Enzymes

Mechanism and its

Regulation.
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Introduction
Enzymes, the catalysts of biological systems are
remarkable feature that determine the patterns of
chemical transformations.
They also mediate the transformation of one form of
energy into another.
The most striking characteristics of enzymes are
their catalytic power and specificity.
Nearly all known enzymes are proteins.
Thats why Enzymes are the functional unit of cell
metabolism.
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Historical Background
In 1835, a Swedish chemist Jon
Jacob Berzelius predict the
existence of biological catalyst.
In 1860 Louise pasture recognize
that, Enzyme were necessary for
fermentation.
In 1878 German physiologist Wilhelm
Khne first used the term Enzyme.

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Naming of enzyme
The common name of the enzyme create a great
confusion, like ATPase is an Enzyme that help in
breaking down ATP, whereas ATP synthase is an
enzyme that help in synthesis of ATP.
Thats why Enzyme commission has create the
systematic basis of Enzyme nomenclature , on the
basis of reaction it catalyzed.
For example, EC 2.7.1.2
The first three number denotes Major class , sub-
class, sub-sub class and last number is the serial
number in the sub class.
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Classification of Enzyme is:
EC 1 Oxidoreductases: catalyze oxidation/
reduction reactions .
EC 2 Transferases: transfer a functional group
(e.g. a methyl or phosphate group).
EC 3 Hydrolases: catalyze the hydrolysis of various
bonds .
EC 4 Lyases: cleave various bonds by means other
than hydrolysis and oxidation.
EC 5 Isomerases: catalyze isomerization changes
within a single molecule.
EC 6 Ligases: join two molecules with covalent
bonds.
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Chemical Nature of Enzymes?
Most enzymes are
Proteins (tertiary and
quaternary structures)
Act as Catalyst to
accelerates a reaction
Not permanently
changed in the process
Are specific for what
they will catalyze
Are Reusable

Enzymes are powerful and highly
specific catalyst
Specificity of an
enzyme is due to the
precise interaction of
the substrate with
enzyme.
This precision is a
result of intricate 3-D
structure of the
enzyme protein.

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Specific Catalytic groups
One substrate is bound to an enzyme, there is a
formation of bonds by variety of mechanisms, i.e.

(1) General Acid-Base catalysis,

(2) Covalent catalysis, and

(3) Metal ion catalysis.
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Acid-Base catalysis
Acid base catalysis involves the proton transfer.
There are two kinds----
(I) Specific Acid-base catalysis Hydronium ion and
hydroxide ion act directly as the acid base group.

(II) General acid-base catalysis- into this donor and
acceptor of proton are other then hydronium and
hydroxide ion.
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Covalent catalysis
In the Covalent catalysis involves the
formation of a reversible covalent bond
between enzyme and substrate.
The proteolytic enzyme Chymotrypsin
provides an excellent example of this
mechanism.
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Metal ion catalysis
It is also called Lewis Acid-base catalysis.
Many enzymes require metal ion for catalytic
activity. For example,

Cu
2+
Cytochrome oxidase
K Pyruvate kinase
Ni Urease
Mg Hexokinase, glucose 6-phosphatase,pyruvate kinase

Etc..

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How do enzymes Work?
Enzymes work by
weakening bonds
which lowers
activation energy
Define the following terms:
1. Catabolic reactions:

2. Metabolism:

3. Catalyst:

4. Metabolic pathway:

5. Specificity:

6. Anabolic reactions:

7. Substrate:

8. Product:
Reactions that build up molecules
Reactions that break down molecules
Combination of anabolic and catabolic
reactions
Sequence of enzyme controlled reactions
Only able to catalyse specific reactions
The molecule(s) the enzyme works on
Molecule(s) produced by enzymes
A substance that speeds up reactions without
changing the produced substances
Reaction coordinate diagram for a chemical reaction

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Enzymes affect reaction rates, not Equilibria
A simple Enzymatic reaction
may be written as:-
E + S ES EP E + P
Where, E = Enzyme,
S = Substrate, and, P = Product.

ES and EP are transient
complexes of the enzyme
with substrate and with the
product respectively.
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Mode of Action
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"Lock and key" model
Enzymes are very specific,
because both the enzyme
and the substrate possess
specific complementary
geometric shapes that fit
exactly into one another.
This is often referred to
as "the lock and key"
model.
However, while this model
explains enzyme
specificity, it fails to
explain the stabilization
of the transition state
that enzymes achieve.

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Induced Fit Model
A change in the configuration of
an enzymes active site (H+ and
ionic bonds are involved).
Induced by the substrate.

Regulation of Enzyme Action
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In cellular metabolism, groups of enzymes work
together in sequential pathways to carry out a
given metabolic process.In such enzyme systems,
the reaction product of one enzyme becomes the
substrate of the next.
These regulatory enzymes exhibit increased or
decreased catalytic activity in response to
certain signals.
The activities of regulatory enzymes are
modulated in a variety of ways..

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1.Allosteric Enzymes Undergo Conformational
Changes in Response to Modulator Binding.

2.Regulatory Enzymes Undergo Reversible
Covalent Modification.

3.Phosphoryl Groups Affect the Structure and
Catalytic Activity of Proteins.

4.Regulation of enzymes action by feedback
inhibition.


Allosteric Enzymes Undergo
Conformational Changes in Response to
Modulator Binding:-
Regulation of enzymes action by
feedback inhibition: -
The rate of production of the pathways end product is thereby
brought into balance with the cells needs. This type of regulation
is called feedback inhibition
Regulation of Enzymes Activity in Plant
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Regulation of NADP-malate degydrogenase in C4
plant:Effect of varying NADPH to NADP ratios
and thioredoxin redox state on enzyme activity in
reconstituted systems.Activation and inactivation
of NADP-malate dehydrogenase purified from
Zea mays leave were folowed in a reconstituted
system provided with thioredoxin poised in
various redox states with dithiothreitol.The
initial rate of activation or in activation of
NADP-malate dehydrogenase was proportional of
reduces or oxidized thioredoxin,respectively.
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As would be predicted from these values, high
proportions of active malate dehydrogenase were
developed only in the presence of very high ratios
of reduced to oxidized thioredixin. Similarly,when
pyridine nucleotide was included,a high degree of
acyivate of malate dehydrogenase was only
observed with high NADPH/NADP ratio as well as
the thioredoxin redox state may be critical in
determining the level of NADPH-malate
dehydrogenase activity in vivo.
Thank YOU..
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