Modeling Enzyme Kinetics

Excel spreadsheet Created by Jon Monroe, Department of Biology James Madison University (monroe

Instructions:
1. Enter values for the apparent Km and Vmax for two enzymes (black and red) in the bllue spaces below.
2. Click on the graphs or save the file to recalculate the graphs.
3. The spreadsheet will then use the Michaelis Menton equation below to plot V vs. [S] curves in the first graph.
4. The spreadsheet will then calculate the inverse data and plot those data in a Lineweaver Burke

Black enzyme Red enzyme

Vmax: 4 4 Michaelis Menton equation:

Apparent Km: 1 5

4.5
4.0
V vs. [S] Plot
[S] rate (V) rate (V) 3.5
0.5 1.3 0.4
3.0
1 2.0 0.7
2.5
V

2 2.7 1.1
4 3.2 1.8
2.0
6 3.4 2.2
10 3.6 2.7 1.5
20 3.8 3.2 1.0
30 3.9 3.4
50 3.9 3.6 0.5
100 4.0 3.8 0.0
0 20 40 60 80 100
[S]
80
Lineweaver Burke Plot 2.
4 0
2.
1/V

1/[S] 1/ (V) 1/ (V)

2.00 0.75 2.75
00
1.00 0.50 1.50 2.
0.50 0.38 0.88
60
0.25 0.31 0.56 1.
0.17 0.29 0.46
20
0.10 0.28 0.38 1.
0.05 0.26 0.31 80
0.03 0.26 0.29 0.
0.02 0.26 0.28 40
0.01 0.25 0.26 0.
-1.00 -0.50 . 000.00 0.50 1.00
0
1/[S]
 8
0.
40
0.
00
-1.00 -0.50 0. 0.00 0.50 1.00

1/[S]
netics
Madison University (monroejd@jmu.edu)

the bllue spaces below.

[S] curves in the first graph.

ta in a Lineweaver Burke plot at the bottom.

Vmax * [S]
Rate = -------------
Km + [S]

0 60 80 100
[S]

0.50 1.00 1.50 2.00

1/[S]
0.50 1.00 1.50 2.00

1/[S]
[S] (moles/litro) V (umol/minuto)
3 10.4
5 14.5
10 22.5
30 33.8
90 40.5
 V (mM/minuto)
[S] (mM) Sin inhibidor Con inhibidor
1.25 1.72 0.98
1.67 2.04 1.17
2.5 2.63 1.47
5 3.33 1.96
10 4.17 2.38