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Learning Objectives

Describe definition of enzyme


Explain several factors that influences
enzymes activity
Explain function of enzyme in metabolism

METABOLISM
Is the sum of all the biochemical
processes that occur within a cell
or organism
2007 Paul Billiet ODWS

Metabolism Function :

Obtain energy for the cell.


Convert nutrients into
macromolecules.
Assemble macromolecules into
cellular structures.
Degrade macromolecules as required
for biological function

Substances of Metabolism :

Enzyme
Energy

(ATP)

What Are Enzymes?

Most enzymes are


Proteins
Act as Catalyst to
accelerates a
reaction
Not permanently
changed in the
process
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Enzymes

Are specific for


what they will
catalyze
Are
Reusable/reversible
End in ase
-Sucrase
- Lactase
- Maltase

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

Enzymes
Without Enzyme
With Enzyme

Free
Energy

Free energy of activation


Reactants

Products

Progress of the reaction


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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

Components of Enzyme :
Apoenzyme + coenzyme =
holoenzyme
Apoenzyme
a protein molecules
Coenzyme
non protein molecules
(NAD and NADP)
Cofactor (ex : Calsium, magnesium
and Calium)

How do Enzymes work ?


Beadle and Tatum conclude that the
work of enzymes very specifik.
Their statement the one gene-one
enzymes

The Lock and Key Theory

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 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

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)

Classify of Enzymes :

Hydrolase enzyme
ex : carbohidrase, proteinase or enzymes in
the digestive system
Desmolase enzyme
ex : peroxinase, dehidroginase, katalase or
enzymes in the respiratory system

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

pH
Enzyme
substrate concentration
temperature
inhibitors

2007 Paul Billiet ODWS

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

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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

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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.

2007 Paul Billiet ODWS

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