SEMINAR TOPIC

ON

ENZYME

Speaker
Mr. Shishir Kumar Behera
Univ. Roll No. : 5905M09003

Under the guidance of

Smt. U. Parida, M.Sc.
Reader in Zoology
UDALA COLLEGE
UDALA, MAYURBHANJ
ORISSA- 757041

ENZYME
INTRODUCTION
The enzymes are biologically produced organic proteins which exhibit catalytic reaction. During reactions enzymes
act in minute quantities and neither themselves used up nor destroyed. The term enzyme was used by Wiely Kunne
(1878) while working on termination.
Enzyme may be defined as organic catalysis which accelerates various biochemical reactions without being
used themselves. It acts as biocatalyst.
NOMENCLATURE

All enzyme name should end with using suffix ase.

Suffix is use to the group of similar chemical reaction. e.g. oxidase, reductase.

Exceptions are some old names like pepsin, ptyalin, trypsin, rennin etc.

To denote enzyme active on group of substrate like lipid it is named as lipolytic, amylolytic.

Some time enzymes are named after the species name like papain (Carica papaya).

CLASSIFICATION
The international union of Biochemistry IUB (1961) classify enzyme into following categories.
a)

Oxido-reductase
They take part on oxidation and reduction reaction reactions. These are three types.

1.

Oxidase

2.

Dehydrogenase

3.

Reductase
E.g.

b)

Isocitric acid

Oxalosuccnic acid

Transferase

It transfers glycosol, methyl, and phosphonyl. They transfer a group foe one molecule to another. It doesnt occur in
Free State.
E.g.

Glucose + ATP

Glucose - 6 phosphate + ADP

c)

Hydrolase
They break up large molecule in presence of H2O. It breaks C-C, C-O, C-N, P-N bond .Hydrolase are
following types.
1. Protease- act on peptide bonds
2. Easterase-catalyse hydrolysis of ester. e.g. Phosphate, Lipase.
3. Carbohydrase - act on glycosylic linkage

E.g. Urea + H2O

d)

CO2 + NH3

Lyases
They cause cleavage, removal of groups without hydrolysis. It breaks C-C, C-O, and C-N bond
E.g. Pyruvic acid

Acetyl phosphate + CO2

Moreover, addition of group to 2C double bonds of formation of double bond by removal of group is
done by this enzyme.
e)

Isomerase
These groups of enzyme catalyze reaction which brings about synthesis of new molecules by the

union of the molecules.

E.g. Glucose - 6 phosphate
f)

Fructose - 6 phosphate

Ligase/Synthetase
They catalyses two chemical with the help of ATP. It breaks C-O, C-S, C-N, C-C bond.

E.g. Acetyl Co A + Oxaloacetic acid

Citric acid

CHEMICAL NATURE
The chemical nature of enzyme could be discovered by J.B. Sumner (1926)
All enzymes are protein but all proteins are not enzyme. These are two types
I.

SIMPLE ENZYME
The whole enzyme is made up of protein without substrate. e.g. Pepsin, trypsin

II.

CONJUGATED ENZYME
It is formed of two parts
Apo-enzyme- it is the protein part
Cofactor

- it is the non-protenious part. It is small and heat stable. It may

be Organic (co-

enzyme) or inorganic (prosthetic group)

PROPERTIES OF ENZYME
The enzymes possess many outstanding physical and chemical characteristics. These are described as below
1.

COLLOIDAL NATURE

The enzymes are giant or large molecules having high molecular weight.

Their molecular weight ranges from 12000 ton to 1 million. Therefore, the enzyme possess extreme low
rate of diffusion and form colloidal system in H2O. More over, due to their colloidal nature they are nondialyzable.

2.

CATALYTIC PROPERTIES

The enzyme action doesnt depend on the quantity of the enzyme because; a small quantity of it is capable
of activating the chemical reaction indefinitely.

But the large quantity of enzyme triggers the faster reaction.

For example one molecule of catalyze is capable of catalyzing the conversion of 5,000,000 molecules of
H2O2 to H2O and oxygen per minute under favorable conditions.
2H2O2

\Catalyse

2H2O + O2

3.

SPECIFICITY OF ENZYME ACTION

Most of the enzymes are specific in their action. Moreover, their specificity lies on substrate molecule.

The key to specificity of enzyme for particular substrates lies in the active site.

For example, amylase enzyme acts on protein but reverse reaction is impossible.

Sucrose

Sucrase

Glucose + Fructose

4.

THERMOLABILITY ( HEAT SENSTIVITY)

Enzyme are stable within the temperature 0-500C

As enzyme are protenious in nature there are very sensitive to heat.

Most of the enzymes are working perfectly between 170C to 500C temperature.

When the temperature above 600C, the enzyme changes their chemical structure.

5.

REVERSIBILITY OF ENZYME ACTION

Enzyme is known to accelerate the chemical reaction in which ever direction, forward or backward.

The enzyme that catalyze both the synthetic and decomposing reactions are said to be reversible in nature.

Enzyme are able to induce reversible reaction for example, the lipase (fat splitting enzyme) catalyses the
synthesis of fat from glycerol and fatty acid. It can also hydrolyse the glycerol and acid to their component units.

E.g. Fat

Lipase

fatty acid + Glycerol

6.

pH SENSTIVITY

Each enzyme acts to its best in a certain pH value,

If the pH value of the medium is changed its activity slows down. The activity of some enzymes in their pH
medium is given below.
Enzyme
Pepsin

1.6

Lipase (stomach)

7.

