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Q: Essays:
1. Classify enzymes. Describe the different types of enzyme inhibition. Add a note on
clinical significance of enzymes. April 99
2. Define enzyme. Write about factors regulating enzymatic activity. Name four
clinically important enzymes. Mention their importance. March 2002
3. What is enzyme inhibition? Describe different kinds of enzyme inhibition with
examples. Feb 2005
4. What are isoenzymes? Write about different isoenzymes and their clinical
importance. Feb 2007
Short notes:
1. Classify enzymes with one example for each class April 2000
2. Types of enzyme inhibition- Oct 99
3. Isoenzymes; Allosteric enzymes-April 2000; Oct 2000
4. Effect of substrate concentration on:
1. Enzyme catalysed reaction and
2. Michelis Menten reaction Oct 2000
3. kM value

Enzymes are protein catalysts synthesized by living cells that increase the
rate of conversion of one or more compounds (substrates) into one or more
different compounds (products) while the enzymes themselves are neither
consumed nor permanently altered as a consequence of their participation in
a reaction.
Properties of Enzymes:
1. Almost all enzymes are proteins. Enzymes follow the physical and chemical
reactions of proteins.
2. They are heat labile. They are water-soluble. They can be precipitated by
protein precipitating reagents (ammonium sulfate or trichloroacetic acid).
3. The enzymes enhance the rates of the reaction by factors of at least 10 6.
4. Enzyme molecules contain a special pocket or cleft called the active site. The
active site contains amino acid chains that create a three-dimensional surface
complementary to the substrate. For the combination with substrate, each
enzyme is said to possess one or more active sites where the substrate can
be taken up.
5. Like all catalysts, enzymes are neither consumed nor permanently altered as
a consequence of their participation in a reaction.
SPECIFICITY OF ENZYMES: Enzymes are specific for their substrate. According
to specificity of enzymes, they are divided into:
1. Absolute specificity: Some enzymes are absolutely specific. E.g. Urea is
the only substrate for urease. Glucose oxidase will oxidise only beta-d
2. Bond specificity: some enzymes show group or bond specificity. Enzymes
that are specific for a bond or linkage such as ester, peptide or glycosidic
belong to this group. Examples:
i. Esterases- acts on ester bonds
ii. Peptidases-acts on peptide bonds
iii. Glycosidases- acts on glycosidic bonds.
iv. Trypsin hydrolyses pepsin bonds of carboxyl group of arginine
and lysine residues only.

3. Group specificity: some enzymes can catalyse reaction of the structurally
similar groups. E.g. Hexokinase can catalyse phosphorylation of glucose,
galactose, and mannose.
4. Stereo specificity: Enzymes are specific for l-amino acids and Dcarbohydrates. E.g.
a. Fumarase can hydrate trans form of fumaric acid and not the cis form.
b. Lactate dehydrogenase acts on pyruvate to form only L-lactate and
not D-lactate.
6. Many enzymes contain small nonprotein molecules and metal ions that
participate directly in substrate binding or catalysis. They are termed
prosthetic groups, cofactors, and coenzymes.
Simple enzyme: enzyme contain only protein
Holo enzyme: protein groups (apoenzyme) + non-protein component
(co factor)
Organic cofactor is called co enzyme; non organic cofactor is called
If cofactor is bound so tightly to the apoenzyme and is difficult to
remove without damaging the enzyme, it is called a prosthetic group.
7. Few enzymes exist in their inactive forms and are called as proenzymes or
zymogens, e.g. pepsin has pepsinogen as its zymogen.
These are enzymes having similar catalytic activity, act on the same
substrate and produces the same product but originated at different
site and exhibiting different physical and chemical characteristics
such as electrophoretic mobilities, amino acid composition and
immunological behavior.
Example: LDH (Lactate dehydrogenase) exists in five different forms
each having four polypeptide chains. H= Heart and M=Muscle.
CLASSIFICATION OF ENZYMES: Enzymes are classified by reaction type; the
International Union of Biochemists (IUB) developed a system of enzyme
nomenclature in which each enzyme has a unique name and code number that
identify the type of reaction catalyzed and the substrates involved. Enzymes are
grouped into the following six classes. The word OTHLIL is used to remember the
six classes.
Class 1: oxidoreductase: Enzymes catalyzing oxidation reduction reactions.
Example: Lactate-dehydrogenase
1- Lactic acid + NAD+ -------- Pyruvic acid + NADH+H+
Class 2: Transferases: Enzymes catalyzing a transfer of a group other than
hydrogen (methyl, acyl, amino or phosphate groups Eg. Hexokinase
E.g. Hexose + ATP --------------------------- Hexose 6 phosphate +
Class.3: Hydrolases: hydrolyse ester, ether, peptide and glycosidic bonds by
adding H2O and breaking the bond. Eg. Acetyl choline esterase
E.g. Acetyl choline + H2O --------------------- Choline + acetate

