Temperature and its effect on Bacillus licheniformis (bacteria) and Aspergillus oryzae (fungal) amylase breakdown of starch
Isidro Planell
Panther ID: 5164085
General Biology lab 1 – U06
Emmanuel Gonzalez
Pablo Gonzalez
Abstract: The experiment’s goal was to study the effects of temperature and the rate of breakdown of starch by fungal and bacterial amylase. In order to conduct this experiment four samples were taken from both specimen and placed in four different temperatures. After a period of time a portion of the sample was taken and added to a portion of starch which was then mixed with iodine. Iodine reacts with starch to produce a change in color in the iodine from a brown to a dark purple. This serves as a method of measuring the breakdown of the starch by the amylase by comparing the color of iodine when mixed with starch and comparing it with the added amylase. It was also noted that because both specimen reacted differently with the temperature change, it was concluded that thy had differing optimal temperatures (Bacillus licheniformis reacting with an optimal temperature of 55 degrees Celsius while Aspergillus oryzae reacted optimally with a temperature of 25 degrees Celsius). In order to conduct the experiment, samples from both specimens were taken and placed in four test tubes all with a different temperature. Iodine was then taken and placed into a spot plate labeled with the type of amylase and corresponding
B. In which test tube(s) did the amylase convert starch to sugar? Why might you expect this outcome?
2. What effect did boiling and freezing have on enzyme activity? Why? How well did the results compare with your prediction?__Boiling caused amylase to be denatured, thus inactivating the enzyme. Freezing has no effect. The function of an enzyme is directly related to its environment, like temperature.__
During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s
The principal objective of this study was to determine how temperature affects the activity of
This experiment consisted of setting up a control group of starch in various temperature and then placing both fungal amylases and bacterial amylases in a mixture of starch and placing the solution of amylase and starch in various temperatures of water. After a certain amount of time- different amount of time needs to be used in order to have reliable results- iodine is added in a well on spot plates, then two drops of the mixture of amylase-starch is added from each temperature used, by adding iodine into the plates the mixture will show how much starch was hydrolyzed, this is used to calculate the amount of
For this experiment, we have to prepare our phosphorylase which extracted from a potato. We prepared by weighed about 250 grams of peeled potato and cut it into cubes. The extracts then blended with 100mL of 0.1M NaF. After filtered the contents into a clean 250mL centrifuge bottle, we centrifuged it for 3 minutes. Then, separated the supernatant into a centrifuge bottle, which is our phosphorylase preparation. The enzyme assay used in this experiment today is the iodine test. As the iodine reacts with starch, it will form a brown, blue or black precipitate due to the iodine ions forcing into a linear arrangement. The endpoint of the enzyme reaction indicates the presence of starch by using the iodine test to determine. The faster the endpoint is reached, the less active the phosphorylase is.
Top of Form Bottom of Form The Effect of Temperature on the Respiration of Yeast
The effects of temperature on fungal amylase Aspergillus oryzae, and bacterial amylase, Bacillus licheniformis ability to break down starch into maltose was studied. The study determined the optimal temperature the Aspergillus oryzae and Bacillus licheniformis was able to break down the fastest. The starch catalysis was monitored by an Iodine test, a substance that turns blue-black in the presence of starch. Amylase catabolizes starch polymers into smaller subunits. Most organisms use the saccharide as a food source and to store energy (Lab Manual, 51). The test tubes were labeled with a different temperature (0°C, 25°C, 55°C, 85°C). Each test tube was placed in its respective water baths for five minutes. After the equilibration process, starch was placed in the first row of the first row of the spot plate. Iodine was then added to the row revealing a blue black color. The starch was then added to the amylase. After every two minute section a pipette was used to transfer the starch-amylase solution to place three drops of the solution into the spot plate row under the corresponding temperature. Iodine drops was placed in the row. Color changes were noted and recorded. The results showed Aspergillus oryzae was found to have an optimal temperature between 25°C and 55°C and Bacillus licheniformis was found to have an
Hypothesis: If we decrease the level of pH in the enzyme Amylase, it will not be able to denature the carbohydrates in the potato starch solution after 10 drops because enzymes are very sensitive to pH levels and lowering it too much will compromise its ability to break them down.
Amylase experiment # 2 was done to see how the pH affected the efficacy of the enzyme. First we collected all of the materials that were necessary to make this experiment. We needed five clean test tubes, the following standard solutions, 1% Starch Solution pH 3,1% Starch Solution pH 5,1% Starch Solution pH 7,1% Starch Solution pH 9,1% Starch Solution pH 11
test the pH of the amylase a drop of the solution should be put on pH
The objective of the lab was to examine the effects of environmental variables on the functions of an enzyme. To the point, an experiment was conducted to test the effect of pH on the function of the enzyme Amylase.
(6.2)Material and Methods in the process or exercise of measuring the starch we were used the following material and how we used them to conduct the experiment. Obtain seven tubes the material to be tested table 6.1 and then add seven to ten drops of iodine to each tube, and then record the color of the tubes contents in table 6.1
How pH Affects the Break Down of Starch by the Enzyme Amylase Hypothesis: The optimum pH for the reaction of starch with amylase is pH 7. PH values lower or higher than this value will result in a slower rate of reaction. Amylase works in the range pH 3 to pH 11.
Amylase is an enzyme that is in human’s saliva as well as the pancreas. Enzymes are biological catalysts that speed up a chemical reaction. They break down complex molecules into simple ones. In this case, amylase converts starches (complex molecule) into simple sugars. That is why foods like potatoes for example, may taste sweet to us, because they contain starch. The optimum pH for pancreatic amylase is the pH of 7. In the experiment I have used buffer solutions with the pHs of 2.8, 4 and 6.5. I have also used iodine and starch. Normally, iodine is orange-yellow, however when you add starch to it, the solution will turn blue-black.