ENZYMES

Prof. Pınar AKSOY SAĞIRLI

 SIGNIFICANCE OF ENZYMES
Enzymes are play important role
on the center of biochemical
prosesses.

They are important biomarkers for

diagnosis

A lots of drug have showed their

thereupatic effect by affecting the
enzymes
History
Enzymes comes from Greek, "leavened" or "in yeast"
In 1877, German physiologist Wilhelm Kühne (1837–
1900) first used the term enzyme
In 1897, Eduard Buchner found that sugar was
fermented by yeast extracts even when there were
no living yeast cells in the mixture.
Buchner, named the enzyme that brought about the Eduard Buchner
fermentation of sucrose"zymase".

In 1907, he received the Nobel Prize in Chemistry for

"his discovery of cell-free fermentation"
The word enzyme was used later to refer to nonliving
substances such as pepsin,

The word ferment was used to refer to chemical activity

produced by living organisms.
In 1926, James B. Sumner showed that the enzyme
urease was a pure protein and crystallized it; he
did likewise for the enzyme catalase in 1937. James Batcheller
Sumner

Pure proteins can be enzymes was definitively

demonstrated by John Howard Northrop and
Wendell Meredith Stanley, who worked on the
digestive enzymes pepsin (1930), trypsin and
chymotrypsin. Wendell Meredith
Stanley

These three scientists were awarded the 1946 Nobel

Prize in Chemistry.

John Howard
Northrop
 The discovery that enzymes could be crystallized
eventually allowed their structures to be solved by x-
ray crystallography. This was first done for
lysozyme; the structure was solved by a group led by
David Chilton Phillips and published in 1965.

 This high-resolution structure of lysozyme marked

the beginning of the field of structural biology
MECHANISM
In 1894, Lock-Key model

Emil Fischer proposed that both the enzyme and the substrate
possess specific complementary geometric shapes that fit exactly
into one another.

In 1930, Haldane suggested that there were weak bonds between

enzyme and substrat
In 1958, Koshland, "induced fit " model
Daniel Koshland suggested a modification to the lock and key
model: since enzymes are rather flexible structures, the active
site is continuously reshaped by interactions with the substrate
as the substrate interacts with the enzyme. As a result, the
substrate does not simply bind to a rigid active site; the amino
acid side-chains that make up the active site are molded into the
precise positions that enable the enzyme to perform its catalytic
function.
In some cases, such as glycosidases, the substrate molecule also
changes shape slightly as it enters the active site. The active site
continues to change until the substrate is completely bound, at
which point the final shape and charge distribution is
determined.
STRUCTURE
Definitions

Enzymes are protein produced by a

living organism which acts as a catalysts
to accelerate biocemical reaction.
The molecules upon which enzymes may
act are called substrates and the enzyme
converts the substrates into different
molecules known as products.
Cofactor is a non-protein chemical
compound or metallic ion that is required
for an enzyme's activity
CLASSIFICATION BASED ON COFACTORS
 ANOTHER CLASSIFICATION

Coenzyme:
-Complex organic coumpounds
-Transfer electrons between enzymes
-NAD, FAD, TPP ect.

Cofactor:
-Metal ions (Mg+2, Fe+2, Mn+2) or simple organic componds
-Bind to enzyme and change its shape
-Essential to function
 OTHER DEFıNıTONS RELATED TO ENZYME
Substrate is the substance on which an enzyme acts.
Cofactor is a substance (other than the substrate) whose
presence is essential for the activity of an enzyme and
identifies biocemical reaction
Apoenzyme is an inactive enzyme, activation of the
enzyme occurs upon binding of an organic or
inorganic cofactor. It identifies substrate..
Holoenzyme (enzyme) is An apoenzyme together with its
cofactor. A holoenzyme is complete and catalytically active
HOLOENZIM
 Some Organic cofactors

Enzymes Cofactors
Oxidoreductase NAD, NADP (Coenzyme)
FAD, FMN (Prosthetic group)
α-Keto asid decarboxylase Tiamine pyrophospate (TPP, coenzyme)
Α-amino asid decarboxylase Pyridoksal phospate (coenzyme)
Transaminase Pyridoksal phospate (coenzyme)
decarboxylase Biotin (Prosthetic group)
Enymes that transfers one carbon Tetrahydrofolic Acid (Prosthetic group)
Enzymes which has metal ions
Ions Enzyme

Copper, Cu+2 Cytocrome Oxidase, Ascorbate

Oxidase, Tyrosinase

Iron , Fe+2 or Fe+3 Catalase, Cytocrome

Peroxidase, Nitrogenase
Hydrogenase

Magnesium, Mg+2 Glucose-6-phospatase,Hekzokinase

Pyruvate Kinase, Creatine kinase,
Phosphofructokinase,
DNA polymerase

Manganese, Mn+2 Arginase

Ribonucleotide reductase Pyruvate
carboxyilase
Molybdenum Nitrate reduktase Nitrogenase

