Enzymes

What are Enzymes?
Enzymes are proteins that function as
biological catalysts. A catalyst is a substance
that increases the rate of a chemical reaction and
is not changed by the reaction. These enzymes
are made in living cells and thus are known as
biological. From this information, we can state
that:
Enzymes are made in the body to speed up
reactions that take place in the body without
getting changed or used up.
As enzymes are made up of proteins, they will
need amino acids as building blocks for them.
Almost all metabolic processes (catabolic –
breaking down, and anabolic – building up
processes) require enzymes.
catalyst. The active site is the point at which a
substrate attaches to an enzyme.
The substrate (molecule on which the enzyme
acts upon) is the key to a lock (the enzyme).
The whole idea of the lock and key mechanism is
known as the lock and key hypothesis derived
by the German scientist Emil Fischer.
In the presence of the enzyme, the substrate
attaches to the enzyme and will either break
(catabolic) or build up (anabolic) into products.
There may be multiple substrates or products or
even 1 of each. The product will separate from
the enzyme. If 2 products are formed (such as
fatty acids and glycerol broken down from fats by
lipase) the products will also separate from each
other.

Enzymes and Metabolism

Metabolism is the sum of all catabolic and

anabolic processes that take place in a living
body.

Catabolic Anabolic
Enzymes break complex Enzymes synthesize
substrates to simple complex products from
products simple substrates
These reactions break These reactions build up
down and release energy and use energy
(exothermic) (endothermic)
Respiration is an Photosynthesis is an
example: breaks down a example: takes in energy
complex sugar (glucose) from sunlight and
into simpler compounds produces a complex Enzymes are specific to one particular substrate
and releases energy sugar (glucose) as the enzyme is a complementary shape to the
substrate.
The production of enzymes will be anabolic as
they are built from amino acids in cells. When the substrate moves into the enzyme’s
active site, they become known as the enzyme-
Where are Enzymes Produced? substrate complex. After the reaction has
occurred, the products leave the enzyme’s
All enzymes are produced within cells. Most are active site as they no longer fit it and it is free to
intracellular: work within the cell but some are take up another substrate.
extracellular: released from cells and work
outside these cells (example: digestive enzymes
such as lipase).

Enzyme Action

Each enzyme is specific to a substrate (protease

is specific to proteins).

An enzyme is a 3D protein molecule with an

active site which allows the enzyme to act as a
Enzymes are specific due to the shape of its
active site. After the reaction takes place, the
enzymes remain unchanged as they are
catalysts.
The diagram below shows the action of both
anabolic and catabolic enzyme action:
Note that anabolic reactions are also known as
synthesis (condensation) reactions and
catabolic reactions may be known as hydrolysis
if water is involved in the breakdown.
Summary of Enzyme Action:
1) Enzymes and substates randomly move
about in solution
2) When an enzyme and its complementary
substrate randomly collide – with the
substrate fitting into the active site of the
enzyme – an enzyme-substrate complex
forms, and the reaction occurs.
3) A product forms from the substrate which is
then released from the active site. The
enzyme is unchanged and will go on to
catalyse further reactions.
Factors Affecting Enzyme Activity
Temperature:
 When the temperature is lower than the
optimum (ideal) temperature, fewer
collisions of enzymes and substrates
occur as there is lesser energy in the
molecules.
 As the temperature increases, substrate
molecules move faster and collide with
greater energy, making it more likely for
binding at the active site.
 The optimum temperature is where
enzymes work best as most collisions
are successful.
 At high temperatures, there is more
energy, but the active site starts to
deform due to vibrations, so fewer
collisions take place (losing ability to
bind). This is called the denaturing of
enzymes.
The optimum temperature for enzymes to work
in:
 Humans: 35 to 40°C – below this, works
slow; above this, denaturing takes place.
 Plants: 25 to 30°C – below this, works
slow; above this, denaturing takes place.
This can be plotted on a graph:
Power of Hydrogen (pH):
The above scale is known as the pH Scale. It
shows the levels of acidity or how alkaline a
substance is. Most enzymes work best around
neutral (6.5 to 8) pH, though there are some
which need acidic or slightly alkaline conditions.
Hydrochloric acid is produced in the stomach for
the enzyme pepsin (a protease) to work at
optimum ph. This acid will pass through the
intestine, thus hydrogen carbonates (alkaline)
are produced in the pancreas and passed on to
make sure the pH in the intestine is neutralized
and to give amylase and lipases its ideal pH.
When it comes to pH, enzymes only work at its
optimum pH and will be denatured below or
above optimum as the active sites shape gets
changed and then fewer collisions take place.
Note that the pH of the enzyme is not 2 (pepsin)
or 7.5 (amylase), but it is the best pH it works at.
Activators and Inhibitors:
These are molecules that affect how an enzyme
binds to a substrate:
 Activators or coenzymes make binding
of enzymes more likely (Chlorine ions
are present and needed along with
salivary amylase).
 Inhibitors make it harder for enzymes to
bind to each substrate (Cyanide ions are
considered as poisonous as they block
the active sites of enzymes involved in
respiration which inhibits this process
and can lead to death).
Biology in Industries
Ammonifying bacteria can process ammonia at
regular temperature and pressure (RTP) rather
than using Haber’s Process of producing
ammonia which requires a higher RTP.
Enzymes are biocatalysts and will work at regular
RTP and pH as found in living organisms.
Therefore:
 If enzymes are used for industrial processes
such as making ammonia, then extreme and
expensive conditions won’t be needed.
 Enzymes can be obtained by microbes.
Note: Microbes secrete enzymes on substrates
during extracellular digestion. Using this
knowledge, scientists have created a device
known as a bio-fermenter or bioreactor to obtain
enzymes and maybe ammonia from microbes as
they secrete them.
microbes from the food/ enzymes and the
microbes may be reused or returned for the
process to occur once again; and preparing
products for sale (involves drying or
crystallization). Sometimes, a tap may be
present to control the flow of output.

