Biology Essay Research Paper Investigation on factors free essay sample

Biology Essay, Research Paper Probe on factors impacting the enzyme Amylase Purpose: An probe into the factors impacting the activity of the enzyme Amylase # 8211 ; Planing Prediction: I predict that as the temperature increases, the velocity of the reaction will increase. When a peculiar temperature is reached, I believe the rate of reaction will dramatically diminish. I believe this because most chemical reaction happens faster when the temperature is higher. At higher temperatures molecules mover around faster, which makes it easier for them to respond together? Normally, a rise of 10 grades celius will duplicate the rate of reaction. This is true for enzymes up to about 40 grades celius. However at 40 grades celius the enzyme begins to be damaged, so the reaction slows. By 60 grades celius, the enzyme is wholly denatured. I predict that the same will go on the further off the pH is from pH 7. I believe this because the optimal temperature for most enzymes is about pH 7 therefore the farther off from pH 7 ( either more alkalic or more acidic ) the less affectional the enzyme. Pilot Experiment: To look into the factors impacting the activity of the enzyme Amylase, I will accommodate a pilot experiment, which investigates if the enzyme amylase interruptions down amylum. The pilot experiment was: 1. Pour amylase solution into a trial tubing to a deepness of 2cm. 2. Half fill another trial tubing with a 4 % starch solution. 3. With a pipette, topographic point a bead of I into each pregnant chad in a dimple tray. 4. With a glass rod lift a bead of the starch solution from the trial tubing and blend it with the first bead of I in the first pregnant chad in the tray. A blue/black coloring material should develop ; this will be used as the control. 5. Rinse the glass rod. 6. Pour amylase solution into the trial tubing of amylum and agitate rapidly. 7. Repeat steps 4 A ; 5 ( for the amylase A ; starch solution mixture ) every 30 seconds until a blue/black coloring material no longer develops. 8. When there is no farther alteration in the coloring material of the I, take the starch-amylase trial tubing, add Benedict s reagent, and topographic point in the H2O bath for 1 minute. Adapted Experiment: I will modify and spread out the pilot experiment in a figure of ways. First, I must make up ones mind what I am traveling to look into. I am traveling to look into the consequence of pH and temperature on the activity of the enzyme amylase. Therefore, I have developed two similar experiments ( one for each factor I am look intoing ) . To look into the consequence of pH on the activity of the enzyme amylase: 1. Pour amylase solution into a trial tubing to a deepness of 2cm. 2. Half fill another trial tubing with a 4 % starch solution. 3. With a pipette, topographic point a bead of I into each pregnant chad in a dimple tray. 4. With a glass rod lift a bead of the starch solution from the trial tubing and blend it with the first bead of I in the first pregnant chad in the tray. A blue/black coloring material should develop ; this will be used as the control. 5. Rinse the glass rod. 6. Add 2cm3 of the appropriate pH buffer to the amylum solution and shingle. 7. Pour amylase solution into the trial tubing of amylum and agitate rapidly. 8. Repeat steps 4 A ; 5 ( for the amylase, amylum A ; pH buffer mixture ) every 30 seconds until a blue/black coloring material no longer develops. 9. Record the consequences in a tabular array. Repeat steps 1-8 for each pH buffer scope until all the provided pH buffers have been used. Temperature: To look into the consequence of temperature on the activity of the enzyme amylase: 1. Pour amylase solution into a trial tubing to a deepness of 2cm. 2. Half fill another trial tubing with a 4 % starch solution. 3. Cool both trial tubing and maintain at 10 grades celius 4. With a pipette, topographic point a bead of I into each pregnant chad in a dimple tray. 5. With a glass rod lift a bead of the starch solution from the trial tubing and blend it with the first bead of I in the first pregnant chad in the tray. A blue/black coloring material should develop ; this will be used as the control. 6. Rinse the glass rod. 7. Add 2cm3 of H2O to starch solution ( see below ) . 8. Pour amylase solution into the trial tubing of amylum and agitate rapidly ( step temperature, seek to keep at 10 grades celius ) . 9. Repeat steps 4 A ; 5 ( for the amylase A ; starch solution mixture ) every 30 seconds until a blue/black coloring material no longer develops. 