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How can we tell if the enzyme has been denatured by a particular temperature treatment.

How can we tell if the enzyme has been denatured by a particular temperature treatment.

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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How can we tell if the enzyme has been denatured by a particular temperature treatment.
oft Word - Lab 5 Enzymat X Download Firefox Browser- Fas X /Downloads/Lab%205%20Enzymatic%20Activity%20(9).pdf Getting Started TV shows ted From Fire Swift - Login Vision Loss Resourc Experiment 3: Determining the Optimal Temperature for Amylase Activity Because enzymes are proteins, and rely on their three-dimensional shape in order to function correctly, changing the three-dimensional shape of the enzyme will alter its ability to catalyze reactions. High temperatures can denature an enzyme by breaking hydrogen bonds, thereby preventing the enzyme from working correctly. In addition, the effectiveness of enzymes at catalyzing reactions is affected by temperature in other ways. Temperatures that are too low might prevent substrate molecules for moving into the active site, reducing the rate of catalysis. In temperatures that are too warm, but not hot enough to denture the enzyme, kinetic energy might be too great, and could cause the substrate to move out of the active site before catalysis can occur. In this experiment, you will try to determine the optimal temperature for amylase activity. In order to investigate this question, you will have to maintain the substrate at a variety of different temperatures for the course of the reaction. To do this, you will place your test tube with substrate into a beaker of water at a set temperature. After two minutes (during which time the temperature of the substrate will increase or lower to the target temperature), you will add your enzyme. As in previous experiments, record the time to reaction completion. It will be important to keep the reaction tube in the beaker of water so its temperature remains constant. To investigate the effect of temperature on amylase activity, we place 6 ml of 0.5% amylose solution and 70ul of 0.5% amylase enzyme solution in test tubes, and incubated them in water baths of various temperatures until they reached equilibrium. The temperatures we investigated were 5 C, 15 C, 25° C, 35° C, 45 C, 55 C, 65' C, 75° C, 85 C, When all fluids were at the target temperature, we mixed the contents of the tubes, but kept the tube at the specified temperature as the reaction progressed. At 15 second intervals, we tested the reaction mixture for the presence of amylose starch by dropping the mixture into a well containing a drop of Lugol's iodine. We compared our results against the following: a well that contained only amylase solution and iodine, a well containing only enzyme solution and iodine, and a well containing completely digested (or broken-down) starch solution and iodine. We recorded the time of completion of the reaction for each of our trials, when no more starch was present, as the first sample that did not show a positive starch reaction. If the reaction did not reach completion in 9 minutes, a time of >540 seconds was recorded, and notes were taken about whether or not some breakdown of starch appeared to have happened.
Transcribed Image Text:oft Word - Lab 5 Enzymat X Download Firefox Browser- Fas X /Downloads/Lab%205%20Enzymatic%20Activity%20(9).pdf Getting Started TV shows ted From Fire Swift - Login Vision Loss Resourc Experiment 3: Determining the Optimal Temperature for Amylase Activity Because enzymes are proteins, and rely on their three-dimensional shape in order to function correctly, changing the three-dimensional shape of the enzyme will alter its ability to catalyze reactions. High temperatures can denature an enzyme by breaking hydrogen bonds, thereby preventing the enzyme from working correctly. In addition, the effectiveness of enzymes at catalyzing reactions is affected by temperature in other ways. Temperatures that are too low might prevent substrate molecules for moving into the active site, reducing the rate of catalysis. In temperatures that are too warm, but not hot enough to denture the enzyme, kinetic energy might be too great, and could cause the substrate to move out of the active site before catalysis can occur. In this experiment, you will try to determine the optimal temperature for amylase activity. In order to investigate this question, you will have to maintain the substrate at a variety of different temperatures for the course of the reaction. To do this, you will place your test tube with substrate into a beaker of water at a set temperature. After two minutes (during which time the temperature of the substrate will increase or lower to the target temperature), you will add your enzyme. As in previous experiments, record the time to reaction completion. It will be important to keep the reaction tube in the beaker of water so its temperature remains constant. To investigate the effect of temperature on amylase activity, we place 6 ml of 0.5% amylose solution and 70ul of 0.5% amylase enzyme solution in test tubes, and incubated them in water baths of various temperatures until they reached equilibrium. The temperatures we investigated were 5 C, 15 C, 25° C, 35° C, 45 C, 55 C, 65' C, 75° C, 85 C, When all fluids were at the target temperature, we mixed the contents of the tubes, but kept the tube at the specified temperature as the reaction progressed. At 15 second intervals, we tested the reaction mixture for the presence of amylose starch by dropping the mixture into a well containing a drop of Lugol's iodine. We compared our results against the following: a well that contained only amylase solution and iodine, a well containing only enzyme solution and iodine, and a well containing completely digested (or broken-down) starch solution and iodine. We recorded the time of completion of the reaction for each of our trials, when no more starch was present, as the first sample that did not show a positive starch reaction. If the reaction did not reach completion in 9 minutes, a time of >540 seconds was recorded, and notes were taken about whether or not some breakdown of starch appeared to have happened.
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