Lecture – 8 – 23.09.

2022
Biochemical Engineering
(CHE F 421)
BITS Pilani
Pilani Campus
Enzyme kinetics
• A characteristic of enzymes reactions is substrate
saturation

Observed effect of substrate concentration on rate of

catalyzed reaction is given below:

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Enzyme kinetics

Saturation kinetics can be obtained from a simple reaction

scheme that involves a reversible step for enzyme–substrate
complex formation and a dissociation step of the ES complex.
Inhibition kinetics

• Certain compounds many bind to enzymes & reduce their

activity
• Such enzymes are called as enzyme inhibitors

• Certain agents may decrease reactivity in which toxic

chemicals can adversely affect a biological treatment
systems
• EI may be reversible or irreversible

• Irreversible EIs may be heavy metals (lead, Hg & Cd); form

a stable complex with enzyme & decrease enzyme activity

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Inhibition enzyme kinetics

• Such enzymes inhibition may be reversed only by using

chelating agents such as EDTA

• Three classes of reversible enzymes (EIs)

• Competitive, Non- competitive, Un-competitive, Substrate

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Inhibition enzyme kinetics

• Such enzymes inhibition may be reversed only by using

chelating agents such as EDTA

• Three classes of reversible enzymes (EIs)

• Competitive, Non- competitive, Un-competitive, Substrate

Inhibition

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Competitive Inhibition
• Since a competitive inhibitor has a strong structural
resemblance to the substrate, both inhibitor and
substrate compete for the active site of the enzyme

• Formation of an enzyme - inhibitor complex reduces the

amount of enzyme available for interaction with the
substrate and, as a result, the rate of reaction
decreases

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Competitive enzyme inhibitors (EIs)
Competitive inhibitors are substrate inhibitors and
compete with substrate for active site of enzymes.

Assuming rapid equilibrium and as per the definition :

derive
Competitive Inhibition

Hence in competitive inhibition, only the Km is

influenced and not the maximum velocity (Vmax).
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• The net effect of competitive
inhibition is an increased
value of Km, and, therefore,
reduced reaction rate

• Competitive inhibition can be

overcome by high
concentrations of substrate

• Figure describes competitive

inhibition in the form of a
double reciprocal plot.

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Non Competitive enzyme inhibitors
(EIs)
• Non Competitive inhibitors are not substrate analogs

• Inhibitors bind on sites other than the active site and

reduce enzyme affinity to the substrate.

Assuming rapid equilibrium and as per the definition :

derive
 Non competitive Inhibition

• 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.
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• Net effect of noncompetitive
inhibition is a reduction in Vm

• High substrate concentrations

would not overcome
noncompetitive inhibition

• Other reagents need to be

added to block binding of the
inhibitor to the enzyme

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3.3.2. Mechanistic Models for Simple Enzyme Kinetics
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

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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

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

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BITS Pilani, Pilani Campus
• Net effect of uncompetitive
inhibition is a reduction in
both Vm and Km values

• Reduction in Vm has a
more pronounced effect
than the reduction in Km ,
and the net result is a
reduction in reaction rate

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Substrate inhibition

• High substrate concentrations may cause inhibition in

certain enzymatic reactions, known as substrate
inhibition

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BITS Pilani, Pilani Campus
BITS Pilani, Pilani Campus
BITS Pilani, Pilani Campus
Substrate inhibition

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BITS Pilani, Pilani Campus