1-Introduction to enzymes

Enzymes are mainly proteins (complex structured proteins) that function

as biological catalysts, and the diversity in their protein structure allow
different enzymes to be specialized in their activity.
substrateenzyme product+enzyme

A catalyst is a chemical that:-

➢ Increase the rate of the reaction ,by lowering the activation energy
{The amount of energy required for the reaction to proceed}
➢ Not consumed by the reaction they catalyze nor alter the equilibrium of
these reactions, i.e.{ the reaction can proceed without the catalyst but at
a slower rate }.

Enzyme activity can be affected by other molecules:-

Inhibitors activators
Molecules that decreases The Molecules that increases The enzyme activity

Enzymes activity Ex. Drugs & Ex. Cofactors (inorganic substances) such
as Ca++, Mg++,Mn++,Cu++ and Zn++ {can be
Poisons.
removed by dialysis

or may not attach tightly to enzyme to give

apoenzymes} & coenzymes (organic

substances)

{derived from soluble vitamins in H2Osuch as

niacin

And riboflavin}.

Note: cofactors that cannot be removed from enzymes are called

prosthetic groups, which are linked tightly to enzymes by covalent bond
OR loosely by H-bond.

Activity of enzymes can also be affected by: ph, temperature,

concentration of the substrate.

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 Enzymes can be classified according to their structure to:

➢ Enzymes composed of Mainly protein structure, ex. Enzymes catalyzing

the hydrolytic reactions of gastric and intestinal tracts
➢ Enzymes with additional substances. {Cofactors &coenzymes}.

Properties of enzymes:-

1-increase the rate of reactions.

2-don’t change the physical activity to give the end result.

3-reactants can work without enzymes, but enzymes fasten the reaction.

4-reactants must have sufficient energy for the reaction to proceed, which
is not needed for all the reaction molecules.

5- The heat increases also the rate of the reaction.

6- Enzymes are essential for the synthesis, breathing, digestion and

metabolic reactions.

7-enymes exhibits a high degree of specificity for their substrate.

8- The biological cell may contain up to 3000 enzymes.

9-they r globular proteins range from 62 a.a. {4-oxalocrotonate

tautomerase monomer} to 2500 a. a. {animal fatty acid synthase }

10-like all proteins, made of long linear chain of a.a. that fold to produce
3D. Structure. Each with specific sequence and properties.

11-activity is determined by the 3-dimensional structure of their protein’s

amino acid.

12-some enzymes are larger than its substrate and only small portion of
enzyme {3-4 amino acids} is involved in catalysis, this portion called the
active site.

13-can be denatured & deactivated by heat or chemical denaturants,

according to enzyme this process can be reversible or irreversible.

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 14-they lower the barriers that normally prevent the chemical reaction
from occurring or slow them down by decreasing the required activation
energy. Thus, in presence of enzyme the reaction proceed in a faster rate.

15-they’re usually named with the suffix –ase at the end of their substrate
name. EX1.sucrose→sucrase.

EX2.lactose→lactase.

The lock and key model

Because both substrate and enzyme possesses specific geometric shapes
that they fits into one another, this is referred to as {the lock and key
model}, but this model failed to explain the transition state of enzyme
reaction.

The induced fit model (Modification of lock and key

model)
➢ As the enzyme is flexible structure, the active site continually reshaped
when interacts with the substrate.
➢ The amino acid side chain which form the active site are molded into the
exact positions that enable the enzyme to function correctly.
➢ Sometimes, the substrate also changes its shape slightly as it enters the
active site, ex. Glycosidase.

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 2-SPECIFICITY
The ability of an enzyme to catalyze the specific reaction and no others is
perhaps its most significant property.
However specificity varies from one enzyme to another, so we have the
following types of enzyme specificity.

