Bioprocess Engineering

22ECEC303

Chapter 3:
Enzyme
Inhibition
Lecture: 19
Reversible Uncompetitive Inhibition
Reversible Uncompetitive inhibition is a scenario where an inhibitor
binds only to the enzyme-substrate complex (ES), not to the free
enzyme (E). This type of inhibition results in a unique kinetic profile:
1.Both the apparent maximum velocity (Vmax​) and the Michaelis
constant (Km​) decrease in the presence of the inhibitor.
2.The inhibitor does not affect the formation of the ES complex but
prevents it from releasing the product, thus lowering the Vmax​.
Figure 3.19 shows how the presence of the inhibitor (I)
decreases the enzyme velocity (v). The curve shifts downwards,
indicating that the maximum rate of reaction (Vmax​) is reduced
when the inhibitor is present.
Figure 3.20 is a Lineweaver-Burk plot, which is a double
reciprocal plot of the Michaelis-Menten equation. It shows two
lines representing enzyme activity with and without the
inhibitor. The line corresponding to the presence of inhibitor
intersects the y-axis below the line without the inhibitor,
reflecting a decrease in 1/Vmax​, and intersects the x-axis to the
left of the line without the inhibitor, indicating a decrease in
−1/Km​.
Substrate Inhibition:
Unlike typical enzyme behavior where an increase in substrate
concentration leads to an increase in reaction rate, substrate inhibition is
a phenomenon where excessive substrate concentration actually
decreases the reaction rate. This happens because additional substrate
molecules bind to an enzyme-substrate complex, inhibiting further
reaction. For instance, invertase or β-fructofuranosidase, enzymes that
hydrolyze disaccharides, exhibit substrate inhibition when two substrate
molecules bind simultaneously to the active site, rendering the enzyme
inactive.
Figure 3.21 is a Lineweaver-Burk plot that departs from
linearity at high substrate concentrations, curving upwards t
show a reduction in velocity, which is indicative of substrat
inhibition.
Figure 3.22 shows a velocity (v) versus substrate concentration
(S) graph, where the curve for substrate inhibition dips down at
higher substrate levels compared to a typical reaction curve,
demonstrating the inhibitory effect.
Cont’d
• Substrate inhibition introduces a secondary enzyme-substrate complex (ES2),
which forms when an additional substrate molecule binds to the ES complex.
The formation of ES2 is governed by a dissociation constant KSI​, which is the
ratio of the concentration of ES2 to the product of the concentrations of ES
and S.
• The enzyme conservation equation accounts for the total enzyme in all forms,
including the secondary complex ES2.
• The Michaelis-Menten equation is modified to account for the formation of
ES2, and different cases are considered:
• Case 1: At low substrate concentrations, substrate inhibition is negligible.
• Case 2: At high substrate concentrations, substrate inhibition is significant and the
reaction rate equation is modified accordingly.