ENZYMOL

OGY– GBSN –
BIOCHEMISTRY
SEMESTER I
SYED MUHAMMAD HASAN

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 WHAT ARE ENZYMES?
• Enzymes are biocatalysts
(typically proteins) that
significantly speed up the
rate of virtually all of the
chemical reactions that
take place within cells.
• They are vital for life and
serve a wide range of
important functions in the
body, such as aiding in
digestion and metabolism.
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 IMPORTANCE OF ENZYMES
• The digestive system - enzymes help the body
break down larger complex molecules into smaller
molecules, such as glucose, so that the body can
use them as fuel.
• DNA replication - each cell in your body contains
DNA. Each time a cell divides, that DNA needs to be
copied. Enzymes help in this process by unwinding
the DNA coils and copying the information.
• Liver enzymes - the liver breaks down toxins in the
body. To do this, it uses a range of enzymes
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 PROPERTIES OF ENZYMES
• Complex macromolecules with high
molecular weight.
• Do not start a chemical reaction,
however they help in accelerating it.
• Affect the rate of biochemical
reaction and not the direction of
reaction.
• Enzymes are specific in action.
• The enzymatic activity is highly
dependent upon several parameters
such as temperature, pH, Substrate
Concentration, Inhibitors.
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 HOW ENZYMES WORK?
• The molecules that an
enzyme works with are
called substrates. The
substrates bind to a region
on the enzyme called the
active site.
• The active site is an
extremely important
region of Enzymes where
the process of catalysis
occurs.
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 ACTIVE SITE
• The binding site is a place of mass chemical
specificity and affinity on protein that forms
chemical bonds with molecules using weak
covalent bonds.
• Only a few residues actually participate in
binding the ligand while the other residues in
the protein act as a framework to provide
correct conformation and orientation.

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 MECHANISM OF ACTION -
ACTIVE SITE
• Because enzymes are made of amino acids,
they can create active sites with a wide variety
of properties that can bind specifically to
different substrates. These binding properties
depend upon:
• Size and Shape of Active Site.
• Polarity or Non Polarity.
• Hydrophobicity and Hydrophilicity.
• Inclusion of Co-factors.
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 Substrate

Transition state Product

Enzyme not only recognizes substrate,

but also induces the formation of transition state 8
Adapted from Nelson & Cox (2000) Lehninger Principles of Biochemistry (3e) p.252
 ENZYME CATALYZED
REACTIONS
• When a substrate (S) fits properly in an active site, an
enzyme-substrate (ES) complex is formed:
E + S  ES
• Within the active site of the ES complex, the reaction
occurs to convert substrate to product (P):
ES  E + P
• The products are then released, allowing another
substrate molecule to bind the enzyme.

The overall reaction for the conversion of substrate to

product can be written as follows:
E + S  ES  E + P
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 FACTORS AFFECTING
ENZYMATIC ACTIVITY

1. pH.
2. Temperature.
3. Substrate concentration
4. Inhibitors.

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 PH AND ENZYME ACTIVITY
• Enzymes are most active at their optimum pH.
• Activity is lost at low or high pH as tertiary structure is disrupted.
• Most enzymes of the body have an optimum pH of about 7.4.

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 TEMPERATURE AND ENZYME
ACTIVITY
• Enzymes are most active at an optimum temperature (usually 37°C in
humans)
• They show little or no activity at low temperatures.
• Activity is lost at high temperatures as denaturation occurs

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 AFFECT OF SUBSTRATE
CONCENTRATION ON ENZYME
• The rate of reaction increases as substrate concentration increases
(at constant enzyme concentration)
• Maximum activity occurs when the enzyme is saturated (when all
active sites are occupied by substrates)
• The relationship between reaction rate and substrate concentration is
exponential and shows linearity when the enzyme is completely
saturated.

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 ENZYME ACTIVATORS AND
INHIBITORS
• Activators are molecules that bind to enzyme’s
active site and increase their activity.
• Many chemical compounds such as Hexokinase
and Fructose 2-6 biphosphate are examples of
activators.
• Inhibitors are molecules that cause a loss of
enzyme activity, reduce the rate of enzymatic
reactions, and work at low concentrations.
• They block the enzyme but they do not usually
destroy it. Many drugs and poisons are inhibitors
of enzymes in the nervous system. 16
 THE AFFECT OF ENZYME
•
INHIBITION
Irreversible inhibitors
• Combine with the functional groups of the amino acids in the active
site, irreversibly.
• Examples: Nerve gases and pesticides, containing organophosphorus,
combine with serine residues in the enzyme acetylcholine esterase.