Class.4: Lyases: remove groups or break bonds by means other than
E.g. Fructose 1,6 biphosphate ----------
Glyceraldehyde-3Phosphate +
dihydroxy acetone
Class.5: Isomerases: produce isomerisation leading to optical, geometric or
positional isomers. E.g. Racemase; epimerase; cis-trans isomerase
E.g. Glyceraldehyde-3-Phosphate ---------------- di-hydroxyacetone-phosphate
Triose phosphate isomerase
Class.6: Ligases: link two substrates (ligate) together, usually with hydrolysis
of ATP.
Example 1. Acetyl CoA + CO2 + ATP ----------------------- Malonyl
CoA + ADP + Pi
Acetyl CoA carboxylase
Example2: Glutamine Synthetase
ATP + L-Glutamate + NH4 - ADP + orthophosphate
+ L-Glutamine
Note: Synthetase belongs to this group and is ATP dependant, while
synthase belongs to different group and ATP non dependant E.g.
Glycogen synthase.
1. Prosthetic groups are distinguished by their tight, stable incorporation into a
proteins structure by covalent or noncovalent forces. Examples include
pyridoxal phosphate, flavin mononucleotide (FMN), flavin dinucleotide (FAD),
thiamin pyrophosphate, biotin, and the metal ions of Co, Cu, Mg, Mn, Se, and
2. Metals are the most common prosthetic groups. The roughly one-third of all
enzymes that contain tightly bound metal ions are termed metalloenzymes.
3. Metal ions promote enzyme action by:
Maintaining or producing the active structural conformation of the
enzyme (e.g. glutamine synthase)
Promoting the formation of the enzyme-substrate complex (Example:
Enolase and carboxypeptidase A.)
Acting as electron donors or acceptors (Example: Fe-S proteins and
Causing distortions in the substrate or the enzyme Example:
4. Metal Enzyme:
Zinc: Carbonic anhydrase, carboxy peptidase, alcohol dehydrogenase
Magnesium: Hexokinase, phospho fructo kinase, enolase, glucose-6phosphatase
Manganese: Phospho gluco mutase, hexokinase, enolase, glycosyl
Copper: Tyrosinase, cytochrome oxidase, lysyl oxidase, superoxide

Iron: Cytochrome oxidase, catalase, peroxidase, xanthine oxidase
Calcium: Lecithinase, lipase
Molybdenum: Xanthine oxidase
1. Cofactors serve functions similar to those of prosthetic groups but bind in a
transient, dissociable manner either to the enzyme or to a substrate such as
2. Unlike the stably associated prosthetic groups, cofactors therefore must be
present in the medium surrounding the enzyme for catalysis to occur.
3. The most common cofactors also are metal ions. Enzymes that require a
metal ion cofactor are termed metal-activated enzymes to distinguish them
from the metalloenzymes for which metal ions serve as prosthetic groups.
1. Coenzymes are mostly derivatives of vitamins without which the enzyme
cannot exhibit any reaction. One molecule of coenzyme is able to convert a
large number of substrate molecules with the help of enzyme
2. Coenzymes serve to transport many substrates from their point of generation
to their point of utilization.
3. Association with the coenzyme also stabilizes substrates such as hydrogen
atoms or hydride ions that are unstable in the aqueous environment of the
4. Other chemical moieties transported by coenzymes include methyl groups
(folates), acyl groups (coenzyme A), and oligosaccharides (dolichol).
5. The water-soluble B vitamins supply important components of numerous
coenzymes. Many coenzymes contain the adenine, ribose, and phosphoryl
moieties of AMP or ADP
6. The coenzyme may participate in forming an intermediate enzyme-substrate
7. Most coenzymes are substances. NAD and ATP are important
coenzymes so also Coenzyme A and FAD.
8. Two group of coenzymes:
1. In Group1: Counterbalancing change occurs in coenzyme along
with changes in substrate.
E.g. Lactate is converted into Pyruvate by lactate
dehydrogenase and the coenzyme Nicotinamide adenine
dinucleotide (NAD) accepts H+
Lactate dehydrogenase
Lactate------------------------------ pyruvate
2. In Group 2: the coenzymes transfer groups other than H+. E.g.
Glucose -------------------- glucose-6-phosphate
1. Lowering of activation energy:

