Michaelis-Menten Kinetics

Michaelis-Menten kinetics is one of the best known models of enzyme kinetics in

biochemistry. It was named after a German biochemist Leonor Michaelis and Canadian
physician Maud Leonora Menten.
In 1912 Michaelis and Menten published their seminal work on enzymes – almost all
of which are proteins. Their research cast new light on these complex compounds that make
possible the chemical reactions of life (Chemical Heritage Foundation, 2015).

Leonor Michaelis
Leonor Michaelis is a medical doctor by profession who
received his degree at the University of Berlin in 1897. He put his
interest in doing a research about embryological study rather than
clinical study to complete his requirements. The year after
graduating he worked as an assistant in a laboratory in Frankfurt,
where he studied the interaction between aniline dyes and the
chemical constituents of living tissues which led him to discover a
dye that specifically stained certain crucial components, later
called the mitochondria, of the cells that make up living tissues
(Chemical Heritage Foundation, 2015).
In 1905, he accepted a position as bacteriologist at one of
berlin’s municipal hospitals. There he and a friend set up a small
research laboratory and conducted studies on the role of
hydrogen-ion concentration in determining the properties of
proteins and enzymes. He also studied how small molecules adhere to the surface of various
proteins and enzymes. He then tried to understand the relation between the rate of
formation of product to the concentrations of enzyme and its substrate – that is, its efficiency
in getting its work done.
Maud Leonora Menten
In the early stages of Michaelis’ work he was joined by Maud
Menten. Maud Menten was one of the 40 coworkers who were attracted
to Michaelis’ modest laboratory during the period of its operation, from
1905 to 1921. Menten had just received her MD in 1911 from the
University of Toronto (Chemical Heritage Foundation, 2015).

The Michaelis-Menten Equation

Michaelis and Menten were able to express mathematically the relationship that they
were investigating, which demonstrated that each enzyme not only has its own substrate but
also that at sufficient concentrations of substrate it has its own rate of causing that substrate
to change chemically (Chemical Heritage Foundation, 2015).
The kinetics of most enzyme reactions are reasonably well represented by the Michaelis-
Menten equation:
𝑣max 𝐶𝐴
𝑣 = 𝑟𝐴 =
𝐾𝑚 + 𝐶𝐴
mol
Where: 𝑣 = 𝑟𝐴 = volumetric rate of reaction, m3 s

𝑣max = maximum rate of reaction

mol
𝐾𝑚 = Michaelis constant, m3

𝑠 = 𝐶𝐴 = concentration of A/substrate
As defined in the equation above, vmax is a volumetric rate that is proportional to the amount
of active enzyme present. The Michaelis constant Km is equal to the reactant concentration at
𝑣
which 𝑣 = 𝑚𝑎𝑥 or we can say that Km is the substrate concentration at which half of the
2
enzyme’s active sites are saturated with substrate. Km is independent of enzyme
concentration but varies from one enzyme to another and with different substrates for the
same enzyme (Doran, 1995). Values of Km for some enzyme-substrate systems are listed in
the table below. Km and other enzyme properties depend on the source of the enzyme.
 Adapted from P. Doran, 1995, Bioprocess Engineering Principles

The simplest reaction sequence that accounts for the kinetic properties of many
enzymes is:
𝑘1 𝑘2
𝐸 + 𝑆 ↔ 𝐸𝑆 → 𝐸 + 𝑃
𝑘−1
Where E is enzyme, S is substrate, and P is product. ES is the enzyme-substrate complex.
Binding of substrate to the enzyme in the first step is considered reversible with forward
reaction constant k1 and reverse reaction constant k-1. Decomposition of the enzyme-
substrate complex to give the product is an irreversible reaction with rate constant k 2; k2 is
known as the turnover number as it defines the number of substrate molecules converted to
product per unit time by an enzyme saturated with substrate. Turnover number (k2) is
sometimes referred to as catalytic constant (Doran, 1995). Analysis of this reaction sequence
yields the relationship:

𝑣𝑚𝑎𝑥 = 𝑘2 𝑒𝑎
Where ea is the concentration of active enzyme. As expected in catalytic reactions, enzyme E
is recovered at the end of the reaction.
Km is considered a relative measure of the substrate binding affinity: lower K m values
imply higher enzyme affinity for the substrate. The catalytic efficiency is defined as the ratio
𝑘2
⁄𝐾 , mol/L ∙ s. Catalytic efficiency is often used to compare utilization of different
𝑚
substrates by a particular enzyme and is a measure of the substrate specificity or relative
suitability of a substrate for reaction with the enzyme. Substrate with higher catalytic
efficiency are more favorable.
Enzyme Inhibitions
Any molecule that reduces the rate of an enzyme reaction is called an inhibitor.
Reaction products, substrate analogues, synthetic molecules, metabolic intermediates, and
heavy metals are examples of enzyme inhibitors; substrates can also act as inhibitors. Binding
between the inhibitor and enzyme may be reversible or irreversible (Doran, 1995).

