Michaelis-Menten Model accounts for the

kinetic properties of many enzymes

Primary function of enzymes is to enhance the
rates of reactions
V0 is reaction velocity
or rate of catalysis:
Number of moles
product/second

Vmax: maximal velocity

Km: Michaelis-Menten
constant = substrate
concentration at which
there is half of
max. velocity
The extent of product formation is determined as
a function of time for a series of substrate
concentrations
 Changes in concentration of
all reaction partners
V0: the rate of increase in product with time
when product concentration is still low (pre-steady
state)
 V0 linear proportional to substrate
concentration

V0 is determined for each substrate concentration by

measuring the rate of product formation at
early times before product accumulates
 The Michaelis – Menten Model
The model proposed, which is the simplest one that accounts for
the kinetic properties of many enzymes, is

We want an expression that relates the rate of catalysis to

the concentrations of substrate and enzyme and the rates of
the individual steps. Our starting point is that the catalytic rate
is equal to the product of the concentration of the ES complex and k2.
 The Michaelis – Menten Model cont.
Now we need to express [ES] in terms of known quantities.
The rates of formation and breakdown of ES are given by:

Assumption: steady – state = concentration of

ES intermediates stays the same even when substrate
and product concentration changes
The Michaelis – Menten Constant Km

KM has the units of a concentration!

Km = [E] [S] / [ES]

The Michaelis – Menten Constant
Km contin.

or
The Michaelis – Menten Constant
Km contin.

Insert ES concentration into:

 The Michaelis – Menten equation

Term for V0
What about Vmax?
(ESconc.=Etotal conc.
Each active site exists as ES complex)

Analogous to:

Michaelis – Menten - Equation

When [S] is much less than
KM: V0 = (Vmax/KM)[S];
that is, the rate is directly
proportional to the substrate
concentration.

When [S] is much greater than

KM: V0 = Vmax; that is,
the rate is maximal,
independent of substrate
concentration.

When [S] is KM: V0 = Vmax/2

 The meaning of KM
1) KM is the concentration of substrate at
which half of the active sites are filled

2) KM is related to the rate constants of

individual steps in the catalytic scheme

If k-1 >>>>k2
 The meaning of KM cont.
The dissociation constant of the ES complex is given by:

KM is equal to the dissociation constant of the ES complex

if k2 is much smaller than k-1.

When this condition is met, KM is a measure of the strength

of the ES complex: a high KM indicates weak binding;
a low KM indicates strong binding.

It must be stressed that KM indicates the affinity of the ES complex

only when k-1 is much greater than k2.
 The meaning of Vmax
The maximal rate, Vmax, reveals the turnover number of an enzyme,
which is the number of substrate molecules converted into product
by an enzyme molecule in a unit time when the enzyme is fully
saturated with substrate.

It is equal to the kinetic constant k2, which is also called kcat.

The maximal rate, Vmax, reveals the turnover number of an enzyme

if the concentration of active sites [E]T is known, because
 Lineweaver – Burk Plot

The plot provides a useful

graphical method for
analysis
of the
Michaelis – Menten
equation:

Taking the reciprocal gives:

Linearisation methods
Protein-Ligand interaction

v/[S] als Funkt. von v.