TOPIC 2.

5 ENZYMES

U.1. Enzymes have an active site to which specific substrates bind.

Enzyme: An enzyme is built from proteins folded in complicated shapes and they are present
throughout the body, the chemical reactions that develop inside of our bodies rely on the
enzymes.

Substrate: A substrate is a molecule that acts with an enzyme. Enzymes catalyse

chemical reactions involving the substrates. The substrate bonds with the enzyme
active site, and that creates an enzyme-substrate complex.

Catalysis process.

1. In the first step the polar regions of amino acids attract the substrate and the active site
of the enzyme.
2. The active site is the surface of an enzyme to which substrates bind and that catalyses
the reaction.
3. Once the substrate has been bind, the reaction is catalysed.
4. The products are released and the enzyme is used again.

The lock and key theory

● This theory says that the substrate and the active site are specific to each other. They
match each other in 2 ways.

Structurally: The structure of the active site is specific

to the substrate. Substrates that don´t fit, don't react.

Chemically: Substrates that are not chemically

attracted to the active site, won't react
Induced-fit model
 ● If the lock and key theory was true then the enzyme could only catalyse one reaction,
but in reality enzymes can catalyse more than one fractions. The induced fit theory says
that as the substrate approaches the enzyme it provoqueis a conformational change in
the active site, this meaning that it changes the shape in order for the substrate to fit.

● This stresses the substrate, reducing the activation energy of the reaction.

U.2. Enzyme catalysis involves molecular motion and the collision of substrates with the active
site.

1 2 3

The substrate binds The substrates bind The products

to the active site. to the active site separate from the
Some enzymes have change into different active site, leaving it
two substrates that chemical making it possible for
bind to different parts substances, which more substrates to
of the active site are the products of bind again.
the reaction
● Enzyme activity is the catalysis of a reaction by an enzyme. These are the three steps

A substrate molecule can only bind to the active site if it moves very close to it. The getting
together of a substrate molecule and an active site is known as collision. With most reactions
the substrates are dissolved in water around the enzyme. Because of water being a liquid, the
particles dissolved in it are in contact with each other and are in constant motion. Both
substrates and enzymes are able to move, either way as substrates are typically known to be
smaller than enzymes they tend to move faster.
Collisions between substrates and active sites occur because of random movement of the
substrate and the enzyme. The successful collisions are those in which the substrate and active
site are in site, allowing the binding to occur.

U.4. Enzymes can be denatured.

Enzymes, like other proteins can have their structure altered by certain conditions. This process
called denaturation can be caused by high temperatures and either high or low pH.

When an enzyme is denatured, the active site is altered, so the substrate can no longer bind,
and if it binds the reaction probably won't occur. In many cases denaturation causes enzymes
that were dissolved in
water to become
insoluble and form a
precipitate.

U.3. Temperature, pH and substrate concentration affect the rate of activity of enzymes.

Enzyme activity is affected by temperature in two ways

1. In liquids, the particles are in constant random motion. When the liquid is heated the
particles in it are given more kinetic energy. Both the enzymes and the substrates move
faster in higher temperature thus increasing the the chance of a substrate molecule
colliding with the active site of the enzyme. Enzyme activity increases.
 2. When enzymes get heated, bonds of the enzyme vibrate, making them more weak,
which increases the probability of the
enzyme´s bonds to break. When the
bonds inside the enzymes break, the
structure of the enzyme changes, and
with it, the active site. This change is
permanent and is called denaturation.
When the enzyme is denatured, it can
no longer get back to its original state,
which means it can’t catalyse reactions.
As more enzymes get denatured
enzyme activity falls, when the enzyme
is completely denatured the activity
stops altogether.

Enzymes are affected by pH

Most enzymes have an optimum pH in which their

activity is in its highest peak. If the pH is increased or
decreased from that optimum, enzyme activity
decreases and then stops altogether, when this
happens the structure of the enzyme is altered,
including its active site. Beyond a specific pH the
structure is irreversible altered, this is another example
of denaturation.
Not all enzymes share the same optimum pH, actually
the range is pretty large. This reflects the wide range of
pH environment in which enzymes work.
The image by the left side shows the wide range of pH
optimus in different places where enzymes function

Enzyme activity is affected by substrate

concentration.

If the concentration of substrates increases, substrate-active site collisions will happen more
frequently and the rate at which the enzyme catalyses the reaction will increase. However, after
the binding of the substrate to an active site, the active site will be unavailable to other substrate
molecules until a product is formed and released. As the concentration rises, more active sites
will be unavailable. This means that the rate in which enzymes catalyse reactions get smaller as
substrate concentration rises.
The graph below shows how the relationship between substrate concentration and enzyme
activity works.

U.5 Immobilized
enzymes are widely used in
industry.

In 1897 the Buchner brothers, showed that an extract of yeast, containing no yeast cells, would
convert sucrose into alcohol. More than 500 enzymes now have commercial uses, the graph
below shows a classification of commercially useful enzymes.

The enzymes used in industries are

normally immobilized. This means the
attachment of the enzymes to another
materia or into aggregations, so that
movement of the enzyme is restricted.
There are many ways of doing this,
attachment of enzymes in glass,
trapping the enzymes in an alginate gel,
or bonding them together.

Advantages of immobilized enzymes

Concentration of substrate can be increased as the enzyme is not dissolved.

Recycled enzymes can be used many times
O-
Separation of products is straightforward
Stability of the enzymes, as changes in the pH and temperature can be controlled
A.1. Methods of production of lactose free milk and its advantages.

Lactose is a sugar that is naturally present in milk, it can be converted into glucose and

galactose by the enzyme lactase: lactose → glucose + galactose.

Reasons for using lactase in the production of food.

1. Some people are lactose intolerant and cannot drink more than 250ml of milk per day,
unless it's lactose reduced.
2. Galactose and glucose are sweeter than lactose, so less sugar needs to be added to
sweet food containing milk, such as milkshakes and yogurts.
3. Bacteria ferments glucose and galactose more quickly than lactose, so the production of
yogurt and cottage cheese is faster.

The image in the left illustrates the

process of producing lactose free milk

S.1. Design of experiments to test the effect of temperature, pH and substrate concentration on
the activity of enzymes.

S.2. Experimental investigation of a factor affecting enzyme activity.

Catalyse is one of the most widespread enzymes, it catalyses the conversion of hydrogen
peroxide, a toxic-by-product of metabolism, into water and oxygen.

1. You are going to investigate the independent variable, you need to decide:
 ● How you are going to vary it, example, with substrate concentration you would
obtain a solution with the highest concentration and dilute it to get lower
concentrations.
● What units will you use to measure the independent variable
● What range do you need for the independent variable, highest and lowest value.

2. The variable you use to find out how fast the enzyme catalyses is the dependent variable
you need to decide:

● How are you going to measure it

● What unit should be used to measure the dependent variable.
● How many repetitions do you need to get reliable results.

3. You also need to be aware of the controlled variables., You need to decide

● What are the control variables

● How can you keep them constant
● What level should they be kept at, for example temperature.