CHAPTER 15

15.1 PROPERTIES OF ENZYME &

MECHANISM OF ENZYME
ACTION
 15.1 PROPERTIES OF ENZYMES &
MECHANISM OF ENZYME ACTIONS
LEARNING OUTCOMES:
At the end of the lesson, students should be able to:
a) State the properties of enzymes
b) State the six classes of enzyme according to IUB
classification.
c) Explain the mechanism of enzyme action
(lowering activation energy)
a) Describe the hypothesis related to its mechanism
of enzyme action :
i. Induced fit Model
ii. Lock and Key Model
e) Explain factors that affect the enzymatic reaction
 WHAT IS ENZYME?
Biological catalyst produce by living cells that speed
up the rate of chemical reaction by lowering the
activation energy
 STRUCTURE OF AN ENZYME

• Has two sites :

 ACTIVE SITE
• Specific site on enzyme
that binds to specific
substrate

 ALLOSTERIC SITE
• Other site than the
active site on enzyme
 SPECIFICITY OF AN ENZYME
• The specificity of an enzyme is due to the
complementary shape of the active site and the
substrate.

• When an enzyme is denatured by heat or a

change in pH, the shape of active site is changed
and the substrate can no longer fit into active
site.
 ENZYME REACTION

E + S E S E + P

Enzyme-Substrate
Complex
(transition state)

E : enzyme
S : substrate (reactant)
ES : enzyme - substrate complex
P : product
 ACTIVATION ENERGY, EA
Minimum amount of energy required by a
reactant (substrate) to start a chemical reaction.

• When substrate
collide with
enzyme at its
active site

• Enzyme facilitate
breaking/formation
of bond in the
substrate to form
product
Enzyme speed up the rate of chemical reactions
by lowering EA
 PROPERTIES OF ENZYME
1. Globular protein
2. Enzyme is highly specific to its substrate
3. Enzyme do not change after the reaction @
reusable
4. Enzyme activity is maximum at optimum
temperature and pH.
- enzyme will be denatured @ destroyed
at extreme temperature and pH.
5. Lowers the activation energy
6. Reversible catalyst ; enzyme catalyse reversible
or forward reaction
6. Required in small quantity to catalyse reaction
7. Act faster
 CLASSIFICATION OF ENZYME
 Recommended by International
Union of Biochemistry (IUB) based on the
type of reaction the enzyme catalyses.

 Naming of enzyme :

Name of the + suffix

substrate “ase”
 CLASSIFICATION OF ENZYME

Lyase

Isomerase Transferase

CLASSES OF
ENZYME
Hydrolase Ligase

Oxidoreductase
 OXIDOREDUCTASE
Catalyse the transfer of oxygen and hydrogen
atoms between substrate (catalyse all oxidation-
reduction reactions)

 need coenzymes FAD , NAD+, NADP+

 e.g. dehydrogenase , oxidase
Example 1:
Malate dehydrogenase

MALATE OXALOACETATE

NAD+ NADH + H+

• Malate is oxidized to oxaloacetate

• NAD+ is reduced to NADH + H+
Example 2:

• Succinic acid is oxidized to fumaric acid

• FAD is reduced to FADH2
 TRANSFERASE

Catalyse the transfers of a functional group

(e.g. methyl, amino, phosphate) from one
substrate to another

 Eg : hexokinase, phosphorylase,
transaminase
Example:

hexokinase
Glucose Glucose -6- phosphate

ATP ADP

Hexokinase
-catalyse the transfer of phosphate group
from ATP to glucose
 HYDROLASE

Catalyse the breakdown of chemical bond by

adding water molecule

 e.g. Digestive enzyme : protease, lipase,

amylase, sucrase

sucrase
sucrose + H2O fructose + α glucose
 HYDROLASE

Catalyse the addition or removal of group of

atoms to a substrate

 e.g. decarboxylase, carboxylase

Example 1:

Pyruvate decarboxylase
Pyruvate ethanal

CO2

Pyruvate decarboxylase
- catalyse removal of CO2 from pyruvate
Example 2:

RuBP carboxylase : catalyse addition of CO2

 ISOMERASE

Catalyse the rearrangement of atoms in a

substrate molecule; conversion of isomer

glucose -1- phosphate

phosphoglucomutase

glucose -6- phosphate

 LIGASE

Catalyse the formation of new bonds between

two substrate molecules by using energy from
ATP

Aminoacyl-tRNA
Amino synthetase
acid Aminoacyl-tRNA
+
ATP AMP
tRNA
+ 2 Pi
 MECHANISM OF ENZYME ACTION
• Enzyme have specific conformation with
active site
• Substrate binds at active site of enzymes
• To form enzyme-substrate complex
• Substrate are brought closer to each other
• Active site of enzymes facilitate the
breaking/formation of chemical bonds between
substrate
• activation energy is lowered & product are formed
 Enzyme Rate Reaction Curve (Hyperbolic curve)

Action of enzymes at the

active site

A : Few substrate
- low reaction rate
B : more substrate
- higher reaction rate
C : more substrate but no
free active site
- no increase reaction
rate (constant)

• Enzyme rate reaction curve showing hyperbolic curve

HYPOTHESIS RELATED TO THE MECHANISM OF
ENZYME ACTION

1) LOCK AND KEY MODEL

2) INDUCED FIT MODEL
(widely accepted)
 LOCK AND KEY MODEL
• The substrate is the ‘key’ that fits exactly the
‘lock’ (enzyme)
• Enzyme + substrate = enzyme-substrate
complex
• Active site of enzyme exactly complementary
to substrate

