Enzymes: Introduction

Dr. Anam Hameed

Assistant Professor
Riphah College of Rehabilitation and Allied Health
Sciences
Riphah International University, Lahore, Pakistan
 Enzymes

•Almost every reaction that happen in the body involves enzymes

•They function as a catalyst from physiological reaction
•Enzymes control in a selective manner reactants in chemical
reactions
•These reactants are called substrate
•Consequently, Enzymes mediate all metabolic interactions in the
body
 Chemical nature
 Enzymes are biological catalysts that speed
up the rate of the biochemical reaction.

 Most enzymes are three dimensional globular

proteins (tertiary and quaternary structure).

 Some RNA also act

enzymesand
special Ribozymes as
hammerhead ribozyme.
species are e.g.
called

Hammerhead enzyme
 STRUCTURE OF ENZYMES
 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.
 Active Sites
Substrate molecule:
Substrate molecules are
the chemicals that an
enzyme acts on. They
Active site:
are drawn into the cleft
The active site contains both binding
of the enzyme.
and catalytic regions. The substrate is
drawn to the enzyme’s surface and the
substrate molecule(s) are positioned in
a way to promote a reaction: either
joining two molecules together or
Enzyme molecule: splitting up a larger one.
The complexity of the active
site is what makes each
enzyme so specific (i.e.
precise in terms of the
substrate it acts on).

This model (above) is an enzyme called

Ribonuclease S, that breaks up RNA molecules.
It has three active sites (arrowed).
Properties of enzymes
Enzymes
Properties: 1. Active site specificity

•Each enzyme has a unique empty space within its structure named
the active site
•The active site contains amino acids portion that participate in the
substrate interactions to form enzyme–substrate (ES) complex
•This binding catalyzes the substrate via structural transformation
 ACTIVE SITE
o Active site can be further divided into:

Active Site

Binding Site Catalytic Site

It chooses the substrate It performs the catalytic

and binds it to active site. action of enzyme.

Enzymes
Properties: 1. Active site
Enzymes
Properties: 2. Catalytic efficiency

•Efficiency is one of the most important properties of enzymes

•When enzymes are not catalyzing a reaction, it takes 108 longer
•The amount of byproduct that results from the catalytic reaction is
called the turnover number
•This number range between 102-104 product per second
Enzymes
Properties: 3. Coenzymes and Cofactors

•Some enzymes can not facilitate a catalytic reaction with a substrate

without the presence of certain molecules
•When these molecules composed of metallic ions such as Zn2+ or
Fe2+, it is called cofactors
•On the other hand, if the extra molecules organic in nature it is
denoted as coenzymes. E.g. vitamins B1
 CO-
o Co-factor is the non FACTORS
protein molecule which carries out chemical
reactions that can not be performed by standard 20 amino acids.

o Co-factors are of two types:

 Organic co-factors
 Inorganic cofactors
 INORGANIC CO-FACTORS
o These are the inorganic molecules required for the
proper
activity of enzymes.
++
 Examples:
Enzyme carbonic anhydrase requires Zn for it‟s activity.
 Hexokinase has co-factor Mg
++

ORGANIC CO-FACTORS
o These are the organic molecules required for
the proper activity of enzymes.
Example:
 Glycogen phosphorylase requires the small
organic molecule pyridoxal phosphate.
 TYPES OF ORGANIC CO-FACTORS
Prosthetic Group Coenzyme

o A prosthetic group is a tightly bind with enzyme … can be organic or

inorganic (heme, metal ion)
o A coenzyme is loosely attached with enzyme….totally organic e.g.

Flavins, NAD+, biotin.

