Team:Imperial College London/Drylab/Enzyme/Analysis

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Assumptions

Enzymatic assumptions:

  • The enzyme is specific only for the substrate and not for any other chemicals.
  • Only one enzyme, our enzyme of interest is present and participating in the reaction.
  • There is negligible formation of product without the enzyme.
  • The rate of enzymatic activity remains constant over time because:
    • There is no co-operativity of the system. Binding of substrate to one enzyme binding site doesn't influence the affinity or activity of an adjacent site.
    • No allosteric regulations from either the product or the substrate.
    • There is no product inhibition of the enzyme.


  • Total [E] does not change with regards to time. Enzyme will not be used up nor degraded over time, thus [E] at the beginning of the reaction will be the same as [E] at the end of reaction.
  • The reaction catalysed is irreversible
  • [S] >> [E], such that the free concentration of substrate is very close to the concentration I added. This also ensures a constant substrate concentration throughout the assay. This allows easy determination of [E].
  • For which

Ek1.jpg

, k1>>k2


Law of mass action assumptions:

  • Free diffusion
  • Free unrestricted molecular motion


Michaelis-Menten assumption:

  • There is a quasi-steady state of [ES] , ie: , during which the enzyme is used for the reaction. This means that the rate of formation of ES complex is equal to the rate of dissociation of ES complex.
  • from the above assumptions, we know that


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