1) Absolute specificity
Catalyze only one substrate not the other.
Ex1. Urase acts only on urea
Ex2.Fumerase acts only on fumrate
⁻OOC.CH=CH.COO⁻ FUMERASE ⁻OOC.CHOH.CH₂.COO⁻
L.Fumerate H2O L.malate

2) Broad specificity
The enzyme is specific to one type of bond possessed by a group of
chemically related substances.
Ex1.Pancreatic lipase hydrolysis alpha ester bonds (at position 1 and 3) of
triglycerides.
Ex2. Salivary amylase attack alpha 1:4 glucosidic bonds of starch.
Ex3.Polypeptidase.

3) Group specificity
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 A- Act on specific chemical group such as
➢ glycosidases on glycosides , pepsin and trypsin on peptide bonds
and esterases on esters.
➢ ii-Lytic enzymes exhibit a higher order of group specificity; e.g
carboxypeptidases and aminopeptidases split off amino acids one at
a time from the carboxy or amino terminal end of polypeptide chains.
B-Linkage specificity is related to group specificity and it act on a
particular type of chemical compound regardless of the rest of the
molecular structure.

4) Optical specificity or Stereochemical specificity

The enzyme is specific for only one isomer of the substrate.
Ex. enzymes oxidizing glucose act on D-forms of glucose only.
➢ Because of high specificity and accuracy the enzymes are involved in the
copying and expression of the genome.
These enzymes have “Proof-reading mechanism”.
Ex. DNA polymerase catalyzes a reaction in first step and then
checks that the product is correct in a second step. This two-step process
results in average error rates of less than I error in 1006 reactions in high-
fidelity mammalian polymerases.
It also found “Proof-reading mechanism” in RNA polymerase, aminoacyl
tRNA synthetases and ribosemes.
➢ Some enzymes that produce second metabolites are described as
promiscuous, as they can act on relatively broad range of different
substrate and it is important for the evolution of new biosynthetic
pathways.

3-NAMING OF ENZYMES

1. Many enzymes have more than one name.

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 Among the first identified and named were the digestive enzyme that
catalyzes the hydrolysis of the glycoside, ester and amide bond of
carbohydrates, lipids and proteins.
2. Enzymes were named for the substrates on which they acted by adding
the suffix -ase to the substrate such as proteinase , lipase.
3. Enzymes given the name of chemical reaction such as oxidases ,
decarboxylases and dehydrogenases.
4. The same enzyme being identified with two different names. One at the
normal physiological reaction and other at artifical conditions.
Ex1. Glucose isomerase industrially to convert glucose into the
sweetner fructose.
EX2. Xylose isomerase in vivo.
5. Naming depends on the types and mechanisms of chemical reactions.
a) Reactions and enzymes that catalyze them divided into 6 classes each
with 4-13 subclasses.
b) The enzyme names has 2 parts , the first is the substrate, second the
ending –ase
c) Each enzyme has a systemic code number (E.C). This number
characterized the reaction type. The Class,Subclass,Sub subclass then
the specific name.
 E.C 2.7.1.1 denotes
Class 2 (transferase)
Subclass 7 (transfer of phosphate)
Sub subclass 1 (an alcohol function as the phosphate acceptor)
The enzyme name (hexokinase)
 E.C 3.4.11.4 (tripeptide aminopeptidases
Class 3 (hydrolases)
Subclass 4 (hydrolases act on peptide bonds)
Sub subclass 11 (hydrolases that cleave off the amino-terminal aminoacid
from a polypeptide)
The enzymes are those that cleave off the amino-terminal end from a
tripeptide.

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 4-CLASSIFICATION OF ENZYMES

Although an international committee has established a uniform naming

system for enzymes, the names that are in common use do not follow a
completely consistent pattern. With the exception of some older enzyme
names (such as pepsin, trypsin), all enzymes names end with the suffix
-ase. Classes of enzymes are named according to their job category.

1) Oxidoreductases

Are biological oxidation and reduction between 2 substrate and there for
are essential in the process metabolism energy releasing, respiration and
fermentation.

