ENZYMES

ENZYME
is a compound, usually a protein, that acts
as a catalyst for a biochemical reaction

ENZYMES
The word enzyme comes from the

Greek words en, which means in, and

zyme, which means yeast.
Enzymes undergo all the reactions of

proteins, including denaturation.

ENZYME STRUCTURE
A

simple enzyme is an enzyme

composed only of protein (amino acid
chains).

A conjugated enzyme is an enzyme

that has a nonprotein part in addition to

a protein part.
An apoenzyme is the protein part of
a conjugated enzyme.
A cofactor is the nonprotein part of a

ENZYME STRUCTURE
A holoenzyme is the biochemically

active conjugated enzyme produced

from an apoenzyme and a cofactor.
Apoenzyme + Cofactor = Holoenzyme

WHY DO APOENZYMES
NEEDS COFACTORS?
Cofactors provide additional chemically
reactive functional groups besides those
present in the amino acid side chains of
apoenzymes.
A cofactor is generally either a small organic
molecule or an inorganic ion (usually a metal
ion).
A coenzyme is a small organic molecule that
serves as a cofactor in a conjugated enzyme.

Typical inorganic ion cofactors include Zn2+,

Mg2+, Mn2+, and Fe2+.
The non metallic ion CI- ion occasionally acts
as a cofactor.
Dietary minerals are an important source of
inorganic ion cofactors.
Many vitamins have a coenzyme functions in
the human body.

NOMENCLATURE AND
CLASSIFICATION OF
ENZYMES
Enzymes are most commonly named by
using a system that attempts to provide
information about the function (rather
than the structure) of the enzyme.
The type of reaction catalyzed and the
substrate identity are focal points for the
nomenclature.
A substrate is the reactant in an enzymecatalyzed reaction.

Three important
aspects of the enzymenaming process:

1. The suffix ase identifies a substance as

an enzyme. The suffix in is still found in

some of the first enzymes studied, many
of which are digestive enzymes.
Trypsin
Chymotrypsin
Pepsin
Carbohydrase
Protease
Lipase

Three important
aspects of the enzymenaming process
2. The type of reaction catalyzed by an
enzyme is often noted with a prefix.
Oxidase
Hydrolase

Three important
aspects of the enzymenaming process
3. The identity of the substrate is often noted
in addition to the type of reaction
Glucose oxidase
Pyruvate carboxylase
Succinate dehydrogenase
In some cases, only the substrate is given
but the
not reaction.
Urease
Lactase

enzymes base on the

type of reaction
catalyzed
1. An oxidoreductase is an enzyme that

catalyzes
reaction.

an

oxidation-reduction

2.

A tranferase is an enzyme that

catalyzes the transfer of a functional
group from one molecule to another.

3.

A hydrolase is an enzyme that

catalyzes a hydrolysis reaction in
which the addition of a water molecule
to a bond causes the bond to break.

4. A lyase is an enzyme that catalyzes

the addition of a group to a double
bond or the removal of a group to form
a double bond in a manner that does
not involve hydrolysis or oxidation.

5. An isomerase is an enzyme that

catalyzes
the
isomerization
(rearrangement of atoms) of a
substance in a reaction, converting it
into a molecule isomeric with itself.

6. A ligase is an enzyme that catalyzes

the bonding together of two molecules
into one with the participation of ATP.

MODELS OF ENZYME ACTION

ENZYME ACTIVE SITE
is the relatively small part of an enzymes
structure that is actually involved in catalysis.
ENZYME-SUBSTRATE COMPLEX
is the intermediate reaction species that is
formed when a substrate binds to the active
site of an enzyme
LOCK-AND-KEY MODEL
in the lock-and-key model, the active site in
the enzyme has a fixed, rigid geometrical
conformation.
Only substrates with a complementary

Lock-and-Key Model for Enzyme Activity

INDUCED-FIT MODEL
allows for small changes in the shape or
geometry of the active site of an enzyme to
accommodate a substrate.

ENZYME SPECIFICITY
is the extent to which an enzymes activity is
restricted to a specific substrate, a specific group of
substrates, a specific type of chemical bond, or a
specific type of chemical reaction.
1. Absolute Specificity the enzyme will catalyze only

one reaction. (e.g. Catalase)

2. Group Specificity the enzyme will act only on

molecules that have a specific functional group,

such as hydroxyl, amino, or phosphate groups. (e.g.
Carboxypeptidase)
3. Linkage Specificity the enzyme will act on a

particular type of chemical bond, irrespective of the

rest of the molecular structure. (e.g. Phosphatases)
4. Stereochemical Specificity the enzyme will act on

FACTORS THAT AFFECT

ENZYME ACTIVITY
ENZYME ACTIVITY
is a measure of the rate at which an enzyme
converts substrate to products in a biochemical
reaction.

