ENZYMOLOGY

THE STUDY OF ENZYMES

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 General consideration
The area of biochemistry that deals with study
of enzymes and various aspects of enzyme
catalyzed reactions is called Enzymology.
The various aspects include-
1.the structure of enzyme
2. the properties of enzyme
3.the classification of enzymes
4. the naming of enzymes
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 General consideration
6.The mechanism of action of enzymes(the
chemistry of catalysis)
7.Study of enzyme inhibitors
8.study of enzyme activators
9. specificity of enzyme
10 regulation/control of enzyme activity.
11. engineering and utilization of
enzymes(applied enzymology/enzyme
technology)
12.Clinicl enzymology etc.
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 General consideration
The study of enzymes will help to
understand How the metabolic pathways
are tacking place in the living system? &
How they are integrated and correlated?.

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 General consideration
Catalysts-are molecules/substances which
help to speed up the rate of reactions.
Two different types
1.Chemical catalyst speed up chemical
reactions(Reactants & products)
2.Biocatalyst-speed up biochemical reactions
and are present in the living system
(substrates and products).
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 General consideration
BIOCATALYST are of three different types.
1. Enzymes-proteins specialised for catalysis

2. Ribozymes- RNA molecules specialised for catalysis

&
3. Abzymes- antibodies specialised for catalysis.

Enzymes are very common and the major.

Ribozymes are less common and
Abzymes are very rare.
The study of all the biocatalyst are coming under
enzymology
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 Definition of enzymes
ENZYMES ARE PROTEINS SPECIALISED TO
CATALYSE BIOCHEMICAL REACTIONS TAKING
PLACE IN THE LIVING SYSTEM

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 General properties
All enzymes are
proteins in nature.
All enzymes possess an
active site-(the region
/cleft /pocket on the
enzyme where the
substrate bind and the
reaction takes place)

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 General properties
Enzyme combines Substrate
with the substrate Enzyme

to form enzyme
substrate complex Active Site
which soon
dissociates to form
enzyme and product
Substrate
Enzyme

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 General properties
Enzyme (E) + Substrate (S)

Enzyme substrate complex (E-S complex)

Enzyme (E) + product (P)

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 General properties
4. Enzymes speed up the reaction rate by
reducing the activation energy of
the reactants (substrate).
Activation energy of substrate is defined as
the minimum energy required by the
reactants to take part in a reaction-ground
state to transition state.
5. The enzyme remains unchanged at the end of
the reaction.

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 General properties
6. Some enzymes require certain non-enzyme /
non-protein factors for the catalysis to take
place. These non protein factors are generally
called as co-factors.
7. The cofactor inorganic molecule (metal ion )
organic molecule(co-enzyme).

i.e. organic cofactors are specifically called as

co-enzymes.

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 General properties
Protein part (apoenzyme)
(Inactive) + coenzyme

Holoenzyme
(Active)
Tightly bound cofactors
called a prosthetic group.

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 General properties
8. Enzymes are highly specific for their
substrate and the reaction catalyzed by them
9. Activity of the enzyme will be maximum at
optimum temperature.
10. Similarly, there is optimum pH at which
the activity will be maximum.

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 General properties
11. Enzyme activity is regulated in the living
body.
12. constitutive enzymes produced
continuously irrespective of the bodys need,
&
inducible enzymes - produced only when it
is required.

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 General properties
13. Extra cellular (Exoenzymes)
Intracellular types(Endoenzymes).
14.Intracellular enzymes soluble type (soluble enzymes)
or
membrane bound type (membrane bound
enzymes).
15. Certain enzymes are secreted in an inactive form
called pro-enzyme form or zymogens which are
then converted to active enzymes.
E.g. Trypsinogen (Trypsin), Pepsinogen (Pepsin).

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 General properties
Single enzyme( Individual enzyme)-only one
activity/one reaction.

Multienzyme complexes-assembly of several

enzymes and
catalyse multiple reactions/several reactions
Eg. Pyruvate dehydrogenase complex

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Naming and classification of enzymes
Early times, the lactase acts on lactose,
enzymes were named urease acts on urea
by adding the suffix amylase acts on starch
ase to the name of (amylose),
the substrate on which
it acts.-trivial naming of sucrase acts on sucrose
enzymes. etc.

