ENZYMES

Introduction to Enzymes
Enzyme Classifications
Role of Coenzymes
Enzyme Activity Relative to Substrate Type
Enzyme-Substrate Interactions
Chemical Reactions and Rates
Chemical Reaction Order
Enzymes as Biological Catalysts
Michaelis-Menton Kinetics
Inhibition of Enzyme Catalyzed Reactions
Regulation of Enzyme Activity
Allosteric Enzymes
Enzymes in the Diagnosis of Pathology

Introduction to Enzymes
Enzymes are biological catalysts responsible for supporting almost all of
the chemical reactions.

The macromolecular components of almost all enzymes are composed of

protein, except for a class of RNA modifying catalysts known as ribozymes.
Ribozymes are molecules of ribonucleic acid that catalyze reactions on the
phosphodiester bond of other RNAs.
Enzymes are found in all tissues and fluids of the body.
Intracellular enzymes catalyze the reactions of metabolic pathways.
Plasma membrane enzymes regulate catalysis within cells in response to
extracellular signals,
Enzymes of the circulatory system are responsible for regulating the clotting
of blood.
Almost every significant life process is dependent on enzyme activity.

KLASIFIKASI ENZIMBerdasarkan fungsi dari enzim the International Union

of Biochemists (I.U.B.). Membagi enzim di dalam 6 grup
Number
ClassificationBio
chemical Properties
.
1
2

Oxidoreductases
Transferases

Act on many chemical groupings to add or

remove hydrogen atoms.2.TransferasesTransfer
Transfer functional groups between donor and
acceptor molecules. Kinases are specialized
transferases that regulate metabolism by
transferring phosphate from ATP to other
molecules.

Hydrolases

Add water across a bond, hydrolyzing it.

Lyases

Add water, ammonia or carbon dioxide across

double bonds, or remove these elements to
produce double bonds.

Isomerases

Carry out many kinds of isomerization: L to D

isomerizations, mutase reactions (shifts of
chemical groups) and others.

Ligases

Catalyze reactions in which two chemical groups

are joined (or ligated) with the use of energy
from ATP.

Enzymes are also classified on the basis of their composition.

Simple enzymes, enzymes composed wholly of protein.
complex enzymes, which are composed of protein plus a
relatively small organic molecule. Complex enzymes are also
known as holoenzymes.
Apoenzyme the protein component
coenzyme or prosthetic group the non-protein component
prosthetic group describes a complex in which the small
organic molecule is bound to the apoenzyme by covalent
bonds;
Coenzyme non-protein components, ,the small organic
molecule is bound to the apoenzyme by non-covalent bond.
Many prosthetic groups and coenzymes are water-soluble
derivatives of vitamins.
Metalloenzymes, enzymes that require a metal in their
composition, if they bind and retain their metal atom(s) under all
conditions, that is with very high affinity.
Those which have a lower affinity for metal ion, but still require the
metal ion for activity, are known as metal-activated enzymes

Role of Coenzymes
The functional role of coenzymes is to act as transporters of
chemical groups from one reactant to another.
The chemical groups carried can be as simple as the
hydride ion (H+ + 2e-) carried by NAD or the mole of
hydrogen carried by FAD; or they can be even more
complex than the amine (-NH2) carried by pyridoxal
phosphate.
Since coenzymes are chemically changed as a consequence
of enzyme action, it is often useful to consider coenzymes
to be a special class of substrates, or second substrates,
which are common to many different holoenzymes.
In all cases, the coenzymes donate the carried
chemical grouping to an acceptor molecule and are
thus regenerated to their original form.

Enzyme Relative to Substrate Type

Although enzymes are highly specific for the kind of reaction
they catalyze, the same is not always true of substrates they
attack. For example, while succinic dehydrogenase (SDH)
always catalyzes an oxidation-reduction reaction and its
substrate is invariably succinic acid, alcohol dehydrogenase
(ADH) always catalyzes oxidation-reduction reactions but
attacks a number of different alcohols, ranging from methanol
to butanol.
Generally, enzymes having broad substrate specificity are
most active against one particular substrate. In the case of
ADH, ethanol is the preferred substrate.
Enzymes also are generally specific for a particular steric
configuration (optical isomer) of a substrate. Enzymes
that attack D sugars will not attack the corresponding L
isomer. Enzymes that act on L amino acids will not employ
the corresponding D optical isomer as a substrate. The
enzymes known as racemases provide a striking
exception to these generalities; in fact, the role of
racemases is to convert D isomers to L isomers and
vice versa. Thus racemases attack both D and L

