The name enzyme was taken from Greek word En = in, and Zymase = ferment/yeast and the name

was coined by F.W.

Kuhne in 1878. J.B. Sumner 1926 for the first time purified the enzyme urease from Canavolia insiformis (Jack bean) in
pure crystalline form.
Enzymes are biological catalyst which accelerate the rate-of biochemical reactions like catalyst. Enzymes are specific
proteins which accelerate all chemical reactions taking place in living organism. There are nonprotein enzymes like metal
ions, H+ and OH-.
SIGNIFICANCE OF ENZYME
 Small amount of enzymes are required to catalyse the chemical reactions in biological systems. The number of substrate
molecules catalysed per enzyme molecule per minute is approximately 103 i.e. they have enormous catalytic power.
 The enzymes remains unchanged in chemical reactions, qualitatively and quantitatively.
 The enzymes form short lived complex with reacting molecules called substrate and then broken into products and free
enzymes.
 They also increase the rate of reaction until it reaches equilibrium.
 Enzymes are highly specific in nature both in the reaction catalysed and in their choice of substrate (reactants). An
enzyme catalyses a single chemical reaction or a set of closely related reactions e.g. urea is converted to ammonia and
carbon dioxide in presence of urease and fumerate is converted into maleate by fumerase enzyme.
 The enzyme enhances rate of chemical reaction by lowering down the energy of activation.
NOMENCLATURE
International union of Biochemists (I.U.B.) classified the enzymes on the basis of type of reaction and reaction
mechanism. Enzymes are named by adding the suffix-ase to the name of substrate on which they act. The enzymes that
act on amylum (starch) and hydrolyse it is called amylase. Similarly fat (lipose) is hydrolysed by lipase and protein by
proteinase. In later cases the name of enzyme indicates the type of chemical reaction catalysed e.g. dehydrogenases,
oxidases. decarboxylases etc.
According to IUB system of nomenclature the name and classification of enzymes is on the basis of type and
mechanism of chemical reaction. The main features are
 The enzyme that catalyse the reaction from six classes in which each of them have different subclass.
 The name of enzymes have two parts. The first is substrate name and second ending in - ase indicate the type of
reaction catalysed.
 Additional information if needed to clarify the reaction is given in parentheses e.g.
L Malate + NAD+ →Pyruvate + NADH + CO2 + H+
is designated as oxidoreductase (decarboxylating)
 Each enzyme has code number (EC) that characterize the type of reaction as to class (the first digit), subclass (the
second digit), sub sub class (the third digit) and the fourth one is for specific enzyme e.g. E.C.=2.7.1.1. denotes class
two, sub class 7 (the phosphate transfer), sub sub class 1 (and alcohol function as phosphate acceptor) and the final
digit 1 denote hexokinase the type of enzyme.
 Oxidoreductases: These enzymes act upon two substrates in which transfer
of e-, atoms or functional groups takes place causing oxidation and
reduction reactions. Example-Various oxido-reductases, dehydrogenases,
NAD oxidases, oxygenises etc.
The enzymes are classified as
1. Oxidoreductases
2. Transferases
3. Hydrolases
4. Lyases Transferases: These enzymes catalyse those biological reactions in which
5. Isomerases transfer of different specific groups from one substrate to another takes
place. The transfer of different groups like methyl, acyl, keto, amino and
6. Ligases acetate is carried out by the enzyme trans-methylase, trans-acylase. trans-
ketolase, trans-aminase and acetate kinase respectively.
Hydrolases: The enzymes which hydrolyse different type of bonds formed by the combination of specific
functional groups of monomers in macromolecules like esters amides, peptides. acid anhydride. polysaccharides
and glycosyl bonds are called hydrolases. Such reactions take place mostly in digestive system. The enzymes of
digestive tract are e.g. Pepsin, trypsin, chymotrypsin hydrolyse protein, amylase hydrolyses starch. Lipase
hydrolyses triglycerides and nucleases hydrolyses nucleic acids.

Lyases: These enzymes catalyse the cleavage of specific covalent bonds like C-C, C-O, C-N, C-S and C-X
without hydrolysis. They also catalyse the addition of groups to double bond or reverse to it.

