15.

1 PROPERTIES OF ENZYME AND MECHANISM

OF ACTIONS

15.2 COFACTORS

15.3 INHIBITION

15.1 PROPERTIES OF ENZYME

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)
d) Describe the hypotheses related to its
mechanism of action: Induced fit model,
lock and key model
e) Explain factors that affect the enzymatic
reaction
Learning Outcomes
Enzyme is a biocatalyst
Bio = life
Catalyse = speed up the rate of reaction.

Definition

Enzyme is a globular proteins which act as
biological catalyst that speed up the rate of
biochemical reaction in living things by lowering
the activation energy.
Learning Outcomes:
15.1 a) State the properties of enzymes
Structure of enzyme
Mostly are globular
proteins.
Consist of one or several
polypeptide chain with
unique amino acids
sequence.
R groups of amino acids
interact with each other
to form hydrogen bonds,
ionic bonds &
hydrophobic interaction.


Learning Outcomes:
15.1 a) State the properties of enzymes

Structure of Enzyme
PROPERTIES OF ENZYME
1. Enzymes are highly specific
One enzyme acts only on a
specific reactions or a few
related reactions.
Since each enzyme has a
unique amino acid sequence,
different interactions between
R groups are formed.
These interactions cause the
polypeptide to fold into a
unique 3 dimensional shape.
Learning Outcomes:
15.1 a) State the properties of enzymes
Structure of Enzyme
1. Enzymes are highly specific
The shape are highly specific especially at small
part of enzyme called active site which binds to a
specific substrate.
Learning Outcomes:
15.1 a) State the properties of enzymes

2. Enzymes remain unchanged at the end of reaction
Enzymes are not altered, destroyed or used by the
reaction it catalyses.
Some enzyme can be used repeatedly.

3. Enzymes are highly efficient
Only small amount of enzyme is needed to react with
large amount of substrate.

Learning Outcomes:
15.1 a) State the properties of enzymes
Properties of Enzyme
4. Most catalysed reaction are reversible
An enzyme can catalyse both the forward and reverse
reactions.

5. Does not influence the direction of reaction and final
concentration of products
It only speeds up the rate of reaction.

6. Enzymes activity is influenced by temperature, pH,
substrate & enzyme concentration and inhibitors.
Enzyme activity are maximum at optimum temperature
or PH
can be denatured by extreme temperature or extreme
pH.

Learning Outcomes:
15.1 a) State the properties of enzymes
15.1 PROPERTIES OF ENZYME

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)
d) Describe the hypotheses related to its
mechanism of action: Induced fit model,
lock and key model
e) Explain factors that affect the enzymatic
reaction
Learning Outcomes
1. In 1964, IUB (International Union of Biochemistry)
introduced a classification guide of enzymes to
differentiate enzymes.

2. Before it is introduced, it is hard to recognize enzyme
according to traditional way of naming.

3. Eg: pepsin, renin, trypsin

4. Enzymes are classified into 6 main classes according
to the general reaction it catalyses.

ENZYME CLASSIFICATION
Learning Outcomes:
15.1 b) State the six classes of enzyme according to IUB classification

The Classification of Enzyme
1. Common naming of enzyme usually has 3
parts
1) name of substrate
2) type of reaction
3) suffix ase
Eg: Alcohol dehydrogenase, RuBP
carboxylase
2. Sometimes, they are named based on the
substrate they act on
3. Eg: sucrase, amylase, lipase, protease

Learning Outcomes:
15.1 b) State the six classes of enzyme according to IUB classification

Enzyme Classification

Enzyme group
Type of reaction
catalysed
Enzyme
examples
1. Oxidoreductase

Transfer of O or H atoms or
electrons between
substances, i.e. all
oxidation-reduction
reactions.
Dehydrogenase
Oxidase
2. Transferase

Transfer of a chemical
group from one substance
to another
Transaminase
Phosphorylase
3. Hydrolase

Breaking a molecule into
simpler molecules by
hydrolysis reactions
(addition of water
molecule)
Peptidase
Lipase
Sucrase
Learning Outcomes:
15.1 b) State the six classes of enzyme according to IUB classification

Enzyme
group
Type of reaction
catalysed
Enzyme
examples
4. Lyase

Breaking of a chemical bonds
without the addition of water
(other than by hydrolysis)
Decarboxylase
(remove CO2)
5. Isomerase

The rearrangement of atoms
within a molecule converting
from 1 isomer to another
Isomerase
Mutase
6. Ligase

Formation of bonds between
two molecules using energy
from the breakdown of ATP
Synthetase

Learning Outcomes:
15.1 b) State the six classes of enzyme according to IUB classification.

15.1 PROPERTIES OF ENZYME

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)
d) Describe the hypotheses related to its
mechanism of action: Induced fit model,
lock and key model
e) Explain factors that affect the enzymatic
reaction
Learning Outcomes
Before any chemical reactions can occur, reactants
must absorb energy to break the existing bonds
before new bonds can be formed.
Definition
The minimum energy required to break the existing
bond for a chemical reaction to take place is called
activation energy.
Learning Outcomes:
15.1 c) Explain the mechanism of enzyme action (lowering activation energy)
Activation Energy (Ea)
The existing bonds start to
break when enough energy is
absorbed.
Reactants become unstable
and very reactive.
This condition is called
transition state.
By lowering the activation
energy, less energy can be
absorbed to start a reaction.
More reactants can reach
transition state and converted to
products within a short time.
Learning Outcomes:
15.1 c) Explain the mechanism of enzyme action (lowering activation energy)
Learning Outcomes:
15.1 c) Explain the mechanism of enzyme action (lowering activation energy)
Activation Energy (Ea)
Learning Outcomes:
14.1 c) Explain the mechanism of enzyme action (lowering activation energy)
Activation Energy (Ea)
Substrate binds to enzymes active site to form an
enzyme substrate complex
Bring substrate molecule closer to each other / in
a correct orientation
Enzyme stretch / distort the existing bonds
Learning Outcomes:
15.1 c) Explain the mechanism of enzyme action (lowering activation energy)
How Enzyme Lowers the Activation Energy
R group of active sites interact with substrates to
increase the reactivity of the substrate.

