Tests were performed to determine the most optimal temperatures for the fungal (Aspergillus oryzae) and bacterial amylase. The tests looked at the effect of the temperatures on the amylase’s ability to breakdown starch to maltose. With the addition of Iodine, the starch catalysis was able to be monitored, which can either be yellow or black depending on the presence of starch. The independent variable of these tests are the temperature and time. While the dependent variable being the breakdown of the starch. 24 samples of both Fungal and Bacterial Amylase were placed in specific Temperatures. The Temperatures being: 0, 25, 65, and 85 degrees Celsius. They were placed in the temperatures for certain ranges of time being: 0, 2, 4, 6, 8, and 10 mins. They were placed in these temperatures for varying amounts of time in order for the amylase reach the temperature required, and to see the effects of the temperature change over time. We gathered that temperatures which were either too low or too high had a negative impact on the brake down of starch. More moderate temperatures allowed for greater starch catalysis. We can gather that cells are more efficient in temperatures around the 65 Celsius mark.
Introduction Section
The human body has to undergo billions of chemical reactions to maintain homeostasis, keep itself alive. These very reactions are what causes the body to produce energy. Within the body there are enzymes which are forms of catalysts for the reactions, and aid in the body’s ability to regulate itself efficiently. Without enzymes, the human body would be much more sluggish and harder to maintain. So, it is fair to say that enzymes are an essential part of human life.Enzymes work as catalysts or something which lowers reaction rate time, and increases the amount of reactions that can be accomplished successfully in a given moment.
The effectiveness of the enzymes depends on the temperature which they have too cold and the enzyme will not be able to aid much in the reaction. Too hot and the enzyme will get denatured and be of no help to the body. Enzymes can also be used to break down molecules instead of helping build them through hydrolysis and dehydration reactions. In this experiment tested for the deconstruction of the starch. The experiment measures the breakdown of starch by the enzyme provided to see the optimal temperature in which the enzyme works best. The predicted ideal temperature for the bacterial amylase is 65 Celsius since it is the temperature of the human body at homeostasis, and for fungal 25 Celsius was determined since fungal plants usually stay low to the ground and cool.
Methods Section
For this experiment, the use of two clean spot plates is required. Place tape or napkin alongside or under plates to label and keep the experiment organize. Label the temperatures Celsius on the top being 0, 25, 65, 85. Down the side write the time increments beginning at 0, 2, 4, 6, 8, and 10. Then label four test tubes and write whether bacterial or fungal and the temperature they will be placed in. Then label another four test tubes in the same way but indicate that these have starch added. Next, use a pipet to retrieve 5ml of 1.5% starch solution and add it to those test tubes which have the starch indicated on them. Then, add 1ml of bacterial amylase to all test tubes. Then place tubes where they belong. The test tube for 0 Celsius should be in an ice bath. The 25 Celsius should be placed in a pot with the same temperature water. The 65 Celsius should be left outside since this is ambient temperature, and the 85 Celsius should be placed in a container with hot water at the temperature. Place a timer set it for every two minutes. When it rings the tubes are to be taken out and a sample put in the according welt. Then two drops of iodine are to be added and record the color. Refer to lab manual for a correct representation of how the colors should be. Record all data once each amount of time runs up. Then the process must be repeated for the fungal amylase.
Results Section
Figure 1: This shows the breakdown of starch
in the bacterial amylase using the stated intervals and temperatures
Figure 2:This shows the breakdown of starch
in the fungal amylase using the stated intervals and temperatures
Figure 3: This Graph shows a visual representation of how temperature and time affected
The bacterial amylase.
Figure 4: This Graph shows a visual representation of how temperature and time affected
The bacterial amylase.
Color #
Temperature °C
0
25
65
85
Time (Mins)
0
4
5
5
5
2
3
3
3
5
4
2
2
2
5
6
2
2
2
5
8
2
2
2
5
10
2
2
2
5
Figure 5: These table shows the data recorded during the experiment for Bacterial Amylase.
Color #
Temperature °C
0
25
55
85
Time (Mins)
0
3
4
4
5
2
2
2
2
5
4
2
2
2
5
6
2
2
2
5
8
2
2
2
5
10
2
2
2
5
Figure 6: These table shows the data recorded during the experiment for Fungal Amylase.
The results of this experiment have been shown in figures one through six. Through the use of iodine, the amount of hydrolysis is able to be detected. Once the iodine is dropped in the welts it could turn either yellow or a black. This all depends in how much starch was broken down. Yellow signifies that the starch has been effectively broken down while black shows that the starch has not been broken down and that there may be something wrong with the amylase. Starch hydrolysis was measured before any of the tubes were submerged in the unique baths to get a baseline as to how the test began and to get a clear starting point for all the amylase. The iodine showed to be somewhat dark at the beginning however this was due to the fact that the amylase was being stored in an environment which may not have been optimal for it. Once the amylase was submerged and the tubes were sampled there was an immediate difference. Showing that our amylase was reaching optimal temperature except for the 95 Celsius amylases since that one seems to have been denatured. After the second sample the amylase ceased to vary in the starch hydrolysis. However, all test show that any warm temperature is good for starch hydrolysis.
Discussion Section
Based off of the experiment it can be concluded that the hypothesis is correct that temperature affects starch hydrolysis. In the first half of the experiment it can be seen that the optimal temperature for the bacterium amylase is 65 Celsius. This is due to the warmth allowing for the enzyme to catalyze with greater efficiency. The fungal amylase seemed to have been most efficient when placed under 65 Celsius which make sense considering fungus thrive in warmer and more humid climates. The enzymes placed in 95 Celsius all suffered low starch hydrolysis. This is due to the high temperature denaturing and destroying the enzymes ability to function. Some sources of errors in this lab were when sampling the lack of mixing the amylase and starch could have led to unbalanced mixtures which could have thrown off how much amylase could have done. Another, error is that the welts may have been previously contaminated with something other than what was going to be tested. This could be prevented by simply cleaning all instrumentation before use. This is a beneficial lab because it provides insight as too why the body functions as it does. The fact why the temperature homeostasis is maintained at is 98.6 Fahrenheit can be determined by this. Through many years pf evolution, the body has seen the temperature in which its enzymes function best and try to maintain a perfect environment for reactions. The experiment also demonstrated why it is so important for individuals with a fever to keep their temperatures from sky rocketing. As well as why they may feel weak during the fever. This experiment shows how important enzymes are, their functions, and the basic understanding as to which environment is best for them. It is important for everyone to understand how their body functions and regulates itself.
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