Activation energy

Activation energy - Initial energy that must be

given to a substrate in order for it to be converted into a
product.
Enzymes decrease the activation energy of the
reactions that they catalyse.
They do this by holding the substrate in such a
way that the molecules can react more easily.

Activation energy
Rxns catalysed by enzymes take place at
a much lower temperature that than they
would without them.

Factors affecting the rate of

enzyme action

Factors affecting the rate of

enzyme activity
Temperature
Four Variables

pH
Enzyme Concentration
Substrate Concentration

Substrate Concentration
As the substrate concentration increases, the
rate increases because more substrate
molecules can collide with active sites, so
more enzyme-substrate complexes form.
At higher concentrations the enzyme molecules
become saturated with substrate, and there are few
free active sites, so adding more substrate doesn't
make much difference (though it will increase the rate
of E-S collisions).

Substrate Concentration

The maximum rate at infinite substrate concentration is called

vmax, and the substrate concentration that gives a rate of half
vmax is called KM. These quantities are useful for characterising an
enzyme. A good enzyme has a high vmax and a low KM.

Enzyme concentration
As the enzyme concentration increases the rate of the
reaction also increases, because there are more
enzyme molecules (and so more active sites),
available to catalyse the reaction therefore more
enzyme-substrate complexes form.

Temperature
Enzymes have an optimum temperature at which
they work fastest. For mammalian enzymes this is
about 40C, but there are enzymes that work best at
very different temperatures, e.g. enzymes from the
arctic snow flea work at -10C, and enzymes from
thermophilic bacteria work at 90C.

Temperature
Up to the optimum temperature the rate increases
geometrically with temperature (i.e. it's a curve, not a
straight line). The rate increases because the enzyme
and substrate molecules both have more kinetic
energy and so collide more often, and also
because more molecules have sufficient energy
to overcome the activation energy.

Temperature
Above the optimum temperature the rate decreases
as more of the enzyme molecules denature. The
thermal energy breaks the hydrogen bonds
holding the secondary and tertiary structure of the
enzyme together, so the enzyme loses its shape
and becomes a random coil - and the substrate can
no longer fit into the active site. This is

Effect of heat on enzyme activty

Denaturing the protein

ACTIVE SITE CHANGES SHAPE
SO SUBSTRATE NO LONGER FITS
Even if temperature lowered enzyme

cant regain its correct shape

Temperature

Enzym
e
Activit
y

OPTIMUM TEMP.

40

Temperature

pH
Enzymes have an optimum pH at which they work
fastest.

For most enzymes this is about pH 7-8 (normal body

pH), but a few enzymes can work at extreme pH, such
as gastric protease (pepsin) in our stomach, which
has an optimum of pH 1.

pH
The pH affects the charge of the amino acids at the
active site, so the properties of the active site change
and the substrate can no longer bind.
For example a carboxyl acid R groups will be
uncharged a low pH (COOH), but charged at high pH
(COO-). The pH affects the charge of the amino acids
at the active site, so the properties of the active site

2. pH

Extreme pH levels will produce denaturation

The structure of the enzyme is changed
The active site is distorted and the substrate molecules will no
longer fit in it
At pH values slightly different from the enzymes optimum
value, small changes in the charges of the enzyme and its
substrate molecules will occur
This change in ionisation will affect the binding of the substrate
with the active site.

pH
Optimum pH values
Enzyme
activity

Pepsi
n

Trypsin

11

pH

Enzyme Inhibitors
Inhibitor - substance that slows down or stops an
enzyme controlled reaction.
Its possible for some other molecule other than
the substrate to bind to an enzymes active site
if its very similar to the enzymes substrate.
This would inhibit the enzymes function.

Competitive Inhibitors
Has a structure that similar to that of the substrate
and so compete with the substrate for the active site.
As a result, it will reduce the no. of enzyme
substrate complexes and so reduce the reaction
rate.
This type of inhibition is reversible by an increase in
substrate concentration: eventually allow substrate to
bind.
So the more substrate available, the less inhibitor
will bind its active site.

Competitive Inhibitors

Competitive Inhibitors

The effect of enzyme inhibition

Succinat
e

Succinate dehydrogenase

CH2COOH

COOH

Fumarate + 2H++
2e-

CHCOOH

CH2
CH2COOH

COOH
Malonate

2007 Paul Billiet ODWS

CHCOOH

Non-Competitive Inhibitors
Does not compete for active site.
It attached to another part of enzyme
causing the overall shape of the enzyme
molecules to change.
Changes the shape of the active site so
substrate can no longer bind.

Increasing the amount of substrate

will not reduce this inhibitors since
there are not in competition for active
site.
Removal of inhibitor will restore normal

Irreversible Inhibitors
Some inhibitors bind irreversibly to an enzyme
and so destroy its catalytic properties
completely.
Usually poisonous to the cells because they
close down some part of the metabolism.
Are always non
competitive inhibitors.