TABARAK PRIVATE SCHOOL
BIOLOGY NOTE (G11)
ENZYMES
Enzymes are simple or conjugated globular proteins that catalyze thousands
of metabolic reactions within cells and organisms.
Enzymes are biological catalysts that increase the rates of reactions in the
cell.
The active site of enzymes
Active site: an area on an enzyme molecule where the substrate can bind.
In a reaction catalyzed by an enzyme, the starting substance is called the
substrate. It is converted to the product.
The way an enzyme works is for the substrate molecule to become attached
to the enzyme at a specially formed pocket in the enzyme, very briefly. This
binding point is called the active site.
Enzymes are highly specific in their action. This means catalyze only one
type of reaction or only a very small group of very similar reactions.
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The activation energy of enzymes
Activation energy: This is an energy barrier that has to be overcome before
the reaction can happen
Enzymes increase reaction rates by lowering the activation energy.
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Enzyme-Substrate Interaction
Lock-and-key model: In this model, the active site of an enzyme is thought
to be shaped to fit a particular substrate like a key fits in a lock. Only
substrates with the right shape and size will fit the active site of the enzyme.
Induced-fit model: In this model, the active site of an enzyme changes
shape due to its flexible nature when approached by substrates that are
close in shape. The substrate is not exactly the same shape as the active
site, so the enzyme changes conformation to accommodate the substrate.
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Studying enzyme-catalyzed reactions
The enzyme catalase catalyzes the breakdown of hydrogen peroxide.
Catalase occurs widely in the cells of living things. It functions as a protective
mechanism for the delicate biochemical machinery of cells. Catalase
inactivates hydrogen peroxide as soon as it is formed, before damage can
occur.
Measuring the rate of reaction
In practical terms, the rate of an enzyme-catalyzed reaction is defined as
The amount of substrate that has disappeared from a reaction mixture
in a given time. Or,
The amount of product that has accumulated in a period.
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Using a colorimeter to measure the progress of enzyme-catalyzed
reactions
In many enzyme-catalyzed reactions, the appearance (or disappearance) of
a colored compound may be observed, indicating the progress of the
reaction. Such reactions can be measured or quantified using a colorimeter.
A colorimeter is an instrument that measures the colour of a solution by
measuring the absorption of different wavelengths of light. This is a light-
sensitive instrument that measures the transmittance or absorbance of light
passing through a liquid sample. The colorimeter measures the intensity of
the color (optical density) that develops in such chemical reactions. The color
of the light can be changed by selecting a particular colored filter – the color
is chosen so that it is the frequency of light that is absorbed by the sample.
Factors Affecting Enzyme Activity
Substrate concentration: For enzyme-catalyzed reactions, increasing the
concentration of substrate increases the rate of reaction only to a point. The
point, that all the enzyme molecules become very much saturated with the
substrate, so that a further increase in substrate concentration does not
affect the rate of the reaction.
Enzyme concentration: The higher the enzyme concentration, the more the
number of collisions and the higher the number of effective collisions, hence
the higher rate of reaction. For a high substrate concentration and constant
temperature, and pH, the rate of reaction is proportional to the enzyme
concentration. For a fixed/limited substrate concentration, the rate of an
enzyme-catalyzed reaction increases with enzyme concentration and then
finally decreases when the substrate is used up.
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Presence of inhibitors: Inhibitors slow down the activity of enzymes, and
activators enhance the catalytic activities of enzymes.
There are two types of inhibitors:
Competitive inhibitor: binds to the same site as the substrate and thus
competes with the substrate for binding. Competitive inhibitors are
structurally similar to the substrate. The binding of the inhibitor can therefore
be reversed by increasing the concentration of the substrate.
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Non-competitive inhibitor: binds to a site on the enzyme other than the
active site of the enzyme. Can bind before or after the substrate.
Noncompetitive inhibitors bind at a site other than the active site of the
enzyme. They cause a conformational change which alters the catalytic
properties of the enzyme.
Michaelis–Menten constant (Kₘ) and its significance
Michaelis–Menten constant (Kₘ): the substrate concentration at which an
enzyme works at half its maximum rate (½Vₘₐₓ).
It is used as a measure of the efficiency of an enzyme by measuring the
degree of attraction or affinity of an enzyme for the substrate. The lower the
value of Kₘ, the more efficient the enzyme.
When the initial rate of reaction of an enzyme is measured over a range of
substrate concentrations (with a fixed amount of enzyme) and the results
plotted on a graph, a typical example of the resulting curve is shown in the
Figure below:
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Kₘ can be experimentally determined at a specified pH and temperature and
is expressed in units of molarity.
Using enzyme in vitro (enzyme immobilization)
Immobilised enzymes: enzymes that have been fixed to a surface or
trapped inside beads of agar gel.
The advantages of using an immobilized enzyme are:
1. It permits the re-use of the enzyme preparation.
2. The product is obtained enzyme-free.
3. The enzyme may be much more stable and long-lasting, due to
protection by the inert matrix.
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