ENZYMES
highly specialized class of proteins. STANLEY – They discovered a complex
GENERAL CHARACTERISTICS OF
ENZYMES
ENZYMES
- Is a compound, usually a protein, that
acts as a catalyst for biochemical
reaction.
- Each cell in the human body contains
thousands of different enzymes
because almost every reaction in the
cell requires its own specific enzyme.
- Enzymes cause cellular reactions to
occur millions of times faster than
corresponding uncatalyzed reactions.
- As catalyst, enzymes are not consumed
during the reaction but merely help in
the reaction occur more rapidly.
- Are chemical substance which hastens
chemical reaction without being affected
in the process.
- Are secreted by living cells and are
complex organic chemical compounds
with definite structure.
- Like the inorganic catalysts, enzymes
have the remarkable property of
speeding up chemical reactions without
being themselves affected in the
process.
As a result, they can be use over and over
again.
- Catalysts or enzymes are responsible
for different biological changes.
- They make up the largest and most
- The catalytic efficiency of enzyme is so
high that per mole is around 10,000 to
10,000,000 moles of substance per
minute.
- The catalytic action of enzymes is the
key to their importance, for by facilitating
chemical changes, enzymes make
possible the continuous replacement
and renewal processes of all living
organisms.
- The word ENZYME comes from a
Greek word EN, which means “IN” and
ZYME, which means “YEAST”.
YEAST
Long before, yeast is used in the production of
bread and alcoholic beverages.
- Yeast reaction to sugar produces the
carbon dioxide gas that causes the
bread to rise.
- Fermentation of sugars in fruit juice
using the yeast enzymes produces
alcoholic beverages.
EARLY ENZYMES DISCOVERIES
JON JAKOB BERZELIUS – 1835 Swedish
Chemist, termed their chemical action –
catalytic.
JAMES B. SUMMER of Cornell University –
Isolate and crystallize the enzymes urease from
the jack bean.
JOHN H. NORTHROP and WENDELL M.
procedure for isolating and purifying pepsin.
ENZYME STRUCTURE
A. Simple Enzymes
 Is an enzyme composed only of
protein (amino acid chains)
B. Conjugated Enzymes
 Is an enzyme that has a
nonprotein part in addition to
protein part.
 APOENZYME is the protein part
of a conjugated enzymes.
 COFACTOR Is the nonprotein
part of the conjugated enzymes.
 HOLOENZYME Is the
biochemically active conjugated
enzymes produced from an
apoenzyme and a cofactor.
Apoenzyme + Cofactor = Holoenzymes
Why do apoenzyme need cofactor?
- Cofactors provide additional chemically
reactive functional group besides those
present in the amino acid side chains of
apoenzymes.
APOENZYME and HOLOENZYME
- The enzyme without its nonprotein
moiety is termed as apoenzyme and it
is inactive.
- Holoenzyme is an active enzyme with
its nonprotein component.
TWO BROAD CATEGORIES OF COFACTOR:
A. Simple Metal Ions
B. Small Organic Molecules
METAL IONS COFACTOR
- Include Zn, Mg, Fe, and Cu.
- All metal ions must be supplied to
human body through dietary mineral
intake.
- Almost any type of diet will provide
adequate amounts of needed metallic
cofactors because they are needed in
very small (trace) amounts.
SMALL ORGANIC MOLECULES
- The second category of cofactors, as a
group, are called COENZYMES.
- CONENZYME is a small organic
molecule that serves as a cofactor in
conjugated enzyme.
- COENZYME is synthesized within the
human body using building blocks from
the nutrients.
o Most often, one of these
building blocks is a B vitamin
or B vitamin derivatives.
o Vitamins must be obtained
through dietary intake.
- Many cofactors are permanently
bonded, via covalent bond, to the amino
acids portion of enzymes.
- Breaking the covalent bond
deactivates the enzymes.
o Ex. Coenzymes FAD which has
riboflavin (a B vitamin)
o Coenzymes NAD which has
niacin (a B vitamin)
TYPES OF COFACTORS
COENZYMES – The nonprotein component,
loosely bound to apoenzyme by non-covalent
bond.