Enzymes lower the energy of activation. Activation is defined as the energy
required to convert all molecules of a reacting substances from the ground state
to the transition state when it will be ready to react.
2. Active site or active centre of enzyme:
Active site refers to the region of the enzyme where substrate binds and
catalysis occurs. During binding, realignment of groups facilitates catalysis. E.g.
Serine in pancreatic protease;
3. Mentens Enzyme-substrate complex
1. Michaelis-Mentens Enzyme-substrate complex theory:
The enzyme combines with substrate to form E-S complex which facilitates
breakdown of substrate into products . E.g. In the following reaction serine
residue is the active site of Alkaline Phosphatase where the E-S complex is
E-serine-OH + Glucose-6-Phosphate E-serine-O-Phosphate
+ glucose
E-serine-O-Phosphate (E-S complex) E-serine-OH + Pi


a. Fishers Template theory: The active site by itself provides a rigid,
pre-shaped template fitting with the size and shape of the substrate
molecule. Substrate fits into active site of an enzyme as the key fits
into the lock and hence it is called the lock-and-key model.
b. Koshlands induced fit theory:
When the substrate binds to the enzyme, conformational changes occur in
the enzyme that facilitates enzyme substrate formation at the active site
leading to facilitated reactions.


1. Exothermic reaction: energy is released during reaction; reaction is
E.g. Urea ammonia+CO2+enegy
2. Isothermic: energy exchange is negligible and the reaction is reversible.
E.g. Glycogen + pi Glucose-1-Phosphate
3. Endothermic: needs energy for the reaction;
E.g. Glucose + ATP Glucose-6-Phosphate + ADP
Enzyme kinetics:
1. The velocity of enzyme reaction is proportional to the concentration of
reacting molecules
2. The reaction id in dynamic state
3. The equilibrium constant (Keq): is the ratio of forward and backward reaction
rate constants
4. At equilibrium forward and backward reaction are equal
5. Enzymes increase the rate of reaction but do not alter the equilibrium
Factors influencing enzyme activity:
1. Enzyme concentration: rate of reaction is directly proportional to enzyme
concentration when sufficient substrate is present. This property is used to
measure enzyme levels for diagnostic purpose.
V= velocity; E= enzyme concentration

2. Substrate concentration:
Initially substrate concentration increases the velocity of reaction but after a
saturation point the velocity remains the same. Vmax refers to the maximum


(Reaction rates increase as substrate concentration increase, but become

saturated at very high concentrations of substrate.)
If velocity is plotted against substrate concentration a typical curve will be

Michaelis constant : Km value: (short notes)

1. Michaelis Theory: It postulated that enzyme (catalyst) and substrate
(reactant) are in fast equilibrium with their complex, which then dissociates to
yield product and free enzyme.
2. The enzyme + substrate reaction is reversible; but the breakdown of the
complex to enzyme+products is irreversible.
E.g. Enzyme + substrate Enzyme-substrate Enzyme + products
3. Definition of Michaelis constant (Km): it denotes 50% of enzymes are bound
to substrate at half-maximal velocity i.e.,1/2 Vmax and the value is known as
Michaelis constant (Km). Irrespective of enzyme concentration, 50% of
enzymes are bound to substrate at that concentration.

