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.

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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.

2007 Paul Billiet ODWS

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

H.PELLETIER, M.R.SAWAYA
ProNuC Database

2007 Paul Billiet ODWS

One part of an 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

Nitrogenase enzyme with Fe, Mo and ADP cofactors

Jmol from a RCSB PDB file 2007 Steve Cook

2007 Paul Billiet ODWS

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

2007 Paul Billiet ODWS

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 binding of
the substrate with the active site.

2007 Paul Billiet ODWS

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

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

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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

2007 Paul Billiet ODWS

CHCOOH

The effect of enzyme inhibition

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.
2.

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