TEMPERATURE
Optimum temperature is the temperature at
which an enzyme exhibits maximum activity.
For humans enzymes, the optimum temperature is
around 37 degrees Celcius.
The destroying effect of temperature on bacterial
enzymes is used in hospital setting to sterilize
medical instruments and laundry. In high
temperature, high vessels called autoclaves, superheated steam is used to produce a temperature

pH
Optimum pH is the pH at which enzyme
exhibits maximum activity
Each enzyme has a characteristic optimum
pH, which usually fall within the physiological
pH range of 7.0 7.5.
Notable exception to this generalization are
the digestive enzymes pepsin and trypsin.
Pepsin pH 2.0
Trypsin pH 8.0

SUBSTRATE CONCENTRATION
Saturation curve enzyme activity increases up to
a certain substrate concentration and thereafter
remains constant.
Turnover number is the number of substrate
molecules transformed per minute by one molecule
of an enzyme under optimum conditions of
temperature, pH, and saturation.
ENZYME CONCENTRATION
If the amount of substrate present is kept constant
and the enzyme concentration is increased, the
reaction rate increases because more substrate
molecules can be accommodated in a given amount
of time. The greater the enzyme concentration, the
greater the reaction rate.

Extremoenzymes
Extremophile is a microorganism that thrives
in extreme environments, environment in
which humans and most other forms of life
could not survive.
acidophiles (3.0 pH or below)
alkaliphiles (9.0 pH or above)
halophiles (salinity that exceeds 0.2 M
NaCL)
hypothermophiles (80 122 degrees
Celsius)
piezophiles (high hydrostatic pressure)
cryophiles (15 degrees Celsius or lower)

The enzyme present in extremophiles are called

extremoenzymes.
Extremoenzymes is a microbial enzyme
active at conditions that would inactivate
human enzymes as well as enzymes present
in other types of higher organisms.

Enzyme Inhibition
Enzyme inhibitor is a substance that slows
or stops the normal catalytic function of an
enzyme by binding to it.
Reversible Competitive Inhibition
Competitive enzyme inhibitor is a
molecule that sufficiently resembles an
enzyme substrate in shape and charge
distribution that it can compete with the
substrate for occupancy of the enzymes
active site.

Reversible Noncompetitive Inhibition

Noncompetitive enzyme inhibitor is a
molecule that decreases enzyme activity by
binding to a site on an enzyme other than the
active site.

Irreversible Inhibition
Irreversible enzyme inhibitor is a
molecule that inactivates enzymes by forming
a strong covalent bond to an amino acid sidechain group at the enzymes active site.

Regulation of Enzyme
Activity
Regulation of enzyme activity within a cell is a
necessity for many reasons.
1. A cell that continually produces large
amounts of an enzyme for which substrate
concentration is always very low is wasting
energy. The production of the enzyme needs
to be turned off.
2. A product of an enzyme-catalyzed reaction
that is present in plentiful amounts in a cell
is a waste of energy if the enzyme continues
to catalyze the reaction that produces the
product. The enzyme needs to be turned
off.

Allosteric Enzymes
Allosteric Enzyme is an enzyme with two or more
protein chains (quaternary structure) and two kind of
binding sites (substrate and regulator).
Characteristics of allosteric enzymes:
Have quaternary structure (composed of two or
more protein chains
Have two kinds of binding sites (for substrates and
for regulators)
Active and regulatory binding sites are distinct from
each other in both location and shape
Binding of a molecule in the regulatory site causes
changes in the overall three-dimensional structure of
the enzyme, including structural changes at the
active site

Regulators substances that bind at the

regulatory sites of an allosteric enzyme.
The binding site of a positive regulator
increases enzyme activity; the shape of the
active site is changed such that it can more
readily accept substrate.
The binding site of a negative regulator
decreases enzyme activity; changes to the
active site are such that substrate is less
readily accepted.

Positive and Negative Allosteric Control

3 mechanisms are considered in which enzyme

within a cell can be turned on or turned off

Feedback Control
is the process in which activation or inhibition of
the first reaction in a reaction sequence is
controlled by a product of the reaction sequence.
As illustrative of the feedback control mechanism,
consider a biochemical process within a cell that
occurs in several steps, each step catalyzed by a
different enzyme. The product of each step is the
substrate for the next enzyme.

