Kurdistan Regional Government – Iraq

Ministry of Higher Education & Scientific Research

University of Duhok
College of health sciences
Department of Medical laboratory sciences

Lect. 1
Enzymology
Biochemistry Theory Lecture

Barhav Issa Abdullah

Asst. Lect. In Biochemistry
Enzyme
• Definition:
• Enzymes can be defined as soluble, colloidal, organic catalyst which
are produced by living cells but are capable of acting independently of
the cells.
• Enzymes are proteins that increase the rate of reaction by lowering
the energy of activation. They catalyze nearly all the chemical
reactions taking place in the cells of the body.

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• Intracellular location of enzymes
Cytoplasm: enzymes of glycolysis (conversion of glycogen and glucose
to pyruvic acid or lactic acid).
Mitochondria: enzymes of citric acid cycle and phosphorylation
(oxidation of acetate molecule in the citric acid cycle yielding energy).
Lysosomes: several hydrolytic enzymes.
Nucleus and Microsome: are also rich in certain enzymes.

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• Extra cellular enzymes
While all enzymes are produced in the living cell, some enzymes are
secreted out of the cell and function out side the cell of origin
• For example: digestive enzymes:
- salivary glands → salivary amylase
- gastric glands → gastric pepsin
- pancreas → pancreatic lipase

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 Importance of Enzyme
• Enzymes play an important role in Metabolism, Diagnosis, and
Therapeutics.
• All biochemical reactions are enzyme catalyzed in the living
organism.
• Level of enzyme in blood are of diagnostic importance e.g. it is a
good indicator in disease such as myocardial infarction.
• Enzyme can be used therapeutically such as digestive enzymes.

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 Enzyme of Diagnostic Importance

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 Enzyme of Therapeutic Importance

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Functions of Enzymes
The enzymes perform a number of functions in our bodies. These
include:
1. Enzymes help in signal transduction
2. They break down large molecules into smaller substances that can be
easily absorbed by the body.
3. They help in generating energy in the body. ATP synthase is the enzyme
involved in the synthesis of energy.
4. Enzymes are responsible for the movement of ions across the plasma
membrane.
5. Enzymes perform a number of biochemical reactions, including
oxidation, reduction, hydrolysis, etc.
6. reorganize the internal structure of the cell to regulate cellular activities
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 • Apoenzyme is the inactive form of the enzyme that activates
upon the binding of a cofactor. Typically, apoenzymes are
present in the conjugate (complex) enzymes.
• Holoenzyme refers to the apoenzyme along with the cofactor
that is complete and catalytically-active.

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CO-FACTORS
• A cofactor is a non-protein chemical compound that is bound (either
tightly or loosely) to an enzyme and is required for catalysis.

Co-factors are of two types:

• Coenzymes
• Prosthetic groups.

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Types of CO-FACTORS
• Coenzyme:
The non-protein component, loosely bound to apoenzyme by non-
covalent bond.
• Examples : vitamins or compound derived from vitamins.
• Prosthetic group
The non-protein component, tightly bound to the apoenzyme by
covalent bonds is called a Prosthetic group.

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Enzyme structure
• The active site of an enzyme is the region
that binds substrates, co-factors and
prosthetic groups and contains residue
that helps to hold the substrate.
• Active sites generally occupy less than 5%
of the total surface area of enzyme.
• Active site has a specific shape due to
tertiary structure of protein.
• A change in the shape of protein affects
the shape of active site and function of the
enzyme.
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Active site
• The area on the enzyme where the substrate or substrates attach to is
called the active site.
• Enzymes are usually very large proteins and the active site is just a
small region of the enzyme molecule.
• Enzyme molecules contain a special pocket or cleft called the active
sites.

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How do enzyme work ?
• The keys to how enzymes control cellular reactions lie in their
catalytic properties and the specificity of their substrate.
• The catalytic properties of an enzyme determine the rate at which a
certain reaction should occur, while its substrate specificity dictates
which starting reactant the enzyme should react with.
• enzymes are made up of a particular sequence of amino acids, whose
intramolecular interactions give proteins their three-dimensional
shape or conformation.