Nickel Ürease
Zinc Alcohol dehydrogenase, Carbonic
anhidrase, DNA polymerase
Enzymes without cofactor
Pepsin
Trypsin
Effect of enzymes on reaction rate
 HOW TO INCREASE CHEMICAL
REACTION RATE
 Increase reactant concentration
 Increase temperature

 Add catalyts
 Change pH
 Enzymes increase reaction rate by 109-1016 times.
 In biological system, energy used by cells expresses as
GIBSS free energy (G).
 A+B ↔C+D The reaction system tends to progress to
equilibrium when not yet in balance.This trend
indicates that there is a driving force for the reaction.
 Driving force, is expressed as “free energy change”
“ΔG” , equal to the energy difference between A+B and
C+D
 Activation Energy (Ea, ΔG+) and change in
free energy (ΔG°)
The figure shows typical energy
profile of the chemical reaction.
ΔG is the change in free energy
of a system as it goes from
some initial state, such as all
reactants, to some other, final
state, such as all products. This
value tells us the maximum
usable energy released (or
absorbed) in going from the
initial to the final state. In
Enzymes increase reaction rate addition, its sign (positive or
by decreasing activation energy. negative) tells us whether a
reaction will occur
spontaneously, that is, without
added energy.
 Reactions that have a negative ∆G release free energy and are
called exergonic reactions. A negative ∆G means that the
reactants, or initial state, have more free energy than the
products, or final state. Exergonic reactions are also called
spontaneous reactions, because they can occur without the
addition of energy.
 Reactions with a positive ∆G (∆G > 0), on the other hand, require
an input of energy and are called endergonic reactions. In this
case, the products, or final state, have more free energy than the
reactants, or initial state. Endergonic reactions are non-
spontaneous, meaning that energy must be added before they
can proceed.
 Transition state is defined as the state corresponding to the
highest potential energy along this reaction coordinate.
 In conclusion, enzymes increase the rate of chemical
reaction by lowering the activation energy barriers
 ENZYME-SUBSTRATE INCORPORATION
When a substrate binds to an enzyme at its active site then
an intermediate is formed which is known as enzyme
substrate complex.

Active site: the active site is the region of an enzyme where

substrate molecules bind and undergo a chemical reaction.

Active site
1.Substrate binding site
(apoenzyme)

2.Catalytic site (cofactor or

some special amino acid
seqences)
KEY-LOCK MODEL
 ENZYME KINETICS
Enzymatic reaction rate
-The rate of a chemical reaction
increases as the substrate
concentration increases.
Enzymes can greatly speed up
the rate of a reaction.

-However, enzymes become

saturated when the substrate
concentration is high.

-The rate of enzyme reaction is

proportional to the substrate
concentration at the low substrate
concentration.
 MICHAELIS-MENTEN KINETICS
In biochemistry, Michaelis–Menten kinetics is one of the best-
known models of enzyme kinetics
Biochemical reactions involving a single substrate are often
assumed to follow Michaelis–Menten kinetics, without
regard to the model's underlying assumptions.
Vmax represents the maximum rate achieved by the system, at
saturating substrate concentration.
The Michaelis constant Km is the substrate concentration at
which the reaction rate is half of Vmax.
 MICHAELIS-MENTEN THEORY
The enzyme (E) combines with the substrate (S), to form an enzyme-substrate (ES)
complex, which immediately breaks down to the enzyme and the product (P).
 MICHAELIS-MENTEN THEORY
Michaelis-Menten Teory

According to the theory, The enzyme (E) combines with the substrate (S), to form
an enzyme-substrate (ES) complex, which immediately breaks down to the enzyme
and the product (P). These reactions are two sided.
MICHAELIS-MENTEN TEORY
 Km, is the concentration of substrate which
permits the enzyme to achieve half of Vmax and
is expressed as Molarity.

 An enzyme with a high Km has a low Leonor Michaelis

affinity for its substrate, and requires a 1875-1949
greater concentration of substrate to achieve
Vmax

 An enzyme with a low Km has a high affinity for

its substrate. A small Km indicates that the
enzyme requires only a small amount of substrate
to become saturated
Maud Menten
1879-1960
 FEATURES OF MICHAELIS-MENTEN
KINETICS

Km is independent of enzyme concentration.

Vmax depends on enzyme concentration
Km and Vmax can be altered by pH, temperature and other
factors.