Ex. Lactic to pyruvic acid

CH3-CH-COOH LDH+NAD CH3-C-COOH + NADH

Lactic acid Lactate dehydrogenase Pyruvic acid
2) Transferases

These enzymes transfer group from one substance,the donor, to another

the acceptor (e.g amino group, pohsphate group etc).

Ex. Transaminases transfer amino groups.

Glutamic acid pyruvic acid α-

ketoglutaric acid alanine

3) Hydrolases

Catalyze hydrolysis reactions when a molecule is split into two or more

smaller molecules by addition of water.

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 Ex. phosphatase catalyze dephosphorylation {removal of phosphate
group}

Glucose + ADP glucose-6-phosphate + ADP+

4) Lyases

Catalyze the cleavage of C-C, C-O, C-S and C-N bonds by

mechanism other than hydrolysis leaving double bond.
Lyase
Ex. L.malate L.fumarate
⁻OOC.CHOH.CH₂.COO⁻ -H2O+Lyase ⁻OOC.CH=CH.COO⁻

5) Isomerases

Catalyze atomic rearrangement within a molecule to give another shape

or structure.

Ex. Rotamase and protein disulfide isomerase which can assist apeptide
chain to fold into a correct 3D structure.

b) D-glucose L-glucose

6) Ligases

Catalyze the reaction which joins two molecules and formation of different
bonds by condensation reaction coupled to ATP cleavage; C-C, C-N, C-S
and C-O.

Coenzymes-5
Definition

Some enzymes are purely protein in nature and depend for activity on
their structure, while certain enzymes requires for their function, one or
more non-protein part, these are termed as Coenzymes, Cofactors or
prosthetic groups.
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 If such a compound is firmly attached to the enzyme protein then it’s
called Coenzyme.

Properties of coenzymes:

➢ They are organic molecules that cooperate in the catalytic action of the
enzyme.

Ex. NADH, NADPH & ATP (that are released from the enzyme’s active site
during the reaction.

➢ Coenzymes can be considered as a special class of substrate, or a

second substrate since they are chemically changes as a consequence
of enzyme action.
➢ Organic prosthetic groups can be covalently attached to enzyme,
Ex. Thiamine pyrophosphate in enzyme (pyruvate dehydrogenase)
➢ They can be used for many different enzymes which mean they’re widely
used and spread in the biological cell reactions.
Ex. About 700 enzymes can use the Coenzyme NADH.
➢ They can increase OR decrease the enzyme activity.
➢ Certain coenzymes exist in Free State in solutions and contact the enzyme
protein, ONLY at the time of reaction.
➢ They’re continuously regenerated which means that even small amounts
of coenzymes are used very intensively.
Ex. Human body turns over its own weight in ATP each day.
➢ Some of those coenzymes are Vitamins, which can’t be synthesized by the
body, but should be acquired from the diet.
Ex. Riboflavin (B₂), Thiamine (B₁) and Folic acid.
(Small amount of those vitamins are produced by the body, EX. Vita. K
and Nicotinic acid)

Different types of Coenzymes:

1. Nicotinamide dinucleotides
NADH, NADPH and their reduced forms are involved in many
dehydrogenase reactions in mitochondrion, cytosol, and endoplasmic
reticulum of the cell.

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 They’re water soluble and diffuse freely away from the enzyme after the
oxidation or reduction reaction is completed to join another
dehydrogenase reaction.
The general form of reactions involving NAD⁺ is:
RH2+ NAD⁺↔NADH + H⁺

2. Flavin nucleotide coenzymes

FAD is another class of dehydrogenases coenzymes known as
(Flavoproteins); its Flavin part is derived from the vit.B₂ (riboflavin).
Unlike the NAD, they remain tightly bound to the protein part of the
coenzyme throughout the reaction which means that FAD never leaves
the enzyme.

3. Adenine nucleotide coenzymes

Such as ATP, ADP and AMP which are influencing the direction of flow of
metabolic pathway.
ATP often joins as a donor of phosphate group to other molecules.

