The enzyme, Amylase is significant to the commercial world and it is important to know the optimal conditions for amylase activity to be able to use amylase efficiently. To determine the optimal temperature for both fungal and bacterial amylase, an Iodine test was used to visually measure starch catalysis. A mixture of starch and amylase, either bacterial or fungal, were placed in four different temperatures, 0⁰C, 25⁰C, 55⁰C, and 85⁰C, and then added to iodine to observe amylase activity. A light yellow color means a weak presence of starch which indicates a high activity rate for amylase while a dark blue-black color means a strong presence of starch which indicates a low activity rate for amylase. Observed was an optimal temperature of 55⁰C for bacterial amylase which showed the lightest yellow color and an optimal temperature around 25⁰C for fungal amylase which showed the lightest yellow color.
Introduction
Enzymes are biological catalysts that regulate chemical reactions which accomplish a constant production of energy that all biological processes require. These proteins are vital to maintaining functionality in people’s daily lives and the absence of an enzyme can cause detrimental harm in the form of illness or even death (Alberte, Pitzer, Calero 49). Enzymes are also used commercially to improve the standard of living. Amylase, for example, an enzyme that is used to break down protein and starch: a storage polysaccharide, is among the most important that have been
0.0375 mg/ml Porcine Pancreatic Amylase Solution (amylase powder in 0.9% NaCl ), Iodine Solution; each solution were pipetted into each of the 5 test tubes with 5 ml of 1% starch. Each tube contained a 1% starch solution with a different pH. All tubes were at room temperature. Room temperature was 22C. 0.2 ml of porcine pancreatic amylase solution was then pipetted into each tube. A timer was started and every 3minutes the starch / amylase mixture were pipetted from each tube and pipetted into the spot plate for every sample tube, then the iodine solution were added to a spot plate cell for each sample. Iodine reacts with starch to change from yellow to deep blue /black in the presence of starch. A lightening of the blue/ black to a brown color will occur as less starch is present. Results were reported as (+) for presence of starch in the sample or (–) for the absence of starch. After every three minute increment had passed, these same
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
amylase enzyme and the optimal temperature for fungal and bacterial amylase. In order to make
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
Enzymes are biological catalysts that lower the activation energy required for a reaction to take place. They speeds up the process of the reaction, while still not being consumed, they can function multiple times. Enzymes are specific to molecules or sets of molecules and only work with these specific substrates. The enzymes cause substrates to be converted to products faster than they would without enzymes. A substrate fits into an enzyme, which then catalyzes the conversion of the substrate into products. The enzyme facilitates reactions, however it can become denatured. When an enzyme is denatured its form changes, thus preventing its function of converting substrates to products faster, and the enzyme does not work. The enzyme used in this experiment is amylase. Amylase is an enzyme that catalyzes the breakdown of the polysaccharide starch into the disaccharide maltose. Amylase is produced by human salivary glands and aids in digestion and breaking down food substances.
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
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.
The purpose of this experiment was to find the optimal temperature and pH of barley alpha-amylase. I hypothesize that the optimal temperature would be 55 degrees Celsius and the optimal pH would be 5.5. In this experiment, the starch is used as a substrate to examine the optimum temperature and pH for the reaction of alpha amylase. It is known that the measuring of disappearance (absorbance) of the substrate starch with iodine using spectrophotometer will show the concentration of the substrate which will also reflect on the reaction rate. Once the reaction rates are figured out, the optimal temperature and pH can be determined. The result concluded that the
The results in Figure 1, supported both the hypothesis as well as the predictions because the graph depicts how the differences in temperature changed the enzyme activity of amylase. The results concluded that as temperature deviates from optimal temperature amylase will hydrolyze starch at a lower rate. At 5 and 70 degrees Celsius amylase took longer to hydrolyze starch compared to 25 and 40 degrees Celsius. The results also supported the prediction that at 40 degrees Celsius, enzymes will break down starch at a faster rate than at 5 degrees Celsius, 25 degrees Celsius, or 70 degrees Celsius. The results showed 40 degrees Celsius as the optimum temperature for the hydrolysis of starch because 40 degrees Celsius is close to the body’s temperature.
Record observations: what color did the solution turn? Orange or blue/ black? This will tell whether the amylase is able to denature the starch solution Orange=Negative Blue/black= Positive
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).
The Effects of Enzyme Concentration on the Activity of Amylase To investigate the effect of Amylase concentration on its activity. the relative activity of Amylase is found by noting the time taken for the starch substrate to be broken down, that is, when it is no longer gives a blue-black colour when tested with iodine solution. This time is referred to as the achromatic point. Equipment: v Amylase solution 0.1% v Starch Solution 1.0% v Distilled water v Iodine in potassium iodine solution v White tile and polythene pipette v Graduate pipettes or syringes v Test tubes in rack v Beaker (used as water bath) v Stopwatch, Thermometer v Eye Protection
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.
In this lab we looked at the role of pancreatic amylase in the digestion of starch and the effect that temperature and pH has on this enzyme. Enzyme’s work as catalysts that increase the rate of chemical reactions within cells (Cooper, 2000). In order to do this, enzymes must show two essential properties: these two fundamental properties of enzymes include increasing the rate of chemical reactions without being eternally altered by the reaction and accelerating the reaction rate with keeping the reactants and products in chemical equilibrium (Cooper, 2000). Enzymatic catalysis is necessary for life. Most biochemical reactions would not occur under the mild temperatures and pressures