Velocity vs. substrate curve depends on substrate concentration

has a hyperbolic shape (rectangle shape)
TWO-SUBSTRATE ENZYME KINETICS
 TWO-SUBSTRATE ENZYME KINETICS

 There are two type of mechanisms for two-substrate

enzyme kinetics. .
1.Single-displacement reaction (sequential
mechanism), In this mechanism, both substrates
must bind to the enzyme before any product is
released (Malate dehydrogenase)
2.Double-displacement reaction (nonsequential
mechanism), in the nonsequential mechanism, do not
require all the substrates to bind before a product is
released thus a substituted-enzyme intermediate, E will
be formed. Such mechanisms are generally referred to
as ping-pong or substituted mechanisms. (aspartate
transaminase).
 ENZIM INHIBITION
Inhibition is - the act of inhibiting : the state of being inhibited
An enzyme inhibitor is a molecule that binds to an enzyme and
decreases/blokes its activity.
 ENZIM INHIBITION

Inhibition of enzymes may be either reversible or irreversible depending on the

specific effect of the inhibitor being used
Enzymes can be inhibited

Competitive inhibition, when the substrate and inhibitor

compete for binding to the same active site
Noncompetitive inhibition, when the inhibitor binds
somewhere else on the enzyme molecule reducing its
efficiency.
Uncompetitive inhibition, binds to a site of the enzyme that
is removed from the active site but only if the substrates
already present.
 Competitive inhibition occurs in four different situations

1. Analog of substrates and substrate compete for the same

binding site on the enzyme
2. In two-substrate reactions, second substrate and first substrate
compete for the same binding site on the enzyme
3. The product forming in the end of the reaction act as an
inhibitor
4. Some reaction required a cofactor such as metal ion can be
inhibited by the metal ions
 COMPETITIVE INHIBITION

Example;
● Inhibition of succinate dehydrogenase by malonic acid
Example;
Methanol poisoning
 NON-COMPETITIVE INHIBITION

In noncompetitive inhibition, a molecule binds to an enzyme

somewhere other than the active site. This changes the enzyme's
three-dimensional structure so that its active site can still bind
substrate with the usual affinity, but is no longer in the optimal
arrangement to stabilize the transition state and catalyze the reation.
On the macroscopic scale, noncompetitive inhibition lowers the
Vmax. Thus, the enzyme simply cannot catalyze the reaction with the
same efficiency as the uninhibited enzyme. Note that noncompetitive
inhibition cannot be overcome by raising the substrate concentration
like competitive inhibition can.
FOR EXAMPLE
For example, the amino acid alanine noncompetitively inhibits the
enzyme pyruvate kinase. Alanine is one product of a series of enzyme-
catalyzed reactions, the first step of which is catalyzed by pyruvate
kinase. j
EXAMPLES;
CO, CN-, H2S: Inhibitors of Fe-porpyrin containing
enymes (cytocroms, catalase, peroxidase).
NaF: Inibitors of enzymes which requires Ca2+ for their
activation (some phosphorilase).
CN-, EDTA: Inibitors of Cu containing enzymes (tirozinase,
askorbate oxidase, urate oxidase).
.
UNCOMPETITIVE INHIBITOR

Uncompetitive inhibitors only recognize and interact with ES.

Inhibitors causes structurel distortion of the active site.
Uncompetitive inhibition typically occurs in reactions with two
or more substrates or products.
Inhibition of enzyme activity is characterized by a decrease in
both substrate Km and Vmax.
Inhibition can’t be reversed by increasing the [S] since İnibitor
does not compete with S for the same binding site.
For example, Inhbition of placental alkaline phosphatase by
phenylalanine.
 IRREVERSIBLE INHIBITION

● Irreversible inhibitors are covalently bound to the target enzyme.

Therefore, they are irreversible.
● Contain reactive functional group such as aldehyde,
haloalkane, phenyl sulfonate, fluorophosphonate
● These groups are covalently bound to –OH or –SH of
amino acid (DIPF on chymotrypsin)
EXAMPLE,
Nitrogen mustards are powerful inhibitors for acetylcholine
esterase
ENZYME ACTIVATORS

 Enzyme activators are molecules that bind to enzymes

and increase their activity.
 These molecules are often involved in the allosteric
regulation of enzymes in the control of metabolism.
 PROENZYMS=ZYMOGENS, PREPROENZYMS

 Most of the enzymes are synthesized in the form of inactive

proenzymes.
 These inactive forms of enzymes are called proenzymes or
zymogens.
 Generally, prefix pro- or suffix -ogen is added to enzyme’s
name to denote its zymogen such as prothrombin,
proelastase, trypsinogen, pepsinogen, fibrinogen etc.
 MEASURING ENZYME ACTIVITY
One international unit (IU), is the amount of enzyme
consuming or forming 1 µmol substrate or 1 µmol product per
minute under standard conditions. The base unit is 1 katal,
corresponding to the amount of enzyme converting 1 mol
substrate per second: Usually U is used instead of IU.