Graphical Methods for Michaelis-Menten Reactions

1. Michaelis - Menten Plot
The simple procedure involves plotting v vs s values. ʋmax and Km can be
estimated roughly from this graph. The accuracy of this method is usually poor
because of the difficulty of extrapolating ʋmax.

The reaction rate v does not increase indefinitely with substrate concentration but
approaches a limit, ʋmax. At high substrate concentrations 𝑠 ≫ 𝐾𝑚 , Km in the
denominator of the original equation of Michaelis-Menten is negligibly small
compared with s sothe equation can now be:
𝑣𝑚𝑎𝑥 𝑠
𝑣= Or 𝑣 ≈ 𝑣𝑚𝑎𝑥
𝑠

Therefore at high substrate concentrations, the reaction rate approaches a constant

value independent of substrate concentration; in this concentration range, the
reaction is essentially zero-order with respect to the substrate. On the other hand, at
low substrate concentrations 𝑠 ≪ 𝐾𝑚 , the value of s in the denominator of the
original equation is negligible compared with Km and can be simplified to:
𝑣𝑚𝑎𝑥
𝑣≈ 𝑠
𝐾𝑚
𝑣𝑚𝑎𝑥
The ratio of constants ⁄𝐾 is, in effect, a first-order rate coefficient for the
𝑚
reaction. Therefore, at low substrate concentrations there is an approximate linear
 dependence of reaction rate on s; in this concentration range, the M-M reactions are
essentially first order with respect to substrate (Murray, 1989)
2. Lineweaver – Burk Plot
This method uses a linearization procedure to give a straight-line plot.
Inverting the original equation of M-M reactions, we can get:
1 𝐾𝑚 1
= +
𝑣 𝑣𝑚𝑎𝑥 𝑠 𝑣𝑚𝑎𝑥

𝟏 𝟏 𝑲𝒎 𝟏
It is a plot of 𝒗 𝒗𝒔 should give a straight line with slope 𝒗 and intercept 𝒗 . This
𝒔 𝒎𝒂𝒙 𝒎𝒂𝒙
double reciprocal plot is found frequently in the literature of enzyme kinetics but this
sometimes gives inaccurate results and is therefore not recommended.
3. Eadie – Hofstee Plot
𝑣
If the Lineweaver-Burk equation is multiplied by 𝑣( 𝐾𝑚𝑎𝑥 ) and then rearranged,
𝑚
another linearized form of the Michaelis-Menten equation is obtained:
𝑣 𝑣𝑚𝑎𝑥 𝑣
= −
𝑠 𝐾𝑚 𝐾𝑚

𝒗 −𝟏 𝒗𝒎𝒂𝒙
It is a plot of 𝒔 𝒗𝒆𝒓𝒔𝒖𝒔 𝒗 gives a straight line with slope 𝑲 and intercept . This
𝒎 𝑲𝒎
linearization distorts errors in the data so that the method has reduced accuracy.
4. Hanes – Woolf Plot
Sometimes called the Langmuir plot. Multiplying the Lineweaver-Burk
equation by s produces a new linearized form of the Michaelis – Menten equation,
and is presented by:
𝑠 𝐾𝑚 𝑠
= +
𝑣 𝑣𝑚𝑎𝑥 𝑣𝑚𝑎𝑥

𝒔
Therefore, a plot of 𝒗 𝒗𝒆𝒓𝒔𝒖𝒔 𝒔 should give a straight line with slope 𝟏⁄𝒗𝒎𝒂𝒙 and
𝑲
intercept 𝒎⁄𝒗𝒎𝒂𝒙 . Linearization of data for the Langmuir plot minimizes distortions
in experimental error. Accordingly, its use for evaluation of 𝒗𝒎𝒂𝒙 and Km is
recommended.
References
1.

Chemical Heritage Foundation. (2015, December 22). Retrieved June 15, 2017, from CHF West:
https://www.chemheritage.org/historical-profile/leonor-michaelis-and-maud-leonora-menten

2. Doran, P. M. (1995). Bioprocess Engineering Principles. San Diego, California: Academic Press
Limited.

3. Murray, J. D. (1989). Mathematical Biology. Springer-Verlag.