These make it easier for substrate to be changed
into product.

Thus, activation energy is reduced without
increasing the temperature in living cells.
Learning Outcomes:
15.1 c) Explain the mechanism of enzyme action (lowering activation energy)
15.1 PROPERTIES OF ENZYME

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)
d) Describe the hypotheses related to its
mechanism of action: Induced fit model,
lock and key model
e) Explain factors that affect the enzymatic
reaction
Learning Outcomes
1. Proposed by Koshland
(1959).
2. Suggested that the
active site is not exactly
complementary to the
shape of substrate.
3. The active site is
flexible and can change
its shape.

Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Induced fit model
4. When collisions occur between
enzyme and substrates,
substrate can bind to the active
site of the enzyme.
5. The binding of the substrate
induces the enzyme to change
its shape slightly.
6. To enable the substrate to fit
precisely to the active site.
7. An Enzyme-Substrate complex
is formed.

Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
8. Enzyme stretched the existing
bond to facilitate the reaction to
occur.
9. Activation energy is reduced,
so it is easier to achieve
transition state.
10. Product is formed and
released from the active site.
11. Enzyme changes back to
original conformation.

Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Induced fit model
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Induced fit model
Learning Outcomes:
15.1d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
1. Enzyme is very specific because its active site
had a specific shape that fits exactly to a
specific substrate.
2. The shape of active site is rigid.
3. This is called lock and key hypothesis which is
proposed by Fischer (1890).
4. Enzyme is bigger and act as lock.
5. Substrate is smaller and act like a key.
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Lock & Key model
Lock & key model
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Lock & key model
1. The shape of active site is
exactly complementary to the
substrate.
2. Substrate binds to the active site
of enzyme to form an enzyme-
substrate complex (ES complex).
3. The shape of substrate slightly
changed.
4. Existing bonds are broken.
5. Transition state is achieved,
reaction occurs.

Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
6. Products are formed.
7. The shape of products differ from substrates.
8. Thus, products no longer fit into active site.
9. Products are released from active site.
10. Free active site binds with other substrate.
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
Lock & key model
Learning Outcomes:
15.1 d) describe the hypotheses related to its mechanism of action: Induced fit model, lock and key model
15.1 PROPERTIES OF ENZYME

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)
d) Describe the hypotheses related to its
mechanism of action: Induced fit model,
lock and key model
e) Explain factors that affect the enzymatic
reaction
Learning Outcomes
1. Substrate concentration
2. pH
3. Temperature
4. Cofactors
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Factors affect the enzymatic reaction
Substrate Concentration
At low substrate
concentration, the rate of
enzyme reaction
increases as substrate
concentration increase.
There are many enzyme
molecules compared to
substrates, not all active
sites are occupied.
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

As substrate concentration
increase, more active sites
are occupied until all of them
bind to its substrate.
The rate of reaction reaches
the maximum rate.
Increasing substrate
concentration does not affect
the rate of reaction since all
active sites are saturated with
the substrate.
The graph reaches a plateau.
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Most enzyme are effective
over a narrow PH range.
Maximum enzymes are
effective over a narrow PH
range.
Different enzyme has
different optimum pH value.
Changes in pH above or
below the optimal pH value
will change the hydrogen
ion (H
+
) concentration.

pH
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

This ion changes the ionic
charge of the acidic & basic
groups of amino acids of the
enzyme.
The ionic & hydrogen bonds
that hold the specific 3D
shape of enzyme are
disrupted.
It changes the conformation of
active sites.
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Substrate cannot fit into the
active site to form Enzyme-
Substrate complex.
Therefore, the rate of reaction
decreases quickly.
If the changes of pH is too
high or too low, the active
sites are destroyed causing
the enzyme to denature.
Therefore the rate of reaction
decreases quickly.
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Enzyme Optimum pH
Pepsin 2.00
Sucrase 4.50
Enterokinase 5.50
Salivary amylase 7.80
Trypsin 8.00
Pancreatic lipase 9.00
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

All enzymes work within
a range of temperature
specific to organism.
Increases in
temperature generally
lead to increases in
reaction rates.
At low temperatures, an
enzyme-catalysed
reaction takes place
slowly.
Temperature
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Hydrogen bond and
hydrophobic interaction
are not flexible enough.
Induced fit mechanism
is not optimum
Movement of molecules
slow
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

As temperature increase, the
movement of molecules
increases.kinetic energy of
the enzyme & substrate
increases.
More collisions occur
between enzyme and
substrates to form ES
complex.
The rate of reaction
increases until it reaches
optimum temperature
(usually 35
0
C-40
0
C).

When temperature
exceeds optimum
temperature, the
bonds which hold the
specific 3D shape of
enzyme are broken.

Active sites are
destroyed, enzyme
denatured.

Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Resulting from the
breakdown of the weak
bond such as hydrogen
bond, hydrophobic
interaction,disulphide
bridge and Van der
Waals
Substrates can no
longer fit into the active
sites of the enzyme.
The rate of reaction
decreases quickly.
Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction

Learning outcome:
15.1 e) explain factors that effect the enzymatic reaction