10. Record the consequences in a tabular array. Repeat steps 1-9 increasing the temperature by 5 grades celius each clip until the enzyme is denatured, this should be around 40 grades celius ( see portion 6 on enzymes. ) To guarantee that the trial is just I will merely change two factors ( at different phases of the experiment non at the same time ) . All the measures will hold to be carefully measured since little fluctuations in the sum of enzyme used can do important fluctuations in the consequences. In the subdivision where temperature is investigated, 2cm3 of H2O is added. This is because the amylum solution is impersonal and in the old probe ( for pH ) 2cm3 of the appropriate buffer was added. If the 2cm3 of H2O was non added so it would non be a just trial since the volumes used in each portion of the probe would be different. If they were different so this would impact the consequences since the solution would be of different concentrations and hence one would respond faster than the other would. Apparatus: I have chosen to utilize a measurement cylinder to mensurate the volumes of substances used since it is more accurate than a pipette. I will utilize an electronic H2O bath for keeping the mixture at a temperature above room temperature since the temperature is more accurate than a H2O bath above a Bunsen burner. I will hold to utilize ice from the deep-freeze to cut down the temperature of the mixture to 10 grades celius. A 100 grades celius thermometer will supply temperature consequences of a sufficient truth ( to 1 grades celius ) . The pH buffer scope will be pre-prepared therefore I do non hold to concern myself with mensurating and keeping pH degrees. Variables: I have chosen to reiterate the experiment 3 times because it hence allows me to cipher an mean clip. This will guarantee that there are no unnatural consequences and it will increase truth. I have decided to get down the temperature at 10 grades celius and increase by 5 grades celius each clip since it will let me to see the addition and lessening of the enzyme activity. It should besides be accurate plenty for me to foretell an optimal operating temperature to an truth of 5 grades celius. I have no control over which pH buffers I will utilize since the school prior to the experiment will supply them. Enzymes: Substances called accelerators speed up many chemical reactions. Catalysts called enzymes command the metabolic reactions in the organic structure. Amylase is an enzyme ; it is present in the digestive systems of many animate beings. Amylase speeds up the dislocation of long concatenation amylum molecules in smaller ironss of malt ose. Enzyme molecules have a really precise 3-dimensional form. This includes a dent, which is called the active site. It is precisely the right size and form for enzyme s substrate to suit into ( in the instance of amylase this is starch ) . When a substrate molecule slots into the active site, the enzyme tweaks the substrate molecule, drawing it out of form and doing it split into merchandise molecules. High temperatures make enzymes inactive: this is because they are proteins, which are damaged by temperatures above approximately 40 grades celius. Most enzymes work best at a pH of about 7. This is besides because they are proteins, which are damaged by really acidic or really alkalic conditions. Due to the enzyme s alone active site it can merely change over one sort of substrate molecule into one sort of merchandise. Consequences: First of all the mean clip taken for the amylum to be digested in each status was calculated. Temperature ( grades celius ) 10 21 32 40 50 55 60 67 80 A ; 90 Average Time ( min ) 9.8 6.7 5.3 5.5 3.5 3.0 3.5 4.5 10 + pH 8 7 6 5 3 2 Average Time ( min ) 9.5 6.7 4.5 2.8 4.7 9.2 The consequences were so plotted on a graph ( see Graphs 1 A ; 2 ) and the points joined together. From graph 1, I can see that as the temperature of the mixture increases, the clip taken for the amylase to digest the amylum decreases. This happens fastest at 55 grades celius, nevertheless if the temperature continues to lift so the clip taken for the amylase to digest the amylum quickly increases. By the clip 80 OC is reached the amylase does non digest the amylum. From graph 2, I can see that as the pH of the mixture increases, the clip taken for the amylase to digest the amylum decreases. This happens fastest at pH 5. At a higher pH degree than pH 5 the clip taken for the amylase to digest the amylum additions. Amylase speeds up the dislocation of long concatenation amylum molecules into smaller ironss of malt sugar. Enzyme molecules have a really precise 3-dimensional form. This includes a dent, which is called the active site. It is precisely the right size and form for the enzyme s substrate to suit into ( in the instance of amylase this is starch ) . When a substrate molecule slots into the active site, the enzyme tweaks the substrate molecule, drawing it out of form and doing it split into merchandise molecules. High temperatures make enzymes inactive: this is because they are proteins, which are damaged by temperatures above approximately 40 grades celius. In