Types of Enzymes

Extracellular Enzymes
- leave the cell & exert actions
outside the cell

Intracelluar Enzymes
- exert action inside the cell
 Control of metabolism
enzyme
A+B C
• Metabolic reactions can be controlled and
speeded up by enzymes
– metabolic reactions would be too slow to
occur if no enzymes are present!
 Investigation 4.1
Detection of the presence of
Catalase in Plant and Animal
Tissues
liver potato meat apple boiled liver growing splint

A B C D E

3
5cm of 3% hydrogen peroxide solution

What is the purpose of setting up tube E ?

Ans: To see whether the catalase in the tissue can still
catalyse the decomposition of hydrogen peroxide
after boiling.
liver potato meat apple boiled liver growing splint

A B C D E

3
5cm of 3% hydrogen peroxide solution

What gas is evolved when fresh tissues are added

into the hydrogen peroxide solution ?
Ans: Oxygen.
liver potato meat apple boiled liver growing splint

A B C D E

3
5cm of 3% hydrogen peroxide solution

Which tissue has the highest catalase activity ?

Ans: Fresh liver tissue.
liver potato meat apple boiled liver growing splint

A B C D E

3
5cm of 3% hydrogen peroxide solution

What conclusion can you draw from the results of

this experiment ?
Ans: Catalase is produced by fresh tissues only.
Boiling kills the cells and destroys the enzyme.
 Nature and properties of
enzymes

✇ Biological
Biological-found
Catalyst
within living organisms
Catalyst-substance which can speed up
a chemical reaction
♎ Proteins in nature Prote
in

♎ Specific in action
– one kind of enzyme will catalyse only one kind
of chemical reaction
 Mechanism of enzyme
action
• Each enzyme has an active site
 Mechanism of enzyme
action

• Active site : the place where

substrate binds with the enzyme
• Each active site can only allow
specific substrate to fit in
 Mechanism of enzyme action
• The enzyme and substrate molecules combine
to form a temporary structure called enzyme-
substrate complex
 Action of enzyme
(Anabolic reaction)
enzyme-substrate enzyme-product
complex complex

product
substrate

enzyme enzyme
 Action of enzyme
(catabolic reaction)
enzyme-product enzyme-substrate
complex complex

products substrate

enzyme enzyme
 Mechanism of enzyme action
• The product molecules are then formed
and they escape from the active site
• The active site is free to attach to other
substrate molecules again ( i.e. it can
be reused )
 This hypothesis to explain the
specificity of enzyme action is called

lock and key hypothesis

 Lock and key hypothesis
product

Substrate

product

Enzyme
Lock and key hypothesis

N ’ T
DO
P ES
SH A T C H
M A
 Nature and properties of
enzymes
* Catalyse reversible reaction in both
directions
* Enzymatic activity is affected by
temperature and pH
– high temperature and extreme pH can
denature enzymes
∵ protein in nature

pH
 Effect of temperature on enzyme
activity

• Low temperature : low kinetic energy possessed

by the substrate and enzyme molecules

The enzyme becomes inactive

\Lower reaction rate (and enzyme

activity)
Effect of temperature on enzyme activity
 Effect of temperature on enzyme
activity

• Higher temperature : more kinetic

energy possessed by the substrate and
enzyme molecules
\Higher reaction rate (and enzyme
activity)
 Effect of temperature on enzyme
activity

• The temperature which allows the

highest enzyme activity is called the
optimum temperature
 Effect of temperature on enzyme
activity

• At temperature higher than the optimum

temperature, the shape of the active site is
changed
The enzyme is said to be denatured
\substrate can no longer bind to the
active site of the enzyme
Effect of temperature on enzyme
activity
 Effect of temperature on enzyme
activity

• Enzymes become inactive but not

denatured at low temperatures
\They can regain catalytic function
when the temperature increases
 Effect of temperature on enzyme
activity

• Denaturation is a permanent process

\When an enzyme is denatured, its
catalytic function is lost permanently,
and cannot be restored ( never become
active again ) even it is put in lower
temperature
Effect of temperature on enzyme
activity
• Different enzymes may have different
optimum temperature
• The optimum temperature of many enzymes
in human body is not 37oC, even though this
is our body temperature!
– Our body temperature can provide a
better environment for all the enzymes in
our bodies to work
 Investigation 4.2

Effect of Temperature on
the action of Amylase
A1 A2 B1 B2 C1 C2 D1 D2

ice bath water bath

at 2℃ at 37 ℃ water at
100 ℃
amylase starch water bath at
solution solution room temperature thermometer

amylase and
starch solution
iodine
solution
spotting tile

Why is it necessary to equilibrate the pair of test tubes

under each condition for 5 minutes before mixing ?
Ans: To ensure the pair of test tubes are in the same
temperature.
A1 A2 B1 B2 C1 C2 D1 D2

ice bath water bath

at 2 ℃ at 37 ℃ water at
100 ℃
amylase starch water bath at
solution solution room temperature thermometer

amylase and
starch solution
iodine
solution
spotting tile

What is the factor affecting the action of amylase in

converting starch into sugar ?
Ans: Temperature.
A1 A2 B1 B2 C1 C2 D1 D2

ice bath water bath

at 2 ℃ at 37 ℃ water at
100 ℃
amylase starch water bath at
solution solution room temperature thermometer

amylase and
starch solution
iodine
solution
spotting tile

At which temperature does all the starch in the tube

disappear first ?
Ans: 37 ℃.
A1 A2 B1 B2 C1 C2 D1 D2

ice bath water bath

at 2 ℃ at 37 ℃ water at
100 ℃
amylase starch water bath at
solution solution room temperature thermometer

amylase and
starch solution
iodine
solution
spotting tile

How does the activity of amylase change with

temperature ?
Ans: At room temperature, amylase works but not as
well as that at 37 ℃. It does not work at 2℃
and 100℃.
A1 A2 B1 B2 C1 C2 D1 D2

ice bath water bath

at 2 ℃ at 37 ℃ water at
100 ℃
amylase starch water bath at
solution solution room temperature thermometer

amylase and
starch solution
iodine
solution
spotting tile

At which temperature does amylase work best ?

Ans: 37 ℃.
 Effect of pH on enzyme
activity
• pH
Too•extreme
Optimum
of mediumpHcan
pH : higher
thedestroy
pH value at which
or lower
the the
enzymes
than enzyme has
optimum
pHthe
canhighest activity
inactivate the enzyme
 Different enzymes has different
optimum pH

Enzyme 2 Enzyme 1
Enzyme 3
Rate of reaction

pH
 Factors affecting
Enzyme Activity & Rate of
Enzymatic Reactions
 Factors affecting enzyme activities :
– Temperature
– pH

 Factors affecting rate of enzymatic

reactions :
– Temperature
– pH
– Enzyme concentration
– Substrate concentration
Factors affecting enzyme action :
Substrate concentration
Factors affecting enzyme action :
Substrate concentration
Max. Rate
Rate of reaction

Substrate
conc.
Application of Enzymes

Biological Washing Powder

~ End ~