ENZYMES

A protein with catalytic

properties due to its power of
specific activation
2007 Paul Billiet ODWS

Chemical reactions
Chemical reactions need an initial input of
energy = THE ACTIVATION ENERGY
During this part of the reaction the
molecules are said to be in a transition
state.

2007 Paul Billiet ODWS

Reaction pathway

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Making reactions go
faster
Increasing the temperature make
molecules move faster
Biological systems are very sensitive to
temperature changes.
Enzymes can increase the rate of reactions
without increasing the temperature.
They do this by lowering the activation
energy.
They create a new reaction pathway a
short cut
2007 Paul Billiet ODWS

An enzyme controlled
pathway

Enzyme controlled reactions proceed 108 to 1011

times faster than corresponding non-enzymic
reactions.
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Enzyme structure
Enzymes are
proteins
They have a
globular shape
A complex 3-D
structure
Human pancreatic
amylase
Dr. Anjuman Begum
2007 Paul Billiet ODWS

The active site

One part of an

H.PELLETIER, M.R.SAWAYA
ProNuC Database

2007 Paul Billiet ODWS

enzyme, the
active site, is
particularly
important
The shape and the
chemical
environment
inside the active
site permits a
chemical reaction
to proceed more
easily

Cofactors
An additional nonprotein molecule
that is needed by
some enzymes to
help the reaction
Tightly bound
cofactors are called
prosthetic groups
Cofactors that are
bound and released
easily are called
coenzymes
Many vitamins are
coenzymes
2007 Paul Billiet ODWS

Nitrogenase enzyme with Fe, Mo and ADP cofactors

Jmol from a RCSB PDB file 2007 Steve Cook
H.SCHINDELIN, C.KISKER, J.L.SCHLESSMAN, J.B.HOWARD, D.C.REES
STRUCTURE OF ADP X ALF4(-)-STABILIZED NITROGENASE COMPLEX AND ITS
IMPLICATIONS FOR SIGNAL TRANSDUCTION; NATURE 387:370 (1997)

The substrate
The substrate of an enzyme are the
reactants that are activated by the
enzyme
Enzymes are specific to their
substrates
The specificity is determined by the
active site

2007 Paul Billiet ODWS

The Lock and Key

Hypothesis
Fit between the substrate and the active site of
the enzyme is exact
Like a key fits into a lock very precisely
The key is analogous to the enzyme and the
substrate analogous to the lock.
Temporary structure called the enzyme-substrate
complex formed
Products have a different shape from the
substrate
Once formed, they are released from the active
site
Leaving it free to become attached to another
substrate
2007
Paul Billiet ODWS

The Lock and Key

Hypothesis
S

E
E
E

Enzymesubstrate
complex

Enzyme may
be used again
P
P

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Reaction coordinate

The Lock and Key

Hypothesis
This explains enzyme specificity
This explains the loss of activity
when enzymes denature

2007 Paul Billiet ODWS

The Induced Fit

Hypothesis
Some proteins can change their shape
(conformation)
When a substrate combines with an
enzyme, it induces a change in the
enzymes conformation
The active site is then moulded into a
precise conformation
Making the chemical environment suitable
for the reaction
The bonds of the substrate are stretched
to make the reaction easier (lowers
activation
energy)
2007
Paul Billiet ODWS

The Induced Fit

Hypothesis

Hexokinase (a) without (b) with glucose substrate

http://www.biochem.arizona.edu/classes/bioc462/462a/NOTES/ENZYMES/enzyme_mechanism.html

This explains the enzymes that can react

with a range of substrates of similar types
2007 Paul Billiet ODWS

Factors affecting
Enzymes

substrate concentration
pH
temperature
inhibitors

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Substrate concentration: Non-enzymic

reactions

Reaction
velocity

Substrate concentration

The increase in velocity is proportional to

the substrate concentration
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Substrate concentration: Enzymic

reactions
Vmax
Reaction
velocity

Substrate concentration

Faster reaction but it reaches a saturation point

when all the enzyme molecules are occupied.
If you alter the concentration of the enzyme then
Vmax will change too.
2007 Paul Billiet ODWS

The effect of pH
Optimum pH values

Enzyme
activity

Trypsin

Pepsin
1
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7
pH

11

The effect of 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
2007
Paul Billiet ODWSof the substrate with the active
binding

The effect of
temperature
Q10 (the temperature coefficient) =
the increase in reaction rate with a 10C
rise in temperature.
For chemical reactions the Q10 = 2 to 3
(the rate of the reaction doubles or triples
with every 10C rise in temperature)
Enzyme-controlled reactions follow this
rule as they are chemical reactions
BUT at high temperatures proteins
denature
The optimum temperature for an enzyme
controlled reaction will be a balance
between the Q10 and denaturation.
2007 Paul Billiet ODWS

The effect of
temperature
Q10

Enzyme
activity

2007 Paul Billiet ODWS

10

20
30
40
Temperature / C

Denaturation

50

The effect of
temperature
For most enzymes the optimum
temperature is about 30C
Many are a lot lower,
cold water fish will die at 30C because
their enzymes denature
A few bacteria have enzymes that can
withstand very high temperatures up to
100C
Most enzymes however are fully denatured
at 70C
2007 Paul Billiet ODWS

Inhibitors
Inhibitors are chemicals that reduce
the rate of enzymic reactions.
The are usually specific and they
work at low concentrations.
They block the enzyme but they do
not usually destroy it.
Many drugs and poisons are
inhibitors of enzymes in the nervous
system.
2007 Paul Billiet ODWS

The effect of enzyme

inhibition
Irreversible inhibitors: Combine
with the functional groups of the
amino acids in the active site,
irreversibly.
Examples: nerve gases and
pesticides, containing
organophosphorus, combine with
serine residues in the enzyme
acetylcholine esterase.
2007 Paul Billiet ODWS

The effect of enzyme

inhibition
Reversible inhibitors: These can
be washed out of the solution of
enzyme by dialysis.
There are two categories.

2007 Paul Billiet ODWS

The effect of enzyme

inhibition
1. Competitive:
These compete
with the substrate
molecules for the
active site.
The inhibitors action
is proportional to
its concentration.
Resembles the
substrates
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structure closely.

E+I
Reversible
reaction

EI
Enzyme inhibitor
complex

The effect of enzyme

inhibition
Fumarate + 2H++ 2e-

Succinate
Succinate dehydrogenase

CH2COOH

COOH

CHCOOH

CH2
CH2COOH

COOH
Malonate

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CHCOOH

The effect of enzyme

inhibition
2. Non-competitive: These are not influenced by
the concentration of the substrate. It inhibits by
binding irreversibly to the enzyme but not at
the active site.
Examples
. Cyanide combines with the Iron in the enzymes
cytochrome oxidase.
. Heavy metals, Ag or Hg, combine with SH
groups.
These can be removed by using a chelating agent
such as EDTA.
2007 Paul Billiet ODWS

Applications of inhibitors
Negative feedback: end point or
end product inhibition
Poisons snake bite, plant alkaloids
and nerve gases.
Medicine antibiotics,
sulphonamides, sedatives and
stimulants
2007 Paul Billiet ODWS