Effect of Temperature on Enzymatic Reaction

As temperature increases so do the rate of enzyme reactions. A ten degree centigrade

rise in temperature will increase the activity of most enzymes by 50% to 100%. Variations
in reaction temperature as small as 1 or 2 degrees may introduce changes of 10% to 20%
in the results. This increase is only up to a certain point until the elevated temperature
breaks the structure of the enzyme. Once the enzyme is denatured, it cannot be repaired.
As each enzyme is different in its structure and bonds between amino acids and peptides,
the temperature for denaturing is specific for each enzyme. Because most animal
enzymes rapidly become denatured at temperatures above 40°C, most enzyme
determinations are carried out some what below that temperature.

Figure 1. Effect of temperature on reaction rate.

enzymes structure
Enzymes are proteins comprised of amino acids linked together in one
or more polypeptide chains. This sequence of amino acids in a
polypeptide chain is called the primary structure. This, in turn,
determines the three-dimensional structure of the enzyme, including
the shape of the active site.A
Types of Enzymes:
The biochemical reactions occurring in the body are basically of 6 types and the enzymes
that bring about these reactions are named accordingly:
Oxidoreductases: These enzymes bring about oxidation and reduction reactions and
hence are called oxidoreductases. In these reactions, electrons in the form of hydride ions
or hydrogen atoms are transferred. When a substrate is being oxidized, these enzymes act
as the hydrogen donor. These enzymes are called dehydrogenases or reductases. When
the oxygen atom is the acceptor, these enzymes are called oxidases.
Transferases: These enzymes are responsible for transferring functional groups from one
molecule to another. Example: alanine aminotransferase which shuffles the alpha‐amino
group between alanine and aspartate etc. Some transferases also transfer phosphate
groups between ATP and other compounds, sugar residues to form disaccharides such as
hexokinase in glycolysis.
Hydrolases: These enzymes catalyze reactions that involve the process of hydrolysis.They
break single bonds by adding water. Some hydrolases function as digestive enzymes
because they break the peptide bonds in proteins. Hydrolases can also be a type of
transferases as they transfer the water molecule from one compound to another.
Example: Glucose-6-phosphatase that removes the phosphate group from glucose-6-
phosphate, leaving glucose and H3PO4.
Lyases: These enzymes catalyze reactions where functional groups are added to
break double bonds in molecules or where double bonds are formed by the
removal of functional groups. Example: Pyruvate decarboxylase is a lyase that
removes CO2 from pyruvate. Other examples include deaminases and
dehydratases.
Isomerases: These enzymes catalyze the reactions where a functional group is
moved to another position within the same molecule such that the resulting
molecule is actually an isomer of the earlier molecule. Example: triosephosphate
isomerase and phosphoglucose isomerase for converting glucose 6-phosphate to
fructose 6-phosphate.
Ligases: These enzymes perform a function that is opposite to that of the
hydrolases. Where hydrolases break bonds by adding water, ligases form bonds by
removal of the water component. There are different subclasses of ligases which
involve the synthesis of ATP.
mechanism of action and factors affecting the activity of them
For any reaction to occur in the universe, there is an energy
requirement. In cases where there is no activation energy provided,
a catalyst plays an important role to reduce the activation energy
and carried forward the reaction. This works in animals and plants
as well. Enzymes help reduce the activation energy of the complex
molecules in the reaction. The following steps simplify how an
enzyme works to speed up a reaction:

Step 1: Each enzyme has an ‘active site’ which is where one of the
substrate molecules can bind to. Thus, an enzyme- substrate
complex is formed.

Step 2: This enzyme-substrate molecule now reacts with the second

substrate to form the product and the enzyme is liberated as the
second product.