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 According to Michaelis-Menten theory:
• The enzymes combines with the substrate on which it acts to form an
enzyme substrate complex.
• From this complex, the enzyme is liberated and the substrate is
broken down into the products of the reaction

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• The molecule is usually more complex than the initial substrate
molecule that entered the chain of reaction, and accordingly, this
sequence of enzyme events is termed anabolic, one example for
anabolism is the conversion of glucose into glycogen.
• Glucose molecule (Monosaccharide) → Glycogen (long-branched polymer)
• Amino acids → Protein
• Nucleotides → Nucleic acids
• Fatty acids and glycerol → Lipids

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• Such sequence of enzymes involved in the breakdown of substrates
are called catabolic, an example of catabolism is the breakdown of
simple hexose sugar glucose to form the smaller compound pyruvate.
• Glucose → Pyruvate → Acetyl COA
• Proteins → Amino acids → Pyruvate → Acetyl COA
• Lipids → Fatty acid → Acetyl COA

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 • Step 1:
• Enzyme and substrate combine to form complex
•E + S ES
• Enzyme Substrate Complex

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 • Step 2:
• An enzyme-product complex is formed.
• ES EP

ES transition EP
state

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 • The enzyme and product separate

• EP E + P
The product
is made

Enzyme is
ready
EP for
another
substrate.

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Enzyme specificity
Specificity of the Enzyme-Substrate Complex
• For enzyme and substrate to react, surfaces of each must be
complementary.
• Enzyme specificity: the ability of an enzyme to bind only one, or a
very few, substrates thereby catalyzing only a single reaction.

• Compare these 2 reactions

Urease is VERY Specific or has
A HIGH DEGREE of Specificity

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Classes of Enzyme Specificity
1. Absolute: enzyme reacts with only one substrate.
2. Group: enzyme catalyzes reaction involving any molecules with the
same functional group.
3. Linkage: enzyme catalyzes the formation or break up of only certain
category or type of bond.

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Classification of Enzymes

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 EC 1. Oxidoreductases
• Biochemical Activity:
• Catalyse Oxidation/Reduction Reactions Act on many
chemical groupings to add or remove hydrogen atoms.
• Examples:
• Lactate dehydrogenase.
• Glucose Oxidase.
• Peroxidase.
• Catalase.
• Phenylalanine hydroxylase.

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Example, 1. Oxidoreductases
Catalyze oxidation-reduction reactions

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 EC 2. Transferases
• Biochemical Activity:
• Transfer a functional groups (e.g. methyl or phosphate)
between donor and acceptor molecules.
• Examples:
• Transaminases.
• Phosphotransferases (Kinases).
• Transmethylases.
• Transpeptidases.
• Transacylases.

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Example, 2. Transferases
Catalyze group transfer reactions

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 EC 3. Hydrolases
• Biochemical Activity:
• Catalyse the hydrolysis of various bonds by Add water across a
bond.
• Examples:
• Protein hydrolyzing enzymes (Peptidases).
• Carbohydrase (Amylase, Maltase, Lactase).
• Lipid hydrolyzing enzymes (Lipase).
• Deaminases.
• Phosphatases.

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Example, 3. Hydrolases
Catalyze hydrolysis reactions where water is the acceptor of
the transferred group

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 EC 4. Lyases
• Biochemical Activity:
• Cleave various bonds by means other than hydrolysis and
oxidation.
• Add Water, Ammonia or Carbon dioxide across double
bonds, or remove these elements to produce double bonds.
• Examples:
• Fumarase.
• Carbonic anhydrase.

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Example, 4. Lyases

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 EC 5. Isomerases
• Biochemical Activity:
• Catalyse isomerization changes within a single molecule.
• Carry out many kinds of isomerization:
• L to D isomerization.
• Mutase reactions (Shifts of chemical groups).
• Examples:
• Isomerase.
• Mutase.

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Example, 5. Isomerases
Catalyze isomerization reactions

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 EC 6. Ligases
• Biochemical Activity:
• Join two molecules with covalent bonds Catalyse reactions in which
two chemical groups are joined (or ligated) with the use of energy from
ATP.
• Examples:
• Acetyl~CoA Carboxylase.
• Glutamine synthetase

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• Example, 6. Ligases (synthetases)
Catalyze ligation, or joining of two substrates
Require chemical energy (e.g. ATP)

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 Any Questions?

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