Spesific activity, is activity of 1mg enzyme protein (U/mg).

Katal, is the unit of catalytic activity in the International

System of Units (SI). One katal is the amonut of enyme that
catalyses the transformation of 1 mole of substrate/second.

1 katal=1mol s-1
 MULTI-ENZYME SYSTEMS

The series of enzymes catalyzing such chains of reactions are said to

form a multi-enzyme system. The product of the enzymatic reaction
uses as a substrate for the next step in multi-enzyme system.
Enymes in the multi enzyme system remain free in the
cytosol as independent entities each interacting with its own
substrate which is also present in -the cytosol Some of the
multi-enzyme systems may operate in a different way, when
the enzymes are closely associated with each other to form a
multi-enzyme complex.

For example,
-Fatty acid synthase (7 enzymes)
-Pyruvate dehydrogenase complex
and 2-Oxoglutarate Dehydrogenase (3 enzymes)
FATTY ACID SYNTHASE
 ISOENZYMES (ISOZYMES)

•Isozymes are enzymes that differ in amino acid sequence but

catalyze the same chemical reaction.
•They are exist in different proportions in different tissues.
• They are usually organ-specific, and therefore exploited for
diagnostic purposes.
•Most enzymes present in plasma are released during normal
cell turnover. However, increased amounts of particular
isozymes appearing in plasma usually reveal trauma or
pathologic processes.
•They are usually separated from each other in the diagnostic
laboratory via electrophoresis.
Example,
Creatine phosphokinase (CPK) and lactate dehydrogenase
(LDH) are two isozymes commonly used for diagnostic
purposes.

Creatine phosphokinase
The CPK enzyme contains two dimers assembled from
muscle (M) and brain (B) subtypes.
Skeletal muscle CPK is almost entirely the MM isozyme,
brain BB, and cardiac muscle 15% MB and 85% MM. Only
cardiac muscle has the MB dimer.
Lactate dehydrogenase is a tetramer, assembled from M
and heart (H) subunits, with 5 different isozymes appearing
via electrophoresis.
Following a myocardial ischemic episode, LDH release into
plasma usually occurs after that of CPK and troponin-I.
ISOENZYMES
FACTORS AFFECTING ENZYMES

 Enzyme concentration
 Substrate concentration
 Temperatre
 pH
 Time
 Reaction products
 Light ve other pysical effects
 Hormons and other substance
ENZYME CONCENTRATION

As the concentration of the

enzyme is increased, the velocity
of the reaction proportionately
increases. This property is used
for determining the activities of
serum enzymes during the
diagnosis of diseases.
FEED-BACK INHIBITION

In some enzymatic reaciton, the product inhibits the

enzyme. Reaction rate decreases even if the enzyme
concentration is increased.
This kind of inibition refers as feed-back inhibition

Generally feed-back inhibition affects allosteric enzymes.

 CONCENTRATION OF SUBSTRATE

In the presence of a given amount

of enzyme, the rate of enzymatic
reaction increases as the substrate Saturation point

concentration increases until a

limiting rate is reached, after which
further increase in the substrate
concentration produces no
significant change in the reaction
rate.
At this point, the enzyme molecules
are saturated with substrate.
 TEMPERATURE
The protein nature of the
enzymes makes them extremely
sensitive to thermal changes.
Enzyme activity occurs within a
narrow range of temperatures
compared to ordinary chemical
reactions. Each enzyme has a
certain temperature at which it
is more active. This point is
called the optimal temperature,
which ranges between 37 to
40C°.
 EFFECT OF pH
Enzymes are protein substances
that contain acidic carboxylic
groups (COOH–) and basic amino
groups (NH2). So, the enzymes are
affected by changing the pH value.
Each enzyme has a pH value that
it works at with maximum
efficiency called the optimal pH.
If the pH is lower or higher than
the optimal pH, the enzyme
activity decreases until it stops
working. For example, pepsin
works at a low pH, i.e, it is highly
acidic, while trypsin works at a
high pH, i.e, it is basic. Most
enzymes work at neutral pH 7.4.
REGULATORY ENZYMES

Regulatory enzymes are

commonly the first enzyme in a
multi-enzyme system: the
product of the reaction catalyzed
by the first enzyme is the
substrate of the second enzyme,
so the cell can control the amount
of resulting product by regulating
the activity of the first enzyme of
the pathway.
Reglatory enzymes are divided in to two groups according
to the way they are affected by signaling molecules.
1) Allosteric enzymes
2) Enzymes affecting by covalent binding
ALLOSTERIC ENZYMES
Allosteric site, The place on an enzyme where a molecule that
is not a substrate may bind, thus changing the shape of the
enzyme and influencing its ability to be active.

