Classification of Enzymes

1.Oxidoreductase
O
CH3 -C- COO - + NADH + H
Pyruvate

lactate
deh ydrogenase

OH
+
CH 3 -CH -COO - + NAD
Lactate

2.Transferase
COOCH2
CH- NH3 +
COO-

COOAspartate amino
C= O
transfer ase
CH2

COOCH2
C= O

CH2

COO-

COOAspartate -Ketoglutarate

COOC-NH3 +
CH2
CH2
COO-

Oxalosuc cinate

G lutamate

3.Hydrolase
O
CH3 -C- OCH 2 CH 2 N( CH3 ) 2 + H O Acetylcholinesterease
2
Acetylcholine

O
CH3 -C- O-

HOCH2 CH2 N( CH3 ) 2

Ace tate

Choline

COOCH2
C-COO - + H O
2
CH
COOcis- Aconitate

4. Lyase

Aconitase

5. Isomerase

CH2 OPO 3 2 Phosphohexose

O
isomerase
OH
OH
HO
OH
- D- Glucose-6-phosphate

COOCH2
C-COO HO C-H
COOIsocitrate

CH2 OPO 3 2 CH2 OH

O
H HO
H
OH
H
HO
- D-Fructose-6-phosphate

6. Ligase
ATP + L -tyros ine + t-RNA

Ty ros ine-tRNA
s yntheta se

L-tyro sy l-tDNA + AM P + PPi

Enzyme controlled reactions proceed 108 to 1011 times

faster than corresponding non-enzymic reactions.

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

Schematic of an Active Site

Apoenzyme: the protein part of an
enzyme.
Cofactor: a nonprotein portion of an
enzyme that is necessary for
catalytic function; examples are
metallic ions such as Zn 2+ and Mg2+.
Coenzyme: a nonprotein organic
molecule, frequently a B vitamin,
that acts as a cofactor.
Substrate: the compound or
compounds whose reaction an
enzyme catalyzes.
Active site: the specific portion of
the enzyme to which a substrate
binds during reaction.

Cofactors (simple vs. complex)

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

Substrate of an
enzyme are the
reactants that are
activated by the
enzyme

The shape and the chemical environment inside the

active site permits a chemical reaction to proceed more
easily.

The Lock and Key Hypothesis

S

E
E
E

Enzymesubstrate
complex

Enzyme may
be used again

P
Reaction coordinate

Fit between the substrate (key) and the active site of the enzyme
(lock) is very precise
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

This explains enzyme

specificity and loss of
activity when enzymes
denature.

This explains the enzymes

that can react with a range of
substrates of similar types.

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)

Factors Affecting Enzyme

Activity

substrate concentration
pH
temperature
inhibitors

Substrate concentration: Non-enzymic

reactions

Reaction
velocity

Substrate concentration

The increase in velocity is proportional to the

substrate concentration

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.

A linear curve

A saturation curve

At constant substrate
concentration, increasing the
enzyme concentration,
increases the rate linearly.
(In practically all enzyme
reactions, the molar conc. of
enzyme is always much lower
than that of substrate.)

At constant enzyme
concentration, increasing the
substrate concentration does
not increases the rate
continuously. A saturation
point is achieved.

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.

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.

The effect of temperature

Q10

Enzyme
activity

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

Organize the tiles!

When you organize these tiles, you will find a phrase describing a feature
of biological catalysts, that in fact is common to all catalysts:

 Allosterism: enzyme regulation based on an

event occurring at a place other than the active
site but that creates a change in the active site.
An enzyme regulated by this mechanism is called
an allosteric enzyme.
Allosteric enzymes often have multiple polypeptide
chains.
Negative modulation: inhibition of an allosteric
enzyme.
Positive modulation: stimulation of an allosteric
enzyme.
Regulator: a substance that binds to an allosteric
enzyme.

 Feedback control: an enzyme-regulation process

where the product of a series of enzymecatalyzed reactions inhibits an earlier reaction in

the sequence.
feedbac kinhibition

E1

E2

E3

The inhibition may be competitive or noncompetitive.

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.

nerve gases and pesticides, containing

organophosphorus, combine with serine
residues in the enzyme acetylcholine esterase.

Two categories of reversible inhibition:

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

E+I

Reversible
reaction

EI

Enzyme inhibitor
complex

2.) Non-competitive: The inhibitor

binds itself to a site other than
the active site (allosterism),
thereby changing the
conformation of the active site.
The substrate still binds but
there is no catalysis.
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.

 Enzyme kinetics in the presence and absence of

inhibitors.
Maximum reaction rate
is the same without an
inhibitor and in the
presence of a
competitive inhibitor
(CI).

Maximum rate is
obtained at high
substrate concentration
for CI but low with no
inhibitor.
If the inhibitor is non
competitive, the
maximum rate of
reaction is lower.