4. The Michaelis equation can be used to demonstrate that at the substrate

concentration that produces exactly half of the maximum reaction rate,
i.e.1/2 Vmax, the substrate concentration is numerically equal to Km.

5. This fact provides a simple yet powerful bioanalytical tool that has been used
to characterize both normal and altered enzymes, such as those that produce
the symptoms of genetic diseases.
6. Km is a constant for the enzyme
7. Km denotes the affinity of the enzyme for the substrate. The lesser the value
of Km the more is the affinity for substrates.
8. Michaelis menten equation: v= velocity; s=substrate conc.; Km= Michaelis
V = Vmax (S)
Lineweaver-Burk plot or Double reciprocal plot: When the substrate
concentration is very low it will be difficult to determine vmax. To overcome
this, the experimental data of lower concentration is plotted as reciprocals.
The straight line graph obtained is extrapolated to get the reciprocal of Km.
From 1/Km , Km can be arrived. This is called Lineweaver-Burk plot or Double
reciprocal plot which uses the Michaelis - Menten equation.

1/v max



Co-operative binding: when some enzymes have many subunits, the

binding of substrate on one unit may enhance the affinity for binding on other
subunits. This is called Co-operative binding. The saturation curve is sigmoid
shaped and Km is determined using Hill equation instead of the Michaelis Menten equation.

9. Effect of concentration of products:

As the concentration of products increases, the reaction is slowed and may
even stopped further. This is seen in inborn errors of metabolism in which if
one enzyme is blocked the products prior to block increase and in turn the
whole pathway is blocked.

A------------B ------------ C------//----------D
In the above equation when E3 is deficient C increases and BC reaction is
slowed or blocked.
10.Effect of temperature:
Enzyme reaction is best at an optimum temperature. Hence will temperature
give a bell shaped curve i.e. as the temp increases the reaction reaches a
maximum and then falls. This is because more heat leads to denaturation of
enzyme proteins. The optimum temp is 37c.

11.Effect of pH:
Enzyme reaction is best at an optimum pH. Hence will pH also give a bell
shaped curve the pH increases the reaction reaches a maximum and
then falls. The optimum pH is 6-8.

12.Enzyme activation:
1. By inorganic ions: chloride activates salivary amylase and calcium
activates lipase.
2. By conversion from proenzyme to active enzyme:
E.g. Trypsinogen to trypsin, the active form and trypsin activates
chymotrypsinogen to chymotripsin. Pro enzyme forms are necessitated
to prevent autolysis by cellular proteins. Series of activation of
enzymes that culminates in one end product is called cascade system.
E.g. Coagulation; complement system etc
Enzyme inhibition:
1. competitive inhibition
1. This is done by certain inhibitor molecules which occupy the active site on
the enzyme and prevent the formation enzyme-substrate complex. They
are structurally similar to substrates. This is called competitive inhibition.
Excess substrates will abolish inhibition and is reversible. In competitive
inhibition Km is increased but Vmax is unaltered. E.g. Malonate inhibits
succinate dehydrogenase.






2. Clinical significance of competitive inhibition:

1. Sulphonamides inhibit folic acid synthesis which is responsible for
its antibacterial activity.
2. Methotrexate inhibits folate reductase and hence prevents cell
multiplication. So it is used as anticancer drug.
3. Dicoumarol inhibits Vit.K and used as an anticoagulant.
4. Due to structural similarity INH can induce pyridoxal deficiency in
chemotherapy of TB.
Non competitive inhibition:
1. Irreversible
2. No structural similarity with substrates and occupy different site on
3. Excess substrates will not abolish inhibition
4. E.g. Cyanide inhibits cytochrome oxidase
5. BAL acts as antidote for mercury poisoning using non competitive
Uncompetitive inhibition:
1. Inhibitor binds to enzyme substrate complex and not to free
enzyme. E.g. Placental alkaline Phosphatase inhibited by phenyl
Suicide inhibition:
1. Special type; irreversible
2. Also called Mechanism based inactivation
3. The inhibitor gets converted into more effective inhibitor with the help of
enzyme itself
4. E.g. Allopurinol gets converted into alloxanthine by xanthine oxidase and
alloxanthine becomes strong inhibitor of xanthine oxidase
5. Aspirin inhibits prostaglandin synthesis by similar mechanism in
controlling inflammation.
Allosteric regulation (short notes)
1. Allosteric enzymes are enzymes that change their conformation upon
binding of an effector.
2. An allosteric enzyme is an oligomer whose biological activity is affected
by altering the conformation(s) of its quaternary structure.
3. Allosteric enzymes tend to have several subunits.