Allosteric effect: The binding of a ligand to one site on a

protein molecule in such a way that the properties of another
site on the same protein are affected. Some enzymes are
allosteric proteins, and their activity is regulated through the
binding of an effector to an allosteric site. "allosteric effect."
Negative allosteric modulation (also known as allosteric
inhibition) occurs when the binding of one ligand decreases
the affinity for substrate at other active sites.
Allosteric enzyme

E1 E2 E3
A B C D

Feed-back inhibition
Effector (signal molecule), is a molecule binding allosteric site of
enyme.
Pozitive effector (activator), increases enzyme activity
Negative efektör (inhibitör), decreases enzyme activity
Allosteric enzymes don’t fit Michaelis-
Menten kinetics
ALLOSTERIC REGULATION OF
PHOSPHOFRUCTOKINASE
 ENZYMES AFFECTING BY COVALENT
BONDING
Some enzymes is regulated by phosphorilation, the most prevalent
reversible covalent modification. Thus, the enzymes can be active or
inactive after this modification by catalysing another enzyme.

Phosphorylase kinase

Phosphorylase b Phosphorylase a
Inactive active

Phosphorylase phosphotase
 ENZYMES IN DIAGNOSIS
Clinical enzymology can
be described as the branch
of science which deals
with the. application of
enzyme analysis to the
diagnosis and treatment of
disease
 CONDITIONS RELATED TO INCREASED SERUM
ENYZME LEVELS

-Cellular membrane damage

-Cell death
-Increasing of enzyme production
-Increased rate of cell turnover
-Proliferation of cells
 CLINICAL SIGNIFICANCE OF ENZYME
ACTIVITY MEASUREMENT
Common clinical samples,
-Serum
-Plasma
-Urine

Enzyme appear in plasma by ways

1.Functional plasma enzymes
2.Non- Functional plasma enzymes
3.Obstruction to secretory pathway
1. FUNCTIONAL PLASMA ENZYMES
2. NON-FUNCTIONAL PLASMA ENZYMES
 3. OBSTRUCTION TO SECRETORY PATHWAY

-Physiological conditions
 Balance between synthesis&release
-Pathological conditions
 Loss of balance between synthesis&release
WHAT IS THE CLINICAL SIGNIFICANCE OF
NON-FUNCTIONAL ENZYMES

1. Diagnosis of disease
2. Prognosis of disease
 WHAT ARE SOURCES OF NON-
FUNCTIONAL ENZYMES
1. Cellular damage
-Myocardial infarction and viral hepatite

2. Obstruction to secretory pathway

-As a results of biliary tract obstruction, alkaline
phosphatase increases in plasma.

3. Increasing enzyme synthesis

-Alkaline phosphatase increases in hepatite diseaase

4. Increasing permeability
-Hypoxia
SIGNIFICANCE OF ENZYMES IN PHARMACY
Drug manufacturers use enzymes or enzyme containing
microorganisms to biocatalyse a chemical reaction.

For example,
Digestive drugs have proteases , cellulase, lipase, lactase
SIGNIFICANCE OF ENZYMES IN PHARMACY

1. Some of enzymes use as a drug

2. Enzyme inibitors can also be a
drug
 ENZYMES IN PHARMACY
1. Some of enzymes are used as a drug
a.In digestive problem related to liver, gall bladder, pancreas disease,
enzymes hydrolysing protein, carbohydrate and lipid are used as a drug
Pancreatine, lipase, amylase, protease
Multanzim
Pankreoflat
Pankrean
Festal
Pancreatin, hemicellulase
Flaton
b.In wound treatment,
Hyaluronidase (lasonil)

2. Enzyme inibitors can also be a drug

 REVERSIBLE INHIBITORS
Drug name Enzyme inhibition Treatment
Lovastatine HMG-CoA reductase Hyperlipidemia
Hypercholesterolemia

Allopurinol Xanthine oxidase Hyperuricemia

Acetazolamide carbonic anhydrase Diuretic

Metazolamide
Methotrexate Dihydrofolate reductase Anti-cancer

Aspirin cyclooxygenase Anti-inflammatory

(COX1 and COX2)
Cytosine arabinoside DNA Antiviral, anticancer
polymerase
RNA
polymerase
ACE inhibitors; angiotensin- Antihypertensive
Enalapril converting enzyme
Cilazapril (ACE)
Lisinopril
 IRREVERSIBLE INHIBITORS

Drug name Enzyme Treatment

Sulfanylamide Dihydrofolate Antibacterial
synthetase
Hydrazine Monoamine oxidase Antidepressant,
(MAO) Psychostimulant
Penisilin Transpeptidase Antibiotic

