This experiment was needed to determine the optimal temperature at which the enzyme amylase best functions. Since an increase in temperature speeds up the movement in molecules, the substrate is supposed to come in contact with the active site of the enzyme more quickly and frequently until the heat becomes an obstacle by denaturing the structure of these enzymes and thus changing its function.
There were a total of four different temperatures at which amylase activity was observed (Table 1). In order to ensure that temperature was the only manipulated variable, four test tubes were prepared with the same contents. Each tube contained about 2mL of the 1% starch solution, 4mL of deionized water, 1mL of a 6.8 pH buffer. After 10 minutes of
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It was predicted that as temperature increases, the chemical reaction would accelerate mainly because the increased temperature would speed up the motion of molecules. “Microorganisms are efficient degraders of starch, chitin, and the polysaccharides in plant cell walls” (Warren, 183). For a variety of biological purposes, plants break down long polysaccharide (starch) molecules into disaccharide (maltose) molecules and monosaccharide (glucose) molecules. There is no reason to assume that amylase is the only enzyme responsible for the hydrolysis of starch (Paleg, 904). As the results show, the reaction that consumes starch and produces maltose would eventually go to completion; however, with the addition of the enzyme amylase, the reaction should significantly speed up. It was also determined via this experiment that the addition of amylase is not the only variable that influenced the speed of this reaction; temperature also played a huge role. The evidence gathered in the experiment supported the hypothesis and matched the prediction: there was a great increase in amylase activity in higher temperature experiments up until the enzymes begin to denature. There were two ways to gather data that would prove the change in the reaction rate in presence of the amylase enzyme: the appearance of maltose or the disappearance of starch. The second test was used for this specific
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
[An active site can be altered by a non-competitive enzyme which encircles the enzyme and alters the shape of the active site which could be very dangerous.]
These results show how temperature of extreme high, or low affects enzyme activity. The highest rate of enzyme activity occurred at 37 Cº. Anything that was hotter or cold than 37 Cº slowed the reaction rate. As I thought, 100 degrees would denature the enzyme, and that was the case. The data provided shows exactly what temperatures enzymes work best, and worst. The objective was achieved as we discovered the different reaction rates under different temperatures. The results are reliable, as we know enzymes do not work well when under extreme heat or denaturation occurs. What I learned in this experiment was that enzymes don’t work well under cold temperatures because they tend to move slower. My hypothesis did not quite match, because I thought they work best at lower temperatures.
1) Amylase is utilized to break down starch molecules into more simple sugars for use by the body. It performs this function by hydrolyzing glycosidic linkages in the polysaccharide chain.
As stated in the introduction, three conditions that may affect enzyme activity are salinity, temperature, and pH. In experiment two, we explored how temperature can affect enzymatic activity. Since most enzymes function best at their optimum temperature or room temperature, it was expected that the best reaction is in this environment. The higher the temperature that faster the reaction unless the enzyme is denatured because it is too hot. Similarly, pH and salinity can affect enzyme activity.
bonds and taking a glucose molecule off of the structure (Ball, Vialle, Alonso-Casajus, Duavillee, Munoz, Baroja-Fernandez, Moran-Zorzano, Eydallin, Pozueta-Romero, 2006). The conditions for the reaction of Phosphorylase and
The purpose of this experiment was to determine (1) the reaction rate of an amylase enzyme in starch and (2) the environmental factors that can affect the enzymatic activity. The hypothesis, in relation to the enzymatic activity by variables such as the substrate concentrations, temperature, PH and chemical interactions on the rate of reaction, stated
Hydrolysis of starch for fungal amylase Aspergillus Oryzae and bacterial amylase Bacillus Licheniformis at different temperatures.
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
Effect of varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and hydrolysis of Starch
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
Finding the optimal temperature for enzymatic activity of bacterial and fungal amylase was the main purpose of the experiment. The effectiveness of an enzyme can be affected by the environment of the organism is in, and to work at its best, the optimal temperature is necessary to breakdown nutrients and produce energy. The results for this particular experiment showed that the optimal temperature for both amylases was 65°C. This is because at this temperature, the breakdown of starch was the most effective, being able to catabolize the starch into minor subunits like maltose (which organisms can then use as energy storage and as a source of food) (Alberte et al., 2012).
Amylase is an enzyme that is located in human saliva. It is solely accountable for breaking down starch as a way to start the breakdown of food and is one of the first steps of digestion. The time at which the enzyme starts the chemical reaction with starch is called the reaction rate. In order to study how amylase works against starch, this experiment consisted of two tests; each testing a different condition of amylase. The first test was to simply study the reaction between saliva and amylase and note the reaction rates. The second test was to see if increasing the pH would decrease the reaction rate or halt it all together. Saliva was collected, diluted, and tested for reactions between starch and amylase. Another sample of saliva was collected, diluted, and had its pH increased and tested for reaction rate. The findings after the experiment was conducted aligned with the original hypothesis. The change in pH did show a significant decrease in the reaction rate.
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.
In this lab our group observed the role of pancreatic amylase in the digestion of starch and the optimum temperature and pH that affects this enzyme. Enzymes are located inside of cells that increase the rate of a chemical reaction (Cooper, 2000). Most enzymes function in a narrow range of pH between 5 through 9 (Won-Park, Zipp, 2000). The temperature for which enzymes can function is limited as well ranging from 0 degrees Celsius (melting point) to 100 degrees Celsius (boiling point)(Won-Park, Zipp, 2000). When the temperature varies in range it can affect the enzyme either by affecting the constant of the reaction rate or by thermal denturization of the particular enzyme (Won-Park, Zipp, 2000). In this lab in particular the enzyme, which was of concern, was pancreatic amylase. This type of amylase comes from and is secreted from the pancreas to digest starch to break it down into a more simple form called maltose. Maltose is a disaccharide composed of two monosaccharides of glucose. The presence of glucose in our experiment can be identified by Benedicts solution, which shows that the reducing of sugars has taken place. If positive the solution will turn into a murky reddish color, where if it is negative it will stay clear in our reaction. We can also test if no reduction of sugars takes place by an iodine test. If starch is present the test will show a dark black color (Ophardt, 2003).