Examples: vitamins or compound derived
from vitamins.
PROSTHETIC GROUP – The nonprotein
component, tightly bound to the apoenzyme by
covalent bonds is called a prosthetic group.
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 enzymes.
- The type of reaction catalyzed, and the
substrate identify are focal points in
nomenclature.
- A substrate is the reactant in an
enzyme-catalyzed reaction. The
substrate is the substances upon which
the enzymes “acts”.
Three important aspects of the enzymes-
naming process are the following:
1. The suffix ASE identifies the
substance as an enzyme.
a. Thus urease, surcease and
lipase are all enzymes
designation.
b. The suffix IN is still found in the
names of some of the first
enzymes studied, many of which
are digestive enzymes.
i. Ex. Trypsin,
chymotrypsin, and
pepsin.
2. The type of reaction catalyzed by an
enzyme is often noted with a prefix.
a. An oxidase enzyme catalyzes an
oxidation.
b. A hydrolase enzyme catalyzes a
hydrolysis reaction.
3. The identity of the substrate is often
noted in addition to the type of
reaction.
a. Enzymes names of this type
include glucose oxidase,
pyruvate carboxylase, and
succinate dehydrogenase.
b. In frequently, the substrate but
not the reaction types are given,
as in the name’s urease and
lactase.
c. In these names, the reaction
involved is hydrolysis; urease
catalyzes the hydrolysis of
 urea, lactase the hydrolysis of
lactose.
Enzymes – are grouped into six major classes
(EC Number Classification) by the
International Union of Biochemists (I.U.R).
On the basis of types of reactions, they
catalyze.
PRINCIPLE OF THE INTERNATIONAL
CLASSIFICATION
Each enzyme has classification number
consisting of four digits:
Example EC: (2.7.1.1) HEXOKINASE
EC: (2.7.1.1) these components indicate the
following groups of enzymes:
2 is class (TRANSFERASE)
7 is subclass (TRASFER OF PHOSPHATE)
1 is sub subclass (ALCOHOL IS PHOSPHATE
ACCEPTOR)
1 specific name: ATP, D-HEXOSE-6-
PHOSPHOTRANSFERASE (HEXOKINASE)
CLASSIFICATION OF ENZYMES
1. OXIDOREDUCTASE – Is an enzyme
that catalyzes an oxidation-reduction
reaction.
An organic oxidation reaction is an oxidation
that increases the number of C-O bonds
and/or decreases the of C-H bonds.
An organic reduction reaction is a reduction
that decreases the number of C-O bonds
and/or increases the number of C-H bonds.
The oxidation-reduction are not independent
processes but linked processes that must occur
together.
An oxidoreductase requires a coenzyme that is
oxidized or reduced as the substrate is reduced
or oxidized.
EX. Lactase Dehydrogenase is an
oxidoreductase that removes hydrogen atom
from a molecule.
2. TRANSFERASE – Is an enzyme that
catalyzes the transfer of a functional
group from one molecule to another.
BIOCHEMICAL ACTIVITY:
 Transfer a functional group (e.g., methyl
or phosphate) between donor and
acceptor molecules.
EXAMPLES:
 Transaminases (ALT & AST)
 Phosphotransferases (Kinases)
 Transmethylases
 Transpeptidases
 Transacylases
Two major subtypes of transferase are
transaminases and kinases.
Transaminase catalyzes the transfer of amino
group from one molecule to another.
Kinases which play a major role in metabolic
energy-production reactions, catalyzes the
transfer of a phosphate group from adenosine
triphosphate (ATP) give adenosine diphosphate
(ADP) and phosphorylated products (a
phosphate containing an additional phosphate).
 3. HYDROLASE – Is an enzyme that
catalyzes a hydrolysis reaction in which
the addition of a water molecule to a
molecule to a bond causes the bond to
break.
Hydrolysis Reactions – are central to the
process of digestion.
Carbohydrates effect the breaking of glycosidic
bonds in oligo and polysaccharides.
Proteases effect the breaking of peptide
linkages in protein.
Lipases effect the breaking of ester linkages in
triacylglycerol’s.