Cephalosporin Transpeptidase Antibiotic

Clavulanic acid Βeta-Lactamase Antibiotic adjuvant
 IRREVERSIBLE INHIBITORS
Sulfanylamide
-an organic sulfur compound
-structurally similar to p-aminobenzoic acid (PABA)
-antibacterial

Sulfanylamide competes with PABA for the enzyme

dihydropteroate synthase, thereby preventing the
incorporation of PABA into dihydrofolic acid, the immediate
precursor of folic acid.
This leads to an inhibition of bacterial folic acid synthesis
and de novo synthesis of purines and pyrimidines,
ultimately resulting in cell growth arrest and cell death.
 FOLIC ACID
 Folic
acid, which plays a key role in one-carbon
metabolism, is essential for the biosynthesis of several
compounds such as purine, pyrimidine.
Folic acid reductase inhibitors
Tetrahydrofolic acid which is a active form of Folic acid transfer one
carbon unit.

Folic acid reductase inhibitors

Aminopterin
Ametopterin

They are competing with folic acid since they are derivative of folic
acid.
 Hydroxymetilglutaril-CoA (HMG-CoA)
redüktaz inhibitörleri
Statins, also known as HMG-CoA reductase inhibitors, are a
class of lipid-lowering medications.They inhibit the enzyme
HMG-CoA reductase which plays a central role in the
production of cholesterol.
 Xanthine oxidase inhibitors

Allopurinol is a xanthine oxidase inhibitor that decreases uric

acid production.
 Angiotensin Converting Enzyme inhibitors
Angiotensin converting enzyme (ACE) inhibitors are high blood
pressure drugs that widen or dilate the blood vessels to improve the
amount of blood the heart pumps and to lower blood pressure
 ENZYMES CLASSIFICATION
Accordingly the enzymes are classified into six
main family classes and many sub-family classes.
The principal classes are given below
Each enzyme has

-A common name

-A systematic name which express its reaction

-The International Union of Biochemistry and Molecular Biology

developed a nomenclature for enzymes. Each enzyme is
described by a sequence of four numbers preceded by "EC".
EC denotes Enzyme Commission and the number of enzyme is
called EC numbers.
For example:

oxalate + O2 + 2 H+ = 2 CO2 + H2O2

 Accepted name:oxalate oxidase

 Systematic name: oxalate:oxygenoxidoreductase
 Classification number: EC.1.2.3.4

EC------Enzyme commission
1--------Class (= oxidoreductase)
2--------Subclass(= it acts on the aldehyde or oxo group of donors)
3--------Sub-subclass(= an acceptor)
4--------Serial number

The first number categorizes the enzyme based on its reaction

For example:
ATP + Creatine ADP + Creatine phosphate

 Accepted name: Creatine Kinase

 Systematic name: ATP: Creatine phosphotranferase
 Classification number: EC.2.7.3.2

EC-----Enzyme commission
1-------Class (= transferase)
2-------Subclass(= phosphotransferase)
3-------Sub-subclass(= acceptor is nitrogen)
4-------Serial number
For example:

 Accepted name: Succinyl-CoA hydrolase

 Systematic name: Succinyl-CoA hydrolase
 Classification number: E.C. 3.1.2.3

EC-----Enzyme commission
3-------Class (=Hydrolase)
1-------Subclass (=hydrolysis of ester)
2-------Sub-subclass (=hydrolysis of thioester)
3-------Serial number
1. Oxidoreductases
They catalyze oxidation-reduction reactions where electrons
are transferred.
These electrons are usually in the form of hydride ions or
hydrogen atoms. When a substrate is being oxidized it is the
hydrogen donor.
The most common name used is a dehydrogenase and
sometimes reductase will be used.
Coenzyms (Donor-Akseptor) NADH, NADPH; FADH2

R-CH2OH + NAD+ R-CHO + NADH+H+

(Primer Alcohol) (Aldehyde)

Enzym : EC.1.1.1.1, Alcohol NAD+ oxidoreductase

SUB-CLASSIFICATION OF OXIDOREDUCTASES

1 Dehydrogenases and cytochromes

a)Nicotinamide nucleotide-linked enzymes
b)Flavin nucleotide-linked enzymes
c)Fe3+ porphyrin-linked enzymes (cytochroms)
d)Lipoate
e)Quinone
2- Oxidases
3- Oxygenases
 1- Dehydrogenases
• They oxidize a substrate by transferring hydrogen to an
electron acceptor.
•Common electron acceptors are NAD+, NADP+ or FAD.
• Dehydrogenase reactions come most commonly in two
forms:
1. the transfer of a hydride and release of a proton
(often with water as a second reactant),

2. the transfer of two hydrogens.

