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Enzyme Kinetics: Michaelis-Menten Equation (Km)

The Michaelis–Menten equation, proposed by Leonor Michaelis and Maud Menton in 1913, is the rate equation for a one-substrate enzyme-catalyzed reaction. While most enzyme-catalyzed reactions have more than one substrate, the Michaelis-Menton equation is still applied if only one substrate concentration is varied.

The Michaelis–Menten equation states that the product formation rate v depends hyperbolically on [S], and is depicted mathematically as follows:

 

Michaelis-Menten Equation

 

where [S] is the substrate concentration and Vmax is the maximum velocity at saturating substrate concentrations. Km is the Michaelis constant, defined as the substrate concentration at which V attains half the value of Vmax.

To implement the Michaelis-Menton plot within CDD:

  • Create readouts for input data such as concentration [S] and initial reaction rate [v]
  • Creating a plot type readout to generate the curve fit
  • Editing and saving the curve

Let’s work through an example implementing this curve fit within our “Michaelis-Menten” Protocol.

Create a numerical readout for the X-axis values:

Create a numerical readout for the Y-axis values:

Create a plot readout, which will create the plot and automatically calculate additional values:

Our protocol now captures our input data and automatically generates a Michaelis-Menten plot for each entity per run of the protocol. 

The following parameters will automatically be calculated by CDD Vault for this plot type:

  • Vmax
  • Km
  • Maximum measured v
  • Minimum measured v
  • N
  • R squared
  • Area under the curve - observed

Area under the curve

  • The CDD Vault AUC is calculated as the (positive area - negative area) under the dose response curve where:
    • The curve is defined as straight line connections between responses, not the fitted curve (Linear Trapezoidal Non-uniform grid method)
    • Units are response units * independent units
    • The baseline is set at the negative control mean or 0 if control data is not available

Additional calculations: 

Please note that all auto-calculated parameters listed above can be used in custom calculations as separate readouts. These parameters can be accessed within our robust formula editor.