4. Thiamine pyrophosphate
Thiamine (B₁) is the precursor of thiamine pyrophosphate, is the
coenzyme for some important oxidation-decarboxylation reactions
Ex. Pyruvate dehydrogenase and Oxoglutarate dehyrogenase.

5. Coenzyme A
Complex group contains sulphahydryl (--SH) group.
This group reacts with carboxyl group.

6. Biotin coenzyme
Coenzyme for carboxylation reactions.
Ex. Pyruvate carboxylase.

7. Ascorpic acid (vit.C)

It prevents poor wound healing in scurvy because it involves in the
hydroxylation of prolin.

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 6-Mechanism of enzymes
Activation energy:

The amount of energy required for the reaction to proceed.

Enzymes act in several ways, all by lowering the ∆G:

➢ Lowering the activation energy by stabilizing the transition state:

Straining the shape of substrate, by binding its transition state
conformation, the enzyme distort the bound substrate into their transition
state form, this reduce the amount of energy required to complete the
transition.
➢ Lowering the energy of the transition state, without distorting
the substrate:
By creating an environment with opposite charge distribution to that of
the transition state.
➢ Providing an alternative pathway:
For example: temporarily reacting with the substrate to form ES
intermediate complex.
➢ Reducing the reaction entropy change:
Entropy is a measure of the number of ways in which a system may be arranged, often taken to be a measure of 'disorder' (the
higher the entropy, the higher the disorder).
By bringing the substrates together in the correct orientation to react, this
entropic effect involves the destabilization of the ground state.
➢ By increasing the temperature:
This helps the enzyme to function and develop the end product faster,
although too much temperature may distort the enzyme’s shape and will
never regain its shape until temperature comes back to normal.

Transition state stabilization:

➢ Enzymes can stabilize its transition state more than the transition state of
uncatalyzed reactions.
➢ Most effective way to reach large stabilization energy is to use
electrostatic effects, by having fixed polar environment which oriented
toward the charge distribution of the transition state.

Dynamics and function:

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 Internal dynamics is the movement of parts of the enzyme’s structure, for
example:
• Individual amino acid.
• Group of amino acids.
• Entire protein domain.

These movements are connected to the enzyme’s mechanism,

whether small and fast movement, or large and slow movement,
depending on the type of the reaction.

7-Enzymes Regulation
Many of enzyme reactions occur in cells involving synthesis, degradation
or interconversion.These reactions regulate by 2 ways

1) Regulation of amount of enzyme “Long term

regulation”
A) Enzyme induction

The inducer is a substrate or sometimes hormone, produce a signal to

specific gene of DNA, this gene starts to synthesis the required enzyme.

Ex1. Enzymes in the liver called cytochrome P450 oxidases, which are
important in drug metabolism. Induction or inhibition of these enzymes
can cause drug interactions.

Ex2.If a new substrate is made available to the cell; it may induce the
synthesis of the enzymes needed to cope with it.

Yeast cells, for example, do not ordinarily metabolize lactose and no

lactase can be detected in them. However, if grown in a medium
containing lactose, they soon begin synthesizing lactase — by transcribing
and translating the necessary gene(s) — and so can begin to metabolize
the sugar.

B) Enzyme repression
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The repressors which may be a product or hormone affect DNA to stop
synthesis of specific protein which is an enzyme.

Ex.If ample quantities of an amino acid are already available to the cell
from its extracellular fluid; synthesis of the enzymes that would enable
the cell to produce that amino acid for itself is shut down.

2) Regulation of catalytic efficiency “short term

regulation”
A) Allosteric modification

In the case if feedback inhibition and precursor activation, the activity of

the enzyme is being regulated by a molecule which is not its substrate.

In these cases, the regulator molecule binds to the enzyme at a different

site than the one to which the substrate binds.

When the regulator binds to its site, it alters the shape of the enzyme so
that its activity is changed.