• Reversible inhibitors
• These inhibitors bind non covalently to enzymes with the help of
hydrogen bonds, hydrophobic interactions and ionic bonds.
• There are two categories.
1. Competitive
2. Non Competitive
3. Uncompetitive. 17
COMPETITIVE INHIBITOR
• A competitive inhibitor resembles the
substrate’s structure closely.
• It is reversible and has a structure like the
substrate. These compete with the substrate
molecules for the active site.
• The inhibitor’s action is proportional to its
concentration, its effect is reversed by
increasing substrate concentration.

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 NON COMPETITIVE INHIBITOR

• A noncompetitive inhibitor has a structure

that is different than that of the substrate. It
binds to an allosteric site rather than to the
active site.
• It distorts the shape of the enzyme, which
alters the shape of the active site and
prevents the binding of substrate.
• The effect can not be reversed by adding
more substrate.
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UNCOMPETITIVE INHIBITOR

• An uncompetitive inhibitor is an inhibitor that

only binds to the enzyme-substrate complex.
• Uncompetitive inhibition requires that an
enzyme-substrate complex must be formed.
• The uncompetitive inhibition does not work
when additional substrates are trying to be
involved.

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COENZYMES AND ITS CLASSIFICATION
• Coenzymes can be defined as a non protein
organic compound that is necessary for the
functioning of an enzyme.
• Co enzymes are classified into:
• NAD/NADP – Derived from Vitamin B3, involved
with dehydrogenases and participate in Redox
reactions.
• FMN/FAD – Also called flavoproteins, involved
with hydrogenases and participate in Redox
reactions.
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COENZYMES AND ITS CLASSIFICATION

• Coenzyme A (CoA) – Involved in the formation of

Acetyl CoA, a major metabolic compound.
• Thiamine Pyrophosphate (TPP) – Involved in
transferring Aldehyde groups and participate in the
oxidative decarboxylation of pyruvate.
• Pyridoxal Phosphate (PAL) – Involved with the
transfer of Amino groups to form keto acids.
• Some other Coenzymes include Tetrahydrofolic Acid
(THF), Biotin, Lipoic Acid and Cobalamine.

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 ENZYMES – MARKERS FOR DISEASE

• There are several enzymes that are typically used

in the clinical laboratory to diagnose diseases.
• There are highly specific markers for enzymes
active in the pancreas, red blood cells, liver,
heart, brain, prostate gland and many of the
endocrine glands.
• Since these enzymes are relatively easy to assay
using automated techniques, they are part of the
standard blood tests that assist in the diagnosis
and prognosis of diseases.
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ALKALINE PHOSPHATASE (ALP)
• Alkaline phosphatase (ALP) is an enzyme found in
several tissues throughout the body. The highest
concentrations of ALP are present in the cells that
comprises of bone and the liver.
• Elevated levels of ALP in the blood are most
commonly caused by liver disease or bone
disorders.
• Normal ranges lie from 37 to 116 U/L in both
males and females.

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 CREATINE KINASE (CK)

• Creatine kinase also known as

phosphocreatine kinase or creatinephospho
kinase.
• They are present in abundance in skeletal
muscles and is associated with myocardial
infarction and muscle diseases such as
myositis and myocarditis.
• Normal ranges in Males are 55-170 U/L and
in Females are 30-135 U/L.
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ALANINE AMINOTRANSFERASE
(ALT)
• It was formerly known as Serum Glutamic
Pyruvate Transaminase (SGPT).
• ALT can be found in the liver, skeletal muscle
and heart and is a well established, sensitive
liver-specific indicator of damage.
• A normal ALT test result can range from 7 to 55
units per liter (U/L). Levels are normally higher
in men.

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ASPARTATE AMINOTRANSFERASE (AST)

• It was formerly called Serum Glutamic

Oxaloacetic Transaminase (SGOT).
• AST is found in skeletal muscle, heart, liver,
kidney and erythrocytes and is associated with
myocardial, hepatic, parenchymal and muscle
diseases in humans and animals.
• The Normal ranges of this enzyme in Males are
10 to 40 units/L and in Females are 9 to 32
units/L.

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LACTATE DEHYDROGENASE
(LDH)
• LDH can be found in the heart, liver,
erythrocyte, skeletal muscle, platelets and
lymph nodes.
• In humans, it is involved in myocardial
infarction, hemolysis and liver disease.
• Normal LDH levels range from 140 U/L to 280
U/L.

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SORBITOL DEHYDROGENASE
(SDH)
• It is also called L-iditol dehydrogenase (IDH).
• SDH is liver specific in humans and all species
of animals and hepatic injury appears to be
the only source of increased SDH activity. 
• In tissues where sorbitol dehydrogenase is low
or absent, sorbitol can accumulate under
conditions of hyperglycemia.
• Normal ranges lie from 0 to 2 U/L.

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