Table 3: Absorbance values per minute for varied substrate concentrations with Inhibitor

	Time (min)
Concentration (mM)		2.0	5.0	8.0	12.0	20.0
	0	0	0	0	0	0
Absorbance	1	0.01	0.03	0.04	0.045	0.05

Table 4: Absorbance values versus substrate concentration at a constant time with Inhibitor

Concentration (mM)	2.0	5.0	8.0	12.0	20.0
Absorbance	0.001	0.02	0.03	0.05	0.08

1. Create a scatter plot of the data in Table 3. This graph compares the absorbance values over time (or product formation over time. The graph should contain a line for each concentration of -galactosidase with inhibitor present that was measured. Obtain the equation for each line and the R² value. Include this graph in your data section.
2. Using concentrations and the corresponding absorption values from Table 7, create a graph of absorbance vs. concentration. Obtain the equation for each line and the R² value. Include this graph in your data section.
3. Determine the rate of reaction for each concentration with inhibitor. You can obtain the rate of reaction (initial velocity) by taking the slope of 2 mM obtained from the graph of Table 3 and dividing by the slope of the line from Table 4 (repeat for each concentration.) This is the rate of reaction for each concentration.
4. Using the information from question 4, create a graph comparing velocity of -galactosidase and ONPG concentration. This is the Michaelis-Menten plot. Remember the units for velocity are mM/min.
5. Create a Lineweaver-Burk plot by taking the reciprocal of both axes from the Michaelis-Menten values from question 3.
6. Based on the graphs constructed in Questions 4 and 5, compare velocity of -galactosidase and ONPG concentrations with 250 M inhibitor present. Determine the K_m and V_max for the reaction by visually observing the Michaelis-Menten plot. Compare this qualitative data by calculating Km and Vmax using the slope from the Lineweaver-Burk plot.
7. Utilizing the data from week 1 (without inhibitor) create a combined Michaelis-Menten plot that includes data for both the uninhibited reaction and the inhibited reaction. Repeat this to make a combined Lineweaver-Burk plot as well.
8. Determine if the inhibitor used is a competitive, uncompetitive, and non-competitive inhibitor and explain.
9. Propose a mechanism under which the type of inhibitor determined in Question 9 could interact with the enzyme. How are you convinced?