CC2

Enzymology
Definition of Terms
• Apoenzyme: The protein part of an enzyme
without the cofactor necessary for catalysis.
• Catalyst: A substance that increases the rate
of a chemical reaction, but is not consumed
or changed by it. An enzyme is a biocatalyst
• Activator: An effector molecule that increases
the catalytic activity of an enzyme when it
binds to a specific site
Definition of Terms
• Catalytic Activity: The property of a catalyst
that is measured by the catalyzed rate of
conversion of a specified chemical reaction
produced in a specified assay system.
• Coenzyme: A diffusible, heat-stable substance
of low molecular weight that, when
combined with an inactive protein called an
apoenzyme ,forms an active compound or a
complete enzyme called a holoenzyme
Definition of Terms
• Enzyme: A PROTEIN molecule that catalyzes
chemical reactions without itself being
destroyed , altered ,consumed .
• Holoenzyme: The functional compound
formed by the combination of a apoenzyme
and its appropriate coenzyme
• Inhibitor: An inhibitor is a substance that
diminishes the rate of a chemical reaction;
the process is called inhibition.
Definition of Terms
• Isoenzyme: One of a group of relate enzymes
catalyzing the same reaction but having
different molecular structures and
characterized by varying physical, biochemical,
and immunological properties.
• International Unit: The amount of enzyme that
catalyzes the conversion of one micromole of
substrate per minute under the specified
conditions of the assay method.
Definition of Terms
• Product: The substance produced by the
enzyme-catalyzed conversion of a substrate.
• Substrate: A reactant in a catalyzed reaction.
Enzyme Nomenclature
• All enzymes are assigned to one of six classes
Characterized by the type of reaction they catalyze
1.Oxidoreductase
2.Transferase
3.Hydrolases
4.Lyases
5. Isomeraseas
6. Ligase
Enzymes based on Type of reaction:
1. Oxidoreductases are a class of enzymes that
catalyze
the transfer of electrons from 1 molecule ( oxidant) to
another molecule (reductant). This process often
requires co-factors such as NAD(P)H.

, A– + B → A + B –

Example: Lactate dehydrogenase

Enzymes based on Type of reaction:
Enzymes based on Type of reaction:
2. Transferases are enzymes that catalyse the
movement of a functional group from one molecule
to another. These functional groups are very diverse
can include phosphate, methyl and glycosyl groups.

AX + B --> A + BX
A= the donor
B= the acceptor
X= the functional group.
Enzymes based on Type of reaction:
Two Important sub-groups
1.Kinases- enzymes are involved in catalysing the
transfer of phosphate groups in a process called
phosphorylation.
Creatine Kinase or Creatine phosphokinase
2. Deaminases- enzymes that catalyse the transfer an
amine group.
Alanine Aminotransferase or Glutamate Pyruvate
Transaminase
Enzymes based on Type of reaction:
3. Hydrolase enzymes simply catalyse hydrolysis; the
breaking of single bonds through the addition of
water.
Example:
Lipase - break down lipids into fatty acids and glycerol
by cleaving ester bonds
 
Enzymes based on Type of reaction:
4. Lyases -catalyse lysis reactions that generate a
double bond.
•Type of elimination reaction but are not hydrolytic or
oxidative.
•The reverse reaction catalyses an addition reaction,
where a substrate is added to a double bond.
•Often referred to as synthase enzymes.
ATP <--> cAMP + Ppi
•1 substrate is required in the forward direction,
whereas 2 are needed for the backward reaction.
Example: Carboxylase
Aldolase
Enzymes based on Type of reaction:
5. Isomerases are enzymes that can catalyse structural
changes within a molecule.
There is only one substrate and one product with
nothing gained or lost, so they represent only a change
in shape.

Alanine racemase
It converts the amino acid alanine between its two
optical isomers. It is used in both alanine and
aspartate metabolism.
D-Alanine <--> L-Alanine
Enzymes based on Type of reaction:
6. Ligases are responsible for the catalysis of
ligation; the joining of two substrates.

• Usually chemical potential energy is required, so

the reaction is coupled to the hydrolysis of a
disphosphate bond in a nucleotide triphosphate
such as ATP.
Enzyme Nomenclature
International Union of Biochemistry- developed a
systematic and trivial name for enzymes.
Systematic name describes the nature of the reaction
catalyzed
L-Lactate:NAD+ oxidoreductase
Enzyme Commission Numerical nomenclature: is
associated with a unique numerical code designation
consisting of 4 numbers separated by periods + EC
( Enzyme Commission) prefixed.
1st number-class of reaction, next 2- subclass
Last 2 – sub sub class
EC1.1.1.27
Enzyme Nomenclature
Trivial or practical name, which may be identical to
the systematic name but is a
•simplification of it, is suitable for everyday use

Lactate dehydrogenase
Enzyme Nomenclature
• The use capital letter abbreviations for the names of
certain enzymes,
Eg. ALT for Alanine aminotransferase
AST for Aspartate aminotransferase
LD for Lactate dehydrogenase
CK for Creatine kinase
Characteristics of Enzymes?
Enzymes
1)speed up chemical reactions
2)are required in minute amounts
3)are highly specific in their action
4)are affected by temperature
5)are affected by pH
6)Some catalyse reversible reactions
7)Some require co-enzymes
8)Are inhibited by inhibitors
 Enzymes Speed up chemical reactions