(LOCK) (KEY)
 LOCK AND KEY MODEL

 An enzyme collides with its substrate molecule

 The substrate binds to the active site of enzyme

 Active site of enzyme exactly complementary

to substrate

 Enzyme-substrate complex formed

 Reaction occur where the substrate reacts

within the complex and product is released
 Enzyme is not change or damage and can
be reused
LOCK AND KEY MODEL
 INDUCED-FIT MODEL

Attachment of substrates induces the

conformation change of active site of enzyme
until its active site fits
with the substrate

* Active site of enzyme NOT exactly complementary to

substrate
 INDUCED-FIT MODEL

 Active site of an enzyme is not exactly

complementary to the shape of substrate

 Enzyme collides with the substrate molecule

 To form enzyme-substrate complex

 The binding induce a slight change in the

conformation of active site of enzyme

Allowing the substrate fit to enzyme precisely

 INDUCED-FIT MODEL

 The active site changes conformation and

becomes fully complementary with the
substrate

 Enable the enzyme to carry out their

catalytic function

 Product is formed and active site of enzyme

changes back to its original conformation
 DIFFERENCES BETWEEN LOCK & KEY AND
INDUCED-FIT MODEL
LOCK & KEY MODEL INDUCED-FIT MODEL
The active site is not flexible The active site is flexible and
and exactly complementary to not exactly complementary to
substrate substrate

The active site is not changed The binding of substrate

after binding with substrate induce the conformational
changes to active site of
enzyme
Enzyme does not change and Enzyme reverts/reconvert to its
can be reused after reaction original conformation when the
product is released
FACTORS AFFECT THE ENZYMATIC REACTION

Substrate concentration

Temperature pH

Cofactor
 EFFECTS OF SUBSTRATE CONCENTRATION
• At low substrate
concentration, reaction
rate is directly
proportional to the
substrate concentration

• Not all active sites bind

with substrates(some
active site remain free).
The active site of an
enzyme molecule can
bind with a certain
number of substrate
molecules at a given
time.
 EFFECTS OF SUBSTRATE CONCENTRATION
• As substrate
concentration increase,
the rate of reaction
increase
collision between
substrates & active
sites of enzymes also
increase
more active sites bind
with substrates
Until there are no more
free active sites //
saturation of active site
 EFFECTS OF SUBSTRATE CONCENTRATION
• At very high substrate
concentration, reaction
rate reaches the
maximum and become
constant
• Any further increase in
substrate concentration
will not increase rate of
reaction
• Because all active sites
are bind with substrates/
enzyme active site is
fully saturated with
substrate
EFFECTS OF SUBSTRATE CONCENTRATION
 EFFECTS OF TEMPERATURE
• At low temperature, the
rate of reaction
• occur slowly
due to the slow
movement of molecules
• collision between
substrate & active site are
decrease

 enzyme also inactive at

low temperature
 EFFECTS OF TEMPERATURE
• As temperature
increase, the rate of
reaction increase
 movement of molecules
increase

 collision between
substrate & active sites
of enzymes are
increase (more
frequently)
 EFFECTS OF TEMPERATURE
• For every 10ºC rise in
temperature,
reaction rate is
doubled up to optimum
temperature at which
reaction rate is maximum

• Optimum temperature:
 Human enzymes
mostly at 35οC – 40οC
 Enzyme of
thermophilic bacteria
(live in hot spring) is
70οC
 EFFECTS OF TEMPERATURE
• Higher temperature
(beyond optimum
temperature), reaction
rate decrease
 > 40°C: activity decrease

 at 60°C/extremely high
temperature: reaction stops
& enzyme is denatured
 EFFECTS OF TEMPERATURE
• Due to denaturation of
protein
 changes in conformation of
active site

 Results from the breaking of

ionic bond / hydrogen
bond/ disulphide bridge/
hydrophobic & van der
Waals interactions

• Substrate can no longer

fit into the active site of
the enzymes
EFFECTS OF TEMPERATURE
 EFFECTS OF pH
• Enzymes function
efficiently within narrow
pH range
• Different enzymes have
different optimum pH
 e.g: pepsin=pH2,
salivary amylase = pH7.2

• Optimum pH:
 pH at which reaction rate
is maximum

• Above or below optimum pH

pH, the rate of reaction Optimal pH for enzyme pepsin
decrease
 EFFECTS OF pH
• Deviation from optimum
pH range results in excess
H+ or OH-
• that alters the acidic /
basic/ functional group /
side chain of amino acids
in the enzyme
• Causing ionic or hydrogen
bonds to be broken
• Change conformation of
the active site // enzyme
denatured
• Substrate cannot bind to
active site pH
• Lower down the rate of Optimal pH for enzyme pepsin
reaction
 EFFECTS OF pH
• If pH is too low or
extreme, enzyme will
denature
 Result from the breaking
of ionic bonds

 Substrate can’t bind to

active site of enzyme

pH
Optimal pH for enzyme pepsin
EFFECTS OF pH
 EFFECTS OF COFACTORS

• An additional non-protein substance

• Present within the active site
• Sometimes needed to activate certain enzyme

It can be permanently bound to the active

site or may bind loosely with the substrate
during catalysis.
 HOTS QUESTION

Why are the products separate from

enzyme at the end of reaction ?