Enzymes
UNIT 5
Learning Objectives
What Are Enzymes?
• Catalysts are substances which speeds up a chemical reaction without being changed
itself
• Enzymes are protein in nature
• Enzymes re biological catalysts (biological because they are made in living cells,
catalysts because they speed up the rate of chemical reactions without being
changed)-speed up metabolic reactions inside the body
• Enzymes are necessary to all living organisms as they maintain reaction speeds of
all metabolic reactions (all the reactions that keep an organism alive) at a rate that
can sustain life
• For example, if we did not produce digestive enzymes, it would take around 2 – 3
weeks to digest one meal; with enzymes, it takes around 4 hours ( reaction times
are increased)
What are Enzymes?
Enzymes are:
• Catalysts that speed up the rate of a chemical
reaction without being changed or used up in
the reaction
• Proteins
• Biological catalysts (biological because they
are made in living cells, catalysts because they
speed up the rate of chemical reactions
without being changed)
• Necessary to all living organisms as
they maintain reaction speeds of all metabolic
reactions (all the reactions that keep an
organism alive) at a rate that can sustain life
• For example, if we did not produce digestive enzymes, it would
take around 2 - 3 weeks to digest one meal; with enzymes, it
takes around 4 hours
The lock and key hypothesis
• The substance which is present at the beginning of the
reaction= substrate
• The substance which is made at the end is called the
product
• E.g. saliva contains amylase which catalyses the
breakdown of starch to maltose;
• Starch=substrate
• Maltose=product
• An enzyme works by allowing the molecule of the substance
(substrate) on which it is acting to fit into it
• The fit has to be perfect
• Enzyme= lock
• Substrate= key
How do Enzymes Work?
• Enzymes are specific to one particular substrate (molecule/s that get broken
down or joined together in the reaction) as the active site of the enzyme has a
complementary shape to the substrate
• The product is made from the substrate(s) and is released
Enzyme specificity: lock
and key model of
enzyme activity
How do enzymes work?
Enzyme substrate
specificity
• Enzymes are specific to one
particular substrate (molecule/s
that get broken down or joined
together in the reaction) as the
enzyme is a complementary
shape to the substrate
• The product is made from the
substrate(s) and is released
•
Enzyme Specificity
Enzymes are specific to one particular substrate(s) as
the active site of the enzyme, where the substrate attaches,
is a complementary shape to the substrate
• This is because the enzyme is a protein and has a specific 3-
D shape
• This is known as the lock and key hypothesis
• When the substrate moves into the enzyme’s active site they
become known as the enzyme-substrate complex
• After the reaction has occurred, the products leave the
enzyme’s active site as they no longer fit it and it is free to
take up another substrate
1. Enzymes and substrates 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 (or products) forms from the substrate(s) which are then released from
Naming Enzymes
• Named according to the reaction that they catalyse
e.g.
enzymes which catalyse the breakdown of
carbohydrates are called carbohydrases
enzymes which break down proteins are called
proteases
if they break down fats/lipids- lipases
Sometimes they are given specific names e.g.
enzyme which breaks down starch is called amylase
One which breaks down maltose is maltase
One which breaks down sucrose is sucrase
Example
• All enzymes have active sites
• Each enzyme has an active site which exactly fits its
substrate
• This means that each enzyme can only act on a
particular kind of substrate
• Amylase cannot break down protein molecules
because they do not fit in its active site
1. The substrate molecule (starch) fits into the active site
2. Each enzyme has an active site where the substrate fits exactly
3. The starch molecule is split into maltose and the enzyme is
unaltered at the end of the reaction, ready to accept another
Properties of Enzymes
1. All enzymes are proteins
2. Enzymes are made inactive by high temperatures
(because they are protein molecules which are damaged
by heat)
3. Enzymes work best at a particular temperature
(enzymes in the human body work best at 37oC)
4. Enzymes work best at an optimum pH
5. Enzymes are catalysts- unchanged in the chemical
reaction which they control; can be used over and over
again
6. Enzymes are specific- each kind of enzymes only
catalyses one kind of chemical reaction
Effect of Temperature on Enzyme
• Most chemical reactions happen faster Function
at higher
temperatures because the molecules have more kinetic
energy (enzyme is likely to collide with substrate more
often)
• Enzymes are proteins and have a specific shape, held in
place by bonds
• This is extremely important around the active site area as
the specific shape is what ensures the substrate will fit
into the active site and enable the reaction to proceed
• Enzymes work fastest at their ‘optimum temperature’ –
in the human body, the optimum temperature is 37⁰C
• Heating to high temperatures (beyond the optimum)
will break the bonds that hold the enzyme together and it
will lose its shape -this is known as denaturation
• Substrates cannot fit into denatured enzymes as the shape
of their active site has been lost
• Denaturation is irreversible – once enzymes are
denatured they cannot regain their proper shape and
Effect of Temperature on
• Enzyme
Increasing the temperature from Function
0⁰C to the optimum increases the
activity of enzymes as the more
energy the molecules have the
faster they move and the
number of collisions with the
substrate molecules increases,
leading to a faster rate of reaction
• This means that low
temperatures do not denature
enzymes, they just make them
work more slowly
Enzymes &
Temperature
• Enzymes are proteins and have a specific shape, held
in place by bonds
• This is extremely important around the active site area
as the specific shape is what ensures the substrate will
fit into the active site and enable the reaction to
proceed
• Enzymes work fastest at their ‘optimum temperature’ –
