Chapter 15

Biocatalysis (Part 2)

Chapter review
biocatalysis
Properties and
mechanisms of
actions
Properties Mechanism
of enzymes of enzyme
action
Classification
Activation
energy

Cofactors
Types &
f(x) of
cofactors

Factors
Models
affecting
enzymatic
2 hypotheses reaction

Lock
and key

Induced
fit

Inhibition
reversible

irreversible

Competitive Non
competitive

LEARNING OUTCOMES
Explain factors that affect the enzymatic
reaction.

Factors affecting the rate of enzyme

reactions
Enzyme Concentration
Substrate Concentration
Temperature
pH
Cofactors

Enzyme Concentration
Rate of reaction is proportional to the enzyme
concentration (pH and temperature kept
constant)
The rate of reaction .... by increasing the
enzyme concentration

[Enzyme]
At low concentration of enzyme
decreases , rate of reaction increases.
As enzyme concentration increases,
more active sites available for
substrate to bind.
Rate of reaction keeps increasing if the
substrate is in excess
If amount of substrate is limited, rate
of reaction reaches its max

Substrate Concentration
The rate of enzyme reaction with increasing
substrate concentration
Any further increase in substrate concentration
- no significant change in rate of reaction.
At high substrate concentration, all the active
sites are fully occupied by substrate molecules.
Any extra substrate molecules will have to wait
for the ES complex to release the product and
become available again.
What is the rate of reaction now?

 At low [substrate], the rate of enzyme reaction

increase as [substrate] ..
There are more enzyme molecules compared
substrates, not all active sites are occupied.

Temperature
Optimum temperature temperature that promotes
max. rate of reaction
At low temp, rate of
reaction is very low
molecules move slowly,
decreasing chances of
substrate molecules to
collide with enzyme
molecules at active sites.

Figure : Effect of
temperature on the
rate of enzyme
controlled reaction.

as temperature increases,
the kinetic Energy of
enzyme & substrate
increases, molecules move
faster, more effective
collision occur between
substrate & active site of
enzyme
Figure : Effect of
temperature on the rate
of and enzyme controlled
reaction.

More ES complexes
form.
Rate of reaction
increases

For every 100C rise in temperature, the

rate of reaction is doubled
until it reaches optimum temperature
at which the rate of reaction reaches its
maximum
beyond the optimum temperature, the
rate of reaction decrease drastically
due to denaturation of enzyme

 High temperature causes the weak bonds such as ionic/

H/ van der Waals/ hydrophobic interaction to break.
Change the conformation of active sites
Substrates can no longer bind to the active sites of the
enzyme

Figure : Effect of temperature on the rate of and

enzyme controlled reaction.

pH
Every enzyme functions most effective/
efficient over a NARROW pH range.
Optimum pH max. rate of reaction
different enzymes have different optimum pH values
Changes in pH above/ below optimum pH change H+
concentration alter ionic charges of functional/ side
chain/ acidic & basic groups of amino acids in the enzyme
Ionic & Hydrogen bonds that hold specific 3D shape of
enzyme disrupted change conformation of active sites
substrate cannot fit into active site to form ES Complex

 The rate of reaction decreases quickly

If the changes in pH is too high or too low, active
sites are distorted causing enzyme to denature
Rate of reaction decreases drastically

Optimum pH values for some

enzymes
Enzyme

Optimum pH

Pepsin

2.00

Sucrase

4.50

Enterokinase

5.50

Salivary Amylase

4.80

Catalase

7.60

Trypsin

8.00

COFACTORS
LEARNING OUTCOME:
Define the meaning of cofactors
Explain the types and functions of
three cofactors: metal ions,
coenzymes, and prosthetic groups.

Chapter review
biocatalysis
Properties and
mechanisms of
actions
Properties Mechanism
of enzymes of enzyme
action
Classification
Activation
energy

Cofactors
Types &
f(x) of
cofactors

Factors
affecting
enzymatic
2 hypotheses reaction

Lock
and key

Models

Induced
fit

Inhibition
reversible

irreversible

Competitive Noncompetitive

Cofactors

Non-protein
substance (organic/inorganic
ions) essential for some enzymes to
function efficiently.
may be bound

tightly to the active site as permanent

residents.
loosely & reversibly along with the substrate.

Three types of cofactors

Cofactors

Metal ions
(activators)

Coenzymes

Prosthetic group

BASIC TERMS
Apoenzyme the enzyme alone
Holoenzyme enzyme + cofactor

Metal ions

Inorganic substances, mostly trace elements

(micronutrients) - required in very small
amounts
Bound tightly & temporarily to the enzyme/
substrate & change its active site/ shape
Easier for substrate molecules to fit into
active site/ assist in the formation ESC
Increase the rate of reaction
May also bind the enzyme & substrate
together

Examples of metal ion/ mineral

cofactors
Salivary amylase requires the presence of
(Cl-) ions before efficiently convert starch
maltose.
The enzyme thrombokinase, which converts
prothrombin into thrombin during blood
clotting, is activated by calcium (Ca2+) ions.

Action of metal cofactors. In this cartoon an enzyme is unable

to become functionally formed unless a metal ion is available.
Once the metal is bound the active site forms and the enzyme
is ready to carry out its function.

Coenzymes

organic non-protein molecule

bind loosely & temporarily to the enzyme.
many coenzymes are derived from vitamins
especially vitamin B.

Vitamin K is found in alfalfa,

fish
livers, leafy/green
vegetables, egg yolks, and
soybean
oil.
It
is a
coenzyme that helps in
blood clotting.

NAD+
acts
as
a
coenzyme to
dehydrogenases by acting as an
electron & hydrogen acceptor.
NAD+

Glyceraldehyde-3-phosphate
1,3-diphosphoglycerate

NADH + H+

dehydrogenase

Coenzymes
Vitamin

Coenzymes

Niacin (Vit B3)

NAD+

Panthothenic acid
(Vit B5)

Coenzyme A (coA)

Coenzymes, consists of small organic molecules, many of

which are derived from vitamins. Hence, vitamins are often
precursors to coenzymes.

Structure of FAD
nitrogens 1 & 5 carry hydrogen atoms in FADH2

Prosthetic groups

organic non-protein molecules

Binds tightly and permanently to the
enzyme a part of the enzyme.
As carriers & transfer grps of atoms/
electrons from 1 molecule to another

Example: haem.

a ring- shaped organic molecule with

iron at its center

Summary
cofactor

3 types
(metal ions)

coenzymes
Prosthetic
group