The diagram below shows a bioreactor:
In a bioreactor, the culture (nutrients and
microbes that are inputted) must be pure (1
microbe type only) and the equipment and
nutrients must be sterile (cleaned) to avoid
competition for expensive nutrients (if any other
microbe manages to get inside) and to reduce
diseases (or spreading of toxic waste products of
microbes which causes microbes to contaminate
the product – decrease in yield).

Batch and Continuous Culture:

Products may be produced in either of the 2
ways.

What is it? Advantages Disadvantages

Batch
Culture:
1 batch of If any
nutrients and contamination
The parts of a bio-fermenter have specific uses: microbes are occurs, only 1
inputted, the batch is
 Microbe inlet/input: The organisms that will It may be
process takes spoiled, the
carry out fermentation (anaerobic) or other expensive as
place and culture is pure
aerobic processes are inputted from here. the fermenter
product is due to
 Nutrient inlet/input: The microbes require needs to be
collected and constant
energy (carbohydrates) and growth materials sterilized and
kept separate sterilization
(amino acids) which are inputted here. replacements
from other and the
are needed
 Paddle stirrer: They continuously mix the product, the fermenter can
which need a
contents of the bioreactor so that: microbes fermenter is be used in
cost, thus not
and nutrients are always in contact; ensures empty and different ways
being efficient.
even temperature throughout and for aerobic sterilized and with different
reactions the mixing occurs from an the process is microbes
airstream which also provides oxygen. allowed to (larger variety
 Hot or Cool Water jacket: This maintains the repeat over of product)
temperature all around the reactor so that and over
enzymes work at its best conditions. The again.
temperature remains constant all throughout.
 Gas outlet: Carbon dioxide and other gases Efficient and Contamination
may be produced from respiration. These Continuous economical as results in
may be harmful for the bacteria so there is a Culture: the fermenter greater losses
gas outlet which allows the gas to escape and In this, is properly and there may
to avoid pressure building up in the reactor. If microbes and used up. be blocks at
this gas is carbon dioxide, then it may be nutrients are inlet or outlet
collected and used to make fizzy drinks. continuously pipes. There
 Probes (Sensors): They monitor conditions inputted so can only be 1
in the reactor as they are connected to there are process taking
computers/ monitors. They will adjust the longer periods place and thus
conditions if there are any non-suitable of running. If a small variety
conditions/ non-optimum conditions. there is a of products.
 Further processing outlet: Further shortage of
processing may be necessary to separate any nutrients
or microbes,
 more can be
placed in.

Recombinant DNA Technology uses

Enzymes

The steps of this process are:

(1) Plasmid take from bacterial cell and cut

open using restriction endonuclease
enzyme.
(2) Gene responsible for production of
insulin is taken out of a healthy human
using the same enzyme.
(3) The gene is combined to the plasmid
using an enzyme known as ligase.
(4) Recombinant plasmid placed back into
the bacterial cell.
(5) Selected bacteria are cultured in a bio-
fermenter under optimum conditions and
are allowed to reproduce (asexually) so
that more insulin is collected.