Maryely Ramirez Professor Theresa Sterner Bio Lab 105 04 November 2013 The Effects Environmental Temperature and pH have on the Activity of Porcine Pancreatic Amylase Introduction There are many types of enzymes and each has a specific job. Enzymes are particular types of proteins that help to speed up some reactions, such as reactants going to products. One of them is the amylase enzyme. Amylases are found in saliva, and pancreatic secretions of the small intestine. The function of amylase is to break down big molecules of starch into small molecules like glucose; this process is called hydrolysis. 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 …show more content…
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 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 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
Enzymes are an organic substance that are made up of polymers of amino acids that help the digestive system and metabolic processes in living organisms (Funk and Wagnalls, 2016). Enzymes are able
The objective of this lab was to develop a protocol to investigate the effect of an environmental variable on the catalytic function of an enzyme. More specifically, the objective was to perform an experiment in order to test the effect of pH on the function of the enzyme catalase.
enzymes that will be used during this lab to test the ability of amylase to break down starch ,a
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
In Table one the solution remained brown since there is no starch present in the mixture in tube 1A.The solution turned blue black due to presence of starch in the solution in tube 2A.The solution remained brown since there is no starch present in the mixture in tube 3A.The solution turned blue black due to presence of undigested starch in the solution. Boiling amylase denatures the enzyme hence it cannot digest amylase in tube 4A.The solution remained brown since there is no starch present in the mixture. All is digested in tube 5A.In tube 6A the solution turned blue black due to presence of undigested starch in the solution. The extremely low temperatures inhibit the action of the enzyme amylase. There was no presence of maltose in tube
The purpose of this experiment is to determine how the change of pH, enzyme concentration, and temperature affect the rate of enzyme reactions. In this experiment, three tests were performed. The first related the effects of different concentrations of the enzyme sucrase on a constant amount of a substrate. The second experiment was used to investigate the changes in enzyme productivity based on the temperature at which the reaction was allowed to occur. The third experiment tests the effect of how the pH of a substance affects the rate at which the enzyme catalyses sucrose.
In this lab, we became familiar with the importance of thermoregulation and the effects of temperature on enzyme functions. In this exercise, we prepared five solutions. Four of the five solutions were mixed with amylase (saliva), HCl, starch, and iodine and potassium iodine. The solutions were placed in different temperatures and we used a spectrophotometer to determine the each absorbance readings. A low absorbance indicates a low presence of starch and high enzymatic function. A high absorbance indicates a high presence of starch and low enzymatic function.
Effect of varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and hydrolysis of Starch
Enzymes are catalysts that affect the rate of a reaction without itself being affected or altered. In this experiment, the enzyme being experimented with is amylase. This enzyme can be found in human saliva and helps to hydrolyze starch into maltose. There are three different parts to this experiment; concentration, pH, and temperature. Each is to test the presence of amylase and the different affects it has on certain substances. Concentration is to determine what effect the variation of will have on the rate of the reaction, pH to determine the optimum pH for the activity of amylase, temperature is manipulated to see how amylase would react in different degrees of temperature and how active it is.
Introduction: Enzymes are specifically folded proteins which catalyse biological reactions in living organisms, they consist of an active site in which specific substrates can enter and a reaction takes place. As the enzyme does not permanently change structure, meaning the active site stays the same it allows multiple substrates to complete the reaction through single enzymes. The substrates are complementary to the active site of the enzyme and can be synthesised or degraded in order to create new products useful to the organism. Enzymes enable the substrates to be orientated in a way that allows for the reaction to take place more readily without relying on the chance of accurately positioned collisions of particles with sufficient energy.
The aim of this experiment was to investigate the effect that different pH levels had on the rate of activity of the enzyme ⍺-amylase. It was hypothesised that pH levels below and above 7 would result in a consistent decrease in the ability of the amylase to break starch down into glucose, and therefore the rate of activity of amylase and the amount of glucose in the solution, until the amylase became denatured to the point at which it no longer could react with the starch, at which point the reaction would stop, and further changes in pH level would have no effect. This is because ⍺-amylase’s natural environment, in saliva or near the pancreas, has a pH level of approximately 7 (Worthington-biochem.com, 2017). As such, any notable shift from a pH level of 7 will result in increasing levels of
The purpose of the experiment was to change the pH of the enzymes and environment to see if there is a significant change in the enzyme’s activity. Test tubes were labeled with their designated pH containing starch ranging from 3 to 8. The amylase enzyme was then added to each test tube and the reaction occurred for ten minutes. Introducing HCL to the solution then stopped the reaction, and its absorbance was taken. The absorbance of the solution determines how active the enzyme was. If the absorbance number is high, then the enzyme was not as active because there is excess starch. When the absorbance reading is low that means the enzyme is being productive and is breaking down the starch at a relatively fast rate. The control is absorbance
For the investigation of enzymes, I am going to conduct an experiment to see how temperature can affect the rate of reaction of enzymes by testing it with starch.
Using a reaction plate, two drops of starch solution were added to three successive walls. To well number one, 8 drops of boiling water was added. To well number 2, 8 drops of room temperature water was added. Lasty, 8 drops of ice water was added to the third successive wall. Two drops of amylase was mixed into each of the three walls. After three minutes, by looking for the presence of starch, one drop of iodine was added to each well. As the water condition was boiling, the solution was clear brown. At room temperature, it was dark brown. When the water was ice, it was simply brown. Temperature does not predominantly influence amylase activity, because the temperature didn’t drastically change the colors. At all three temperatures, all of the solutions remained some sort of brown. The boiling whatever, however should have denatured the amylase; therefore, the amylase should contain starch and turn dark black. If someone were to wait too long before adding amylase, temperature of the solution could have changed. This could have been a possible defect of the boiling water. The optimal temperature of amylase is 37 degrees C. The optimum pH is the neutral one because your mouth has a 7 pH. As the reaction progresses, less starch will be present and more sugar (maltose) will be
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.