POST-LAB QUESTIONS 4. Facilitating Chemical Reactions with Enzymes 93 Name 1. Would an enzymatic reaction occur faster if you heated the substrates and enzyme? Explain your answer. 2. Predict what effect boiling has on the structure and active site of an enzyme? Explain. 3. Why can't a denatured enzyme convert substrate into product? 4. Explain why a denatured enzyme in a cell could be detrimental. 5. If you did not stop the reaction, would products continue to be produced indefinitely? Explain. 6. Cells often use the product as an inhibitor to slow or stop enzymatic activity. What is advantage to doing so? 7. You used 1 mL of buffer throughout today's experiments. How many liters is 1 mL 8. The spectrophotometer weighs 3.2 kg. How many grams is that?
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Convert 3.2 kg to grams: 1 kg = 1000 g, so 3.2 kg = 3.2 * 1000 g. Show more…
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Temperature Effect on Enzyme Activity When you are done, use the test tube brush provided to wash test tubes. Thoroughly wash them with soap, rinse with deionized water, and place them in the test tube rack so they may drain. Finally, tidy up your work area, making certain all equipment and materials are there for the next class. Questions: 1. Over what temperature range does catechol oxidase remain active? 2. What is the optimum (best) temperature for activity of this enzyme? 3. What happens to enzymes exposed to very high temperatures?
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PART 1: MEASUREMENT OF ENZYME ACTIVITY What color change happened when you mixed the first two tubes? Was the color change instantaneous or gradual? What did the blank tube for this experiment contain; what was missing, and why? When you mixed the enzyme and substrate tubes together the second time, did the absorbance continue to increase with time; or did it plateau? Explain why. Part 2: FACTORS THAT AFFECT ENZYME ACTIVITY Which factor will your group be investigating? Based on your understanding of enzyme activity, predict how this factor will influence the activity of the turnip peroxidase. Table 4: Absorbance Factor tested Time Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 0 sec 10 sec 20 sec 30 sec 40 sec 50 sec
8. Kinetic assays were performed to better understand the activity of the enzyme 245C in the absence and presence of a molecule B. The enzyme follows Michaelis-Menten kinetics: (a) (2 marks) Calculate the Vo for this enzyme when [S] = 0.03 mM and no molecule B is present. (b) (4 marks) Using a properly labeled Lineweaver-Burk plot with accurate X- and Y-intercepts, describe the effect of molecule B on the enzyme. Briefly explain how the molecule could be causing these effects.
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