4. Whereas enzymes with single active sites display normal Michaelis-Menten
kinetics, allosteric enzymes have multiple active sites and show
cooperative binding.
5. As a result, allosteric enzymes display a sigmoidal dependence on the
concentration of their substrates, allowing them to greatly vary catalytic
output in response to small changes in effector concentration.
6. Effector molecules, which may be the substrate itself (homotropic
effectors) or some other small molecule (heterotropic effector), may cause
the enzyme to become more active or less active.

The binding sites for heterotropic effectors, called allosteric sites, are
separate from the active site.

8. An enzyme may have two active sites:

i. catalytic site where substrate binds;
ii. Allostreic site where a regulatory molecule binds.
9. Regulatory molecule may enhance (Allosteric activation) or inhibit
(Allosteric inhibition) the activity of the enzyme.
10.Salient Features, Allosteric Inhibition:
1. Inhibitor enzyme is not substrate analogue
2. It is partially reversible
3. Km decreased; Vmax increased
4. Allosteric inhibitor modifies catalytic site to make it
5. Body uses Allosteric enzymes to regulate metabolic pathways
and they are called rate limiting or key enzymes
6. Allostreic regulatory enzymes are more effective with low
substrate concentration hence a biological advantage
11.Examples of Allosteric enzymes:
1. Sucinyl coA+ glycine ---------------------- deltaamino-levulinic acid (ALA) in heme synthesis
ALA synthase
2. HMGco A reductase in cholesterol synthesis
6. Feedback inhibition:
The activity of the enzyme is inhibited by the final product.
Threonine E1 E2 E3 E4 E5 Isoleucine
Isoleucine acts as a negative Allosteric regulator, turning off E1 when
there is enough isoleucine
7. Induction:
This means substances that increase synthesis of enzyme proteins by
removing the block or repression. E.g. Insulin induces glucokinase;
glucocoticoids induce tryptophan pyrrolase and transaminase. Induction is
switching on the gene to produce more enzymes.
8. Repression:

Repression is similar to inhibition but by different mechanism at gene
level. After a lag period the repressor molecule reduces the no of enzyme
molecules. Repression is switching off the gene to stop producing enzyme
E.g. ALA synthase is auto regulated by Heme through repression.
9. Covalent modification:
1. It refers to addition or removal of a group to the enzyme protein in
covalent bond
2. The examples of the covalent modification strategy are acetylation /
deacytilation; phosphorylation/Dephosphorylation;
3. The phosphorylation reaction is used in almost every metabolic process;
moreover, approximately 30% of the known proteins are phosphorylated.
The enzyme which catalyzes the phosphorylation process is protein
4. Protein phosphotases are another type of enzyme which catalyze the
process of the removal of phosphate group from the protein molecule. This
is also known as dephosphorylation.
5. ADP ribosylation is another Covalent modification. Cholera toxin and
pertusis toxin act through ADP ribosylation.
Ribosomes are RNA which can catalyse cutting of mRNA. This is an exception
of a nonprotein having an enzymatic action.
Enzyme Units:
One standard unit or international unit (U) of enzyme activity is the amount of
enzyme that will convert one micromole of substrate per minute per litre of
sample and written as U/L.
Modern expression of enzyme activity is in Katals (kat). One katal is defined
as the no of mole of a substrate transformed per second per litre of sample.
60 U= 1 .kat
Specific activity is the no of enzyme units present per mg of protein
Turnover number refers to no of substrate molecules transformed per unit
time by single enzyme molecule or by a single catalytic site.
1. It refers to physically distinct forms with similar enzyme activity and they are
synthesised from various tissues as multiple molecular forms of the same
2. Homomultimer: proteins with similar subunits produced by the same gene
3. Heteromultimer: proteins with different subunits produced by the different
4. Formation of isoenzymes:
1. Formed by different genes - true isoenzymes e.g. Salivary and
pancreatic amylase