Dehydrogenases Example Group
Class

NAD-linked Glycerol phosphate dehydrogenase hydrogen atom

dehydrogenase Lactate dehydrogenase (H+ + e-)
Malate dehydrogenase
L-Glutamate dehydrogenase
glucose-6-phosphate dehydrogenase

FAD-linked succinate dehydrogenase hydrogen atom

dehydrogenase Choline dehydrogenase (H+ + e-)
Acyl CoA dehydrogenase

Cytochromes (Fe3+ cytochrome a ve cytochrome a3, cytochrome b, electron

porphyrin) c1; CYP450 and cytochrome b5 bulunur.

Lipoate Dihidrolipoil asetil transferaz Dihidrolipoil 2H+ +2 e-

süksiniltransferaz (Decarboxylation of α-
keto acids)

Quinone Coenzim Q
NAD-linked dehydrogenases
Trigliserid biosentezi
Glycolysis

TCA
FAD-linked dehydroenase

Succinate
dehydrogenase
ΒETA-OXIDATION
LIPOATE
QUINONE

Coenzyme Q, ubiquinone
 2- OXIDASE
They catalyze removal of hydrogen atoms from substrate and
oxygen accepts these hydrogen atoms, as a result; either H2O or
H2O2 is produced.
Örn Detoxification reaction
Catalase peroxide radical
Peroxidase peroxide radical
Superoxide dismutase superoxid radical
Glutation peroxidase peroxide radical
 Oxidase classification based on the cofactors
1. Copper containing oxidase (ascorbate oxidase, urat oxidase)

2. Iron-porphrin oxidase (Catalase, Peroxidase, Cytocrome oxidase)

3. Flavin-linked oxidase (L-amino acid ve D-amino acid
oxidases, ksantin oxidases, , Diamin oxidase)
L- amino acid oxidases (cofactor FMN):
-catalyze oxidative deamination of L-amino acids

D- amino acid oxidase (cofactor FAD):

-catalyze oxidative deamination of D-amino acids
(L-amino acids present in human)
3- OXIGENASE
They catalyze the introduction of oxygen molecule to the substrate
a) Dioxigenases

a) Monooxigenases (hydroxylases)
A) DIOXIGENASE
They catalyze the introduction of oxygen molecule to
carbon atoms, usually existing next to each other in an
aromatic ring. Thus, they cause splitting of the ring.

Example, tryptofan-2,3-dioxigenase.
B) MONOOXIGENASES (HYDROXYLASES):
They catalyze the introduction of one oxygen atom of the
oxygen molecule (O2) as hydroxy group to the substrate and
the other oxygen atom to be reduced to produce water (H2O).
For reduction of oxygen atoms, reducing agents (cosubstrates)
are necessary.

Reducing factors: Ubiqinon, glutation, Fe2+-2-oxoglutaric

acid, ascorbate, NADH, NADPH etc.
For example, fenylalanine-4-monooxygenase.
For example,
fenylalanine-4-monooxygenase.
Kynurenine-3-monooxygenase,
Tyrozine-3-monooxygenase,
Proline monooxygenase,
Lysine monooxygenase,
steroid monooxygenase
 ENZIM SISTEMLERI HALINDE IŞLEV YAPAN
MONOOKSIJENAZLAR
Örneğin
sitokrom b5 (sit b5), doymuş yağ asidlerinin doymamış şekle
dönüştürülmesinde görev yapan monooksijenaz enzim sisteminde rol
alır.
sitokrom P450 (sit P450), steroid maddelerin biosentezindeki
hidroksilasyon reaksiyonlarıyla ksenobiyotiklerin metabolizmasındaki
(ilaç ve toksik maddelerin zehirsizleştirilmesinde) hidroksilasyon
reaksiyonlarında (Faz I) yer alan monooksijenaz enzim sisteminde
vardır.
Sit b5 ve sit P450 ile katalizlenen reaksiyonlar karaciğerin düz yüzeyli
endoplazmik retikulumunda gerçekleşir.
 2. TRANSFERASES
1. One carbon atom
2. Aldehyte or ketone (Transaldolase, transketolase),
3. Acyle (Acetyl acyl transferase),
4. Glycosyle (glycogen synthetase),
5. Nitrogen (Glutamat-Oxalacetat-Transaminase,
glutamine-pyruvate transaminase)
6. Phosphotransferase
1.phosphomutase; Phosphoglucomutase
2.Phosphokinase; Glucokinase, Fructokinase,
Galactokinase
7. Sulfur
8. Selenium
Enzyme Group Example
Transacylases, Acetyl or acyl Acetyl acyl transferase
Transaminases, Amino Glutamat-Oxalacetat-
Transaminase, glutamine-
pyruvate transaminase
Phosphotransferases Phosphate Glucokinase,
(Kinases) Fructokinase,
Galactokinase
 3. HYDROLASES

They catalze hydrolyzing reaction. Thus, they cleave the

molecules.