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Naming and classification of enzymes
In 1964, Enzyme Commission of the
International Union of Biochemistry and
Molecular Biology (IUBMB) suggested a more
systematic method for naming the enzymes ,
known as IUBMB system of Enzyme
Nomenclature .

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Naming and classification of enzymes
IUBMB classify the enzymes in to six major
classes according to the types of reaction
catalyzed.
Each class is further divided in to subclass and
sub-subclass according to the specification of the
substrate, the reactive group coenzyme involved.
As per IUBMB system each enzyme is represented
by a 4 digit number which is known as E.C
number (E.C Enzyme commission).
It starts with E.C which followed by four digits
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Naming and classification of enzymes
First digit indicate the Ethyl alcohol+ NAD+
class.
Second digit indicate Alcohol dehydrogenase
the subclass.
Third digit indicate the
sub-subclass or Acetaldehyde+ NADH
subgroups. alcohol dehydrogenase
Fourth digit indicate the is represented as
number of particular E.C.1.1.1.1.
enzyme in the list. (IUMB name is alcohol:
NAD+ oxido-reductase)

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 Classification of enzymes
A-H2 + B

Class- I : Oxido-reductase oxidoreductase

catalyzes the oxidation of one

substrate with the A

simultaneous reduction of +
B-H2

another substrate or co-

enzyme. Lactate dehydrogenase

This type of reaction is generally Lactate + NAD+ Pyruvate + NADH+H+

called as oxidation reduction

reactions.
Oxidation / reduction take place
by the transfer of either
hydrogen or electrons.

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 Classification of enzymes
Class II: Transferases R +B A + B-R

catalyses the transfer of Glucose +ATP

Hexokinase
Glucose 6- P +ADP
atoms or groups other than
H-atom from one substrate
to another. Alanine + alpha keto-glutarate

-phosphotransferases( Alanine transaminase (ALT )

kinases),
-aminotransferases etc.

Pyruvate + Glutamate

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 Classification of enzymes
Class-III: Hydrolases e.g. Trypsin, pepsin ,
catalyzes the cleavage of amylase, lipase etc.)
ester, ether, peptide,
and glycosidic linkages Protein
in biomolecules by the trypsin/pepsin
addition of water
molecule. .

Amino acids

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 Classification of enzymes
Class- IV : Lyases pyruvate
catalyses the removal of a
group from a compound
by cleavage of a chemical
bond (by mechanisms
other than hydrolysis).
Pyruvate decarboxylase
These are non- hydrolytic
and non-oxidative
elimination reactions.

acetaldehyde + CO2

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 Classification of enzymes
Class -V: Isomerases
catalyzes isomerization
reactions i.e. the
introversion of optical, Glucose -6-P
geometrical and position
isomers of a substrate .
They are concerned with the
structural changes within a Phosphoglucoisomerase
single molecule.

Fructose-6-P

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 Classification of enzymes
Class -IV: Ligases (Ligare Small DNA fragments
(Latin word = to bind )

join or link two molecules

together to form a DNA ligase
bigger molecule which
is usually associated
with the simultaneous
hydrolysis of ATP.
larger DNA

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 Mechanism of Enzyme catalyzed reactions
In 1913, Lenor Michaelis and his associate Moud
Menten proposed the general mechanism of
enzyme action.
According to them the enzyme combines with
the substrate to form enzyme substrate complex
which soon dissociates to give rise to enzyme
and product.
The enzyme becomes free and active for further
catalysis.

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 Mechanism of Enzyme catalyzed
reactions
Active site-specific location or site on the enzyme
molecule where the substrate binds and the catalysis
takes place. The active site is in the form of a cleft on
the enzyme molecule.
Active residues-amino acid residues which are
actively involved in substrate binding and catalysis.
Allosteric site certain sites other than the usual
substrate binding site (active site) to which certain
substanes can bind and regulate the enzyme activity.
Allosteric enzymes.-Enzymes possessing allosteric sites

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 Mechanism of Enzyme catalyzed
reactions
Two different hypotheses explain the nature
of interaction between the enzyme and
substrate that takes place during an enzyme
catalyzed reaction.
1. Lock-key hypothesis
2. Induced fit hypothesis

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 Types of Enzyme catalyzed reactions

1.Mono substrate reactions

2. Bi substrate reaction

AND

1.Exergonic or Exothermic reaction

2.Isothermic reactions
3.Endergonic reactions

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 Factors affecting velocity of an
enzyme catalyzed reaction
1.Temperature
2.PH
3.Substrate concentration

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 ENZYME SPECIFICITY

One of the important characteristic features of enzyme catalyzed

reaction is the specificity shown by the enzyme.
Enzymes show varying degree of specificity towards the substrate and
the reactions catalyzed by t
1.Absolute specificity:.
E.g. The enzyme urease acts only on urea.

