Unit-2 Part-2

Enzymes
 Characteristics of Enzymes
• Some characteristics of enzymes are as follows:
• (1) Enzymes are unique in nature and show varying degree of specificities, while they are
highly specific for a particular substrate. The active site of an enzyme shows a strong affinity
for a specific substrate, and further slightly changes the conformation of substrate. It is
known as an "induced fit" to accommodate the reacting substrate, and this induced fit
further leads to a stabilized transition state to lower the activation energy of the reaction.
The same catalyst cannot be applied in more than one reaction. Stereo specificity means that
enzymes can detect the different optical isomers and react to only one type of isomer. Group
specific enzymes refer to those catalyzing a group of substances with specific groups.

• (2) Enzymes are extremely efficient and possess great catalytic power, transforming about
100 to 10,000 substrate molecules into product per second and proceeding from 103 to 108
times faster than the uncatalyzed reaction. Enzymes do not affect the equilibrium constant
in spite of the increase in the conversion rate of substrate into product. The number of
substrate molecules converted to product is called the turnover number. Only a small
number of enzymes are typically required under normal cellular conditions to catalyze a
chemical reaction.
• (3) Enzymes itself remain unchanged during the reaction. Although
the amino acid residues of enzyme may get broken or form
covalent bonds with the substrate, it can typically reform those
broken bonds or separate with substrates, allowing the enzyme to
bind with more substrates.

• (4) Enzymes can be allosterically regulated by a variety of means,.

Sometimes, activators and coenzymes are necessary for enzymatic
catalysis, which are helpful to increase enzyme potency due to the
presence of a weak bond between the enzyme and a metal ion.
Inorganic substances are often known as activators. Additionally,
there are also molecules that can allosterically inhibit enzyme
function by modifying its conformational shape.
• (5) The effectiveness of an enzyme catalyst could
achieve maximum at its optimum temperature. Both
the enhancement and decrease of the optimum
temperature result in a decrease in catalytic activity.
• (6) Biochemical catalysis of enzyme also depends on
the pH of the solution. An enzyme exerts its full
potential at an optimum pH ranging between 5-7 pH
values.
• (7) The catalytic activity of enzymes can be inhibited by
competitive inhibitors, noncompetitive inhibitors or
irreversible inhibitors. Competitive inhibitors reversibly bind
to the active site of the enzyme, thus blocking the substrate
from binding to the enzyme. Noncompetitive inhibitors bind to
any site of the enzyme other than the active site, enabling the
enzyme less active or inactive. Irreversible inhibitors form
bonds with enzymes to make them inactive.

• (8) Increase in the concentration of the reactants could

increase the reaction rate until the enzyme become
saturated with the substrate, while increase in the amount of
enzyme will continuously enhance the rate of the reaction.
 Effect of various factors on rate of enzyme catalyzed reaction

