NPTEL – Biotechnology - Systems Biology

Michaelis Menten Kinetics- Enzyme Inhibition

Dr. M. Vijayalakshmi
School of Chemical and Biotechnology
SASTRA University

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Table of Contents
1 INTRODUCTION .............................................................................................. 3
1.1 COMPETITIVE INHIBITION ............................................................................. 3
1.2 UNCOMPETITIVE INHIBITION ......................................................................... 5
1.3 NON-COMPETITIVE INHIBITION ..................................................................... 7
1.4 MIXED INHIBITION ....................................................................................... 8
2 REFERENCES .................................................................................................. 9
2.1 TEXT BOOKS ................................................................................................. 9

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1 Introduction
Enzyme inhibition can happen when the inhibitors (structural analogs of substrate)
binds to the active site of the enzyme and prevents the catalysis. Enzyme inhibition
is specific and is different from the alteration of structure of enzyme and reduction of
reaction rate by environmental factors such as pH, temperature, presence of
hydrophobic compounds, detergents etc., which are non specific. For e.g. consider
sudden addition of an acid or base to the reaction mix which changes the pH and
thereby influences the rate of the reaction. These are often confused with inhibition
as they also reduce the turnover rate of enzymes.

In general, the binding of enzyme to the inhibitor is reversible but few of them bind
covalently and become irreversible. The reversible and irreversible inhibitors have
different kinetics. Michaelis-Menten kinetics explains the inhibition of enzyme in a
single substrate complex, but the complexity increases with the number of
substrates. The inhibitors also depend on their homology with the substrate apart
from the nature of binding site and binding affinity.

Some inhibitors even bind stronger than the natural substrate because of specific
interactions and act as antagonists. Most therapeutic drug molecules act in this way.
Yet there are different forms of inhibition based on the affinity of inhibitor to the
enzyme and substrate.

1.1 Competitive inhibition

Competitive inhibition occurs when the inhibitor is highly homologous to the

substrate molecule and competes with substrate to bind to the free enzyme. So,
either one of them can bind with an enzyme and not both together Fig 1. In this
condition, there is a need for excess substrate to overcome the competition with
inhibitor. Classical example for competitive inhibition is the molecule methotrexate
which inhibits the action of dihydrofolate reductase to convert dihydrofolate to
tetrahydrofolate.

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E+S E.S E+P

+I

E.I

Fig 1. Schematic representation of competitive inhibition

The rate of product formation in the reaction is given by,

And from the Michaelis-Menten kinetics,

Similarly, rate of formation of enzyme – inhibitor complex will give,

The total enzyme concentration in the system will be the sum of the concentration of
three forms in which the enzymes exists:

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Hence in competitive inhibition, only the Km is influenced and not the maximum
velocity (Vmax).

1.2 Uncompetitive inhibition

Uncompetitive inhibition occurs when the inhibitor does not bind to the free enzyme
and instead binds to the already formed enzyme substrate complex and makes the
complex inactive Fig 2. This phenomenon of inhibition is commonly observed in
multimeric enzymes.

E+S E.S E+P

+I

E.S.I

The rate of product formation in the reaction is given by,

And from the Michaelis-Menten kinetics,

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Similarly, rate of formation of enzyme-substrate-inhibitor complex will give,

Fig 2. Schematic representation of uncompetitive inhibition

The total enzyme concentration in the system will be the sum of the concentration of
three forms in which the enzymes exists:

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In uncompetitive inhibition, both Km as well as maximum velocity (Vmax) is influenced.

1.3 Non-Competitive inhibition

Non-Competitive inhibition is where; the inhibitor binds to the different site in the
enzyme. So, in contrast to competitive inhibition, they can bind along with substrate
to the enzyme and here both EI and ESI is inactive Fig 3.

E+S E.S E+P

ki +I Kii + I

+S

E.I E.S.I

Fig 3. Schematic representation of non competitive inhibition

The rate of product formation in the reaction is given by,

And from the Michaelis-Menten kinetics,

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Similarly, rate of formation of enzyme-substrate-inhibitor complex will give,

The total enzyme concentration in the system will be the sum of the concentration of
four forms in which the enzymes exists:

Non competitive inhibition influences the maximum velocity while the Km does not
changes. Vmax is changed because high substrate concentration cannot prevent the
binding of inhibitor.

1.4 Mixed inhibition

Another mode of inhibition which is similar to that of non competitive inhibition but
with an active ESI complex is termed the mixed inhibition. Such inhibition is common
in metabolic feedback pathways. Enzymes showing this form of inhibition are
generally allosteric in nature.
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2 References

2.1 Text Books

1. Bisswanger H, Enzyme Kinetics, Principles and Methods, WILEY-VCH, (2002).

2. J. D. Murray, Mathematical Biology, Springer-Verlag, (1989).
3. Berg JM, Tymoczko JL, Stryer L. Biochemistry, 5/e, W H Freeman, (2002).

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