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CHAPTER 10

ENZYME

1. DEFINITION

Enzymes are globular proteins.


Enzymes lower the activation energy needed for
reaction take place.
Enzymes act as biological catalysts and speed up
the rate of metabolic reactions.
Enzymes are highly specific in action.
Enzymes possess active sites and will only catalyse
reaction when substrate and active site have
complementary shapes.
The enzyme combine with its substrate to form an
enzyme-substrate complex.
The complex then breaks up into product and
enzyme.

Every enzyme is specific for a particular reaction.


Every step in a metabolic pathway is catalysed by
a different enzyme.
Reaction begins with reactants and end with
products.
A+B
reactants

C+D
products

If substrate and active site are complementary,


the enzyme will catalyse.

substrate + active site = enzyme-substrate


complex = products
Substrate
Active site

Enzyme

Products

2. Enzymes Reduce
Activation Energy

In a chemical reaction, the reactants must first


reach a high-energy intermediate state called the
transition state before the products are formed
The amount of energy required for reactants to
reach the transition
Transition state
state before
Activation energy
changing into
without enzyme
product is called
Substrate
Activation energy with
activation energy
enzyme
(reactants)
Energy of system

Products (final state)

Progress of a reaction

Figure 1

Every chemical reaction involves the breaking and


forming of chemical bonds.
Bond breaking requires external energy while
bond formation releases energy.
Catabolic reactions are reactions that release
energy (exergonic).
Anabolic reactions are reactions that absorb
energy (endergonic).
The initial energy required to break a bond is
referred to activation energy.
An enzyme provides an alternate reaction pathway.

The enzyme can bind to the substrate (reactant)


to form an enzyme-substrate complex which
corresponds to the transition state.
The shape of the substrate is slightly changed,
existing bonds are broken and new ones are
formed.
This makes it easier for the substrate to be
changed into the product.

3. Mechanism of enzyme
action

A substrate molecule binds to an enzyme


molecule at its active site.
Forming an enzyme-substrate complex.
The substrate is converted into products while
still being attached to the enzyme.
Once reaction has occurred, the products are
released and the enzyme available to catalyse
another cycle of reaction.

4. Types of enzymes
1.

Intracellular enzyme

Enzymes that are produced in certain cells


and remain to react in the cell

These enzymes could exist either in


cytoplasm (in organelles) or nucleus.

2.

Extracellular enzyme

Some enzymes are produced by cell but then


transported out of the cell for action outside
the cell

5. Characteristics
1.

2.

3.
4.

Enzymes generally act quickly

the speed of reaction is usually stated in


turnover number (refer to the number of
substrates)
Enzymes are not damaged

however, this does not mean that enzymes can


be used repeatedly forever without replacement
Enzymes can react in both direction
Enzymes are specific

each enzymes limited to one specific reaction


that involved one specific substrate only

5.

6.

7.

An enzymes molecule is usually bigger than


its
substrates
All enzymes are proteins and not all protein
are enzymes
Enzymes are complex globular protein and
three dimensional.

Main function of enzymes

Increase the rate of chemical reaction by


lowering activation energy

How an
enzyme
works

Enzyme
(sucrase)

Glucose

Fructose

Active
site

Substrate
(sucrose)

4
Enzyme available
with empty active
site

Products are
released

3
Substrate is
converted to
products
Figure 5.6

2
Substrate
binds to
enzyme with
induced fit

The enzyme is unchanged and can repeat the


process

6. Mechanisms of Enzymes
There are 2 main hypothesis explaining the
mechanism of enzyme action :

The lock-and-key hypothesis

The induced-fit hypothesis

Simple mechanism

5.1 Lock-and-key
hypothesis

The hypothesis proposed that the active site and


substrate are exactly complementary
An enzyme is a large globular protein with specific three
dimensional shape.
It has a site called the active site containing amino acid
that are complementary to the substrate.
In the lock-and-key hypothesis, the shape of the
substrate (key) fits into the active site of the
enzyme (lock), forming an enzyme-substrate complex
Reaction takes place and products are formed and
released.