B) Feedback inhibition (F.B.I.)

The end product(s) of a metabolic pathway are often inhibitors for one of
the first enzymes of the pathway (usually the first irreversible step, called
committed step), thus regulating the amount of end product made by the
pathways.

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The amount of the end product produced is regulated by its own
concentration.

Negative feedback mechanism can effectively adjust the rate of synthesis

of intermediate metabolites according to the demands of the cells.

This helps allocate materials and energy economically, and prevents the
manufacture of excess end products.

The control of enzymatic action helps to maintain a stable internal

environment in living organisms.

C) Inactive precursors (zymogens)

The enzymes are produced in inactive form and when it reaches the site
of reaction it is activated.

Ex1. Chymotrypsin, a digestive protease, is produced in inactive form as

chymotrypsinogen in the pancreas and transported in this form to the
stomach where it is activated.

This stops the enzyme from digesting the pancreas or other tissues before
it enters the gut.

Ex2.Some enzymes may become activated when localized to a different

environment (e.g. from a reducing (cytoplasm) to an oxidising (periplasm)
environment, high pH to low pH etc).For example, (hemagglutinin) in the
influenza virus is activated by a conformational change caused by the
acidic conditions, these occur when it is taken up inside its host cell and
enters the lysosome.

Ex3.Pepsin is synthesized within the chief cells (in gastric glands) as an

inactive precursor, pepsinogen. Only when exposed to the low pH outside
the cell is the inhibiting portion of the molecule removed and active
pepsin produced.

D) Regulatory protein

The activity of an enzyme is regulated by “calmodulin” which is a protein

very rich in glutamic and aspartic amino acid that form a Ca+2 binding site.

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A Ca+2 bind to calmodulin induce a co-formational change that convert it
from inactive to active form.

Activated calmodulin bind to many enzymes and other proteins in the cell
and modify their activity either by stimulation or inhibition.

E) Covalent modification

Some key enzymes exist in forms, phosphorylated and a non-

phosphorylated form, which differ greatly in enzyme activity. The total
activity of the enzyme depends on the ration of these 2 forms.

Protein kinase makes phosphorylation.

Phosphatase makes dephosphrylation.

Phosphorylation activate some enzymes and inactive other responsible for

the opposite reaction.

8-Factors affecting the activity (velocity) of

enzyme
1) Concentration of substrate and enzyme:

Rate of the reaction increase when the concentration of substrate or the

enzyme concentration increases.

If the enzyme concentration is constant the rate of the reaction will

increase up to maximum rate and will not increase upon further addition
of the substrate.

As the substrate concentration increase more substrate molecule fills the

active site for longer period of time and more products.

Enzyme rate

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 Conc. Of sub. And enzyme

2) Temperature:

When the temperature increase the rate of the reaction will increase even
without enzyme but with low rate .But when add enzyme it accelerates
the reaction, because it removes barriers.

Enzymes work at optimal temperature. Under or above this temperature

the reaction is altered. For most animal enzymes the optimum
temperature is between 38oc-40oc .

Like most chemical reactions, the rate of an enzyme-catalyzed reaction

increases as the temperature is raised. A ten degree Centigrade rise in
temperature will increase the activity of most enzymes by 50 to 100%.
Variations in reaction temperature as small as 1 or 2 degrees may
introduce changes of 10 to 20% in the results. In the case of enzymatic
reactions, this is complicated by the fact that many enzymes are
adversely affected by high temperatures. As shown in Figure, the reaction
rate increases with temperature to a maximum level, then abruptly
declines with further increase of temperature. Because most animal
enzymes rapidly become denatured at temperatures above 40°C, most
enzyme determinations are carried out somewhat below that
temperature.

Over a period of time, enzymes will be deactivated at even moderate

temperatures. Storage of enzymes at 5°C or below is generally the most
suitable. Some enzymes lose their activity when frozen.