2.Group specificity:-
E.g. The enzyme Hexokinase can catalyze the phosphorylation of all hexoses.
( Glucose,Fructose ,mannose and Galactose ).

3.Bond specificity:
E.g. Trypsin hydrolyze peptide linkages involving Arginine and Lysine.

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 ENZYME SPECIFICITY
4.Reaction specificity: Protein kinases can
phosphorylate a number of proteins.
5. Stereo chemical specificity
D- amino acid oxidases act only on D- amino
acids whereas L- aminoacid oxidases acts only
on L- amino acids.

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 ENZYME INHIBITION

Any substance that stops or lowers the enzyme activity is called an

enzyme inhibitor and the phenomenon is called enzyme inhibition.
Enzyme inhibition can be of two different types irreversible
inhibition and reversible inhibition.
1. Irriversible inhibition -In irreversible inhibition the inhibitor binds
tightly to the enzyme protein at the active site to form an enzyme-
inhibitor complex.
The binding is achieved by strong covalent linkage and will not
dissociate easily.

E.g. Inhibition of acetyl choline esterase enzyme by di-

isopropyl fluorophosphate (DIFP).
Inhibition of cytochrome oxidase enzyme by cyanide.

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 ENZYME INHIBITION

2. Reversible inhibition -Here the inhibitor binds

with the enzyme to form a temporary enzyme
inhibitor complex and is reversible.
The inhibitor binds with the enzyme by non-
covalent forces.
Enzyme- inhibitor complex dissociate to re-form
the active enzyme. Based on the nature of
interaction between the enzyme, inhibitor and
the substrate, reversible inhibition is classified in
to three types.
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 ENZYME INHIBITION

a) Competitive inhibition-In competitive inhibition the

inhibitor competes with the substrate for entering the
active site of the enzyme. The inhibitors in this case are
compounds having structural similarity with the usual
substrate. So there is the formation of Enzyme
substrate complex and enzyme-inhibitor complex.
E+S E-S Complex
E+ I E I complex
E.g. Malonate (a structural analogue of the usual
substrate ,succinate) is the competitive inhibitor of
Succinate dehydrogenase enzyme of citric acid cycle.

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 ENZYME INHIBITION

b) Uncompetitive inhibition -In uncompetitive inhibition the inhibitor

binds only to the enzyme substrate complex and not to the free
enzyme, forming Enzyme substrate inhibitor complex.
E+ S E S complex
E S -Complex + I E S- I- Complex

c) Non -competitive inhibition-In uncompetitive inhibition, the

inhibitor binds to both free enzyme and enzyme substrate complex
to form enzyme-inhibitor complex and enzyme- substrate -
inhibitor complexes.
E+ I E I -Complex
E+ S E S -Complex
E S -Complex + I E S- I- Complex

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 ENZYME REGULATION
In the body systems, the activity of all enzymes is regulated by one
or another mechanism. There are different mechanisms.
1.Covalent modification
The covalent binding of certain atoms or groups to the enzyme
molecule lead to either their activation or inactivation. This type of
regulation achieved by the chemical modification of the enzyme
protein is called covalent modification.
Phosphorylation is an example for covalent modification.
Phosphorylation activates enzymes such as glycogen
phosphorylase, phosphorylase b kinase citrate lyase etc. Whereas
phosphorylation inactivates enzymes like glycogen synthase

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 ENZYME REGULATION
2.Allosteric modulation -Some enzymes posses a second site called
allosteric site to which certain substances can bind and modulate
the enzyme activity. Such substances are called allosteric
modulators and the phenomenon of regulation is called allostreic
modulation.
When the modulator increases the activity, such modulators are
called allosteric activators or positive allosteric modulators and
the phenomenon is called allosteric activation.
When the modulator decreases the enzyme activity, such
modulators are called allosteric inhibitors or negative allosteric
modulators and the phenomenon is called allosteric inhibition.

