SPECIFICITY

OF
ENZYMES
 Introduction
• A group of very important proteins,
known as enzymes, play a critical role in
breaking down food in your body.
• Enzymes are biological catalysts— this
means they help to accelerate the rate of
metabolic reactions in the cells by
reducing the activation energy of the
reactants.
• Enzymes are proteins, and hence, they
have a specific 3D conformation as a
result of the unique twisting and folding
of the polypeptide chain. Therefore,
enzymes are highly specific.
 What is Enzyme Specificity ?
• The ability of an enzyme to bind to a particular
substrate only, and thus, it will only be able to catalyse a
particular reaction .
• This specificity is a molecule recognition mechanism and
it operates through the conformational and structural
complementarity between the enzyme and substrate.
Types Of Enzyme Specificity
Bond Specificity:
Enzymes that show bond specificity are
specific to substrates having similar bonds
and structure.
Therefore, they bind to substrates that
contain specific bonds only, such as ester
bonds, glycosidic bonds and peptide bonds.

Example
α-amylase can only hydrolyses α-1,4-
glycosidic bonds in starch and glycogen, and
not any other types of bonds.
Group Specificity:
Group specificity is that the enzyme will act only on
molecules that have specific functional groups, such
as amino, phosphate and methyl groups.
Exopeptidase and Endopeptidase are classical
examples.
 Substrate Specificity:
• Substrate specificity is also known as absolute specificity.
• Enzymes showing substrate specificity are only specific to
one substrate and one reaction.
• This can be explained through the lock and key theory,
which states that the enzyme has a rigid active site which
can only fit substrates with the complementary 3D
configuration, explaining the high specificity of such
enzymes.
• Enzyme urease catalyses the
decomposition of urea. This
specificity is so pronounced that a
given enzyme may catalyse a
particular reaction of one
stereoisomer without affecting the
other.
• Glucokinase is specific for its
substrate glucose only.
Cofactor Specificity
Co-factors are the non-protein parts of
some enzymes, and are required for the proper
functioning of these enzymes.
Enzymes which require co-factors for their activity
show co-factor specificity.
The enzyme can only catalyze a reaction in the
presence of the correct substrate and co-factor.
In the absence of the specific co-factor, the enzyme
will be inactive, even if many substrates are
available.
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Geometrical Specificity
In geometrical specificity, an enzyme can bind
to different substrate that have similar
molecular geometry.
Therefore, specificity is lower in this case. An
example of such an enzyme is alcohol
dehydrogenase, which is able to catalyze both
methanol and ethanol since they have similar
molecular geometry.
Optical Specificity
Optical specificity of enzymes is also known as stereo-
specificity.
Enzymes showing optical specificity are not only specific
to the substrate, but also to its optical configuration
For such enzymes, the spatial arrangement of the
substrate is critical in determining if the enzyme will be
able to catalyze the reaction.
An example of such an enzyme showing optical specificity
is the amino acid oxidase.
L-amino acid oxidase can only bind specifically to L-amino
acids.
Conversely, D-amino acid oxidase can
only catalyze reactions involving D-amino acids.
Another Good example for optical and stereo
specificity is amylase α -glycosodic bonds of starch and
glycogen are hydrolyzed only by α -glycosidase (α -
amylase ).
Similarly β-glycosidic bonds of cellulose are
hydrolyzed only by β-glycosidase (β-amylase ).
• Enzymes react stereo
specifically with chiral
compounds
Enzymes are chiral reagents and display stereospeficity
In an enzyme catalyzed reaction, the substrate specifically
fits and binds to the active site in an enzyme. The following
points may be noted:
Enzymes are chiral and enantiomerically homogeneous and
provide a chiral cavity at the active site. Optically inactive
starting materials can give optically active products if they
are treated with an optically active reagent or if the reaction
is catalyzed by an optically active substance.
Recall that enzymes being chiral (biological catalyst) typically
catalyze reactions of only one of the stereoisomers of a
given molecule e.g., only one of the enantiomers. Thus
enzymes recognize only D-sugars and not L-sugars for
example. This discrimination is termed as stereospecificity.
Chirality: Three dimensional structure of
Molecules.
In chemistry context – chirality is applicable to three
dimensional structure of molecules.

1. A pair of isomers having identical physical and chemical

properties,
2. Rotate the plane polarized light equally but in opposite directions,
3. And their molecules are non-superimpossible mirror images

They have identical chemical and physical properties in the absence

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of an external chiral influence. i.e. same m. p.; b. p.; solubility;
CONFIGURATION: The spatial arrangement of atoms or
groups of atoms around a chiral carbon is known as its
configuration.
Ex: R & S

CONFORMATION: Different spatial arrangements of

atoms or groups of atoms in a molecule that can be
readily interconverted by rotation around C-C single
bond are called conformers.
Ex: Butane: Anti, Eclipsed, Gauche or skew, Fully
eclipsed.

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 Preparation of chiral compounds:
A) Resolution of enantiomers:

• Unlike enantiomers , diastereomers have different physical

properties.
• A racemic compound can be resolved if converted to a
separable pair of diastereomers by reacting it with naturally
occurring enantiomerically pure compound.

Diastereomers are formed as:

1. Ionic salts : from carboxylic acids and chiral tert-amines
such as brucine, cinchona alkaloids.
2. Amides : carboxylic acids/derivatives with chiral amines
or vice-versa.
3. Esters : carboxylic acids with chiral alcohols or vice-
Enantiomers have similar chemical properties and react with
achiral reagents at the same rate. For example, hydroxide
ion an achiral reagent, reacts with (R)-2-bromobutane at the
same rate that it reacts with (S)-2-bromobutane.