Activation velocity

Plato

16
 Inactivation

Activation temperature

3)PH:

Enzymes are affected by changes in PH The most favorable pH value - the

point where the enzyme is most active - is known as the optimum pH and
is between 2-10 PH
The PH optimum of enzyme usually reflects the PH of the body fluid in
which the enzyme is found.
Extremely high or low pH values generally result in complete loss of
activity for most enzymes. PH is also a factor in the stability of enzymes.
As with activity, for each enzyme there is also a region of pH optimal
stability.The optimum pH value will vary greatly from one enzyme to
another, as shown in this table :

Enzyme PH Optimum

Lipase (pancreas) 8.0

Lipase (stomach) 4.0 - 5.0

Lipase (castor oil) 4.7

Pepsin 1.5 - 1.6

Trypsin 7.8 - 8.7

Enzyme Inhibition-9
Enzymes can be inhibited by many substances and reduce the initial
velocity of such enzymes. Accordingly there are 2 types of enzymes
inhibition.

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1) Irreversible inhibitors
React with the enzyme and form a covalent adducts with the protein. The
inactivation is irreversible. These compounds include eflornithine a drug
used to treat the parasitic disease sleeping sickness. Penicillin and Aspirin
also act in this manner. With these drugs, the compound is bound in the
active site and the enzyme then converts the inhibitor into an activated
form that reacts irreversibly with one or more amino acid residues.

2) Reversible inhibitors
Reversible inhibitors bind and dissociate with their enzyme in equilibrium
process. It can be classified as

a) Competitive inhibition
b) Non competitive inhibition
c) Uncompetitive inhibition
d) Mixed inhibition

It classified according to where on the enzyme the inhibitor binds, or the

order with which it binds, relative to substrate.

A) Competitive inhibition

Competitive inhibitors compete with substrate for an enzyme’s active site,

lowering the enzyme’s likelihood of binding substrate and slowing the
observed reaction velocity.

Competitive inhibitors bind reversibly to the enzyme, preventing the

binding of substrate. On the other hand, binding of substrate prevents
binding of the inhibitor. Substrate and inhibitor compete for the enzyme.

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B) Non competitive inhibition

Can bind to the enzyme at the same time as the substrate, i.e. they never
bind to the active site. Both the EI and EIS complexes are enzymatically
inactive. Because the inhibitor cannot be driven from the enzyme by
higher substrate concentration (in contrast to competitive inhibition), the
apparent Vmax changes. But because the substrate can still bind to the
enzyme, the Km stays the same.

Occur in cells when specific molecule bind to an enzyme site other than
the active site called Allosteric site causing a shift in the enzyme
configuration which prevent the substrate from binding to the active site.

C) Uncompetitive inhibition

In uncompetitive inhibition the inhibitor cannot bind to the free enzyme,

but only to the ES-complex. The EIS-complex thus formed is enzymatically
inactive. This type of inhibition is rare, but may occur in multimeric
enzymes.

D) Mixed inhibition

This type of inhibition resembles the non-competitive, except that the EIS-
complex has residual enzymatic activity.

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In many organisms inhibitors may act as part of a feedback mechanism. If
an enzyme produces too much of one substance in the organism, that
substance may act as an inhibitor for the enzyme at the beginning of the
pathway that produces it, causing production of the substance to slow
down or stop when there is sufficient amount. This is a form of negative
feedback. Enzymes which are subject to this form of regulation are often
multimeric and have Allosteric binding sites for regulatory substances.
Their substrate/velocity plots are not hyperbolar, but sigmoidal (S-
shaped).

Uses of inhibitors:
Since inhibitors modulate the function of enzymes they are often used as
drugs and poisons.

Drugs A common example of an inhibitor that is used as a drug is

aspirin. This inhibits the COX-1 and COX-2 enzymes that produce the
inflammation messenger prostaglandin, thus suppressing pain and
inflammation.

Poisons the poison cyanide is an irreversible enzyme inhibitor that

combines with the copper and iron in the active site of the enzyme
cytochrome c oxidase and blocks cellular respiration.