 Continued…
Types of co-factors

 An with it‟ co-factor removed is designated

enzyme s as
apoenzyme.
 The complete complex of a protein with all necessary small organic
molecules, metal ions and other components is termed as
holoenzyme of holoprotein.
Enzymes
Properties: 4. Location

•Enzymatic reactions take place in specific location within a cell

•This aspect of enzyme compartmentalization provide the specify of
enzymes
•Since there are thousands of enzymes within a cell, isolating
enzymes from their substrates into specific organelles helps in
organizing the huge number of pathways
 INTRACELLULAR AND
EXTRACELLULAR ENZYMES
o Intracellular enzymes are synthesized and retained in the cell for
the use of cell itself.
o They are found in the cytoplasm, nucleus, mitochondria and
chloroplast.
Example :
 Oxydoreductase catalyses biological oxidation.
 Enzymes involved in reduction in the mitochondria.

o Extracellular enzymesare in the

synthesized secreted from the cell to work cell but
externally.
 Example
Digestive :
enzyme produced by the pancreas, are not used by the
cells in the pancreas but are transported to the duodenum.
Enzymes
Properties: 4. Location
 SUBSTRAT
E
 The reactant in biochemical reaction is termed as substrate.

 When a substrate binds to an enzyme it forms an enzyme- substrate

complex.

Substrate Joins Enzyme

 SITES OF ENZYME SYNTHESIS
o Enzymes are synthesized by ribosomes which are attached to the
rough endoplasmic reticulum.

o Information for the synthesis of enzyme is carried by DNA.

o Amino acids are bonded together to form specific

enzyme
according to the DNA‟s codes.
 NOMENCLATURE OF ENZYMES
o An enzyme is named according to the name of the substrate it
catalyses.
o Some enzymes were named before a systematic way
of
naming enzyme was formed.
Example: pepsin, trypsin and rennin
Recommended name
o By adding suffix -ase at the end of the name of
the
substrate, enzymes are named.
o Enzyme for catalyzing the hydrolysis is termed as hydrolase.
Example :
maltase
maltose + water glucose + glucose
 EXAMPLES
substrate enzymes products
lactose lactase glucose + galactose
maltose maltase Glucose
cellulose cellulase Glucose
lipid lipase Glycerol + fatty acid
starch amylase Maltose
protein protease Peptides +
polypeptide
Nomenclature “Systematic name”

•According to this nomenclature, all enzymes are classified under 6

classes
•For any enzyme, the suffix “ase” is part of the chemical modification
initialed by the enzyme
Nomenclature
• synthetase (requires ATP),

• synthase (no ATP required);

• phosphatase (uses water to remove phosphoryl group),

phosphorylase (uses Pi to break a bond and generate a
phosphorylated product);

• dehydrogenase (NAD+/FAD is an electron acceptor in a

redox reaction),

• oxidase (O2 is the acceptor, and oxygen atoms are not

incorporated into substrate),

• oxygenase (one or both oxygen atoms are incorporated).

CLASSIFICATION
 CLASSIFICATION OF ENZYMES
 A systematic classification of enzymes has been developed by
International Enzyme Commission.

 This classification is based on the type of reactions catalyzed by

enzymes.

 There are six major classes.

 Each class is further divided into sub classes, sub sub-classes and so
on, to describe the huge number of different enzyme- catalyzed
reactions.
 Classification of enzymes Continued……..

ENZYME CLASS REACTION TYPE EXAMPLES

Oxidoreductases Reduction-oxidation Lactate
(redox) dehydrogenase
Transferases Move chemical group Hexokinase
Hydrolases Hydrolysis; bond Lysozyme
cleavage with transfer
of functional group of
water
Lysases Non-hydrolytic bond Fumarase
cleavage
Isomerases Intramolecular group Triose phosphate
transfer isomerase
(isomerization)
Ligases Synthesis of new RNA polymerase
covalent bond
between substrates,
using ATP hydrolysis
Classification
 CHARACTERISTICS
 Enzymes speed up the reaction by lowering the
activation energy of the reaction.
 Their presence does not effect the nature and properties of
end product.
 They are highly specific in their action that is each enzyme
can catalyze one kind of substrate.
 Small amount of enzymes can accelerate chemical reactions.
 Enzymes are sensitive to change in pH, temperature
and substrate concentration.
 Turnover number is defined as the number of
substrate molecules transformed per minute by one enzyme
molecule.