BIOCHEMICAL ACTIVITY:
 Catalyze the hydrolysis of various
bonds add water across a bond.
EXAMPLES:
 Protein Hydrolyzing Enzymes
(Peptidases)
 Carbohydrases (Amylase, Maltase,
Lactase)
 Lipid Hydrolyzing Enzymes (Lipase)
 Deaminases
 Phosphatases
4. LYASES – 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 monomer that does
not involve hydrolysis or oxidation.
A dehydratase effects the removal of the
components of water from a double bond.
Hydratase effects the addition of the
components of water to a double bond.
BIOCHEMICAL ACTIVITY:
 Cleave various bonds by means other
than hydrolysis and oxidation.
 Add water, ammonia or carbon dioxide
across double bonds, or remove these
elements to produce double bonds.
EXAMPLES:
 Fumarase
 Carbonic Anhydrase
5. ISOMERASE – Is an enzyme that
catalyzes the isomerization
(rearrangement of atoms) of a
substrate in a reaction, converting it into
a molecule isomeric with itself.
There is only one reactant and one product in
reactions where isomerases are operative.
Isomerases – are a general class of enzymes
which convert a molecule from one isomer to
another. The general form of such a reaction is
as follows: A-B – B-A.
BIOCHEMICAL ACTIVITY:
 Catalyze isomerization changes within a
single molecule.
 Carry out many kinds of isomerization:
o L to D isomerization
 Many kinds of isomerization
o L to D isomerization
o Mutase reactions (shifts of
chemical groups)
EXAMPLES:
 Isomerase
 Mutase
6. LIGASES – Is an enzyme that catalyzes
the bonding together of two molecules
into one with the participation of ATP.
ATP involvement is required because such
reaction is generally energetically unfavorable,
and they required the simultaneous input of
energy obtained by a hydrolysis reaction in
which ATP is converted to ADP.
BIOCHEMICAL ACTIVITY:
 Join two molecules with covalent bonds
catalyze reactions in which two
chemical groups are joined (or ligated)
with the use of energy from ATP.
EXAMPLES:
 Acetyl-CoA Carboxylase
 Glutamine Synthetase
 Ligases (Synthetases)
 Catalyze ligation or joining of two
substrates.
 Require chemical energy (e.g., ATP)
7. TRANSLOCASES – catalyzing the
translocation of hydrogen ions,
inorganic cations and anions, amino
acids, carbohydrates, or other
compounds.
BASIC ENZYME REACTIONS
Explanation of how enzymes functions as
catalyst in biochemical systems are based on
the concepts active site and enzyme-substrate
complex formation.
In enzymatic reactions, the substance at the E + S → ES → E + P
beginning of the process, on which an enzyme ENZYME ACTIVE SITE
begins its action is called substrate.
- Enzyme molecules contain a special
S+E–P+E
pocket or cleft called the active sites.
- The active site is the relatively small
THE ENZYMES – SUBSTRATE COMPLEX part of an enzyme's structure that is
actually involved in catalysis.
An enzyme-substrate complex is the
- The active site in an enzyme is a three-
intermediate reaction species that is formed
dimensional entity formed by groups
when a substrate binds to the active site if the
that come from different parts of the
enzymes.
protein chain(s); these groups are
Within the enzyme-substrate complex, the brought together by the folding and
substrate encounters more favorable reaction bending of the protein. The active site is
conditions than if it were free. The results are usually a "crevicelike" location in the
faster formation of product. enzyme.
ENZYME CATALYZED REACTIONS TWO MODELS FOR FORMATION OF ES
COMPLEX
When a substrate (S) fits properly in an active
site, an enzyme-substrate (ES) complex is
1. Lock- and Key Model
formed:
2. Induced-Fit Model
E (enzymes) + S (substrate) ES (complex)
Lock- and Key Model
Within the active site of the ES complex, the
In the lock- and Key Model enzyme action:
reaction occurs to convert substrate to product
(P): ESE + P(product) - The active site has a rigid shape.
- Only substrates with the matching
The products are then released, allowing
shape can fit.
another substrate molecule to bind the enzyme.
- The substrate is a key that fits the lock
- this cycle can be repeated millions (or of the active site.
even more) times per minute.
