Enzymes

Definition
• Enzymes are biological catalysts.
• They increase the rate of chemical reactions (speeds it up)
• Enzymes enables a chemical reaction to proceed under different conditions (e.g., lower temperature),
• Substrates are changed into products
• Enzymes are not used up or changed in a reaction (they can be used over and over again)
Chemical nature
•Almost all enzymes are proteins:
•Small number are RNA in nature (ribozymes): They are involved in:
1) RNA splicing: They catalyze cutting and formation of phosphodiester bond of other RNAs.
2) Protein synthesis: They catalyze the formation of peptide bonds during p
☆Protein enzymes may be simple proteins or conjugated proteins☆
• Conjugated protein enzymes are called holoenzymes.
• Holoenzyme is formed of: 1. Protein part called apoenzyme.
2. Non protein part called cofactor.

Enzyme specificity
I. Reaction specificity

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2-Substrate specificity: 4 types
Absolute Group Bond Stereospecificity
specificity (broad) specificity specificity
Enzyme act only on Enzyme act on a Enzyme act only on Enzyme act only on one
one substrate group of related certain bond of the isomers of the
substrates substrate
e.g. e.g. Hexokinase e.g.Proteolytic e.g.L-amino acid oxidase
Glucokinase Phosphorylates enzymes act on Acts on only L-a.as
phosphorylates glucose, fructose & peptide bonds D-amino acid oxidase
only glucose other hexoses Lipases act on ester Acts on only D-a.as
bond

Mechanism of action
•Michaelis and Menton proposed a hypothesis for enzyme action.
•According to their hypothesis, the following steps occurs:
1. Combination of enzyme with substrate on the active site of enzyme to form enzyme-
substrate complex which is a transient complex .
2. This complex immediately dissociates to form products and enzyme.
¤Active Site: The site in the three-dimensional enzyme structure at which the substrate
binds and is converted to product.
¤Transition state : It is the state in which the substrate is bound to the enzyme
● 2 theories to explain the substrate enzyme binding:
1. Fischer's template theory (Key& lock theory):
► The 3 dimensional structure of the
active site of the enzyme is
complementary to the substrate
(even in the absence of substrate).
► The substrate fits in the active site
of the enzyme as the key fits in its lock.
1. Induced fit theory:
► The active site of the enzyme and the substrate are not complementary
to each other in shape.
► Binding of the substrate to the active site, induces a change in shape
of the active site so that complete binding occurs.

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● Lowering of activation Energy:
► All chemical reactions need energy to get started.
► Even reactions that release energy need a boost
of energy in order to begin.
► The energy needed to start a chemical reaction
is called activation energy. Enzyme works by
reducing the amount of activation energy
needed to start the reaction. Since less energy is required to start the reaction, the rate of the
reaction is increased.
¤Activation energy: is the energy needed to convert all the molecules of reacting
substances from the ground state (enzyme unbound) to the transition state (enzyme bound).
Classification
Classification according the type of reactions catalyzed by the enzyme.
1- Oxidoreductases:
o Catalyze oxidation reduction reactions.
o Oxidation is the addition of O2 or loss of electrons or H.
o Reduction is the loss of O2 oraddition of electrons or H.
o Include four subtypes

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2- Transferases: Transfer groups between donar & acceptor molecules
e.g: Aminotransferases (Transaminases).
3. Hydrolases: Break a bond using water
(hydrolytic cleavage of bonds).
e.g: Glycosidases: hydrolyze glycosidic bond using H2O .
4. Lyase: Add or remove water, ammonia or CO2 by
cleavage of C-C,C-S 7 some C-N bonds
e.g. 1. Lyases as Citrate lyase: cleaves C-C bond.
2. Decarboxylase: removes CO2 from the substrate.
5. Isomerases: Transfer groups within the same molecule
e.g. Epimerase: catalyze interconversion of epimers
6. Ligases Form new covalent bond using high energy
from ATP
e.g. Synthetase: Enzyme that link 2 molecules in ATP- dependant
Factors affecting enzyme activity
1- substrate concentration:
● ↑ substrate conc. →↑ reaction velocity (V), till the maximum velocity
(Vmax) is reached, where any further ↑ in the substrate conc. Will not
cause increase in the reaction velocity.
● Vmax is reached when all the enzyme molecules are in the form of E-S
complex.
● Michaelis Constant (km) :
It is the substrate conc. at which the reaction velocity is 1/2 the max. velocity.
• It is constant for each enzyme.
• It is: *High → when the enzyme has lower affinity for the substrate
*Low → when the enzyme has higher affinity for the substrate
[S] α V until Vmax is reached
2- Products concentration [P]:
In reversible reaction, ↑product conc.→slowing, stoppage or even reversal of
the reaction occurs
3- Enzyme concentration [E]:
↑Enzyme conc. →↑ reaction velocity when enough substrate present
[E] α V
4- Coenzyme or activators concentration
↑Coenzyme conc. →↑ reaction velocity
[C] α V
5- Enzyme inhibitors concentration
↑enzyme inhibitor conc. →↓ reaction velocity
[I] α 1/V
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6- Temperature:
● Optimum temperature is the temperature at which the enzyme is optimally active (enzyme
velocity reaches Vmax)..
● The optimum temp. of most enzymes is 37°C.
● Any ↑ or ↓ of temp. away from the optimal temp. → ↓ enzyme activity.
● At 0°C→there is no activity of the enzyme
● Above the optimal temp, (usually above 56°C)→ Denaturation occurs with loss of enzyme
activity
7. PH:
● Optimum PH is the PH at which the enzyme is optimally active.
● Each enzyme has optimal PH.
● Any ↑ or ↓ of PH away from the optimal PH. → ↓ enzyme activity.
● Exposure to strong acidic or alkaline conditions )→ irreversible enzyme denaturation with
loss of enzyme activity

8- Allosteric regulation
● Some enzymes contain an allosteric site (other than the
active site).
● Allosteric enzymes catalyze irreversible reactions
● Allosteric enzyme frequently catalyze the early rate limiting
step (irreversible reaction) in a metabolic pathway (i.e. is the
key enzyme or the rate limiting enzyme of the metabolic
pathway)
● Allosteric effectors are usually the end products of the
pathway, that causes feed back inhibition of the allosteric
enzyme.

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