As enzymes have a more or less broad range of

substrate specificity, it follows that a given substrate
may be acted on by a number of different enzymes,
each of which uses the same substrate(s) and
produces the same product(s).
The individual members of a set of enzymes sharing
such characteristics are known as isozymes.
These are the products of genes that vary only
slightly; often, various isozymes of a group are
expressed in different tissues of the body.
The best studied set of isozymes is the lactate
dehydrogenase (LDH) system. LDH is a tetrameric
enzyme composed of all possible arrangements of
two different protein subunits; the subunits are
known as H (for heart) and M (for skeletal muscle).
These subunits combine in various combinations
leading to 5 distinct isozymes.
The all H isozyme is characteristic of that from heart
tissue, and the all M isozyme is typically found in
skeletal muscle and liver. These isozymes all catalyze
the same chemical reaction, but they exhibit
differing degrees of efficiency. The detection of

Enzyme-Substrate Interactions
Complementary shape, charge and hydrophilic/hydrophobic
characteristics of enzymes and substrates are responsible for
this specificity.
Lock and key" model (by Emil Fischer in 1894 )
Induced fit model (Daniel Koshland in 1958)

Michaelis-Menton Kinetics
The catalytic event that converts substrate to product involves the
formation of a transition state, and it occurs most easily at a specific
binding site on the enzyme.
This site, called the catalytic site of the enzyme, has been
evolutionarily structured to provide specific, high-affinity
binding of substrate(s) and to provide an environment that
favors the catalytic events.
The complex that forms when substrate(s) and enzyme
combine is called the enzyme substrate (ES) complex. Reaction
products arise when the ES complex breaks down releasing free
enzyme.

The Michaelis-Menten equation can be used to demonstrate

that at the substrate concentration that produces exactly
half of the maximum reaction rate, i.e.,1/2 Vmax, the
substrate concentration is numerically equal to Km.
This fact provides a simple yet powerful bioanalytical tool
that has been used to characterize both normal and altered
enzymes, such as those that produce the symptoms of
genetic diseases.

Inhibition of Enzyme Catalyzed Reactions

To avoid dealing with curvilinear plots of enzyme catalyzed
reactions, biochemists Lineweaver and Burk introduced an
analysis of enzyme kinetics based on the following
rearrangement of the Michaelis-Menten equation:
[1/v] = [Km (1)/ Vmax[S] + (1)/Vmax]
Plots of 1/v versus 1/[S] yield straight lines having
a slope of Km/Vmax and an intercept on the
ordinate at 1/Vmax.

Regulation of Enzyme Activity

While it is clear that enzymes are responsible for the catalysis
of almost all biochemical reactions, it is important to also
recognize that rarely, if ever, do enzymatic reactions proceed
in isolation. The most common scenario is that enzymes
catalyze individual steps of multi-step metabolic pathways, as
is the case with glycolysis, gluconeogenesis or the synthesis of
fatty acids.
As a consequence of these lock- step sequences of reactions,
any given enzyme is dependent on the activity of preceding
reaction steps for its substrate.
In humans, substrate concentration is dependent on food
supply and is not usually a physiologically important
mechanism for the routine regulation of enzyme activity.
Enzyme concentration, by contrast, is continually modulated in
response to physiological needs. Three principal mechanisms
are known to regulate the concentration of active enzyme in
tissues:
1. Regulation of gene expression controls the quantity
and rate of enzyme synthesis.
2. Proteolytic enzyme activity determines the rate of
enzyme degradation.
3. Covalent modification of preexisting pools of

Thermodynamics

Regulation of Enzyme Activity

While it is clear that enzymes are responsible for the catalysis of
almost all biochemical reactions, it is important to also recognize
that rarely, if ever, do enzymatic reactions proceed in isolation.
The most common scenario is that enzymes catalyze individual
steps of multi-step metabolic pathways, as is the case with
glycolysis, gluconeogenesis or the synthesis of fatty acids. As a
consequence of these lock- step sequences of reactions, any
given enzyme is dependent on the activity of preceding reaction
steps for its substrate.
In humans, substrate concentration is dependent on food supply
and is not usually a physiologically important mechanism for the
routine regulation of enzyme activity. Enzyme concentration, by
contrast, is continually modulated in response to physiological
needs. Three principal mechanisms are known to regulate the
concentration of active enzyme in tissues:
1. Regulation of gene expression controls the quantity
and rate of enzyme synthesis.
2. Proteolytic enzyme activity determines the rate of
enzyme degradation.
3. Covalent modification of preexisting pools of inactive
proenzymes produces active enzymes.