Examples: ATP and citrate are the allosteric inhibitors for

phosphofructokinase enzyme
Whereas AMP is an allosteric activator.

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 ENZYME REGULATION
3.Feed back inhibition - Feed- back inhibition is
also known as end product inhibition. This type
of regulation is seen in the case of long
metabolic pathways where the end product
of the pathway acts as a specific inhibitor of one
of the enzyme early in the pathway.
Example: During the biosynthesis of the amino
acid L- Isoleucine from L- Threonine, the end
product Isoleucine acts as an inhibitor for
Threonine deaminase the first enzyme in the
pathway.

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 ENZYME REGULATION
4. Activation by structural changes-Zymogens are
the inactive forms of certain enzymes. Zymogens
are converted to active enzyme by structural
modification
E.g. Pepsinogen (zymogen)

HCL (hydrolysis)

pepsin (active enzyme)

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CLINICAL ENZYMOLOGY

Enzymes in disease diagnosis

( marker enzymes)
Enzymes in therapy( therapeutic
enzymes)
Enzymes as drug targets
Diagnostic enzymes( enzymes as
diagnostic reagents)

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 GENERAL ASPECTS
Clinical enzymology is a specific area of Medical Biochemistry which
deals with the application of enzymology in clinical practice includes
disease diagnosis, disease treatment, disease management ,patient
evaluation etc.
There are certain enzymes termed marker enzymes whose
level will be indicative of a particular disease state or
conditions.
There are certain enzymes called therapeutic enzymes
which are very useful as drugs in the treatment of several
diseases.
There are certain enzymes which are target of certain
drugs used in the treatment of certain diseases.
There are certain enzymes diagnostic enzymes which are
very useful in diagnostic techniques as a reagent tool .

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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Classification of plasma enzymes based on their
source and the site of action:
Plasma contain a wide verity of enzymes which can
be categorized in to two classes based on the source
and the location of its action.
1. Plasma derived enzymes &
2. Cell derived enzymes

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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Plasma derived enzymes are enzymes which acts
on the substrate present in the plasma and its
activity is higher in plasma than in the cell.
Eg. Enzymes involved in blood coagulation
(coagulation factors) and Ceruloplasmin
( ferroxidase activity).
Cell derived enzymes are having higher activity
in the cells than in the plasma and are overflow
in to the plasma from tissues and organs.

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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Cell derived enzymes can again be classified
in to two types based on their specific
function
a) Secretory:- Mainly derived from secretory
glands and function in the extracellular
space.
b) Metabolic :- They are concerned with the
cellular metabolism and functions within the
cells.

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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Classification of plasma enzymes based on their function in the
plasma:
1)Functional &
2)Non- functional enzymes
Functional enzymes: These are having very specific / highly essential
well defined functions in the blood plasma.
They are present in higher concentration in the plasma than in
the cells.
They are mainly synthesised by the liver and then released in to
the plasma.
They are clinically very significant when the serum level is reduced
below the normal range.
Eg. Pseudocholine esterase, Lipoprotein lipase ceruloplasmin and
blood coagulation factors /enzymes.

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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Non functional enzymes: have no specific /
essential functions in the plasma.
They are derived from cells of various organs /
tissues and released in to the plasma in minute
quantities.
Their activity will be higher in cells than in the
plasma.
They are present in the plasma in very small
concentration under normal conditions.
They become clinically significant when the plasma
level is increased above the normal range.
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 ENZYMES IN BLOOD PLASMAAS MARKERS
(indicators) OF CERTAIN DISEASES
Possible causes / reasons for the increase in the levels of
non functional enzymes in the plasma.
1. Hypoxic / infective insults
2.Disease states of the tissue
3.Excessive synthesis or induction of the enzyme followed by
overflow in to the plasma.
4. Vigorous exercise.
5. Decreased renal clearance
Therefore, these enzymes(functional and non functional)
can serve as indicators(markers) of the functional status of
an organ or a tissue that produce it and they are called
marker enzymes.

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ISOZYMES- AS MARKER ENZYMES
Iso-enzymes or Isozymes are multiple forms of the
same enzyme that catalyze the same chemical
reaction.
They differ in their physico- chemical properties such
as,
Electro-phoretic mobility.
Kinetic properties(opt. temp/opt.PH, Km value).
Amino acid sequence.
Amino acid composition.
There are a number of examples for Isozymes which
are useful in disease diagnosis as marker enzymes.