2. Formed by same gene with different alleles allozymes; one form will
be seen in one individual but different forms may be seen in total
population. E.g.G6PD
3. Enzyme may attain difference in molecular structure after formation;
this is called post translational modification. These are called iso-forms.
E.g. Sialic acid content in Alkaline Phosphatase may vary in the same
5. Identification of isoenzymes:
1. LDH, CK, ALP etc can be identified by electrophoresis
2. ALP can be denatured by heat
3. One of the isoenzymes may be sensitive to inhibitor e.g. ACP
4. Km or substrate specificity may be variable. E.g. Glucokinase has
high Km and Hexokinase has low Km for glucose.
5. Cofactors requirement may be different. E.g. Mitochondrial isocitrate
dehydrogenase depends on NAD and the cytoplasmic isoenzymes
depends on NADP
6. Tissue localization may be different. LDH is in H4 form in heart and
M4 in skeletal muscle.
7. Specific Antibodies can also identify isoenzymes. E.g. CK isoenzymes
Lactate dehydrogenase:
1. The enzyme lactate dehydrogenase (also known as lactic dehydrogenase, or
LDH) is found in the cells of almost all body tissues. It converts pyruvate into
2. LDH is a tetramer with 4 subunits with H and M chains in 5 combinations ; M4
is seen in skeletal muscles and H4 in heart muscles.
3. Five components or fractions are labelled by number: LDH-1, LDH-2, LDH-3,
LDH-4, and LDH-5. Each of these fractions, called isoenzymes, is used mainly
by a different set of cells or tissues in the body. For this reason, the relative
amounts of a particular isoenzyme of LDH in the blood can provide valuable
diagnostic information. The enzyme is especially concentrated in the heart,
liver, red blood cells, kidneys, muscles, brain, and lungs.
i. LDH-1 - heart muscle- 30 %
LDH-2 -red blood cells- 35 %
LDH-3 brain- 20%
LDH-4 liver- 10%
LDH-5 -skeletal muscle- 5 %
4. Normal value in serum is 100-200 U/L; more in children
5. Increase slightly after exercise and 100 fold in haemolysis (false positives)
6. There is 10 fold increase in myocardial infarction. Therefore, LDH provides a
valuable late indicator of infarction. Assessment of LD1 and LD2 provide a
more specific index of infarction.
7. Following a myocardial infarction:

8. Levels increase after 12 to 24 hours
9. Peak at 48 to 72 hours
10.Return to normal after 8 to 10 days, providing there are no complications
11.The peak reached is from two to ten times the upper reference limit
12.The normal plasma LD1:LD2 isoenzymes ratio of less than 1.0 is reversed
(flipped pattern)
13.Plasma LDH may also be raised in:
b. Liver disease
c. Skeletal muscle disease e.g. Muscular dystrophies
d. Haemolysis,
e. Leukaemia - to very high levels
f. Malignancy of all types
g. Renal infarction
h. Pulmonary embolus
Creatine Kinase(CK):
1. Normal value: Normal value for CK is 15-100 U/L for males and 10-80 U/L for
2. Old name is creatine phophokinase and the reaction is as follows:
3. Creatine ------------------------------ Creatine phosphate
1. ATP
b. CK and heart attack:
4. CK is increased in heart attack; as the level increases within 3-6 hours of
infarction it has early diagnostic value.
5. As it does not increase in haemolysis and CCF it is more useful than LDH.
6. The pattern of rise will give the severity of infarction.
a. CK is increased in (500 1500 U/L) in muscular dystrophy.
b. CK is also elevated in crush injury and CNS stroke.
c. The CK enzyme consists of two subunits, which can be either B (brain
type) or M (muscle type). There are, three different isoenzymes are
i. CK 1-BB - 1 %
ii. CK2-MB - 5 %
iii. CK3-MM - 80%
Cardiac troponins:
1. They are not enzymes but accepted as markers of myocardial infarction.
Troponin is a complex of three regulatory proteins that is integral to muscle
contraction in skeletal and cardiac muscle, but not smooth muscle. Both
cardiac and skeletal muscles are controlled by changes in the intracellular
calcium concentration. When calcium rises, the muscles contract, and when
calcium falls the muscles relax.
2. Troponins are seen in skeletal and cardiac muscles, but not in smooth