Some lyases add hydroxyl or amino or carboxyl group to larger molecules and remove them.
Isomerases: The enzymes which catalyse the transfer of groups within the molecules to yield different
isomeric forms like optical, geometrical or positional isomers. The interconversions include cis-trans, keto-
enol and aldose-ketose. The examples of isomerases are mutases, epimerases and racemases.

Ligases: Such enzymes catalyses formation of C-C, C-O, C–N and C–S bonds by condensation reactions
coupled to cleavage of ATP and other nucleoside phosphate. They link two molecules at the cost of
breakdown of third molecule and known as synthatase.
FACTOR AFFECTING ENZYME ACTIVITY:
SUBSTRATE CONCENTRATION: The velocity of an enzyme catalysed
reaction is directly proportional substrate concentration. Finally, a point
reaches when there is no increase in the velocity of reaction with an
increase in substrate concentration. At this point the velocity of reaction is
maximum (Vmax) when the enzyme becomes saturated with substrate.
In few cases the enzyme activity decreases at a very high concentration of
substrate and is known as substrate inhibition.
EFFECT OF pH: Since all the enzymes are proteins hence, they are
extremely sensitive towards pH of the medium in which the reaction
takes place. All the enzymes have a characteristic range of pH at which
velocity of reaction will be maximum. When the pH is plotted against
enzymes activity a bell shape curve is obtained. The optimum pH range
of an enzyme depends upon the proton donating or accepting group of
active sites. The optimum pH range of most of the enzymes lies between
4-9. However, the digestive enzymes are active at a pH value 2.0. The
denaturation and conformational changes occur at high or low pH values.
EFFECT OF TEMPERATURE:
In most of the enzymatic reactions velocity of reactions increases with the
increase of temperature, due to increase in kinetic energy of reacting
molecules. The rate of most of the biological reactions becomes double
with the 10°C rise in temperature. This value is known as temperature
coefficient and denoted by Q10. The range of enzymatic reaction lie in
between 5°-40°C. They are damaged irreversibly at 50-60°C because of
their proteinaceous nature. However, few enzymes continue to act above
60°C also. The enzyme in thermophilic bacteria and fungi are also active at
high temperature.
EFFECT OF IONS
Some enzymes need certain cations like Ca2+, Mg2+, Zn2+ or K+ for their activation. These metal ions either act as
a coenzyme or loosely bound with enzyme. In some cases, they only assist the process without being a part of the
enzyme. Some anions like Cl- is also effective in enhancing enzyme activity of salivary amylase.
REDOX POTENTIAL
Enzymes are also sensitive towards redox potential of a cell. The oxido reductase enzyme are involved in
changing the redox potential of a cell and influencing the activity of such type of enzymes.
ENZYMES KINETICS deals with the rate of enzyme catalysed reactions.
 The standard unit of enzyme activity (U) is defined as an amount of activity that catalyses the
transformation of one micromole substrate per minute. A new unit, the katal or kat has been proposed and
one kat denotes the conversion of one mole substrate per second.
 Another terminology in this context is the maximum velocity (Vmax) which is defined as the rate of
enzymatic reaction obtained under condition of substrate saturation of the enzyme under a given set of pH
and temperature.
MICHAELIS MENTEN EQUATION & MICHAELIS CONSTANT
 It is the relation between substrate concentration and enzyme activity. When enzyme concentration is
fixed, the velocity of reaction is linearly proportional to the substrate concentration [s] when its value is
small.
 At high value of substrate concentration, the velocity of reaction is independent of [s].
In 1913 L. Michaelis and M. Menten proposed a simple model to account for kinetic characteristic
of these reactions. They defined a constant called Michaelis constant and denoted by Km.
 This constant is defined as concentration of the substrate at which a given enzyme yield one half of its
maximum velocity.
ACTIVATORS The substances which either initiate or enhance the rate of enzyme catalysed reactions are
called as activators and the phenomenon is called as activation. These activators are of two types-
 Metallic ion or inorganic activators. Metallic ion activators require specific metallic ions for better activity
e.g.- arginase requires CO, Mn or Fe ions, leucyl peptidase require Mn or Mg ions, and α-amylase requires
Cl- ions.
 Proenzyme activators. It is considered that the activators combine with the enzyme to give an altered
protein which is better catalyse than the original one. Sometime in few cases metallic ion activators acts as a
bridge between the substrate and the enzyme.