There are many theories that explain how enzymes work. But, there
are two important theories that we will discuss here.
Theory 1: Lock and Key Hypothesis
This is the most accepted of the theories of
enzyme action.
This theory states that the substrate fits exactly into the active site of the enzyme to
form an enzyme-substrate complex. This model also describes why enzymes are so
specific in their action because they are specific to the substrate molecules.
Theory 2: Induced Fit Hypothesis
This is similar to the lock and key hypothesis. It says that the shape of the enzyme molecule
changes as it gets closer to the substrate molecule in such a way that the substrate
molecule fits exactly into the active site of the enzyme.
What factors affect enzyme activity in the cell?
Concentration of Enzymes and Substrates: The rate of reaction increases with increasing
substrate concentration up to a point, beyond which any further increase in substrate
concentration produces no significant change in reaction rate. This occurs because after a
certain concentration of the substrate, all the active sites on the enzyme are full and no
further reaction can occur.
Temperature: With the increase in temperature, the enzyme activity increases because of
the increase in kinetic energy of the molecules. There is an optimum level when the
enzymes work at the best and maximum. This temperature is often the normal body
temperature of the body. When the temperature increases beyond a certain limit, enzymes,
which are actually made up of proteins, begin to disintegrate and the rate of reaction slows
down.
pH: Enzymes are very sensitive to changes in the pH and work in a very small window of
permissible pH levels. Below or above the optimum pH level, there is a risk of the enzymes
disintegrating and thereby the reaction slows down.
Inhibitors: Presence of certain substances that inhibit the action of a particular enzyme. This
occurs when the inhibiting substance attaches itself to the active site of the enzyme thereby
preventing the substrate attachment and slows down the process.
Diagram displaying the structure of an enzyme, including its
active site to which a substrate molecule binds to form an
enzyme–substrate complex. When the reaction has
occurred, products are released from the active site.
Diagram displaying the difference between a
normal enzyme and denatured enzyme.
The temperature changes induced by evaporation
Use a thermometer strip to examine temperature changes when drops
of different liquids evaporate

In this practical, students will take three liquids, ethoxyethane, dionised water and ethanol,
and discover the rate of evaporation for each in relation to temperature.

This experiment should take 10 minutes.

Equipment
Apparatus
Student worksheet
Temperature strip
Pipettes
Chemicals
Ethoxyethane (diethyl ether)
Deionised water
Ethanol
(NB Do not use propanone for this experiment – it attacks the temperature strip)
Health, safety and technical notes
Read our standard health and safety guidance.
Students must wear eye protection.
Ethoxyethane (diethyl ether) and ethanol are both highly FLAMMABLE (see CLEAPSS
Hazcard HC042, HC040a).
Ethoxyethane is also a respiratory irritant and harmful if swallowed.
Procedure
Put a row of drops of water along the strip. Note the shape of the drops and note whether
there are any temperature changes over the next few minutes.
Repeat using ethanol.
Repeat using ethoxyethane.
Record your observations in a table and try to give explanations. Bear in mind what you
know about intermolecular forces when you interpret your findings.
Observations
Water, which forms well-defined droplets, produces very little, if any, change in
temperature since the rate of evaporation is slow due to the high degree of
hydrogen bonding.
With ethanol, the drops spread out and a fall in temperature will be noted due to
the higher rate of evaporation. With ethoxyethane the drops evaporate very quickly
and a marked drop in temperature is observed.
This is consistent with the low boiling point and absence of hydrogen bonding
between the molecules. The energy changes accompanying changes of state are an
important concept in science. One example is the addition of ice to cool drinks. Here
it is the melting of ice that cools the drink rather than the contact of ice with the
liquid.
Effects of pH
Enzymes are affected by changes in pH. The most favorable pH value - the
point where the enzyme is most active - is known as the optimum pH. This is
graphically illustrated in Figure 14.

Extremely high or low pH values generally result in complete loss of activity for most
enzymes. pH is also a factor in the stability of enzymes. As with activity, for each enzyme
there is also a region of pH optimal stability.
 The optimum pH value will vary greatly from one enzyme to another, as Table II
shows:

In addition to temperature and pH there are other factors, such as ionic strength, which can
affect the enzymatic reaction. Each of these physical and chemical parameters must be
considered and optimized in order for an enzymatic reaction to be accurate and reproducible.