark blue brown at 85 °C at all times. According to The Iodine test, dark color indicates the starch is present and not catalyzed.
The average of starch concentration of all groups at each temperature for fungal amylase demonstrates that there are no outsiders in our data and that the results’ standard deviation is not significantly big for the experiment concluded. The results obtained for all the trials suggest that amylases (bacterial or fungal) have a respective optimum temperature. It is also evident that if these amylases are not placed on their ideal temperature, it loses their ability to break starch down.