3. Troponin is a component of thin filaments of muscle protein, and is the protein
to which calcium binds to accomplish this regulation. Troponin has three
a. Tn- C, calcium binding
b. Tn- I, actomyosin ATPase inhibitory element
c. Tn -T, tropomyosin binding element
4. Troponin I is released within 4 hrs after infarction; peaks at14-24 hrs; remains
elevated for 3-5 days. It is not elevated in muscle injury and is a useful
marker for myocardial infarction.
5. Troponin T increases within 6 hrs and peeks at 72 hrs and remains elevated
upto 7-10 days.
6. Cardiac troponin elevations at lower concentrations than the 99th percentile
value used for MI diagnosis may identify patients who have not had an MI but
still have a risk of having an adverse cardiac event.
Aspartate amino transferase (AST) :
1. It is known as SGOT in old literature.
2. It needs VIt.B6 as coenzyme
3. Normal serum level is 8-20 U/L
4. It is significantly elevated in myocardial infarction and is moderately
elevated in liver disease; very marked increase in hepatoma.
Brain Natriuretic Peptide:
This consists of 3 peptides:
1. atrial Natriuretic Peptide (ANP) - produced in cardiac atria
2. Brain Natriuretic Peptide (BNP) - present in brain and cardiac ventricles
3. C- type Natriuretic Peptide (CNP) significance not known
These peptides are seen in circulation and defend against salt and water
retention. Patients with CCF have high levels of ANP and BNP. The level correlates
with the degree of ventricular dysfunction.

BNP helps to differentiate CCF and obstructive lung disease.


1. Alanine Amino Transferase (ALP) :
a. Old name is SGPT
b. Needs pyridoxal phosphate as coenzyme
c. Normal level is M 13-35 and F 10-30 U/L
d. 300-1000 u/l in acute hepatitis. AST also increase but lesser extent.
1. Moderate increase in chronic liver disease.
2. Alkaline Phosphatase:
1. Non specific enzyme
2. Hydrolyses aliphatic, aromatic and heterocyclic compounds
3. Contains zinc; it is activated by magnesium and manganese.
4. It is produced by osteoblasts and is associated with bone formation
5. It is also associated with transport mechanism in liver, kidney and
intestinal mucosa.
6. Normal value is 40-125 U/L ;upper level in children
7. Moderate increase in liver disease

8. Very high levels are seen in obstructive jaundice as ALP is secreted by
epithelial cells of biliary canal.
9. Very high levels in bone diseases such as Pagets disease, rickets,
osteoblastoma, metastatic carcinoma and hyperparathyroidism.
10.Isoenzymes of ALP:
i. Alpha 1 ALP: increased in obstructive jaundice
ii. Alpha 2 heat labile ALP: 25 % of total ALP; produced by liver cells.
iii. Alpha 2 heat stable ALP: placental origin; 1% of total ALP; seen
elevated n carcinoma lung, liver and intestines. It is also known as
Regan iso-enzyme.
iv. Pre-beta-ALP: bone origin; heat labile; 50 % of ALP; elevated in
bone diseases.
v. Gamma-ALP: originates from intestinal cells; 10 % of total ALP;
increased in ulcerative colitis;
vi. The leukocyte ALP: increases in lymphoma; decreases in chronic
myeloid leukaemia.
3. Nucleotide Phosphatase (NTP):
1. Known as 5 nucleotidase; hydrolyses 5neucleotides to nucleosides
2. Present on cell membrane
3. Normal level is 2-10 IU/L
4. Moderately increased in hepatitis
5. Highly elevated in biliary obstruction
6. No change in bone diseases.
4. Gamma Glutamyl Transferase (GGT):
1. Old name is gamma glutamyl transpeptidase
2. Used in the synthesis of glutathione
3. Has 11 isoenzymes

