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LABORATORY REPORT (Click on the Save a Copy button on the panel above to save your report) Activity: Enzyme Activity Predictions 1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 60 ??C (140 ??F) 3. Sucrase activity decreases with increasing sucrose concentration. Materials and Methods Effect of pH on Enzyme Activity. 1. Dependent Variable. amount of product (glucose and fructose) produced 2. Independent Variable. pH 3. Controlled Variables. temperature; amount of substrate (sucrose) present; sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity. . 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.
Amount of product (glucose and fructose) produced is what is being measured for this lab. This is an indicator of sucrase activity because the amount of product produced is a result of the expiriment itself. 2. Explain why denatured sucrase was used as a control. Denatured sucrase was used as a control because activated sucrase would increase the activity. 3. Alkaline DNS denatures sucrase. Explain why it is important to denature the enzyme. It is important to denature the enzyme because leaving it active would increase the activity of the enzyme. Results
Table 1: Effect of pH on Sucrase Activity Optical Density ph 3 ph 4 ph 5 ph 6 ph 7 ph 8 ph 9 1 0. 156 0. 292 0. 653 0. 935 0. 782 0. 429 0. 151 2 0. 165 0. 321 0. 666 0. 92 0. 763 0. 429 0. 151 3 0. 188 0. 322 0. 674 0. 942 0. 821 0. 421 0. 151 average 0. 17 0. 312 0. 664 0. 932 0. 789 0. 426 0. 151 Effect of pH on Sucrase Activity 1. State the optimum pH for sucrase activity and how sucrase activity changes at more acidic and alkaline pH values. Glucose of sucrase is the highest at pH value 6. When acidic or basicsucrase activity becomes less active.
Table 2: Effect of Temperature on Sucrase Activity Optical Density 10 ??C (50 ??F) 20 ??C (68 ??F) 30 ??C (86 ??F) 40 ??C (104 ??F) 50 ??C (122 ??F) 60 ??C (140 ??F) 70 ??C (158 ??F) 1 0. 01 0. 276 0. 757 0. 964 0. 906 0. 835 0. 636 2 0. 009 0. 291 0. 777 0. 959 0. 942 0. 835 0. 64 3 0. 009 0. 263 0. 764 0. 944 0. 933 0. 8 0. 648 average 0. 009 0. 277 0. 766 0. 956 0. 927 0. 823 0. 641 Effect of Temperature on Sucrase Activity 2. State the optimum temperature for sucrase activity and how sucrase activity changes at lower and higher temperatures.
Optimum temperature for sucrase activity is 40 degrees C. Sucrase activity decreases and becomes denatured at lower and higher temperatures. Table 3: Effect of Sucrose Concentration on Sucrase Activity Optical Density 35 g/L 30 g/L 25 g/L 20 g/L 15 g/L 10 g/L 5 g/L 0 g/L 1 0. 991 1. 015 0. 942 0. 892 0. 83 0. 777 0. 524 0. 002 2 0. 993 0. 998 0. 944 0. 898 0. 83 0. 753 0. 507 0. 003 3 0. 97 1. 004 0. 948 0. 899 0. 856 0. 756 0. 53 0. 002 average 0. 985 1. 006 0. 945 0. 896 0. 839 0. 762 0. 52 0. 002 Effect of Sucrose Concentration on Sucrase Activity 3.
State how sucrase activity changes with increasing sucrose concentration. Sucrase activity increases with increasing sucrose concentrations. Discussion 1. Explain how pH and temperature affect enzyme activity. Increasing temperature means the molecules gain more kinetic energy and move faster resulting in more chances of succesful collisions of enzymes and substrates forming enzyme-substrate molecules. Since pH is one of the factors for the denaturation of proteins, if an enzyme is submitted to a pH level under which it is denatured there will be no enzymatic activity. 2.
Compare optimal temperature for sucrase activity to body temperature. Optimal temperature for sucrase activity is 104 degrees F and normal body temperature is 98. 6 degrees F. The human body would not be able to sustain a temperature of 104 degrees F for any length of time. 3. Specifically state where in the intestine sucrase is likely to be most active (pH along GI tract). Small intestine (duodenum) is where the sucrase is likely to be most active. 4. Explain how sucrase activity in the intestine is affected by the sucrose content of food. Why is this important? Sucrase activity in the ntestine is affected by sucrose content in food as most foods contain some sucrose. Adding sucrose will increase sucrase activity. 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. Heat treatment was most likely used as temperatures too high would cause the enzymatic reaction to decrease.
The probable temperature that was used would most likely be 70 degrees C. 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. I predicted that sucrase will have the greatest activity at pH 6 based on Table 6 of data collected I was correct. I predicted that sucrase will have the greatest activity at 60 degrees celcius, but based on my results for Table/graph 2 it will be at 40 degrees.
I predicted that sucrase activity will decrease with increasing sucrose concentrations; based on the results on Table/graph 3 it will inrease with increasing sucrose concentrations. Conclusion Briefly state how pH, temperature, and sucrose concentration affect sucrase activity. ( type below ) Temperature, pH, and sucrose concentration affect sucrase activity in similar ways. Temperatures and pH, caused sucrase activity to decrease. Sucrose concentration affects sucrase activity as increasing amounts of sucrose will increase 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. Athletes do warm-up exercises prior to an event to increase blood flow and they stretch to prevent any muscle tearing. 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. Metabolic acidosis would cause the enzyme-catalyzed chemical reaction to decrease.