Enzymes

• Describe the major components of an enzyme (STEM_BIO11/12-II-j-17).

• Determine how factors such as pH, temperature, and substance affect
enzyme activity.
• Recognize the importance of enzymes
Chemical reaction is a process that changes one set
of chemical into another set of chemical and some
chemical reactions occur slowly, other reaction
occur quickly. Also you have learned elements or
compounds that enter into a chemical reaction are
known as reactants. The elements or compounds
produced by a chemical reaction are known as
products.
Fig. 8-13

Sucrose (C12H22O11)

Sucrase

Glucose (C6H12O6) Fructose (C6H12O6)

The Activation Energy Barrier
• Every chemical reaction between molecules
involves bond breaking and bond forming
• The initial energy needed to start a chemical
reaction is called the free energy of activation, or
activation energy (EA)
• Activation energy is often supplied in the form of
heat from the surroundings
• Enzymes catalyze reactions by lowering the E A
barrier
• Enzymes do not affect the change in free energy
(∆G); instead, they hasten reactions that would
occur eventually
Fig. 8-15

Course of
reaction
without EA
enzyme without
enzyme EA with
enzyme
is lower
Free energy

Reactants

Course of
∆G is unaffected
reaction
by enzyme
with enzyme

Products

Progress of the reaction

What is an enzyme?
• globular protein which
functions as a biological catalyst,
speeding up reaction rate by
lowering activation energy
without being affected by the
Active
site
reaction it catalyze.
• biologically active proteins that
accelerate the breakdown of
food that is eaten
• large proteins with complex,
three-dimensional structures.
• work in an aqueous environment
in our body so that the protein
chain folds such that the polar
amino acids are on the surface.
Enzymes speed up metabolic reactions
by lowering energy barriers

• A catalyst is a chemical agent that speeds up a

reaction without being consumed by the reaction
• An enzyme is a catalytic protein
• Hydrolysis of sucrose by the enzyme sucrase is an
example of an enzyme-catalyzed reaction
 Substrate Specificity of Enzymes
• The reactant that an enzyme acts on is
called the enzyme’s substrate
• The enzyme binds to its substrate,
forming an enzyme-substrate
complex
• The active site is the region on the
enzyme where the substrate binds
• Induced fit of a substrate brings
chemical groups of the active site into
positions that enhance their ability to
catalyze the reaction
 The Active Site
The folded structure for hexokinase is shown here.
• When in its proper three-
dimensional shape,
hexokinase has an indentation
on one side of the structure.
• This indentation is known as
the active site, and it is lined
with amino acid side chains. It
is a region on the surface of an
enzyme to which substrates
will bind and catalyzes a
chemical reaction.
• The active site is the functional
part of an enzyme where
catalysis occurs.
• Glucose, the reactant for
hexokinase, fits snugly in
the active site. In an
enzyme reaction, the
reactant is called the
substrate.
• Enzymes have specific
substrates, a property
known as substrate
specificity. For example,
the active site of hexokinase
reacts with
D-glucose, but will not react
with L-glucose.
• Enzymes are specific for
one enantiomer of the 12
Enzyme–Substrate Models

• A substrate is drawn into the active site by

intermolecular attractions like hydrogen bonding.

• Hydrogen bonding orients the substrate properly

within the active site.

• The initial interaction of the enzyme with the

substrate is called the enzyme–substrate
complex (ES). This complex forms prior to
catalysis.
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There are two enzyme–substrate models:
1. In the Lock-and-key model, the active site is
thought to be a rigid, inflexible shape that is an exact
complement to the shape of the substrate. The
substrate fits in the active site much like a key fits in
a lock.
2. In the induced-fit model, the active site is flexible,
has a shape roughly complementary to the shape of
its substrate, and undergoes a conformational
change, adjusting to the shape of the substrate when
the substrate interacts with the enzyme.
"Lock-and-key model"
A good example of an induced-fit model is when
hexokinase and glucose form an enzyme–
substrate complex as shown.
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 Some enzymes, like hexokinase, have non-protein
helpers. Two categories of helpers are as follows:
– Cofactors are inorganic substances like magnesium ions
but sometimes they can be organic which is consist of
organic groups that are permanently bound to the enzyme
(prosthetic groups) and cations - positively charged
metal ions (activators), which temporarily bind to the
active site of the enzyme, giving an intense positive charge
to the enzyme's protein
– Coenzymes are small organic molecules derived from
vitamins which are not permanently bound to the enzyme
molecule, but combine with the enzyme-substrate complex
temporarily. Riboflavin found in the coenzyme flavin
adenine dinucleotide (FAD) is a coenzyme.
 Mechanism of enzyme action
The enzymatic reactions takes place by binding of
the substrate with the active site of the enzyme
molecule by several weak bonds.

Formation of ES complex is the first step in the

enzyme catalyzed reaction then ES complex is
subsequently converted to product and free
enzyme.
• In this equation, the enzyme name is written
above or below the reaction arrow.

• The phosphate group is represented by a P in a

circle.