1 Ester bound,
a. Carboxylesterase (Lipase, Cholesterol esterase )
b. Thiol Ester Hydrolase (Acetyl-CoA hydrolase, Succinyl-
CoA hydrolase)
c. Phosphotases (Phosphomonoesterase,
Phosphodiesterase)
d. Sulfatase
2 Glycosidic linkage (ether bound),
a. O-glycosyle
α-glucosidase, β-galactosidase
β-D-fructofuranosid fruktohydrolase
β-glucoronydase,
oligodextrin-6-glocanohydrolase,
α-amylase, hyaluronidase, heparinase
a. N-glycosyle
Nucleosidase
a. S-glycosyle
3 Peptid bond,
Carboxypeptidase A ve B, Pepsin
Rennin, Trypsin
Chymotrypsin
Catepsin
Trombin
4 Other C–N bonds,
Urease, Glutaminase, Asparaginase, Arginase
5- Acid anhydrid bonds,
ATP phosphohydrolase
Enzyme Oligo, polisaccarid Bond
α-Glucosidase maltose Glycosid bond
β-Galactosidase lactose Glycosid bond
β-D-Fruktofuranozid saccarose Glycosid bond
fruktohydrolase
α-Amylase Glycosid bond
starch
EFFECTIVE HYDROLASES DURING PROTEIN DIGESTION
Trypsin,
Chymotrypsin,
Pepsin
Rennin
also called chymosin, protein-digesting enzyme that
curdles milk by transforming caseinogen into insoluble
casein;

EFFECTIVE HYDROLASES TO ESTER BONDS DURING

DIGESTION OF LIPIDS

Lipase
Phosphatase hydrolyzes phosphate bonds
4. LYASES
They catalyze the breaking (an "elimination" reaction) of various
chemical bonds by means other than hydrolysis and oxidation, often
forming a new double bond or a new ring structure.
1. C–C Lyases
a. Carboxy Lyases, Decarboxylyases
Pyruvate decarboxylase
2-oxoglutarate decarboxylase
α-aminoasid decarboxylase
tyrosine, histidine, DOPA decarboxylase
b. Aldehyd lyases
Fructose-1,6- bisphosphate D gliseraldehit-3-
fosfat- lyase
c. Keto acid Lyases
Citrate oxalasetat lyase
2. C–O Lyases,
Carbonic anhydrase
Fumarat hydratase
3. C–N Lyases,
Arginino succinate Lyases
4. C–S Lyases,
5. C–halogene Lyases,
6. P–O Lyases,
7. Other Lyases
Exp.(keto acid lyases) citrate oxaloacetate lyase

Dekarboksilazlar

They catalyze removing CO2 from a molecule

Exp. Tyrosin deckarboxylase

 5. ISOMERASES

They catalyze intramolecular changes.

1 Racemases
2 Epimerases
3 Cis-trans isomerase,
Epimerases,
They catalyzes the stereochemical inversion of the configuration
about an asymmetric carbon atom in a substrate having more than
one center of asymmetry.
Example;

Racemerase.
Type of isomerase that catalyzes the conversion of one enantiomer
to another (one chiral center)
6. LIGASES
They catalyze the joining of two large molecules by forming a new
chemical bond.
Energy is required for the reaction (ATP, ADP)

1. C–O ligases (Amino acid-RNA ligase),

2. C–S ligases (Acetyl CoA synthetase, Acyl CoA synthetase),
3. C–N ligases (Asparagien synthetase, Glutation synthetase),
4. C–C ligases (Pyruvate carboxylase, Acetyl CoA carboxylase),
5. Phosphate-esther (DNA Ligase)
6. N–metal
Carboxylases
They add CO2 to molecule. Their cofactor is Biotine.

Exp.
Pyruvate carboxylase,
Asetyl-CoA carboxylase
1. Oxydoreductases 3. Hydrolases
• dehydrogenase • esterases
• oxydase 2. Transferases • glicosidase
• reductase • transaldolase • peptidase
• peroxidase • transketolase • phosphotase
• catalase • acyl, metil, glicosyl and • tiyolase
• oxygenase phosphotransferases • phospholyase
• hydroxylase • kinase • amidase
• phosphomutases • deaminase
• ribonuclease
4.Lyases
• decarboxylase
• aldolase 5. Isomerases
• racemase 6. Lyase
• hydratase • synthetase
• dehydratase • epimerase
• isomerase • carboxylase
• Synthase
• lyase • mutase