INHIBITORS The activity of most of the enzymes

can be inhibited by specific molecules called
inhibitors. They are classified on the basis of their
site of actions.
IRREVERSIBLE INHIBITORS Such inhibitors
either combine with or destroy a functional group
of enzymes molecule which is necessary for the
catalytic activity e.g. Di-isopropyl fluorophosphate
(DFP) inhibits the enzyme cholinesterase.
REVERSIBLE INHIBITOR The inhibitor which do not cause a
permanent damage in the functional group of enzymes. The
enzyme becomes fully active after the removal of such inhibitor.
They are of two types.
A. COMPETETIVE INHIBITOR In competitive inhibition, an
inhibitor resembles the normal substrate & binds to the enzyme,
usually at the active site, and prevents the substrate from binding.
Here inhibitor combine reversibly with the enzyme forming
enzyme inhibitor complex rather than enzyme substrate complex.
In this case substrate and inhibitor both compete for the same
binding site of enzyme,
If (I) binds very tightly to E then a little enzyme is available to bind with S to form [ES]
complex which finally dissociate to product and enzyme. Hence the rate of reaction will be
very slow. Many antibacterial and sulpha-drugs work on the principle of competitive inhibitor.
A competitive inhibitor raises the value of Km but no effect on the value of Vmax.
Structure
An enzymes is a protein that facilitates a cellular metabolic
process by lowering activation energy levels, in order to catalyse
the chemical reactions between biomolecules. Some enzymes
reduce the activation energy to such low levels, that they actually
reverse cellar reactions. But in all cases, enzymes facilitate
reactions without becoming altered.
Composition
An enzyme's typical molecular weight ranges from about 10,000
to more than 1 million. A small number of enzymes are not
actually proteins, but instead consist of small catalytic RNA
molecules. Other enzymes are multiprotein complexes that
comprise multiple individual protein subunits.
While many enzymes catalyse reactions by themselves, some
require additional nonproteins components called "cofactors",
which may be inorganic ions such as Fe2+, Mg2+, Mn2+, or Zn2+,
or they may consist of organic or metallo-organic molecules
known as "coenzymes".
(B) NON-COMPETITIVE INHIBITOR Such inhibitors bind at the
site other than its active site which is called allosteric site. The
inhibitors usually bear little or no structural resemblances to
substrate. It lowers the value of maximum velocity but does not
affect the km value. The most important non-competitive inhibitors
are naturally occurring metabolic intermediates. The inhibition is
not reversed by increasing concentration of substrate. Many heavy
metal ions, cyanide act as a non-competitive inhibitors of enzymes.
ACTIVE SITE The active site consists of residues that form temporary bonds with the substrate (binding site) and
residues that catalyse a reaction of that substrate (catalytic site). The active site is very small relative to the whole
volume of the enzyme (10~20% of the total volume). It usually consists of three to four amino acids, while other
amino acids within the protein are required to maintain the tertiary structure of the enzyme.

The unique amino acids present in an active site promote specific interactions
that are necessary for proper binding and resulting catalysis. Complementary
shapes between enzyme and substrate(s) allow a greater amount of weak
non-covalent interactions including electrostatic forces, Van der Waals
forces, hydrogen bonding, and hydrophobic interactions. Specific amino acids
also allow the formation of hydrogen bonds. That shows the uniqueness of the
microenvironment for the active site.
THE MECHANISM OF ENZYME ACTION
The enzyme binds reversibly with the substrate through their active site and enzyme substrate complex is
formed. Two models have been proposed to explain the type of linkage between enzyme and substrate.
a. Lock and key model
b. Induced fit model

Lock and key model: was proposed by German

chemist Emil Fischer in 1890's. The enzymes are
larger molecules as compared to substrate. The
substrate bound at the active or catalytic site of
enzyme. The catalytic site forms a complimentary
structure to that of substrate molecule in terms of
lock and key analogy. This is a rigid template model
and converted to products by distortion of bonds
which increases the speed of reactions.
Induced fit model: This mechanism was proposed by D. Koshland in 1959. According to this mechanism the
active site is flexible and substrate induces a conformational change in the flexible active site of enzyme so that
there is an induced fit between the two. This active site bears buttressing and catalytic group. Buttressing group
supports the substrate whereas catalytic group, weakens the bond between enzyme and substrate by electrophilic
or nucleophilic processes. This concept explains why enzymes are proteins and have larger area than most of the
substrate.