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Catalysis in the Enzyme’s Active Site

• In an enzymatic reaction, the substrate binds to

the active site of the enzyme
• The active site can lower an EA barrier by
– Orienting substrates correctly
– Straining substrate bonds
– Providing a favorable microenvironment
– Covalently bonding to the substrate
The active site and catalytic cycle of an
enzyme
e.g. H2O2

e.g. O2 + H2O

Progress of Reaction
Classes of Enzyme
Classes of Enzyme
Effects of Local Conditions on Enzyme Activity

• An enzyme’s activity can be affected by

– General environmental factors, such as
temperature and pH
– Concentration of substrate and enzyme
– Inhibitors
Effects of Temperature
• Each enzyme has an optimal temperature in which it can
function. As the temperature rises, reacting molecules
have more and more kinetic energy. This increases the
chances of a successful collision and so the rate increases.
There is a certain temperature at which an enzyme’s
catalytic activity is at its greatest. This optimal temperature
is usually around human body temperature for the
enzymes in human cells.
• Above this temperature the enzyme structure begins to
break down (denature) since at higher temperatures intra-
and intermolecular bonds are broken as the enzyme
molecules gain even more kinetic energy.
• Optimum temperature, the temperature at which enzymes
activity is greatest
• At high temperatures, enzymes denature, which
modifies the active site.

• At low temperatures, enzyme activity is low due

to a lack of energy for the reaction to occur.

• Food is stored in a refrigerator or freezer to slow

spoilage brought on by enzymes.

• Boiling contaminated water will destroy enzymes

in bacteria that are present in the water.
Effects of pH
• . Each enzyme works within quite a small pH range. There
is a pH at which its activity is greatest (the optimal pH). This
is because changes in pH can make and break intra- and
intermolecular bonds, changing the shape of the enzyme
and, therefore, its effectiveness.
• Changes in pH can disrupt weak interaction that hold
proteins together and can cause an enzymes to denature
(lose their precise shape). Once an enzymes has
denatured, it will not function again.
Effects of pH
• Each enzyme has an optimum pH. Above or below an
enzyme’s optimum pH, its activity is lower. The optimum pH
of a particular enzyme corresponds to the pH of its natural
environment. For many enzymes, this corresponds to pH
values of around 7. For pepsin, which is active in the
stomach, the optimum pH is 2 (the pH of the stomach).
Trypsin, which is active in the small intestine, has an
optimum pH of 8 that matches the pH of the small intestine.
• Changes in pH will also affect the nature of the
amino acid side chains in the active site.

• The optimum pH for enzymes is based on the

location of the enzymes as shown:
Effects of Concentration of enzyme and substrate
• The rate of an enzyme-catalyzed reaction depends on the
concentrations of enzyme and substrate.
• For a given enzyme concentration, the rate of reaction
increases with increasing substrate concentration up to a
point, above which any further increase in substrate
concentration produces no significant change in reaction
rate. This is because the active sites of the enzyme
molecules at any given moment are virtually saturated with
substrate.
• The enzyme/substrate complex has to dissociate before the
active sites are free to accommodate more substrate.
Provided that the substrate concentration is high and that
temperature and pH are kept constant, the rate of reaction is
proportional to the enzyme concentration.
Effects of Concentration of enzyme and substrate

• Increasing substrate concentration will

not affect the rate of the reaction.

• A condition known as steady state is

when an enzyme is operating under
maximum activity.
Effect of Inhibitors
• Some substances reduce or even stop the catalytic
activity of enzymes in biochemical reactions. They
block or distort the active site. These chemicals are
called inhibitors, because they inhibit reaction.
• Inhibitors that occupy the active site and prevent a
substrate molecule from binding to the enzyme are
said to be active site-directed (or competitive, as
they 'compete' with the substrate for the active
site).
• Inhibitors that attach to other parts of the enzyme
molecule, perhaps distorting its shape, are said to
be non-active site-directed (or noncompetitive).
 Enzyme Inhibition

• Any substance that can diminish the velocity

of an enzyme catalyzed
• These include drugs, antibiotics, poisons,
and anti-metabolites.
• Useful in understanding the sequence of
enzyme catalyzed reactions, metabolic
regulation, studying the mechanism of cell
toxicity produced by toxicants.
• Forms the basis of drug designing.
• Reversible inhibition occurs when the
inhibitor causes a temporary loss of
activity. However, activity is regained if
the inhibitor is removed.

• Reversible inhibitors can be competitive

or noncompetitive.

• Competitive inhibitors are molecules

that compete with a substrate for the
active site, and have a structure similar
to the substrate.
• As long as an inhibitor remains in the
active site, the enzyme cannot react with
the substrate to form product.
• An example of a medical therapy that involves a
competitive inhibitor involves liver alcohol
dehydrogenase (LAD). This enzyme oxidizes
ethanol, the alcohol found in alcoholic
beverages.

• This enzyme will also react with ethylene glycol

and methanol, which are found in antifreeze,
and will compete with ethanol for the active site.

• If a pet is poisoned by drinking antifreeze, a slow

intravenous infusion of ethanol is administrated.

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• Administration of ethanol slows the production of
the toxic metabolites of ethylene glycol and
methanol, giving the kidneys time to eliminate
these two substrates.
• Noncompetitive inhibitors do not resemble the
substrate. They do not compete for the
enzyme’s active site. They bind to another part
of an enzyme, causing the enzyme to change
shape and making the active site less effective
• Noncompetitive inhibitors bind at a site on the
enzyme that is usually remote to the active site.
• Examples of inhibitors include toxins, poisons,
pesticides, and antibiotics
•
• When a noncompetitive
inhibitor binds to an enzyme, it
causes a conformational
change in the enzyme. This
change in shape causes the
active site to no longer interact
with the substrate.

• As long as this type of inhibitor

is bound to the enzyme, it will
no longer function effectively.
• This figure diagrams how a
noncompetitive inhibitor
functions.

41
Antibiotics Inhibit Bacterial Enzymes

• Enzyme inhibitors are used to fight bacterial

infections.

• Penicillin is an example of an irreversible inhibitor.

It binds to the enzyme that bacteria use to
synthesize cell walls, and slows the growth of cell
walls.

• Without a cell wall, bacteria cannot survive and the

infection stops.