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2.5 Eadie (1942) measured the initial reaction rate of hydrolysis of acetylcholme (substrate) by
dog serum (source of enzyme) and obtained the following data:
Substrate Concentration Initial Reaction Rate
mol/L mol/L min
0.0032 0.111
0.0049 0.148
0.0062 0.143
0.0080 0.166
0.0095 0.200
Evaluate the MIchaelis – Menten kinetic parameters by employing (a) the Langmuir plot, (b) the
Lineweaver – Burk plot, (c) the Eadie – Hofstee plot, and (d) non – linear regression procedure.
Solution:
a. Langmuir Equation
Cs r Cs/r
0.0032 0.111 0.028829
0.0049 0.148 0.033108
0.0062 0.143 0.043357
0.008 0.166 0.048193
0.0095 0.2 0.0475
Langmuir Plot
0.06
0.05
0.04
Cs/r
0.01
0
0 0.002 0.004 0.006 0.008 0.01
Cs
Vmax = 0.301813901 mol/L min
Km = 0.005764645509 mol/L
Cs 1/Cs V 1/V
0.0032 312.5 0.111 9.009009
0.0049 204.0816 0.148 6.756757
0.0062 161.2903 0.143 6.993007
0.008 125 0.166 6.024096
0.0095 105.2632 0.2 5
Lineweaver-Burk Plot
10
8
y = 0.0172x + 3.6342
1/V
7 R² = 0.9146
4
100 150 200 250 300
1/Cs
1 1 K 1
m
V Vmax Vmax C s
0.17
0.12
r
0.02
15 20 25 30 35
-0.03
r/Cs
r
r K m rmax
Cs
Langmuir Plot
25
20
15
Cs/R
10 y = 2.3722x + 6.2429
R² = 0.9937
0
0 1 2 3 4 5 6 7 8
Cs
4
1/V
y = 6.7758x + 2.2168
3 R² = 0.9922
2
0
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
1/Cs
1 1 K 1
m
V Vmax Vmax C s
0.3
0.25
0.2
r
0.15
y = -2.8096x + 0.4336
0.1 R² = 0.9566
0.05
0
0 0.02 0.04 0.06 0.08 0.1
r/Cs
r
r K m rmax
Cs
Michaelis-Menten Plot
0.35
0.3
0.25
0.2
rp
0.15
0.1
0.05
0
0 1 2 3 4 5 6 7 8
Cs
Vmax C s
V
K m Cs