Enzymes

Integrated Programme
CEDAR GIRLS SECONDARY SCHOOL
Extra reading: Biology in Context pp.46-53

Overview
Definition
Enzyme hypotheses
Types of enzymes
a. proteases
b. carbohydrase
c. lipases
Mode and specificity of action
Factors affecting enzyme activity
Enzyme inhibitors

Definition
Enzymes are biological catalysts that speed up the
rate of reaction by lowering its activation energy by
remaining chemically unaltered at the end of the
reaction.

Properties of enzyme
Enzymes are proteins
Enzymes are biological catalysts.They speed up
a reaction by lowering the activation energy.
Enzymes are not chemically changed after a
reaction. They can be used over and over again. A
small amount of enzyme can effect the change of
a large amount of chemical.
Enzyme activities can be affected by temperature
and pH
Enzymes are substrate-specific

Enzyme structure
Globular proteins
Have specific three-dimensional shape
determined by the sequence of amino acids
Special site known as active site allows substrate
molecule to bind
Substrate molecule binds temporarily with
enzyme active site forming enzyme-substrate
complex

Enzyme-substrate
complex

E-S
complex

Activation Energy
Minimum amount of energy required for a
chemical reaction to proceed
The lower the activation energy, the easier the
substrates can be turned into products
Enzyme helps to lower the energy barrier for the
reaction to proceed

Enzyme hypotheses
Lock and key theory
Induced-fit model theory

Lock and Key hypothesis

suggested that both a substrate and an enzyme
have specific geometric shapes that fit exactly into
each other.
This idea of both substrates and enzymes having a
natural geometric fit has been called the lock and
key hypothesis.
The problem with this hypothesis is that it doesnt
explain the stabilization of the enzyme.
When an enzyme has a substrate enter into its
active site, the enzyme will change its shape slightly
to match the substrate.
If the enzymes were to be specifically designed to fit
a substrate, then there would be no need for it to
have to adjust its shape.

Induced fit model

since enzymes were so flexible, the active site is
constantly being reshaped by its interaction with
the substrate.
It was suggested that substrate doesnt bind to an
active site as if it were specifically the right
shape, but that the amino acid side-chains that
are a part of the active site are molded into a
specific position.
This position allows the enzyme to start the
catalyzing process.

Types of enzymes

Mode and Specificity of

action
Enzyme catalyses reaction through the
complementary binding of substrate to the active
site of an enzyme, forming an unstable enzymesubstrate complex (E-S complex)
Breaking down of E-S complex releases the
products.
Formation of E-S complexes help to lower the
activation energy and thereby increases the rate
of reaction.
Enzymes are specific and binds only with a
specific substrate

Measuring enzymecatalysed reactions

Time needed for the substrates to disappear or
Time needed for the products to be formed

Volume of oxygen produced/

cm3

Formation of products

time/
min

Mass of starch/
g

Disappearance of
substrate

time/
min

 Initially there are a lot of substrate (hydrogen

peroxide/ starch) but no product (water and
oxygen/ maltose)
Substrates can enter into the unoccupied enzyme
active sites
As time pass by, all of the active sites are filled
and products formed. The amount of substrates
decreases as they start to disappear.
The amount of products formed becomes more
and more as the reaction proceeds
The reaction rate starts to slow down as the
number of empty active sites get lesser and lesser
Graphs starts to flatten out

Rate of reaction can be calculated using the

gradient method:

Measuring rate of
reaction at a
point in time.
Eg. Time = t

Factors affecting Enzyme

Activity
Any factor which affects the enzyme and hinder or
slow down the rate of formation of E-S complex
will reduce the speed of the reaction.
Any factors which increases the formation of E-S
complex speeds up the rate of reaction

Temperature
When temperature is low, kinetic energy of both
enzyme and substrate molecules are low
Rate of effective collision is low
Rate of E-S complex formation is low
Enzyme is inactivated
When temperature increases, kinetic energy
increases resulting in increase in effective
collision between enzyme and substrates to form
E-S complexes

 At optimum temperature, enzyme is most active

and most number of E-S complexes formed
Beyond optimum temperature, enzymes
denatured. High temperature breaks down the 3D
conformation of active sites.
Active sites no longer complementary to substrate

pH
Enzyme works best within a narrow pH range
Any deviation from the optimum pH reduces the
rate of reaction
Enzymes are denatured at extreme pHs
Extreme pH interferes with the ionic bonds that
help to maintain the 3-dimensional conformation
of the active sites.

Substrate and Enzyme

conc.
At the fixed enzyme concentration, the rate of
reaction increases with increasing substrate
concentration.
Higher substrate concentration means that the
chance of enzyme-substrate effective collision
increases.
More E-S complexes formed and as a result the
amount of products formed increases.
At very high substrate concentration, the rate of
reaction remains unchanged as substrate
concentration is no longer the limiting factor.
Rate of reaction can only increase if more
enzymes are added.

Enzyme inhibitors
Affect enzyme activity resulting in a decrease in
the rate of enzyme-catalysed reaction
Lesser E-S complexes formed and lesser products
Used to reduce the rate of enzyme activity (eg. To
kill off bacteria cells as a form of antibiotic)
Two main forms of inhibition Competitive, Noncompetitive

Competitive Inhibitor
(active site directed)
Inhibitor is structurally similar to substrate
Binds reversibly to enzymes active site and
compete with the substrate
Substrate unable to bind with active site
Lesser E-S complexes formed
Can be overcome by increasing the concentration
of the substrate so that there is a higher chance
for the substrate to bind with the active site

Non-Competitive Inhibitor
(non-active site directed)
Does not bind to active site of the enzyme
Bind reversibly or non-reversibly at a site that is
not the active site, the allosteric site.
Once bound, the conformation of the active site is
altered and substrate unable to fit into the active
site
Lesser E-S complexes formed
Increasing the substrate concentration does not
help to overcome the inhibition since the active
sites are no longer complementary to the
substrate due to a comformation change.

Specific application of non-competitive

inhibition (allosteric deactivation)

Application: End product

Inhibition
In a series of metabolic reactions in which the
product of one reaction acts as an inhibitor for the
previous/ earlier reactions
This is to ensure that resources are conserved and
no overproduction of a certain products
Product D can inhibit the enzyme reaction A, so
that there will be lesser B and C in the subsequent
reactions.
This in turn will decrease the amount of D
produced.

Co-factors & Co-enzymes

assist the enzyme and tend to increase its overall
efficiency.
Many vitaminsare components of coenzymes
and aid in various metabolic reactions within the
cell.