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Amylase Investigation Plan (D) Aim: To investigate how Enzymes (bacterial Amylase) behave under varied temperature conditions during enzyme reaction with a substrate solution of starch. Hypothesis: I predict that the rate of enzyme reaction will increase as the temperature rises but after optimum temperature is reached the reaction will no longer produce maltose because the enzyme's bonds will break and lose its shape causing it to denature. I also predict that when the temperature is high (approximately 40 degrees Celsius), the rate of enzyme reaction is twice as faster than it is when the temperature is low (approximately 5 degrees Celsius) because of its activation energy. The heats in the enzymes surroundings supply it with energy so it becomes activated so that its reactions can start off. At increasing temperature it provides more energy for collisions with the substrate and for reactions to occur at a high rate. But after optimum temperature is reached, the iodine will no longer show any differences that would indicate that there's a lack of starch present and no maltose is indicated through its colour indication. Beyond optimum temperature, the rate of reaction denatures the enzyme because the structure loses its shape and the substrate can't fit into the enzyme.
* Add the bacterial amylase into the solution and start timing immediately * After every 15 seconds a single droplet of the solution should be transferred onto the dimple tray with a pipette * This process should be done consecutively in rows (for every 15 seconds) of the dimple tray until the iodine indicator turns from blue/black to a chocolate brown colour indicating that there is a lack of starch present. * Use three trials for each temperature value. Data collection and processing (DCP): Temperature at which the starch indicator shows the presence of lack of starch average Temperature at 5?C average Temperature at 15?C average Temperature at 25?C average Temperature at 35?C average Temperature at 45?C average Temperature at 55?C average Temperature at 65?C average Temperature at 75?C average Temperature at 80?C time (s) 415 330 235 210 150 130 65 50 15 Average was found finding the total of each three trials and then dividing it by three. Trends and pattern from Graph: The general trend seen from this graph is that when temperature is increased, the rate of bacterial amylase reaction is much faster and works more effective. The substrate and enzyme is more likely to attach to each other as the temperature increases.
The results obtained could be tested for 15 seconds per result and this was slightly ineffective because it had not shown the exact time at which the enzyme had been processing with the active site. A more effective procedure would be if the iodine was directly put into the test tube of starch and amylase solution right after the amylase is put into the test tube. The amount of time taken for the reaction to occur can be measured exactly from reaction when the indicator changes color at the given temperature. Improving the investigation To measure the exact amount of time for the reaction to happen in order get more accurate results, a different method would have been more useful like the one indicated above. However, a color indication comparison paper (just like pH scales) for iodine solution would have been more effective in the process of comparing the color after every 15 seconds for the experiment. If I did the experiment again, I would also have used more than 3 trials even though the error bars from the graphical results obtained were close together from the maximum and minimum values. Philip Mannas 11/30/2008 IA: Biology HL --1--
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