This is an older model, however, and does not
work for all enzymes.
The overall reaction for the conversion of
substrate to product can be written as follows:
In lock- and key model, the active site in the
enzymes has a fixed, rigid geometrical
conformation.
Only substrate with the complementary
geometry can accommodated at such a site,
much as a lock accepts only certain keys.
Induced- Fit Model
The Induced- Fit Model allows for small
changes in the shape or geometry of the active
site of an enzyme to accommodate a substrate.
The induced fit is a result of the enzymes
flexibility, it adapts to accept the incoming
substrate.
The active site adjusts to fit the shape of the
substrate more closely.
At the same time the substrate adjusts its 3. LINKAGE SPECIFICITY
shape to better adapt to geometry of the active 4. STEREOCHEMICAL SPECIFICITY
site.
ABSOLUTE SPECIFICITY
As a result, the reacting a section of the
The enzyme will catalyze only one reaction.
substrate becomes aligned exactly with the
This most restrictive of all specificities is not
groups in the active site that catalyzes the
common.
reaction.
Catalase is an enzyme with absolute
A different substrate could not induce these
specificity.
structural changes and no catalysis would
occur. It catalyzes the conversion of hydrogen
ENZYME SPECIFITY
peroxide to oxygen and water.
Hydrogen Peroxide is the only substrate it will
- Is the extend to which an enzyme’s
accept.
activity is restricted to a specific
substrate. GROUP SPECIFICITY
- A specific group of substrates, a specific
type of chemical bond, or specific type The enzyme will act only on molecules that
have a specific functional group, such as a
of chemical reaction
- The degree of specificity is hydroxyl, amino, or phosphate groups.
determined by active site. Some active Carboxypeptidase is group specificity, it
sites accommodate only one particular cleaves amino acids, one at a time, from the
compound, whereas others can carboxyl end of the peptide chain.
accommodate a “family” of closely
LINKAGE SPECIFICITY
related compound.
Enzymes have varying degrees of specificity The enzyme will act on a particular type of
chemical bond, irrespective of the rest of the
for substrates.
molecular structure.
Enzymes may recognize and catalyze:
Phosphatases hydrolyze phosphate – ester
- A single substrate bond types of phosphate esters.
- A group of similar substrates
- A particular type of bond Linkage Specificity is the most general of the
common specificities.
TYPE OF SPECIFITY
STEREOCHEMICAL SPECIFICITY
1. ABSOLUTE SPECIFICITY
2. GROUP SPECIFICITY
The enzyme will act on a particular - A region high temperature in which the What limits enzymatic activity to a certain
stereoisomer. Chirality is inherent in an enzyme rate decreases with increased maximum value?
active site because amino acid are chiral temperature due to the thermal
- As substrate concentration increases,
compounds. inactivation of the enzyme
the point is eventually reached where
(denaturation).
An L-amino acid oxidase will catalyze the enzyme capabilities are used to their
- Optimum temperature 37℃ – 40℃ . At
oxidation of the L-form of an amino acid but not maximum extent. The rate remains
temperature above 50 degrees, the
the D-form of the same amino acid. constant from this point.
tertiary structure and thus the shape of
- The rate of reaction increases as
most proteins is destroyed, which
substrate concentration increases (at
causes a loss in enzyme activity.
constant enzyme concentration)
- For these reasons, equipment in
- Maximum activity occurs when the
hospitals is sterilized in autoclaves
FACTOR THAT AFFECTS THE ENZYME enzyme is saturated (when all enzymes
where high temperature denatures the
ACTIVITY are binding substrate).
enzymes in harmful bacteria.
- Each substrate must occupy an enzyme
- Optimum Temp. is the temperature at
Enzyme activity is a measure of the art at active site for a finite amount of time,
which an enzyme exhibits maximum
which an enzyme converts substrate to and the products must leave the site
activity.
products in a biochemical reaction. before the cycle can be repeated.
pH - When each enzyme molecule is working
FACTORS:
at a full capacity, the incoming substrate
- The pH of an enzyme’s environment molecules must “wait their turn” for an
1. Temperature
can affect its activity. empty active site. At this point, the
2. pH
- Enzymes are most active at their enzyme is said to be under saturation
3. Substrate concentration
optimum pH, the pH that maintains the conditions.