4. Seen in liver, kidney, pancreas, intestinal cells and prostate
5. Normal value is 10-30 U/L
6. Moderate increase in infective hepatitis and prostate cancer
7. Increases after alcohol intake and sensitive test for alcohol abuse.
Other clinical disorders:
1. Acid Phosphatase (ACP):
1. Hydrolyses phosphoric acid ester
2. Normal value is 2.5-12 U/L
3. It is secreted by prostate, RBC, platelets and WBC
4. One isoenzymes is Inactivated by Tartarate - its level is 1 U/L
5. Increase in prostatic cancer
6. Tartarate labile fraction is highly elevated in bone metastasis from prostate
cancer and acts as a tumour marker
7. Note: haemolysis and prostatic massage may alter the value
2. Prostate specific antigen (PSA):
1. Produced by epithelial cells of prostate
2. Seen in seminal fluid
3. It is a serine protease
4. It is bound to alpha-i-antitrypsin and alpha-2-macroglobulin
5. Normal value is 1-5 microgram/ L
6. >10 is indicative of prostate cancer
3. Cholin esterase (ChE):
1. Acts on acetyl choline
2. Present in nerve endings and in RBCs esp. young RBCs

3. Organo- phosphorous compounds irreversibly inhibit cholinesterase; ChE in
rbcs is a measure of toxic levels industrial workers
4. Psuedocholinestrase or type II ChE is a nonspecific substance; can
hydrolyse acyl esters; produced by liver cells
5. Succinyl choline is used as a muscle relaxant in anaesthesia. This is
removed by liver ChE. Those who lack genetically liver ChE may go in for
scoline apnoea and death.
4. Glucose-6-phosphatase dehydrogenase:
1. Dimer with identical subunits
2. Important enzyme in hexose monophosphate shunt pathway of glucose.
3. Main role is to produce NADPH
4. NADPH is necessary for removing hydrogen peroxide in RBCs and thereby
maintains the integrity of RBCs.
5. G6PD deficiency will lead to tendency for haemolysis esp. After exposure
to drugs like aspirin, primaquin, sulpha etc
6. Favabin will induce similar haemolysis and is called favism.
7. The gene for G6PD is x-linked and hence hemizygous males and
homozygous females manifest the disease.
8. G6PD deficiency protects individuals from falciform malaria as the pentose
pathway necessary for malaria parasite is defective.
9. G6PD deficiency may lead to methaemoglobinemia
10. Nearly 400 iso forms of G6PD are described.
5. Amylase:
1. Produced by pancreas and salivary glands
2. Activated by calcium and chloride ions
3. Normal value is 50-10 IU/L
4. Increased to 100 times in acute pancreatitis
5. Moderate increase in chronic pancreatitis, mumbs and obstruction of
pancreatic duct.


Urinary amylase is also increased acute pancreatitis.

6. Lipase:
1. Present in pancreatic secretion
2. Hydrolyses triglyceride to monoglycerides and fatty acid
3. Elevated in acute pancreatitis but not in mumbs
4. Moderately increased in Ca pancreas, biliary disease and perforating peptic
7. Enolase:
1. Glycolytic enzyme
2. Neuron specific enolase (NSE) is seen in nural tissues and apudomas
3. NSE is a tumour marker for nuro-endocrain tumours, small cell lung cancer,
nuroblastoma, pheochromocytoma, medullary carcinoma of thyroid etc
4. It is measured by RIA or ELIZA
8. Enzymes as therapeutic agents:
1. Streptokinase from streptococcus, urokinase from urine is used as
thrombolytic agents in myocardial infarction.
2. Pepsin and trypsin are used as digestive enzymes
3. Asparaginase is an anti cancer drug.

4. Paper strip coated with glucose oxidase is used for testing glucose urine .