Analysis Assignment 6
1. Using the data of measured absorbance vs. time (min) for each substrate concentration, determine the slope of the linear portion of this curve. The value of the slope is the initial velocity (V_{o}) in Abs/min. Convert AU/min to mM/sec using the extinction coefficient you calculated for 5TNB. Use these values to construct a table in Excel with the following column titles.
Trial # 
Sample # 
[E] mM 
[S] mM 
1/[S] mM^{1} 
V_{o} (mM/sec) 
1/V_{o} (sec/mM) 
2. Plot the initial velocity as a function of substrate concentration. This is a MichaelisMenton plot. Fit the curve the best way you can. Estimate V_{max} and K_{M} from this plot. Note: The curve may not level off to give an accurate V_{max}, but please try your best. Print a copy of this plot with proper labels. Add additional curves for each additional substrate.
3. Plot (1/V_{0}) as a function of (1/[S]). This is a Lineweaver  Burk plot or a double reciprocal plot. The data should fit a straight line. Calculate V_{max} and K_{M} from this plot. Print a copy of this plot. Complete plots and calculations for each additional substrate.
4. Complete the table on the back of this page for easy comparison. Use additional tables if needed.
5. Write a descriptive paragraph interpreting your results. Relate the numbers to your hypothesis (as best you can) and to the mechanism for specificity and chemistry of the enzyme substrate interaction. Remember our goal is not to just generate numbers, but to give these numbers meaning in terms of how the enzyme shows specificity and enhances the rate of the reaction.
6. Turn in your analysis by Friday at 5:00.
Name
Substrate:

MichaelisMenton 
Lineweaver Burke 

Enzyme 
Conc. (mM) 
pH 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 





















Substrate:

MichaelisMenton 
Lineweaver Burke 

Enzyme 
Conc. (mM) 
pH 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 





















Substrate:

MichaelisMenton 
Lineweaver Burke 

Enzyme 
Conc. (mM) 
pH 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 
Km (mM) 
Vmax (mM/sec) 
kcat (sec^{1}) 




