this probe it was found that amylase operated fastest at 55 OC and was damaged above this temperature. The enzyme is damaged because the molecules are traveling faster, these bombard the active site of the enzyme altering its form ; when it s form has been changed so the amylum will no longer suit in the active site and hence can non be digested. This is 15 OC above the suggested typical temperature, see Measuring Evidence for possible grounds why it is higher than expected. By 80 grades celius the amylase was wholly denatured. From this information I can reason that the optimal operating temperature for the enzyme amylase is 55 grades celius. Most enzymes work best at a pH of about 7. This is besides because they are proteins, which are damaged by really acidic or really alkalic conditions. However in this experiment the amylase worked fastest at pH 5. This pH is lower than suggested above because the amylase provided by the school was a bacterial enzyme which had an optimal pH between pH 4 and pH 5. Decision: In my design, I predicted that as the temperature increases, the velocity of the reaction would increase. When a specific temperature was reached, I believed that the rate of reaction would dramatically diminish. I believed this because most chemical reactions happen quicker when the temperature is higher. At higher temperatures molecules move about faster, which makes it easier for them to respond together. Normally, a rise of 10 grades celius will duplicate the rate of reaction. This is true for enzymes up to about 40 grades celius. However at 40 grades celius the enzyme begins to be damaged, so the reaction slows down. By 60 grades celius the enzyme is wholly denatured. I predicted that the same would go on the further off the pH is from pH 7. I believed this because the optimal temperature for most enzymes is about pH 7 therefore the farther off from pH 7 ( either more alkalic or more acidic ) the less effectual the enzyme. My consequences support most of the anticipation since as the pH/temperature increases the rate of reaction additions until a point is reached and so the rate of reaction lessenings. However, the optimal temperature was non 40 OC but 55 OC ( remarkably high ) and the optimal pH was pH 5 non pH7. I besides predicted that a rise of 10 OC would duplicate the rate of reaction ; this was non right. As is apparent in the tabular array above, the clip taken does non halve for every 10 grades celius addition in temperature. I could non happen a additive relationship between the temperature and rate of reaction, nevertheless with adequate consequences ( from a really big scope of temperatures ) a parabola could be made on Graph 1, it would so be possible to happen a mathematical expression for the building of the parabola and hence find the relationship between the temperature and rate of reaction. At current I do non hold plenty dependable grounds to pull any decisions more specific than the rate of reaction is relative to the temperature until 55 grades celius is reached and after this point the temperature is reciprocally relative to the rate of reaction. Note: # 8220 ; Optimum runing temperature/pH # 8221 ; means this is the temperature/pH where the enzyme ( amylase ) operates the fastest. I believe that the experiment was successful but some of the consequences were unexpected/unreliable. The clip taken for the amylase to digest the amylum at 40 OC was far excessively fast ( see Graph 1 ) it should hold been between 4 and 4.5 proceedingss. All the other consequences seemed to suit into the tendency on the graphs. I believe that the experiment was designed good but there were a few jobs. The optimal temperature for the amylase was excessively high. I believe that all the consequences were skewed because the enzyme was non given adequate clip at each peculiar temperature to be to the full affected before it was added to the amylum. Because of clip restraints, they were merely left in the H2O bath for 10 proceedingss before get downing the experiment. However, they should hold been left in the H2O bath for about 30 proceedingss so that the amylase had been wholly affected by the temperature before the experiment was started. I decided to carry on the experiment at 10 OC intervals alternatively of 5 OC because there was non adequate clip. When the consequences were collected, I plotted them on a unsmooth graph to happen the optimal temperature and so carry on the experiment at this temperature to guarantee it was the optimal temperature. I besides conducted all three experiments for each status at the same clip to salvage clip. Additional work, which could be carried out, is to reiterate the experiment utilizing, a wider scope of temperatures and pH degrees, and a scope of different amylum solution concentrations or utilizing different enzymes such as peptidase with a protein. 316