• 1. Temperature
• As with many chemical reactions, the rate of an enzyme-catalysed reaction increases as the
temperature increases. However, at high temperatures the rate decreases again because the
enzyme becomes denatured and can no longer function. This is shown in the graph below.
• As the temperature increases so does the rate of enzyme activity. An optimum activity is
reached at the enzyme's optimum temperature. A continued increase in temperature results
in a sharp decrease in activity as the enzyme's active site changes shape. It is now denatured.
• 2. pH
• Changes in pH also alter the shape of an enzyme’s active site. Each enzyme work bests at a
specific pH value. The optimum pH for an enzyme depends on where it normally works. For
example, enzymes in the small intestine have an optimum pH of about 7.5, but stomach
enzymes have an optimum pH of about 2.
• In this graph , as the pH increases so does the rate of enzyme activity. An optimum activity is
reached at the enzyme’s optimum pH, pH 8 in this example. A continued increase in pH
results in a sharp decrease in activity as the enzyme’s active site changes shape. It is now
denatured.
• 3. Substrate concentration
• Enzymes will work best if there is plenty of substrate. As the concentration of the substrate
increases, so does the rate of enzyme activity. However, the rate of enzyme activity does not
increase forever. This is because a point will be reached when the enzymes become
saturated and no more substrates can fit at any one time even though there is plenty of
substrate available.
• As the substrate concentration increases so does the rate of enzyme activity. An optimum
rate is reached at the enzyme’s optimum substrate concentration. A continued increase in
substrate concentration results in the same activity as there are not enough enzyme
molecules available to break down the excess substrate molecules.
 How enzymes catalyze a reaction?
• Enzymes are catalyst that accelerate the rate of biochemical reaction by
decreasing the energy of activation.
• Every chemical reaction have energy barrier that must be crossed by the reactant
molecules in order to convert itself into the product.
• The amount of energy supplied to reactant molecules in order to cross the energy
barrier to from product is known as Energy of activation.
• If energy of activation is higher, rate of reaction is slower and if it is lower, the rate
of reaction is faster.
• The role of enzyme in biochemical reaction is to reduce the amount of energy of
activation such that the rate of reaction increases.
• During enzyme catalysis, active site of enzyme binds with substrate molecules to
form Enzyme-substrate (ES) complex. During this binding some binding energy is
released which is utilized to activate the substrate (reactant) molecules to form
product. Thus the requirement of the amount of activation energy is decreased
such that rate of reaction increases. The amount of activation decrease is equal to
the amount of binding energy released during binding of enzyme and substrate.
 Mechanism of enzyme action
• 1. Lock and Key model:
• According to this model, shape of active site of enzyme is complementary
to the shape of substrate molecules. Ie. the substrate is like a key whose
shape is complementary to the enzyme which is supposed to be lock and
they fit perfectly.
• Enzymes catalyze only those substrates which fit perfectly on the active
site of that enzyme.
• Most enzymes are far larger than the substrates molecules that act on
and the active site is usually a very small portion of the enzyme, between
3 and 12 amino acids. The remaining amino acids which make the bulk of
the enzyme, function to maintain the correct globular shape of the
enzyme.
• Once the product is formed, they no longer fit into the active site and
escape into surrounding medium.
• According to lock and key model, enzymes behave as rigid molecules.
However, most enzymes are globular and are flexible with varying shape.
 2.Induced-fit model
• In 1959, Koshland suggested a modification to the ‘Lock and Key’
hypothesis which is known as ‘Induced fit’ hypothesis.
• Working from evidence that suggested that some enzymes and
their active site are more flexible. To this, he proposed that the
active site can modify its shape as the substrate interact with the
enzyme.
• The amino acids which make up the active site are moulded into
precise shape which enable the enzyme to perform its catalytic
function most efficiently.
• For instance, a suitable analogy to describe Induced fit model
would be that of a hand changing the shape of the glove as the
individual put on the glove. Therefore in this case, glove is the
active site of enzyme and the hand is substrate.
• However, in some cases, the substrate molecules changes slightly
as it enters the active site before binding.
Mechanism of enzyme action
 Coenzymes and cofactors
• Cofactors are molecules that attach to an enzyme during chemical
reactions. In general, all compounds that help enzymes are called cofactors.
However, cofactors can be broken down into three subgroups based on
chemical makeup and function:
• Coenzymes
These are reusable non-protein molecules that contain carbon (organic).
They bind loosely to an enzyme at the active site to help catalyze reactions.
Most are vitamins, vitamin derivatives, or form from nucleotides.
• Metal ions
These cofactors are reusable non-protein molecules that do not contain
carbon (inorganic). Usually, cofactors are metal ions such as iron, zinc,
cobalt, and copper that loosely bind to an enzyme’s active site. They must
also be supplemented in the diet as most organisms do not naturally
synthesize metal ions.
• Prosthetic group
• These can be organic vitamins, sugars, lipids, or inorganic metal ions.
However, unlike coenzymes, these groups bind very tightly or covalently
to an enzyme to aid in catalyzing reactions. These groups are often used in
cellular respiration and photosynthesis. Eg. Porphyrin moiety of
peroxidase,
 Enzyme kinetics
(Micheaelis-Menten equation)
 Nomenclature and classification of enzymes

• NC-IUBMB Enzyme List, or, to give it its full title, “Recommendations of

the Nomenclature Committee of the International Union of Biochemistry
and Molecular Biology on the Nomenclature and Classification of Enzymes
by the Reactions they Catalyse.

• EC numbers :Enzymes are identified by EC (Enzyme Commission)

numbers. It is unique for each enzyme.

• The EC number is made up of four components separated by full stops.

Scheme of classification of enzyme

• The first digit identifies the class of reaction

catalysed. . Enzymes were divided into 6 major
classes according to the type of reaction catalysed
and a seventh, the translocases,was added in 2018.
• The second digit(the subclass) generally contains
information about the type of compound or group
involved.
• The third digit, the sub-subclass, further
specifies the type of reaction involved.

• The fourth is a serial number that is used to

identify the individual enzyme within a sub-
subclass.
Seven classes of Enzymes
Illustration of 4 digits assignment