Catalase turnover number = 6 x106/min

 Two models have been proposed to explain how an enzyme
binds its substrate: the lock-and –key model and the
induced-fit model.

Lock-and-Key Model of Enzyme-

Substrate Binding In this model, the
active site of the unbound enzyme is
complementary in shape to the substrate.

"lock and key model" accounted for the

exquisite specificity of enzyme- substrate
interactions, the implied rigidity of the
enzyme's active site failed to account for the
dynamic changes that accompany catalysis.
Induced-Fit Model of
Enzyme-Substrate Binding In
this model, the enzyme changes
shape on substrate binding. The
active site forms a shape
complementary to the substrate
only after the substrate has
been bound. When a substrate
approaches and binds to an
enzyme they induce a
conformational change, a
change analogous to placing a
hand (substrate) into a glove
(enzyme).
  Enzymes are catalysts and increase the speed of a chemical
reaction without themselves undergoing any permanent chemical change. They
are neither used up in the reaction nor do they appear as reaction products.
 The basic enzymatic reaction can be represented as follows

 where E represents the enzyme catalyzing the reaction, S the substrate, the
substance being changed, and P the product of the reaction.
FACTORS AFFECTING RATE OF ENZYME
CATALYZED REACTIONS

 Temperature
 Hydrogen ion concentration(pH)
 Substrate concentration
 Enzyme concentration
EFFECT OF TEMPERATURE

 Raising the temperature increases the rate of enzyme catalyzed reaction

by increasing kinetic energy of reacting molecules.
 Enzymes work maximum over a particular temperature known as
optimum temperature. Enzymes for humans generally exhibit stability
temperature up to 35-45 ᵒ C.
 The temperature coefficient is a factor Q₁₀ by which the rate of biological
processes increases for a 10 ᵒ C increase in temperature.
 For most biological processes Q₁₀ = 2.
 However some times heat energy can also increase kinetic energy to a
point that exceed the energy barrier which results in denaturing of
enzymes.
 5- 40oC Temperature
Increase in Activity

40oC - denatures

Rate of Reaction

0 10 20
30 40
50 60

<5oC - inactive
The rate of almost all enzyme-catalyzed reactions
exhibits a significant dependence on hydrogen ion
concentration
 Most intracellular enzymes exhibit optimal
activity at pH values between 5 and 9.
 The relationship of activity to hydrogen ion
concentration reflects the balance between enzyme
denaturation at high or low pH and effects on the
charged state of the enzyme, the substrates, or both.
EFFECT OF PH

 Rate of almost all enzymes catalyzed reactions depends on pH

 Most enzymes exhibit optimal activity at pH value between 5
and 9
 High or low pH value than optimum value will cause ionization of
enzyme which result in denaturation of enzyme
Except for Pepsin, acid phosphatase and alkaline phosphatase, most
enzyme have optimum pH between 5 to 9
As the amount of enzyme is
increased, the rate of reaction
increases. If there are more
enzyme molecules than are
needed, adding additional enzyme
will not increase the rate. Reaction
rate therefore increases as enzyme
concentration increases but then it
levels off.
 At lower concentrations, the active sites on most of the
enzyme molecules are not filled because there is not much
substrate. Higher concentrations cause more collisions
between the molecules. The rate of reaction increases (First
order reaction).
 The maximum velocity of a reaction is reached when the
active sites are almost continuously filled. Increased
substrate concentration after this point will not increase
the rate. Reaction rate therefore increases as substrate
concentration is increased but it levels off (Zero order
reaction)
The shape of the
curve that relates Zero order
activity to substrate reaction
concentration is
hyperbolic.

First order
reaction