ERT 211/1

Biochemical Engineering
CHAPTER 1:
THE KINETICS OF ENZYME
CATALYZED REACTIONS
B Y:
EN. MOHD. FA HR UR R A ZI TOMPA NG

Classification of Enzyme
Enzymes fall into 6 classes based on function
1.
2.
3.
4.
5.

Oxidoreductases: which are involved in oxidation,

reduction, and electron or proton transfer reactions
Transferases : catalysing reactions in which groups
are transferred
Hydrolases : which cleave various covalent bonds by
hydrolysis
Lyases : catalyse reactions forming or breaking
double bonds
Isomerases : catalyse isomerisation reactions

6. Ligases : join substituents together covalently.

Enzyme Reaction
For design and analysis of a reacting system, we

must have a mathematical formula which gives the

reaction rate
in terms of

composition,
temperature,
and pressure of the reaction mixture.

Enzyme Kinetics
Enzymes are protein catalysts that, like all catalysts,

speed up the rate of a chemical reaction without

being used up in the process.

Enzyme Kinetics
Synthetic catalysts and enzymes use the common

technique for modeling reaction kinetics.

The rate expressions eventually obtained for both
types of catalysts are very similar and sometimes of
identical forms.
This is because, in both cases, the reacting molecules
form some sort of complex with the catalyst.

Enzyme Kinetics
Most synthetic catalysts are not specific; i.e. they will

catalyze similar reactions involving many different

kinds of reactants.
While some enzymes are not very specific, many will
catalyze only one reaction involving only certain
substrates.

 Explain why enzyme catalysis are nonspecific but

enzyme catalysis are specific?

 Define
Cofactors
Apoenzyme
Holoenzyme
coenzyme

Enzyme Kinetic
Enzyme is a catalyst which increases the rate of a

chemical reaction without undergoing a permanent

chemical change.
While a catalyst influences the rate of a chemical
reaction, it does not affect reaction equilibrium.
Equilibrium concentrations can be calculated using
only the thermodynamic properties of the substrates
and products.

Enzyme

Enzyme Kinetics
Both synthetic and biological catalysts can gradually

lose activity as they participate in chemical reactions.

However, enzymes are in general far more fragile.
Enzymes contorted shapes in space often endow
enzymes with unusual specificity and activity
It is relatively easy to disturb the native
conformation and destroy the enzyme's catalytic
properties.

Enzyme Kinetics
It is often asserted that enzymes are more active, i.e.,

allow reactions to go faster, than nonbiological catalysts.

At the ambient temperatures where enzymes are most
active they are able to catalyze reactions faster than the
majority of artificial catalysts.
When the reaction temperature is increased, solid
(synthetic) catalysts may become as active as enzymes.
The enzyme activity does not increase continuously as
the temperature is raised. Instead, the enzyme usually
loses activity at quite a low temperature, often only
slightly above that at which it is typically found.

Enzyme Kinetics

Enzyme reaction rates are determined by several

factors.
Concentration of substrate molecules

The more of them available, the quicker the enzyme molecules

collide and bind with them).
The concentration of substrate is designated [S] and is expressed in
unit of molarity.
Temperature.
As the temperature rises, molecular motion - and hence collisions
between enzyme and substrate - speed up.
But as enzymes are proteins, there is an upper limit beyond which
the enzyme becomes denatured and ineffective.

Enzymes cont.
the presence of inhibitors.

competitive inhibitors are molecules that bind

to the same site as the substrate - preventing the
substrate from binding as they do so - but are not
changed by the enzyme.
noncompetitive inhibitors are molecules that
bind to some other site on the enzyme reducing its
catalytic power.
pH. The conformation of a protein is influenced by pH
and as enzyme activity is crucially dependent on its
conformation, its activity is likewise affected.

How we determine how fast an enzyme

works
We set up a series of tubes containing graded

concentrations of substrate, [S] . At time zero,

we add a fixed amount of the enzyme
preparation.
Over the next few minutes, we measure the
concentration of product formed. If the
product absorbs light, we can easily do this in
a spectrophotometer.
Early in the run, when the amount of
substrate is in substantial excess to the
amount of enzyme, the rate we observe is the
initial velocity of Vi.

THE ENZYME-SUBSTRATE
COMPLEX AND ENZYME ACTION
There is no single theory which accounts for the

unusual specificity and activity of enzyme catalysis.

However, there are a number of plausible ideas
supported by experimental evidence for a few
specific enzymes.
Probably, all or some collection of these phenomena
acting together combine to give enzymes their
special properties.

THE ENZYME-SUBSTRATE
COMPLEX AND ENZYME ACTION
The x-ray crystallography, spectroscopy, and

electron-spin resonance showed the existence of a

substrate-enzyme complex.
The substrate binds to a specific region of the
enzyme called the active site, where reaction occurs
and products are released.
Binding to create the complex is sometimes due to
the type of weak attractive forces.

THE ENZYME-SUBSTRATE
COMPLEX AND ENZYME ACTION
The complex is formed when the substrate key joins

with the enzyme lock.

The hydrogen bonds formed between the substrate
and groups widely separated in the amino acid chain
of the enzyme.

THE ENZYME-SUBSTRATE COMPLEX

AND ENZYME ACTION
The protein molecule is folded in such a way that a

group of reactive amino acid side chains in the

enzyme presents a very specific site to the substrate.
The reactive groups encountered in enzymes include
the R group of Asp, Cys, Glu, His, Lys, Met, Ser, Thr,
and the end amino and carboxyl functions.
Since the number of such groups near the substrate
is typically 20, only a small fraction of the enzyme is
believed to participate directly in the enzyme's active
site.

 Large enzymes may have more than one active site.

Many of the remaining amino acids determine the

folding along a chain of amino acids (secondary

structure) and the placement of one part of a folded
chain next to another (tertiary structure), which help
create the active site itself

THE ENZYME-SUBSTRATE COMPLEX

AND ENZYME ACTION

 Enzymes can hold substrates so that their reactive

regions are close to each other and to the enzyme's

catalytic groups.
This feature, which quite logically can accelerate a
chemical reaction, is known as the proximity effect.

 Reaction will occur only when the molecules come together at

the proper orientation so that the reactive atoms or groups are

in close juxtaposition.
Enzymes are believed to bind substrates in especially
favorable positions, thereby contributing an orientation effect,
which accelerates the rate of reaction.
Also called orbital steering, this phenomenon has qualitative
merit as a contributing factor to enzyme catalysis. The
quantitative magnitude of its effect, however, is still difficult
to assess in general.