10-measurement of enzymes activity

To measure the amount of the enzyme in a sample of tissue extract or
other biologic fluid, the rate of the reaction catalyzed by the enzyme in
the sample is measured.

The measured rate is proportional to the quantity of enzyme present. This

rate is compared with rate catalyzed by a known quantity of the highly
purified enzyme.

Enzyme units:

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Enzyme units are expressed in micromoles (10-6 mol), nanomoles (10-9
mol) or picomoles (10-12 mol) of substrate reacting or product produced 1
minute.

Intracellular distribution of enzymes-11

The enzymes of glycolysis are located in the cytoplasm, where enzymes of
the citric acid cycle are in the mitochondria.

The distribution of enzymes among sub cellular organelles may be studied

fraction of cell homogenates by high-speed centrifugation. The enzyme
content of each fraction is then examined.

Isozyme

It is the presence of enzyme in different forms but all have the same
reaction differ from each other in some chemical or physical properties.
m-dehydrogenase
EX1.Malate oxaloacetate

Malate dehydrogenase is a generic name that includes all proteins

(enzymes) which catalyze the oxidation of malate to oxaloactate.

Ex2.Lactate dehydrogenase (LDH) enzyme presents in blood in a form of 5

fractions (isozymes) LDH1, LDH2, LDH3, LDH4, and LDH5.

Proenzymes

Many proteins are manufactured and secreted from cells in the form of
inactive precursor or proteins known as "proproteins". When the proteins
are enzymes, they are termed proenzymes or zymogens.

Conversion of a proprotein to mature protein involves a process known as

limited selective proteolysis.

Ex1.Insulin proinsulin

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Ex2.Pepsin pepsinogen

Enzymes in the Diagnosis of Pathology-12

The measurement of the serum levels of numerous enzymes has been
shown to be of diagnostic significance. This is because the presence of
these enzymes in the serum indicates that tissue or cellular damage has
occurred resulting in the release of intracellular components into the
blood. Hence, when a physician indicates that he/she is going to assay for
liver enzymes, the purpose is to ascertain the potential for liver cell
damage. Commonly assayed enzymes are the amino transferases: alanine
transaminase, ALT (sometimes still referred to as serum glutamate-
pyruvate aminotransferase, SGPT) and aspartate aminotransferase, AST
(also referred to as serum glutamate-oxaloacetate aminotransferase,
SGOT); lactate dehydrogenase, LDH; creatine kinase, CK (also called
creatine phosphokinase, CPK); gamma-glutamyl transpeptidase, GGT.
Other enzymes are assayed under a variety of different clinical situations
but they will not be covered here.
The typical liver enzymes measured are AST and ALT. ALT is particularly
diagnostic of liver involvement as this enzyme is found predominantly in
hepatocytes. When assaying for both ALT and AST the ratio of the level of
these two enzymes can also be diagnostic. Normally in liver disease or
damage that is not of viral origin the ratio of ALT/AST is less than 1.
However, with viral hepatitis the ALT/AST ratio will be greater than 1.
Measurement of AST is useful not only for liver involvement but also for
heart disease or damage. The level of AST elevation in the serum is
directly proportional to the number of cells involved as well as on the time
following injury that the AST assay was performed. Following injury, levels
of AST rise within 8 hours and peak 24–36 hours later. Within 3–7 days
the level of AST should return to pre-injury levels, provided a continuous
insult is not present or further injury occurs. Although measurement of
AST is not, in and of itself, diagnostic for myocardial infarction, taken
together with LDH and CK measurements (see below) the level of AST is
useful for timing of the infarct.
The measurement of LDH is especially diagnostic for myocardial infarction
because this enzyme exists in 5 closely related, but slightly different
forms (isozymes). The 5 types and their normal distribution and levels in
non-disease/injury are listed below:

