Team:Imperial College London/Drylab/Enzyme/Analysis/Detailed

From 2009.igem.org

Revision as of 05:21, 22 September 2009 by Tywang (Talk | contribs)



The behaviour of the system can be described with ODEs. The first 4 equations describe the single-substrate mechanism for an enzyme reaction without Michaelis-Menten assumptions, while equation 5 describes modification with the Michaelis-Menten assumption.

The Enzyme Mechanism

Ek1.jpg

k1, k2 ,k3 are the kinetic rate constants for each step.

In substrate binding, 1 molecule of Enzyme (E) binds with 1 molecule of Substrate (S) in a reversible reaction to form the Enzyme-Substrate complex (ES). We are unable to measure this step.

In catalysis, the Enzyme-Substrate complex (ES) produces the unchanged enzyme (E) and the product (P) in an irreversible reaction. This step can be measured.

When [S] becomes high enough, the second step of the enzyme kinetics becomes the rate determining step, controlled by k3.
This enzyme mechanism scheme can be described by the following differential equations describing rate of change of each component. Each component is described by a simple rate term for either its production or its decomposition.



Equation 1: Rate of change of Enzyme

Ekk1.jpg
Equation 2: Rate of change of substrate

Ekk2.jpg
Without MM assumption

Equation 3: Rate of change of enzyme-substrate complex

Ekk3.jpg
Equation 4: Rate of change of product

Ekk4.jpg
With Michaelis-Menten Assumptions

Consider the enzyme mechanism again:

Ek1.jpg

From the model, we want to derive an equation that describes the rate of enzyme activity (amount of product formed per time interval) as a function of substrate concentration.

Consider equation 4: the rate of product formation. This equation is essentially useless as we cannot measure [ES] experimentally, therefore, to correctly determine d[P]/dt, we need to solve for ES in terms of the other quantities. Therefore, we need to make some approximations as mentioned previously. (hyperlink to assumptions part) At steady state, equation 3 is reduced to: Ek2.jpg When this result is used together with equation 4, a new differential equation for rate of change of product is obtained:

Equation 5: New rate of change of product

Ekk5.jpg

Mr. Gene   Geneart   Clontech   Giant Microbes