Enzyme Activity

November 13, 2017 General Studies

LABORATORY REPORT (Click on the Save a Copy button on the panel above to save your report) Activity: Name: Instructor: Date: Enzyme Activity Pam Campbell Id 0002337 Dr. Murphy Nmezi August 9, 2011 Predictions 1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 40 °C (104 °F) 3. Sucrase activity increases with increasing sucrose concentration until a plateau is reached. Materials and Methods Effect of pH on Enzyme Activity. 1. Dependent Variable. amount of product (glucose and fructose) produced 2. Independent Variable. H 3. Controlled Variables. temperature; amount of substrate (sucrose) present; sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity. 1. Dependent Variable. amount of product (glucose and fructose) produced 2. Independent Variable. temperature 3. Controlled Variables. pH; amount of substrate (sucrose) present; sucrase + sucrose incubation time Effect of Substrate Concentration on Enzyme Activity. 1. Dependent Variable. amount of product (glucose and fructose) produced 2. Independent Variable. amount of substrate (sucrose) present 3.

Controlled Variables. temperature; pH; sucrase + sucrose incubation time 1. Describe what is measured as an indicator of sucrase activity and why this is an indicator of sucrase activity. Glucose is measured as an indicator of sucrase activity, as glucose is a producut produce by sucrase activity. 2. Explain why denatured sucrase was used as a control. Denatured sucrase was used as a control to determine if a true effect was made on the active sucrase by pH, temperature, and sucrose, or if affect would be seen in both the denatured and active sucrase. . Alkaline DNS denatures sucrase. Explain why it is important to denature the enzyme. It is important to determine if pH, temperature, and sucrose do have a true effect on active sucrase. Results Table 1: Effect of pH on Sucrase Activity 1 2 3 average ph 3 0. 171 0. 159 0. 166 0. 165 ph 4 0. 309 0. 319 0. 302 0. 31 ph 5 0. 646 0. 658 0. 652 0. 652 Optical Density ph 6 0. 964 0. 925 0. 964 0. 951 ph 7 0. 762 0. 775 0. 778 0. 772 ph 8 0. 401 0. 43 0. 435 0. 422 ph 9 0. 176 0. 15 0. 173 0. 166 Effect of pH on Sucrase Activity 1.

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State the optimum pH for sucrase activity and how sucrase activity changes at more acidic and alkaline pH values. The optimum pH for sucrase activity is 6 and decrease with greater acidic and alkalline values. Table 2: Effect of Temperature on Sucrase Activity 10 °C (50 °F) 0. 006 0. 006 0. 007 0. 006 20 °C (68 °F) 0. 251 0. 285 0. 273 0. 27 30 °C (86 °F) 0. 767 0. 789 0. 799 0. 785 Optical Density 40 °C 50 °C (104 °F) (122 °F) 0. 969 0. 93 0. 947 0. 922 0. 945 0. 947 0. 954 0. 933 60 °C (140 °F) 0. 843 0. 827 0. 829 0. 833 70 °C (158 °F) 0. 641 0. 623 0. 632 0. 632 1 2 3 average

Effect of Temperature on Sucrase Activity 2. State the optimum temperature for sucrase activity and how sucrase activity changes at lower and higher temperatures. The optimum temperature for sucrase activity is 40 degrees Celcius. Sucrase activity decrease at greater or lower temperature and may become denatured. Table 3: Effect of Sucrose Concentration on Sucrase Activity 1 2 3 average 35 g/L 0. 988 1. 001 0. 987 0. 992 30 g/L 0. 983 0. 977 1. 012 0. 991 25 g/L 0. 946 0. 94 0. 943 0. 943 Optical Density 20 g/L 15 g/L 0. 93 0. 861 0. 922 0. 86 0. 908 0. 848 0. 92 0. 856 10 g/L 0. 79 0. 759 0. 774 0. 771 5 g/L 0. 513 0. 523 0. 506 0. 514 0 g/L 0. 003 0. 002 0. 002 0. 002 Effect of Sucrose Concentration on Sucrase Activity 3. State how sucrase activity changes with increasing sucrose concentration. Sucrase activity increase with increasing sucrose concentration until it reaches a level of plateau at 30 g/L. Discussion 1. Explain how pH and temperature affect enzyme activity. Temperature and pH can affect enzyme activity by increase or decreasing the enzymes activity. Optimum enzyme activity occurs at it’s optimum pH and temperature levels. 2.

Compare optimal temperature for sucrase activity to body temperature. Optimal temperature for sucrase activity is 37. 5 degrees Celcius which is the normal body temperature in the small intestine where sucrase is produced. 3. Specifically state where in the intestine sucrase is likely to be most active (pH along GI tract). Sucrase activity is most likely to most active in the duodenum where there is a pH level of 6. 4. Explain how sucrase activity in the intestine is affected by the sucrose content of food. Why is this important? Sucrase activity in the intestine is increase by the sucrose content of food.

This is important due to the formation of glucose which is used by the body for energy. 5. Heat treatment was used to denature the sucrase that was added to the control test tube. In the experimental test tube, alkaline DNS was added to denature sucrase to stop the enzymatic reaction. Discuss what temperature was probably used to denature sucrase and what the probable final pH was of each solution after DNS was added. The probable final pH was a level of 9 due to the alkaline nature of DNS and the temperature was 10 degrees Celcius as Sucrase shows no glucose formation at this temperature. 6.

Restate your predictions that were correct and give the data from your experiment that supports them. Restate your predictions that were not correct and correct them, giving the data from your experiment that supports the corrections. 1. Correct – Optimum pH for sucrase activity was achieved at a pH of 6. 2. Correct -Optimum temperature for Sucrase activity was obtained at 40 degrees Celcius. 3. Correct – Increased sucrose concentration did increase sucrase activity until it reached a plateau at 30 g/L. Conclusion Briefly state how pH, temperature, and sucrose concentration affect sucrase activity. type below ) Temperature, pH and sucrose concentration all have the ability to increase or decrease sucrase activity. Application 1. Myosin ATPase is an enzyme that is involved in muscle contraction. Athletes do warm-up exercises prior to an event to increase athletic performance. Explain why. undefined 2. Metabolic acidosis is a condition that occurs when the body cannot maintain a normal blood pH between 7. 35 and 7. 45, and pH falls below pH 7. 35. Describe the effect of metabolic acidosis on enzyme-catalyzed chemical reactions that occur in the blood. undefined

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