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ISOZYMES- AS MARKER ENZYMES
1. Lactate dehydrogenase(LDH)
Lactate dehydrogenase(LDH) is an enzyme
present in our body systems, distributed in
different tissues.
LDH catalyzes the reversible conversion of
pyruvate in to lactate (Anaerobic glycolysis).
LDH is a tetrameric enzyme consisting of four
subunits.
The subunits are of two different types-
M( muscle) type and H( heart) type.

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ISOZYMES- AS MARKER ENZYMES
There are 5 different isomeric forms of LDH-
LDH1, LDH 2 , LDH 3 , LDH 4 and LDH 5.
They differ in their subunit composition.
LDH isozymes can be separated by cellulose
acetate electrophoresis at PH 8.6.
The serum normal value: 100- 200 U/L
CSF normal value: 7- 30 U /L

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 ISOZYMES- AS MARKER ENZYMES
Clinical significance of LDH
LDH 2 is the predominant isozyme in plasma and
LDH 5 is present in least concentration.
In the plasma:
During Myocardial infarction( MI) LDH 1 level will be
greater than LDH 2.
In Megaloblastic anaemia almost LDH 1 and LDH 2
will be 50 times of normal.
In muscular dystrophy LDH 5 level will be elevated.
In toxic hepatitis and jaundice LDH 5 will be almost 10
times high.

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ISOZYMES- AS MARKER ENZYMES
In renal diseases-Tubular necrosis,
pyelonephritis, and in pulmonary embolism LDH
3 level will be increased.
Total LDH value will be increased in neoplastic
diseases.
LDH 5 will be elevated in breast cancer, CNS
malignancy and prostatic carcinoma.
LDH 2 and LDH 3 will be increased in leukemia.
In malignant tumours of testes and ovaries LDH2
, LDH 3 and LDH 5 levels will be elevated.

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ISOZYMES- AS MARKER ENZYMES
In Cerebro-spinal fluid( CSF):
In Bacterial meningitis LDH 4 and LDH 5
levels will be elevated.
In viral meningitis LDH 1 level will be higher.
In metastatic tumours LDH 5 level will be
higher.

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ISOZYMES- AS MARKER ENZYMES
2.Creatine kinase
is a marker isozyme that catalyses the
conversion of creatine to creatine Phosphate.
CPK has three isozyme forms.
CPK is a 40 KD molecular weight enzyme.
There are two subunits( dimeric protein).
There are two different types of subunits
present in CPK B ( brain ) type and M( muscle )
type.

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ISOZYMES- AS MARKER ENZYMES
Depending on the combination of different
subunits there are 3 isozyme forms
CPK 1( CPK BB), CPK 2 ( CPK MB) and CPK 3 (
CPK MM).
CPK is an important enzyme in energy
metabolism.
CPK is important for the formation of
creatine P the immediate source of energy
for muscle contraction.

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ISOZYMES- AS MARKER ENZYMES
Clinical significance:
CPK and Myocardial infarction (MI) or heart
attack:
CPK 2 is very negligible in the normal plasma(
2% of the total CPK activity) and is usually
undetectable in the plasma.
During MI ,CPK 2 activity is increased within 4
hours and then falls rapidly.
Total CPK activity is elevated up to 20 times in
MI.

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ISOZYMES- AS MARKER ENZYMES
CPK in muscle diseases and brain damage:
CPK-3 activity is specifically elevated in
muscular dystrophies (500 1500 U/L).
CPK-1 level is highly elevated in crush injury
fracture and in acute cerebro-vascular
accidents.
Usually, estimation of total CPK is usually
employed in muscular dystrophy CPK-1 in
Brain diseases and specifically CPK MB ( CPK
2) isozyme is employed during MI.
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ISOZYMES- AS MARKER ENZYMES
Atypical forms of CPK: There are two unusual
forms of CPK.
1. Macro CPK and 2. CPK Mi
Macro CK is formed by the association of CPK
MB with Ig G or Ig A.
Also formed by the association of CPK MB with
lipoproteins.
Electrophoretically migrated between CK MB
and CK MM
Usually occurs in women above 50 years.

