UNIVERSIDAD DEL ATLÁNTICO

FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

ENZIMATIC KINETIC
Authors: Yourladys Acosta – Roberto Altamar - Stephanie Vargas
Bioprocesses
Delivery date: 07/03/2022

1. SIMULATION OF MICHAELIS-MENTEN KINETIC MODEL

Parameter value
k1 20
k-1 1
k2 10
S0 10
[E]0 2

The following systems of equations are solved in the program R studio.

𝜕𝑆
= 𝑘−1 [𝐸𝑆] − 𝑘−1 [𝑆][𝐸]
𝜕𝑡
𝜕𝐸
= (𝑘2 + 𝑘−1 )[𝐸𝑆] − 𝑘1 [𝑆][𝐸]
𝜕𝑡
𝜕𝐸𝑆
= 𝑘1 [𝑆][𝐸] − [𝐸𝑆](𝑘2 + 𝑘−1 )
𝜕𝑡
𝜕𝑃
= 𝑟2 = 𝑘2 = [𝐸𝑆] = 𝜈
𝜕𝑡

SOLUTION

First you must have the desolve library installed, then enter the equations to be solved,
the parameters, the time range where the equations will be evaluated and then use the
ode function to solve the equations.
 UNIVERSIDAD DEL ATLÁNTICO
FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Then the concentrations of the substrate, enzyme, enzyme-substrate and product are
plotted with respect to time.

Figure 1. Profile of concentrations of enzyme reaction.

 UNIVERSIDAD DEL ATLÁNTICO
FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

2. EXAMPLE 1 MICHAELIS-MENTEN ENZYME KINETICS

Lactase, also known as β-galactosidase, catalyses the hydrolysis of lactose to produce

glucose and galactose from milk and whey. Experiments are carried out to determine
the kinetic parameters for the enzyme. The initial rate data are as follows.

Calculate the kinetic parameters of Michaelis-Menten.

SOLUTION
First we add the data that give us the concentration and initial velocity of the lactose, then
we plot the graph the profile of initial reaction velocity vs substrate concentration.
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Figure 2. Profile of initial reaction velocity vs substrate concentration.

Lineweaver - Burk
For the method Lineweaver- Burk, the inverse of the velocity is plotted with respect to the
inverse of the subtract concentration, then the parameters for km and vm are obtained from
the plots. Taking into account that the slope corresponds to km/vm and the intercept to the
inverse of vm, the following equation is also obtained.
1 𝑘 1 1
= (𝜈𝑚 ) [𝑆] + 𝜈
𝜈 𝑚 𝑚
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Figure 3. Lineweaver-Burk
Lineweaver Eady – Hofstee
For the method we plot v vs v/[S], from the graphs we obtain the parameters to find km and
vm. Taking into account that the slope corresponds to -km and the intercept to vm.
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Figure 4. Lineweaver Eadie – Hofstee.

Hanes-Woolf or Langmiur
For the method Hanes-Woolf or Langmiur, we plot [S]/v vs [S], from the plots we obtain the
parameters to find km and vm. Taking into account that the slope corresponds to the inverse
of vm and the intercept to km/vm.
[𝑆] [𝑆] 𝑘𝑚
= +
𝜈 𝜈𝑚 𝜈𝑚
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Figure 5. Hanes-Woolf or Langmiur.

.
3. PROBLEM 1: REVERSIBLE INHIBITION
The initial rate of reaction of an enzyme catalyzed reaction was determined in the presence
of several inhibitors A, B, and C at various concentrations of substrate S. Determine the type
of inhibition and their kinetic parameters (including Ki)
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Solution
Initially find the inverse of the substrate concentration and the inverse of the rate of product
formation of the three types of inhibitor and no inhibitor and plot this values

Fig. 6: 1/V vs 1/S

Regression is calculated to find the values of the slope and the intercept, this is done
without inhibitor type and inhibitor A, B and C
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Being the first coeficcientthe values of intercept and second one the values of slope

After that, we calculated the kinetic parameters of the different types of reversible
inhibition
Competitive Inhibition
𝑣𝑚 𝑆
𝑣=
𝐼
𝐾𝑑 (1 + 𝐾 ) + 𝑆
𝑖

𝐼
𝐾𝑑 (1 + ) = 𝐾𝑑𝑎𝑝𝑝
𝐾𝑖
𝑣𝑚 𝑆
𝑣=
𝐾𝑑𝑎𝑝𝑝 + 𝑆
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Uncompetitive Inhibition
𝑣𝑚𝑎𝑝𝑝 [𝑆]
𝑣= ′
𝐾𝑑𝑎𝑝𝑝 + [𝑆]

′
𝐾𝑑
𝐾𝑑𝑎𝑝𝑝 =
𝐼
(1 + )
𝐾𝑖′
𝑣𝑚
𝑣𝑚𝑎𝑝 =
𝐼
1+
𝐾𝑖′
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FACULTAD DE INGENIERIA
INGENIERIA QUÍMICA

Non-Competitive Inhibition
𝑣𝑚𝑎𝑝𝑝 [𝑆]
𝑣=
𝐾𝑑 + [𝑆]
𝐾𝑖 = 𝐾𝑖′
𝑣𝑚
𝑣𝑚𝑎𝑝𝑝 =
𝐼
(1 + 𝐾 )
𝑖