Laredo Community College

BIOLOGY 1308

Enzyme Kinetics

1)What is the function of a spectrophotometer?

2. Which wavelength is optimal to measure NADH?

3. Define the absorbance value displayed on the spectrophotometer?

4. What is included in the master mix?

5. Why don’t we add the enzyme into the mix?

6. What role does ethanol play in this reaction? What is the function of NAD+ in this reaction?
7. Why do we need buffer in the master mix?

8. Enzymes are biological catalysts, what aspect of a reaction do they lower, thereby increasing the reaction rate dramatically.

9. What is the substrate concentration in this mix?

10. What is the function of the NAD+ factor?

11. What was just shown in the animation?

12. When the reaction produces no intermediates, which of the following rates constants will be equal to k(eat)?

13. You are given ethanol with a stock concentration of 50 (lrm). Calculate the ethanol colume for two masters mixes with final substrates concentrations of 0mM and 0.0053mM in a final volume of 1mL.

14. What is the optimal enzyme concentration?
15. What is the optimal pH for the alcohol dehydrogenase enzyme?

16. What is the optimal temperature for the alcohol dehydrogenase enzyme?

17. The y-axis of the process curve shows the absorbance value. How can we convert this into product concentration?

18. What values do we need to calculate to plot a Michaelis-Menten curve? What is the estimated V(max) for wild type ADH?
19. And what is the estimated for mutant ADH?  Looking at the plot, which one is Km? What is the Km for wild type ADH?
20. And what would be the Km for the mutant ADH?
21. What could explain the difference observed in the two enzymes?
22. Which plot is useful in determining the type of inhibition taking place?

23. What kind of inhibition is Hydroxylamine?
24. In competitive inhabitation how are the V(max)app and the Km(app) affected?

25. What is the value for the hydroxylamine inhabitation?

26. How can you calculate the Ki for competitive inhibition?

27. What is the Ki value for Hydroxylamine?  

28. What kind of inhibitors is cimetidine?
29. In noncompetitive inhibition, the is … and the Km(app) is….