Optimum pH
4.0 to 5.0

Nature of medium
Highly acidic
Acidic

Lipase (pancreas)

8.0

Alkaline

Urease

7.0

Neutral

Arginase

10.0

Highly alkaline

ACTIVE ENZYME
Certain enzyme requires specific catalyst for their action. For example - the prosthetic group of nucleotide act as
catalyst in the break down of nuclein some metallic ion Zn, Co, Mn, Mg, also activates peptidase enzyme during
its catalysis.
Enzyme are unstable compounds being soluble in water, glycerol, sodium chloride solution and dilute alcohol,
these may be participated by saturation with Ammonium sulphate or by excess alcohol.

KINETICS OF MULTISUBSTRATE REACTION

Kinetics recognizes three general mechanisms for multi-substrate enzyme system.

Two of three mechanisms, termed ordered and random mechanism which requires that all substrates
can be released.

In third mechanism called Ping pong mechanism one or more products is released from the enzyme
before all the substrates have reacted with in the enzyme.

1)

ORDERED MECHANISM

In this mechanism there is a definite order in which substrates associated with active sites of
enzyme.

All the products are released in a specific order and only after all the substrate have reacted.

This reaction is also called ordered Bi, Bi mechanism.

EA1

E + A1
2)

EA1A2

EP1P2

EP2

E + P2

RANDOM MECHANISM
When two substrates S1 & S2 add to an enzyme in random order, two products P1 & P2 are released in
random order, such a mechanism is known as Random Bi, Bi mechanism.
E + S1

ES1

EP1
ES1S2

E + S2
3)

E + P1

EP1P2

ES2

EP2

E + P2

PINGPONG MECHANISM
In this mechanism when two substrates S1 & S2 are participated in bio chemical reactions , first substrate S1
is added to the enzyme to form ES1 complex and then it converted to EP1 complex . Product P1 release and
the enzyme added to the substrate S2 to form EP2 complex, the product released and the enzyme free.
E + S1

ES1

EP1

ES2

EP2

In this reaction the enzyme complexes formed are ES1, EP1, EP2, and ES2. Here S1 convertedtoP1and S2
converted to P2. E designates a modified enzyme.

THEORY OF MECHANISM OF ENZYME ACTION

To explain the mechanism of enzyme action two theories have been put forwarded. These are
1. Lock and key theory
2. Induced fit theory
1. LOCK AND KEY THEORY

It is also known as complimentary hypothesis and was given by Emil Fischer (1898)

According to this hypothesis both enzyme and substrate molecules have specific geometrical shape.

The active site of enzyme (E) held the particular substrate (S) form (ES) complex. Here the enzyme
is analogous to lock (ES) complex undergo chemical changes and form enzyme product (EP) complex.

Very soon product (P) is released from (EP) complex leaving the enzyme free.

The active site on the surface of the enzyme molecule has some special groups like
-COOH, -NH2, -SH etc that bind the substrate molecule to form an intermediate complex. The substrate
molecule undergoes chemical changes and form product molecule.
2.

INDUCED FIT THEORY

This theory was proposed by Koshland (1959)
This is a modification to lock and key theory. It is a flexible model.

According to this theory the

enzyme molecule is not a rigid structure; the active site of enzyme contains two groups such as
buttressing group and catalytic group d. As soon as the substrate comes in contact with the
buttressing group, the active site of
converts to product.

enzyme undergoes conformational changes. The substrate

ENZYME CATALYSIS
1.

ACID BASE CATALYSIS

Generally acid base catalysis is related to proton donor and acceptor during enzymatic reaction .The bond
present in substrate is broken during catalysis is known as scissile bond. During enzymatic catalysis
substrate recognizes enzyme but not the reverse.
2.

COVALENT CATALYSIS

Some enzyme may enhance the rate of chemical reaction to form highly reactive covalent intermediate
between enzyme and substrate.
Enzyme

Covalent intermediate

Chymotrypsin

Acyl enzyme

Transaldolase

Schiffs base

INHIBITION OF ENZYME
The compounds that reduce or inhibit the rate of enzyme action are called enzyme inhibitors and the
phenomenon is known as enzyme inhibition. It is three types.
(a)

Competitive inhibition

(b)

Un competitive inhibition

(c)

Non competitive inhibition

(a)

COMPETATIVE INHIBITION
In this case substrate(S) & inhibitor (I) complete some active site on the enzyme (E) so both the enzyme
substrate (ES) & (EI) enzyme inhibitor complex are formed.
E+ S

(b)

ES

E+P

In this inhibition the dose not combine with the free (E) but (I) combine with ES complex to give
an in active (ESI) complex. It is also known as allosteric inhibition.
E+S

ES
+

E+P

I
ESI

(c)

NON COMPETATIVE INHIBITION

A non competitive inhibitor combining either the free enzyme or ES complex and interfere their action. In
this case the reaction with inhibitor yields to inactive forms such as EI & ESI. It is also known as mixed
inhibition.
E+S

ES

EI + S

E+P

ESI

FACTOR AFFECTING ENZYME ACTIVITY

TEMPERATURE
The rate which an enzymatic reaction precedes is influenced by temperature. In general the initial velocity
of enzymatic reaction is accelerated with increase in temperature until a certain optimum is attained. Above
400 c the destructive effect of temperature most plant enzyme increases.

CONCENTRATION
The rate of reaction increases with the increasing concentration of enzyme up to some extent. But when all
the substrate molecules engaged with the enzymes in that situation of the enzyme concentration increases
the rate of reaction remain unchanged. Enzyme reactions like all other chemical reaction are subject to lows
of chemical equilibrium and mass action. So as the end product of the reaction accumulate the apparent rate
of the reaction decreases. The velocity of the enzymatic action usually increases with the increase in the
substrate concentration up to certain maximum after which the rate of reaction decreases due to high
substrate concentration.