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LDH 1 – Found in heart and red-blood cells and is 17% – 27% of the normal
serum total.
LDH 2 – Found in heart and red-blood cells and is 27% – 37% of the normal
serum total.
LDH 3 – Found in a variety of organs and is 18% – 25% of the normal
serum total.
LDH 4 – Found in a variety of organs and is 3% – 8% of the normal serum
total.
LDH 5 – Found in liver and skeletal muscle and is 0% – 5% of the normal
serum total.
Following a myocardial infarct the serum levels of LDH rise within 24-48
hours reaching a peak by 2–3 days and return to normal in 5-10 days.
Especially diagnostic is a comparison of the LDH-1/LDH-2 ratio. Normally,
this ration is less than 1. A reversal of this ration is referred to as a
“flipped LDH”. Following an acute myocardial infarct the flipped LDH ratio
will appear in 12–24 hours and is definitely present by 48 hours in over
80% of patients. Also important is the fact that persons suffering chest
pain due to angina only will not likely have altered LDH levels.
Diagnostic and prognostic value of specific enzymes
The determination of the activity of the following enzymes can provide the
physician with valuable diagnostic evidence.
1-Lipase
The plasma lipase level may be low in liver disease, vitamin A deficiency,
some malignancies and diabetes mellitus. It may be elevated in acute
pancreatitis and pancreatic carcinoma.
2-Amylase
The plasma amylase may below in liver disease and increased in acute
pancreatitis and diabetes mellitus.
3-Trypsin
Elevated level of trypsin in plasma occurs during acute pancreatic
disease. The elevation is more sensitive and considers indicator of
pancreatic disease than plasma amylase or lipase.
4-Cholinesterase

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In general, low levels are found in patients ill with liver disease
malnutrition, acute infection disease and anemia. High levels occur in
nephritic syndrome.
Since the content of cholinesterase in young red blood cells is
considerably higher than in the adult red blood cells, the cholinesterase
titer of erythrocytes in peripheral blood gives an indication of
hematopoietic activity.
5-Alkaline phosphatase
The level of enzymes capable of catalyzing the hydrolysis of various
phosphate esterase at alkaline PH (alkaline phosphatase activity) may be
increased in hyper parathyrodisim and osteoblastic sarcoma.
Isozymes of alkaline phosphatase are present in body fluid. These include
specific isozymes originating from bone, liver, placenta and intestine.
Measurement of specific alkaline phosphatase isozymes may therefore
improve the diagnostic value of this test.
6. Acid phosphatase
The level of enzymes capable of catalyzing the hydrolysis of various
phosphate esters at acidic PH (acid phosphatase activity) may be elevated
in the metastatic prostatic carcinoma.
7. Transaminases
Two transaminases are of clinical interest
a) Glutamic oxaloacetic transaminases (GOT) which catalyzes the transfer
of the amino group of aspartic acid to alpha-ketoglutaric acid, forming
glutamic and oxalo acetic acids.
b) Glutamic pyruvic transaminase (GPT) transfers the amino group of
alanine to alpha-ketoglutaric acid, forming glutamic and pyruvic acids.
Liver tissue rich in both transaminases are elevated in sera than GOT.
While both transaminases are elevated in sera of patients with acute
hepatic disease, GPT, which is only elevated by cardiac necrosis, is a
more specific indicator of liver damage.
8. Lactate dehydrogenase
In myocardial infarction, the concentration of serum lactate
dehydrogenase (LDH) rises within 24hrs after the infract and return to the
normal range within 5-6 days. High levels of LDH also occur in patients
with acute and chronic leukemia in relapse.
9. LDH isozymes

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Cardiac muscle contains a preponderance of LDH-1.
Measurements of the serum isozyme pattern following myocardial
infarction is a more sensitive and lasting indication of myocardial necrosis
than the simple measurements of the total serum or plasma LDH activity.

Y
oussra Sayed Othman.
M
arwa Mohamed Amin.
Di
na Gamal Tawfik.

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