in the human body, the optimum temperature is 37⁰C
• Heating to high temperatures (beyond the optimum)
will break the bonds that hold the enzyme together
and it will lose its shape -this is known as denaturation
• Substrates cannot fit into denatured enzymes as the
shape of their active site has been lost
• Denaturation is irreversible - once enzymes are
denatured they cannot regain their proper shape and
activity will stop
Effect of pH on Enzyme
• Function
The optimum pH for most enzymes
some that are produced in acidic
is 7 but
conditions e.g. protease, such as the
stomach because of HCl has a lower
optimum pH (pH 2) and some that are
produced in alkaline conditions, such as
the duodenum, have a higher optimum pH
(pH 8 or 9)
• If the pH is too high or too low, the bonds
that hold the amino acid chain together to
make up the protein can be destroyed
• This will change the shape of the active
site, so the substrate can no longer fit into NEVER USE THE WORD “KILLED” for
it, reducing the rate of activity enzymes as they are not living organisms
• Moving too far away from the optimum pH
will cause the enzyme to denature and
activity will stop
Effect of pH on Enzyme
In the graph above, as the pH
Function
increases so does the rate of
enzyme activity. An optimum
activity is reached at the
enzyme’s optimum pH, pH 8
in this example. A continued
increase in pH results in a
sharp decrease in activity as
the enzyme’s active site
changes shape. It is now
denatured. Graph showing the effect of pH on rate of activity for an enzyme
from the duodenum
Effect of substrate
concentration on enzymatic
activity
Enzymes will work best if there is plenty of
substrate. As the concentration of the
substrate increases, so does the rate of
enzyme activity. However, the rate of enzyme
activity does not increase forever. This is
because a point will be reached when the
enzymes become saturated (there are not
enough enzyme molecules available to break
down the excess substrate molecules) and no
more substrates can fit at any one time even
though there is plenty of substrate available.
Enzyme Investigations
• Amylase is an enzyme that digests starch (a polysaccharide of glucose) into
maltose (a disaccharide of glucose).
• Starch can be tested for easily using iodine solution.
Effect of temperature
on enzyme activity
• Increasing the temperature from 0⁰C to the
optimum increases the activity of enzymes
as the more energy the molecules have the
faster they move and the number of
collisions with the substrate molecules
increases, leading to a faster rate of reaction
• This means that low temperatures do not
denature enzymes, they just make them work
more slowly
Effect of pH on
Enzyme Activity
The optimum pH for most enzymes is 7 but some
that are produced in acidic conditions, such as the
stomach, have a lower optimum pH (pH 2) and
some that are produced in alkaline conditions,
such as the duodenum, have a higher optimum pH
(pH 8 or 9)
If the pH is too high or too low, the bonds that hold
the amino acid chain together to make up the
protein can be destroyed
This will change the shape of the active site, so the
substrate can no longer fit into it, reducing the
rate of activity
Moving too far away from the optimum pH will
cause the enzyme to denature and activity will
stop
Investigating the Effect
of Temperature on
Amylase
• Starch solution is heated to a set temperature
• Iodine is added to wells of a spotting tile
• Amylase is added to the starch solution and mixed
well
• Every minute, droplets of solution are added to a
new well of iodine solution
• This is continued until the iodine stops turning
blue-black (this means there is no more
starch left in the solution as the amylase has
broken it all down)
• Time taken for the reaction to be completed is
recorded
• Experiment is repeated at different temperatures
• The quicker the reaction is completed, the faster the
enzyme is working
Investigating the Effect
of pH on Amylase
Place single drops of iodine solution in rows on the tile
Label a test tube with the pH to be tested
Use the syringe to place 2cm3 of amylase in the test tube
Add 1cm3 of buffer solution to the test tube using a syringe
Use another test tube to add 2cm3 of starch solution to the
amylase and buffer solution, start the stopwatch whilst mixing
using a pipette
After 10 seconds, use a pipette to place one drop of mixture on
the first drop of iodine, which should turn blue-black
Wait another 10 seconds and place another drop of mixture on
the second drop of iodine
Repeat every 10 seconds until iodine solution remains orange-
brown
Repeat experiment at different pH values - the less time the
iodine solution takes to remain orange-brown, the quicker all
the starch has been digested and so the better the enzyme
works at that p
Thermostatically controlled
water bath
Error in step 1: Uneven
temperature/ temperature does
not remain constant throughout
the experiment
Improvement: Thermostatically
controlled Water bath
Effect of pH on Amylase
• activity
The different pHs under investigation will be
produced using buffer solutions. Buffer
solutions produce a particular pH, and will
maintain it if other substances are added.
• A series of test tubes containing a mixture of
starch and amylase is set up at different pHs.
• A sample is removed from the test tubes every
10 seconds to test for the presence of
starch. Iodine solution will turn a blue/black
colour when starch is present, so when all the
starch is broken down, a blue-black colour is no
longer produced. The iodine solution will remain
orange-brown.
• A control experiment must be set up – without
the amylase – to make sure that the starch
would not break down anyway. The result of the
control experiment must be negative – the
colour must remain blue-black – for results with
How enzymes work
1. Enzymes and substrates 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 (or products) forms from the
substrate(s) which are then released from the
active site. The enzyme is unchanged and will go
on to catalyse further reactions
List four chemical elements that are found in proteins. 1. 2. 3. 4. [4]
0610 S 2014/ Paper 31
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