Amylase lab report Essays

Amylase lab report Essays Amylase lab report Paper Amylase lab report Paper Enzymes are very specific; for example, amylase is the only enzyme that will break down starch. It is similar to the theory of the lock and the key. The enzyme is the lock and the key is the substrate; only the correct key could fit into the keyhole of the lock. Porcine pancreatic amylase is The purpose of this lab experiment is to investigate factors that can affect the porcine pancreatic amylase enzyme activity in different environments such as the temperature, pH and also how being stored in extreme temperatures can affect the activity of the amylase. The activity of the amylase is going to be determined by the presence or absence of starch in the samples over time. There are some hypotheses on the Effects of temperature and pH; as I add the amylase to the starch in different temperatures the reactions rate increases in high temperatures; I believe that the amylase will work better. As the environment grows warmer, the amylase is going to become more energetic and more effective. Amylase is affected by environmental PH. Predict that the amylase activity will work best at a pH 7. As the pH changes from this point I predicted that the amylase activity is going to decrease and eventually stop. If I boiled and rose some amylase solution, and try to digest starch with at it at room temperature, I predict the previously-boiled and frozen amylase will not work quite as well as amylase that has not been previously boiled and frozen. Experiment #1: Investigating the Effect of Environmental Temperature on the Activity of Porcine Pancreatic Amylase Materials and Methods Amylase experiment #1 was done to see how the temperature affected the efficacy of the enzyme. First we collected all of the materials that were necessary to perform this experiment. We needed five clean test tubes, a test tube rack, a beaker of 1 % starch solution (pH 7. ), a test tube of porcine amylase solution which was previously prepared by mixing porcine pancreatic amylase powder with 0. 9% Nasal concentration to create an amylase concentration of 0. Magma/ ml, one I-ml pipette, one 5-ml pipette with pipette pump, five transfer pipettes, two spot plates, a bottle of iodine solution, a timer, one beaker half way filled with ice and a small amount of water, two thermometers, and three water baths each with a different temperature. In this experiment, we first labeled the test tubes. We then added 5-ml of 1% starch solution with a pH of 7. 0 to each of them. After each tube was filled, the first one was placed in the freezer at O degrees Celsius, the second one was placed in room temperature with a temperature of 20 degrees Celsius, the third one was placed in warm water at 40 degrees Celsius, the fourth one was placed in hot water at 60 degrees Celsius, and fifth one was placed in extremely hot water at 80 degrees Celsius. Each test tube was then left in the selected place for ten minutes so that the starch solution could acclimate to its surrounding temperature. After ten minutes, we then used a 1 -ml pipette to add 0. Ml of the amylase mixture to each of the tubes and then the timer was set. After three minutes, we put 3 drops of the starch/amylase mixture from each of the five tubes into the spot plate, and then added two drops of iodine to each spot plate and noted the result. Iodine reacts with starch to change from yellow (not starch) to deep blue black in the presence of starch. After every three minutes had passed, these same steps were repeated, until 30 minutes had passed and noted all the result. Experiment #2: Investigating the Effect of Environmental pH on the Activity of Porcine Pancreatic Amylase Materials and Methods Amylase experiment # 2 was done to see how the pH affected the efficacy f 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 0. 0375 MGM/ml Porcine Pancreatic Amylase Solution (amylase powder in 0. % Nasal ), Iodine Solution; each solution were pipettes 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 ICC. 0. 2 ml of porcine pancreatic amylase solution was then pipettes into each tube. A timer was started and every minutes the starch / amylase mixture were pipettes from each tube and pipettes into the spo t 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 steps ere repeated, until 30 minutes had passed and all the results were noted. Experiment #3: Investigating the Effect of Freezing and of Boiling on the Activity of Porcine Pancreatic Amylase Materials and Methods Amylase experiment #3 was made to see if boiling or freezing the amylase would have an effect on the enzyme. For this experiment, we collected three clean test tubes, porcine pancreatic amylase solution, a test tube containing frozen amylase solution, two 600-ml beakers each containing 200-ml of room temperature water, a hot plate, two transfer pipettes, two spot plates, a bottle of iodine solution, First we took the frozen amylase solution out of the freezer and warmed it to room temp. Then we put about 0. Ml of amylase solution in a test tube and boiled it in at beaker for two minutes. Then we let the beakers sit in room temperature. We labeled each test tube, one boiled and the other frozen. We placed them in water and allowed them to sit for at least ten minutes until it cooled down to room temperature. Then we put 5-ml of starch solution pH 7. 0 in two test tubes and labeled them boiled amylase, or frozen amylase. We used a ml pipette to add 0. Ml of the boiled and frozen amylase solutions to their exceptive test tubes. The solutions were then thoroughly mixed together with a transfer pipette and the timer was set. After three minutes, we did the first test. We took out approximately three drops of starch/amylase solution from all two of the test tubes and put them on a spot plate into two separate wells. Then about two drops of iodine solution were added to each one. This same routine was repeated every three minute intervals until thirty minutes all together had passed. All of these tests were noted prior to each one. Discussion In the first experiment my hypothesis was that the amylase is going to work deter. As the environment gets warmer, and is going to become more active and get more effective. The amylase was put in different temperatures and the results were that amylase best works at a temperature near body temperature. While when the temperature starts decreasing to successes or increase to Celsius the amylase will work, but not at its 100 %, but when temperature reach Celsius or Celsius amylase dont work at all. The results of the experiments showed a significant effect of temperature on the amylase activity. The best temperature for porcine pancreatic amylase activity was ICC; it takes 6 min to gets the starch, as the temperature changed, the amylase activity decreased. When ICC was reached the amylase takes 12 min digesting the starch, also when ICC was reached the amylase took mini digesting the starch. Amylase is affected by environmental PH. I predict that the amylase activity will work best at pH 7. As the pH changes from this point I believe that the amylase activity is going to decrease and eventually stop. In experiment #2 the amylase was added into different pH to determine the optimum pH range. The result was that porcine pancreatic enzymes have best efficiency in the neutral pH 7 range, and does to work at its 100% potential at either between pH 9 and phi. The amylase did not work at all in extremely high or low pH such as phi and pH 11 . The results indicate that in pH 7 the enzyme works at its best. This can indicate that the amylase activity is optimal at pH ranges of the saliva and the small intestine. The two extreme pH values of pH 3 and pH 11 showed no amylase activity. This could be due to the anticipated competition at the enzyme active sites with increased hydrogen ion or hydroxide ion concentrations. Also the attraction of H or OH ions to the protein functional groups can alter the shape of the enzyme and thereby emit its activity. If I boiled or froze some amylase solution, and tried to digest starch with at it at room temperature, I predict the previously-boiled or frozen amylase will not work quite as well as amylase that has not been previously boiled or frozen. In experiment #3 the amylase activity was zero in the boiled sample. This confirms the prediction that freezing the amylase is not going to stop it from functioning, but is not going to work quite as well. In contrast the boiled sample results weakened my hypothesis because the amylase never showed any activity during the thirty min of the experiment. This can be imparted to the results in experiment #1 because amylase had no activity before being boiled, and neither when being in temperatures too high such as ICC.