UNIVERSITY OF MINDANAO

COLLEGE OF ENGINEERING EDUCATION

BIOCHEMICAL ENGINEERING

(ChE 544)

ENZYME INHIBITION

A Written Discussion

Submitted to:

Engr. Arjan C. Lingaya

Submitted by:

Mary Jane C. Jumawan

June 2017
 Introduction

Enzymes are biological catalysts that are protein molecules in nature. They are

produced by living cells (animal, plant, and microorganisms) and are absolutely essential as

catalysts in biochemical reactions. Almost every reaction in a cell requires the presence of a

specific enzyme. A major function of enzymes in a living system is to catalyse the making

and breaking of chemical bonds. Therefore, like any other catalysts, they increase the rate

reaction without themselves undergoing permanent chemical changes.

The catalytic ability of enzymes is due to its particular protein structure. A

specific chemical reaction is catalyzed at a small portion of the surface of the enzyme, which

is known as the active site. Some physical and chemical interactions occur at this site to

catalyze a certain chemical reaction for a certain enzyme.

Enzyme kinetics deals with the rate of enzyme reaction and how it is affected

by various chemical and physical conditions. Kinetic studies of enzymatic reactions provide

information about the basic mechanism of the enzyme reaction and other parameters that

characterize the properties of the enzyme. The rate equations developed from the kinetic

studies can be applied in calculating reaction time, yields, and optimum economic condition,

which are important in the design of an effective bioreactor.

Knowledge of basic enzyme kinetic theory is important in enzyme analysis in

order both to understand the basic enzymatic mechanism and to select a method for enzyme

analysis. The conditions selected to measure the activity of an enzyme would not be the same

as those selected to measure the concentration of its substrate. Several factors affect the rate

at which enzymatic reactions proceed - temperature, pH, enzyme concentration, substrate

concentration, and the presence of any inhibitors or activators.

A modulator (or effector) is a substance which can combine with enzymes to

alter their catalytic activities. An inhibitor is a modulator which decreases enzyme activity.

Enzyme inhibitors are substances which alter the catalytic action of the enzyme and

consequently slow down, or in some cases, stop catalysis. It can decrease the rate of reaction

either competitively or noncompetitively.

The enzyme inhibitors are low molecular weight chemical compounds. They

can reduce or completely inhibit the enzyme catalytic activity or permanently (irreversibly).

Inhibitor can modify one amino acid, or several side chain(s) required in enzyme catalytic

activity. To protect enzyme catalytic site from any change, ligand binds with critical side

chain enzyme.

Reversible Inhibition
Reversible inhibition is the process by which the inhibitor binds to the

enzyme non-covalently and can dissociate from the enzyme with great ease. Reversible

inhibitors are those that can be readily removed from the enzyme to which they bind.

Sometimes they bind covalently; more often they operate via electrostatic or van der Waals

interactions.

Competitive Inhibition
A competitive inhibitor is any compound which closely resembles the

chemical structure and molecular geometry of the substrate. The inhibitor competes for the

same active site as the substrate molecule. The inhibitor may interact with the enzyme at the

active site, but no reaction takes place. The inhibitor is "stuck" on the enzyme and prevents

any substrate molecules from reacting with the enzyme. However, a competitive inhibition is
usually reversible if sufficient substrate molecules are available to ultimately displace the

inhibitor. Therefore, the amount of enzyme inhibition depends upon the inhibitor

concentration, substrate concentration, and the relative affinities of the inhibitor and substrate

for the active site.

Illustration:

Example:

Sildenafil (Viagra)

Nitric Oxide (NO) binds receptors in the smooth muscle cells of the

penis. This results in increased levels of cyclic guanosine monophosphate (cGMP) which

increases vasodilation. An enzyme called PDE5 degrades cGMP. Sildenafil fits into the same

active site of PDE5 as cGMP, thus competitively inhibiting PDE5 from working.

Non-Competitive Inhibition
A noncompetitive inhibitor is a substance that interacts with the

enzyme, but usually not at the active site. The noncompetitive inhibitor reacts either remote

from or very close to the active site. The net effect of a non competitive inhibitor is to

change the shape of the enzyme and thus the active site, so that the substrate can no longer

interact with the enzyme to give a reaction. Non competitive inhibitors are usually reversible,
but are not influenced by concentrations of the substrate as is the case for a reversible

competitive inhibitor.

Illustration:

Example:

Deoxycyclin (an antibiotic), which inhibits collagenase (a proteolytic enzyme

involved in periodontal diseases).

Uncompetitive Inhibition
Uncompetitive inhibitors are those that bind to a site distinct from the

active site, but only in the presence of bound substrate; when they bind, they reduce the

reaction velocity, because they convert some molecules of the enzyme-substrate complex into

the nonproductive enzyme-substrate-inhibitor form. But they also decrease maximum

concentration because the equilibria for the formation of both the enzyme-substrate complex

and the enzyme-substrate-inhibitor complex are shifted toward the complex by the binding of

inhibitor.
 Illustration:

Example:

A few pesticides are uncompetitive inhibitors, the best-known

example being the herbicide N-phosphonomethylglycine, commonly known as glyphosate or

Roundup, an uncompetitive inhibitor of 3-phosphoshikimate 1-carboxyvinyltransferase.

Irreversible Inhibition
Irreversible Inhibitors form strong covalent bonds with an enzyme. These

inhibitors may act at, near, or remote from the active site. Consequently, they may not be

displaced by the addition of excess substrate. In any case, the basic structure of the enzyme

is modified to the degree that it ceases to work.

Suicide Inhibition
A subset of irreversible inhibitors called suicide irreversible inhibitors,

are relatively inactive compounds. They get activated upon binding with the active site of a

specific enzyme. After such binding, the suicide irreversible inhibitor is activated by the first
few intermediary steps of the biochemical reaction - like the normal substrate. However, it

does not release any product because of its irreversible binding at the enzyme active site.

Inhibitors make use of the normal enzyme reaction mechanism to get activated and

subsequently inactivate the enzyme.

Suicide Inhibitors are like modified substrates. Inhibitors are initially

processed by enzyme as if it were the normal substrate. However, a reaction intermediate

covalently and irreversibly binds the enzyme, leading to its inhibition.

Example :

The antibiotic penicillin is another transition state analog suicidal

inhibitor that binds irreversibly covalently to serine at the active site of the bacterial enzyme

glycopeptides transpeptidase. The enzyme is a serine protease required for synthesis of the

bacterial cell wall and is essential for bacterial growth and survival. It normally cleaves the

peptide bond between two D-alanine residues in a polypeptide. Penicillin structure contains a

strained peptide bond within the β-lactam ring that resembles the transition state of the

normal cleavage reaction, and thus penicillin binds very readily to the enzyme active site. The

partial reaction to cleave the imitating penicillin peptide bond activates penicillin to bind

irreversibly covalently to the active site serine.

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