5.2 Induced-fit hypothesis

It is a modified version of the lock-and-key hypothesis


The hypothesis suggested that active site is flexible and is
not exactly complementary to the shape of the substrate
An enzyme collides with the substrate molecule. The
substrate binds to the active site
The binding induces a slight change in the shape of the
enzyme to enclose the substrate, making the fit more
precise.

6. Factors affecting enzyme

Any factors that affect the activity of an enzyme will


change the rate of the reaction catalysed by that
enzyme
Enzyme characteristic are affected by several factors
temperature
pH
Substrate concentration
enzyme concentration.

6.1 The effect of


Temperature

At lower temperature, enzymes


are not active
As the temperature rises, the
substrate and the enzyme
molecules move rapidly and are
more likely to collide
The increase in temperature
to a specific level can increase
the rate of the enzyme
activity until the optimum
temperature rate.
After the optimum
temperature the enzymes
activities decreased and totally
stopped at temperature of
60oC

6.2 The effect of pH

All enzymes have a specific


optimum pH at which they function
most efficiently
Most enzyme act at a pH in range
of 5 9, and reactions most
efficiently at a pH of 7
The small change in pH can
produce a large effect on enzyme
activity
A pH that is extreme usually
destroy enzymes

However, there are exceptions for certain enzymes


such as:
Pepsin pH 1.5 2.5 (acidic)
Renin pH 8.5 (alkaline)

6.3 The effect of Enzyme


Concentration

The rate of an enzyme-catalysed reaction is directly


proportional to the concentration of the enzymes if
substrates are present in excess concentration and
no other factors are limiting

6.4 The effect of Substrate


Concentration

Increasing the substrate concentration can give an


increase in reaction rate
At lower concentrations the rate increase in direct
proportion to the substrate concentration
At higher substance
concentration the rate of
reaction becomes
constant

7. Classification of Enzyme

Many enzymes have a name derived from the


name of their substrate, with the ending ase
added. Example, lactase hydrolyse lactose
Other enzymes have been given less informative
names. Example include many of the enzyme of
digestion, such as pepsin, renin
Systematic naming of enzymes is based on an
agreed classification of enzyme and on the
name of the substrate of the reaction catalysed

Enzymes can be classified by the kind of


chemical reaction catalyzed:
1. Hydrolase
2. Lyase
3. Isomerase
4. Ligase
5. Transferase
6. Oxidoreductase

This classification system introduced in 1961


by IUB (International Union of Biochemistry)

7.1 Hydrolase

These enzyme catalyse the hydrolysis of a


substrate by the addition of water
Example:
Lipase, amylase, sucrase, peptidase
sucrase

Sucrose + water
+ fructose

glucose

7.2 Lyase

These enzyme catalyse the breaking of chemical


bonds without the addition of water
Example:
Pyruvate decarboxylase decarboxylation of
pyruvic acid
Pyruvate decarboxylase

Pyruvate
carbon dioxide

ethanol +

7.3 Isomerase

These enzyme catalyse the rearrangement of atoms


within a molecule converting one isomer to another
Example:
phosphoglucoisomerase

Glucose-1-phosphate
glucose-6-phosphate

7.4 Ligase

These enzymes catalyse the joining together of


two molecules with the simultaneous hydrolysis
of ATP
Example:
synthetase

amino acid + specific tRNA + ATP


amino acid-tRNA complex + ADP + Pi

7.5 Transferase

These enzymes catalyse the transfer of a


chemical group from one substrate to another
Example:

Glucose

glucose-6-phosphate
ATP

ADP

7.6 Oxireductase

These enzyme catalyse redox reactions


(biological oxidation and reduction reactions) by
the transfer of hydrogen, oxygen or electron from
one molecule to another
Example:
Oxidase catalyses the addition of oxygen to
hydrogen, forming water
Glucose oxidase

Glucose + oxygen
acid + water

gluconic