Proteolytic Enzymes and

Zymogens
Proteolytic enzyme is an enzyme that catalyzes the
breaking of peptide bonds that maintain the primary
structure of an enzyme.
Zymogen is an inactive precursor of a proteolytic
enzyme.

Covalent Modification of
Enzymes

Covalent modification is a process in which enzyme

activity is altered by covalently modifying the
structure of the enzyme through attachment of a
chemical group to or removal of a chemical group from
a particular amino acid within the enzymes structure.

The most commonly encountered type of

covalent modification involves the process by
which a phosphate group is added to or
removed from an enzyme. The source of the
added phosphate group is often an ATP
molecule. The process of addition of the
phosphate group to the enzyme is called
phosphorylation, and the removal of the
phosphate group from the enzyme is called
dephosphorylation.
This two processes are the off/on or on/off
switch for the enzyme.

The preceding covalent modification

processes are governed by other enzymes.
Protein kinases effect the addition of
phosphate groups
Phophatases catalyze the removal of the
phosphate groups
Usually, the phosphate group is added to
(or removed from) the R group of a serine,
tyrosine, or threonine amino acid residue
present in protein. The R groups of these
three amino acid have a common structural
feature, the presence of a tree OH group. The
hydroxyl group is the site where
phosphorylation or dephosphorylation occurs.

Prescription Drugs That

Inhibit Enzyme Activity
ACE INHIBITORS

ACE stands for angiotensin-converting

enzyme.
Angiotensin is an octapeptide hormone
involve in blood pressure regulation. It
increases blood pressure by narrowing blood
vessels. Until neeeded, angiotensin is present
in the body in an inactive form as the
zymogen angiotensinogen, which is a
decapeptide.

ACE converts the inactive decapeptide zymogen

to the active octapeptide form (angiotensin) by
cleaving two amino acids from the zymogen
structure.

Ace inhibitor medications block the action of ACE

in converting angiotensinogen to angiotensin.
The effect of this is a lower blood pressure than
if the zymogen activation had occurred.

SULFA DRUGS

antibiotics is a substance that kills bacteria

or inhibits their growth.
Sulfanilamide inhibits bacterial growth
because it is structurally similar to PABA (paminobenzoic acid).
Many bacteria need PABA in order to produce
an important coenzyme, folic acid.
Sulfanilamide acts as a competitive inhibitor
to enzymes in the biosynthesis pathway for
converting PABA into folic acid in these
bacteria. Folic acid deficiency retards growth
of the bacteria and can eventually kill them.

PENICILLINS

Penicillins inhibit transpeptidase, an enzyme

that catalyzes the formation of peptide across
links between polysaccharide strands in
bacterial cell walls. These cross links
strengthen cell walls. A strong cell wall is
necessary to protect the bacterium form lysis
(breaking open). By inhibiting transpeptidase,
penicillin prevents the formation of a strong
cell wall. Any osmotic or mechanical shock
then causes lysis, killing the bacterium.

FOOD-ENZYME INTERACTIONS THAT

AFFECT PRESCRIPTION METHOD

Cytochrome P450 liver enzymes involved in
the process by which many prescription
medications are metabolized in the body.
The most well-known and most studied of the
unknown food-enzyme-drug-interactions
involves grapefruit, and grapefruit juice, an
interaction which is now called the grapefruit
effect.

Medical Uses of
Enzymes
Enzymes can be used to diagnose certain
diseases. Although blood serum contains
many enzymes, some enzymes are not
normally found in the blood but are produced
only inside cells of certain organs and tissues.
The appearance of these enzymes in the
blood often indicates that there is tissue
damage in an organ and that cellular content
are spilling out into the bloodstream.

Enzymes can also be used in the treatment of

diseases. A recent advance in treating heart
attacks is the use of tissue plasminogen activator
(TPA), which activates the enzyme plasminogen.
When so activated, this enzyme dissolves blood
clots in the heart and often provides immediate
relief.
Selected Blood Enzyme Assays Used In Diagnostic Medicine

Another medical use for enzymes is in clinical

laboratory chemical analysis. For example, no
simple direct test for the measurement of
urea in the blood is available. However, if the
urea in the blood is converted to ammonia via
the enzyme urease, the ammonia produced,
which is easily measured, becomes an
indicator of urea.
Blood urea nitrogen (BUN) test is a
common clinical laboratory procedure. High
urea levels in the blood indicate kidney
malfunction.