Energy

Activation
Activation Energy Energy without
with enzyme enzyme

Substrate

Products:
Time

• By lowering the activation energy needed to

start the reaction.
 Enzymes are required in minute
amounts
Chemically unchanged

Sucrase
Sucrose Glucose + Fructose

• They remain chemically unchanged after

catalysing the reactions.
• The same enzyme molecules can be reused
over again.
• Therefore, only a small amount of enzyme is
required to catalyse a large number of reactions
 Enzymes are highly specific
Amylase
Starch Maltose

Maltose Maltase Glucose + Glucose

• Each chemical reaction is catalysed by its own

specific, unique enzyme.
• This is due to every enzyme’s specific 3-d
configuration.
• How the shape of an enzyme affects its function can
be explained by the “LOCK & KEY HYPOTHESIS”.
• Enzymes are proteins and each has an active site.
• Active sites are depressions on the surface of
enzyme molecules with specific shape and
specific charges.
• Active sites are formed due to the way the proteins
are folded and held together by H-bonds.
• Every active site has it’s own specific shape and
charges.
• It only allows substrates with a
COMPLEMENTARY shape and charge to bind
with it to form an Enzyme-Substrate Complex
Enzyme

Substrate Enzyme-Substrate Complex Products + Enzyme

Anabolic Reaction - Synthesis

Catabolic Reaction - Hydrolysis

• The subcomplementary active site to form the
enzyme-substrate complex
• The enzyme-substrate complex brings about the
necessary reactions
• The product separates from the enzyme, leaving
the enzyme molecule unchanged and free to strate
binds to a free enzyme with a combine again with
more substrate molecules
 Enzymes are affected by temperature

• Enzymes can function • But all enzymes have

over a range of their own optimum
temperatures. temperature.
 Optimum temperature
• The optimum temperature is the temperature at
which the enzyme is most active, catalysing the
largest number of reactions per second.
• Different enzymes have different optimum temp.
• Example:
most enzymes in the human body
functions best at about 37oC, near body
temperature.
Enzymes of thermophilic bacteria that
live in hotsprings will have very high
optimum temperatures.
 How enzyme activity is affected by
NOTE:
temperature
Describe the • Beyond
Increasing temp beyond
the optimum optimum
temperature, ratetemp.
of
enzyme activity with enzyme activity deceases.
respect to the Rate- • Until it is completely denatured by the extreme
Temperature Graph heat

• Increase in temperature increases the

vibrations of the atoms in the enzymes.
• Beyond the OT, the vibrations are so
violent that they break the hydrogen bonds
that hold the 3-D structrure in place.
• The enzyme loses its shape and active site
• The enzyme is DENATURED.
 How enzyme activity is affected by
temperature
• Beyond the optimum temperature, rate of
• Most enzymes are
enzyme activity deceases.
INACTIVATED at 65C
• Until it is completely denatured by the extreme
LOW temperature or
heatFREEZING does NOT usually
destroy enzymes.

Except:
LD – Freezing at -20 destroys activity BUT freezing
at -70 DOES NOT.

CK- Storing at 4C lose its activity.

Enzymes are affected by pH

• Different enzymes have

different optimum pH.
• Enzymes are affected
by the acidity or
alkalinity of the solutions
• M indicates the optimum
pH when the rate of
reaction is the highest
 Different enzymes have different
optimum pH
Examples:
•Enzymes that work best at ACIDIC conditions
 Renin and Pepsin: Found in the stomach

•Enzymes that work best at ALKALINE conditions

 Intestinal Enzymes

•Enzymes that work best at NEUTRAL conditions

 Amylase
Different enzymes have different
optimum pH
 How enzyme activity is affected by pH
At pH slightly above or below the optimum

• Enzyme activity reduces when the

conditions are slightly more acidic or
alkaline than the optimum pH

• Slight changes in the pH brings about reversible

changes
• Changes can be restored by bringing enzyme back to
the optimum pH
 How enzyme activity is affected by pH
At extreme pH conditions

• Enzymes are DENATURED

• Extreme changes in the pH causes:

 A change in the charges at the active sites which
repels the substrate molecules, preventing them from
binding.
 The irreversible alteration to the bonds that holds the
shape of the enzyme. Enzyme thus loses its original
3-D structure. The active site loses its shape.
 Some enzymes catalyze reversible
reactions
Reactants Products

A + B C + D
Products Reactants
• Some enzymes catalyse both reactions until equilibrium
is reached.
• Reactions will proceed in the direction where the products
are constantly being removed
• E.g formation of glucose during photosynthesis
 Some enzymes require co-enzymes

• Some enzymes require co-enzymes(organic co-

factor) to be bound to them in order to ENHANCE
before they can catalytic reactions
Co-enzymes
• Small, non-protein, organic “helper” molecules
• Example of Co enzyme- NAD
Co-enzyme –is an inorganic co factor.
When BOUND tightly to ENZYME, the co enzyme
is called PROSTHETIC GROUP.
The enzyme( apoenzyme)+ coenzyme=
HOLOENZYME
 Enzymes are inhibited by inhibitors

Type 1 (Competitive)
Substrate is PREVENTED
from binding to active site
by inhibitor
Type 2 (Non-competitive)
Binding of inhibitor does not
prevent the binding of substrate
but SLOWS DOWN reaction speed