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 CPK Mi ( mitochondrial CK isozyme):
It is found attached to the outer surface of the
inner mitochondrial membrane of muscle liver
and brain.
In electrophoretogram it is present behind CK
MM band.
It is not present in normal plasma.
Clinical significance of CK Mi:
It occurs in the plasma when there is extensive
tissue damage that cause the breakdown of
mitochondrial membrane.
Its presence in serum also indicates cellular
damage in malignancies.
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 ISOZYMES- AS MARKER ENZYMES
3.Alkaline phosphatase ( ALP):
Alkaline phosphatase is a nonspecific enzyme it hydrolyzes
aliphatic, aromatic and heterocyclic compounds.
Produced mainly by osteoblasts of bone and is associated with bone
calcification process.
Little amounts of ALP is also produced by epithelial cells of the biliary
canaliculi( there are also many other sources-liver ,intestine and
WBC).
It is localised in the cell membrane- ectoenzyme.
It is associated with the transport of phosphates in liver kidney and
intestinal mucosa.
Normal range in the plasma is 40- 125 U/L( in children increased
levels are observed because of increased osteoblast activity.

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 ISOZYMES- AS MARKER ENZYMES
Moderate(2-3 times) increase in ALP activity is seen in
hepatic diseases such as infective hepatitis, alcoholic
hepatitis and in hepato-cellular carcinoma.
Very high activity( 10-12 times) is observed in extra-hepatic
obstruction( obstructive jaundice) caused by gall stones or
by pressure on bile duct by carcinoma of the head of the
pancreas.
ALP is also produced by epithelial cells of the biliary
canaliculi, therefore obstruction of the bile duct with
consequent irritation of epithelial cells will lead to secretion
of ALP in to the plasma.
Drastically high levels( 10-25 times) is seen in bone diseases ,
rickets , osteomalacia, osteoblastoma, metastatic carcinoma
of bone and also in hyper parathyroidism.

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 ISOZYMES- AS MARKER ENZYMES
Isozymes of ALP.
Alpha -1 ALP( epithelial cells of biliary canaliculi): 10 %
of the total increased in bile duct obstruction (
obstructive jaundice)
Alpha-2 ALP- heat labile ( produced by hepatic cells: 25
% of total ALP- increased in liver diseases.
Alpha-2 ALP- heat stable( produced by placenta) : 1% -
observed in the blood only during pregnancy.
Pre -beta ALP (produced by osteoblasts) 50 % of the
total ALP- elevated in bone diseases.
Gamma ALP( originates from intestinal cells): 10 % of
total activity- increased in ulcerative colitis.
Lekocyte ALP (LAP)- decreased in chronic myeloid
leukemia and increased in lymphomas.
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 MARKERS OF CARDIAC DISEASE
1.Cardiac troponin
They are not enzymes.
Now accepted as a reliable marker of MI.
Significant in the early detection of ischemia and
in monitoring the patient.
2.Brain Natriuretic Peptides ( BNP)
Produced by the ventricles of heart in response
to the excessive stretching of the heart muscle.
Reliable marker of ventricular dysfunction.
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 MARKERS OF CARDIAC DISEASE
3.Aspartate amino transferase (AST)
Also called Glutamate oxaloacetate transaminase ( SGOT).
It is a marker of liver as well as heart diseases.
4.Gamma glutamyl transferase (GGT)
It is used in the synthesis of glutathione , a reductant in liver and RBC.
It is seen in liver kidney , pancreas intestinal cells and prostate gland.
Normal plasma level is 10- 30 U /L.
It is moderately elevated in infective hepatitis and in liver cancer.
GGT is a very sensitive marker of alcohol abuse ( increased
level is observed in the plasma of alcoholics).
The activity level is usually proportional to the amount of alcohol
consumed.

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 MARKERS OF CARDIAC DISEASE
5.Nucleotide phosphatase ( NTP):
It is also known as 5nucleotidase that convert
nucleotide to nucleoside.
It is a marker enzyme for plasma membrane(
membrane damage).
It is an ecto-enzyme.
Normal level in plasma : 2- 10 U/L.
Moderately increase in hepatitis and highly
elevated in billiary obstruction.
Unaffected by bone diseases.