4. Enzyme concentration
proper tertiary structure of proteins.
5. Enzyme inhibition
- Enzymes in most cells have optimum The change in enzyme activity with a change in
Temperature pH values at physiological pH values substrate concentration at constant
around 7.4. temperature, pH, and enzyme concentration.
- Temperature is a measure of the kinetic Enzyme activity remains constant after a
energy (energy of motion) of Substrate Concentration certain substrate concentration is reached.
molecules.
- When the concentration of an enzymes Enzyme Concentration
- At higher temperatures molecules are
is kept constant and the concentration
moving faster and colliding more
of substrate is increased, the enzyme - Because enzymes are not consumed in
frequently.
activity pattern is called a saturation the reactions they catalyze, the cell
- Enzymes are very sensitive to
curve. usually keeps the number of enzymes
temperature.
- Enzyme activity increases up to a low compared with the number of
- On enzyme reaction is two.
certain substrate concentration and substrate molecules.
thereafter remains constant.
The change in reaction rate with a change in
enzyme concentration for an enzymatic
reaction. Temperature, pH, and substrate
concentration are constant. The substrate
concentration is relative to enzyme
concentration.
Enzyme Inhibition
The rates of enzymes-catalyzed reactions can
be decreased by a group of substances called
INHIBITORS.
An Enzyme Inhibitor is a substance that slows
or stops the normal catalytic function of an
enzyme by binding.
A. Competitive Enzyme Inhibitor
B. Noncompetitive Enzyme Inhibitor
C. Irreversible Enzyme Inhibitor
Competitive Enzyme Inhibitor
- A molecule closely resembling the
substrate. Binds to the active site and
temporarily prevents substrates from
occupying it, thus blocking the reaction.
- A competitive inhibition has a
structure that is so similar to the
substrate it competes for active site on
the enzyme.
- As long as the inhibitor occupies the
active site, the substrate cannot bind to
the enzyme and no reaction takes
place.
- As long as the concentration of the
inhibitor is substantial, there is a loss of
enzyme activity.
- However, increasing the substrate
concentration displaces more of the
inhibitor molecules. As more enzyme
molecules bind to substrate (ES),
enzyme activity is gained.
Ex.
 Antibiotic – competitive inhibitor
 Antihistamine – are competitive
inhibitor of histidine decarboxylation, the
enzymatic reaction that converts
histidine to histamine. Histamine
causes the usual allergy and cold
symptoms: watery eyes and runny
nose.
Noncompetitive Enzymes Inhibitor
- A molecule that binds to a site on an
enzyme that is not the active site. The
normal substrate still occupies the
active site, but the enzyme cannot
catalyze the reaction due to the
presence of the inhibitor.
- The structure of a noncompetitive
inhibitor does not resemble the
substrate and does not compete for the
active site. Instead, it binds to a site of
the enzyme that is not the active site.
- When the noncompetitive inhibitor is
bonded to the enzyme, the shape of the
enzyme is distorted. Inhibition occurs
because-the substrate cannot fit in the
active site.
Ex.
 Heavy metals ions lead silver, mercury
that bond with amino acid side groups
such as -COO or OH.
 Antibiotics produced by bacteria, mold,
or yeast are inhibitors used to stop
bacterial growth.
 Penicillin inhibits an enzyme needed for
the formation of cell walls in bacteria,
but not human cell membranes. With an
incomplete wall, bacteria cannot
survive, and the infection is stopped.
Irreversible Enzyme Inhibitor
- A molecule that forms a covalent bond
to a part of the active site, permanently
preventing substrate from occupying it.
- Is a molecule that inactivates enzymes
by forming a strong covalent bond to
 amino acid side-chain group at the
enzyme's active site.
- In general, such inhibitors do not have
structures similar to that of the enzymes
normal structure.
- The inhibitor-active site bond is
sufficiently strong that addition of
excess substrate does not reverse the
inhibition process. Thus, the enzyme is
permanently deactivated.