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 MARKERS OF OTHER
ORGANS/TISSUES
1.Alanine aminotransferase ( ALT)
It is also called serum Glutamate pyruvate transaminase (
SGPT).
Serum level is : 13- 35 U/L.( female ( 10- 30 U/L).
Very high values ( 300- 1000 U/L) are seen in acute
hepatitis( toxic or viral).
Both AST and ALT levels are increased in liver disease.
Rise in ALT will be noticed even several days before the
development of the signs of jaundice.
Moderate increase( 50- 100 U/L) is seen in chronic liver
diseases such as cihrrosis, hepatitis C and non alcoholic
steatohepatitis( NA SH).

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 MARKERS OF OTHER ORGANS/TISSUES
2.Acid phosphatase(ACP).
Hydrolyses phosphoric acid esters.
Secreted by prostate cells , RBC ,platelet and WBC.
ACP level is increased in prostate cancer and also in bone
metastasis of prostate cancer.
3.Prostate specific antigen( PSA):
It is produced by the secretory epithelium of the prostate
gland .
It is a serine protease enzyme.
Normally secreted in to the seminal fluid and is essential for
the liquification of the semen coagulum.
Normal level in the serum: 1- 5 micro gm / L.
Values above 10 micro gram/L indicates prostate cancer. 90
 MARKERS OF OTHER ORGANS/TISSUES
4.Aldolase :
It is a glycolytic enzyme.
Normal level in the plasma is 1.5- 7 U/L.
It is drastically elevated in the serum during muscle
damage progressive muscular distrophy, polyomyelitis,
myesthemia gravis (muscle wasting disease) and in
multiple sclerosis.
It is a very sensitive early marker for muscle wasting
diseases.
5.Enolase:
It is also a glycolytic enzyme .
Neuron specific enolase ( NSE) is an isoenzyme in nervous
tissue and is a marker for cancer associated with
neuroendocrine origin, neuroblastoma etc.

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 MARKERS OF OTHER ORGANS/TISSUES
6.Beta glucuronidase:
It is abundant in liver,spleen, endometrium
breast and adrenals.
Leukocyte contain very high amounts.
Increased levels are seen in cancer of urinary
bladder.
Very high levels are seen in carcinoma of the
head of the pancreas and in 50 % of the
breast cancer.

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 MARKERS OF OTHER ORGANS/TISSUES
7.Amylase :
Starch and glycogen digesting enzyme produced by pancreas
and salivary glands.
Normal serum value is : 50- 100 I U /L.
1000 times increase is noticed in acute pancreatitis.
Moderate increase is seen chronic pancreatitis and in
obstruction of the pancreatic duct.
8.Lipase :
Lipase is produced mainly by pancreas .
It hydrolyzed dietary TAG ( neutral fat) in to MAG and free
fatty acids.
Moderately elevated in the serum in pancreatic carcinoma,
biliary disease and perforating peptic ulcers.
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 MARKERS OF OTHER ORGANS/TISSUES
9.Choline esterase( Ch E) :
Present in nerve endings and in RBC s. Newly formed RBC will have high
levels of the enzyme and decrease according to the age of the RBC.
Organo-phosphorus pesticides irreversibly inhibit choline esterase in the
RBC. Measurement of Ch E is useful in determining the magnitude of
exposure to pesticides.
10.Glucose 6- P dehydrogenase ( GPDH):
Enzyme of HMP shunt pathway.
Involved in the production of NADPH ( a co- enzyme for many
antioxidant enzymes and also useful in many of the biosynthetic
pathways ) .
Normal value in RBC : 6- 12 U / gm Hb.
Activity will be decreased in drug induced hemolytic anaemia.
RBC life span will be decreased in GPDH deficient individuals.

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Enzymes useful as diagnostic reagents
Urease Urea estimation.
Uricase Uric acid estimation.
Glucose oxidase Glucose estimation.
Hexokinase Glucose estimation.
Lipase TAG estimation.
ALP- ELISA.
Restriction Endonuclease RFLP(restriction
fragment length polymorphism).
Reverse transcriptase PCR( polymerase chain
reaction).

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 Enzymes as drug targets
1.Cyclo-oxigenase aspirin(analgesic: pain and
inflammation).
2.HMG COA reductaselovastatin( hypocholestremic
drug).
3.Reverse transcriptase anti retroviral drugs.
4.Xanthine oxidase allopurinol ( hypourecemic
drug).

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THANK YOU
FOR YOUR ATTENTION....

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