5.
Enzymes
5.1 Enzymes
5.1.1 Enzymes
What Are Enzymes?
Enzymes are:
o Catalysts that speed up the rate of a chemical reaction without being
changed or used up in the reaction
o Proteins
o Biological catalysts (biological because they are made in living cells,
catalysts because they speed up the rate of chemical reactions without
being changed)
o 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
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: lock and key model of enzyme activity
5.1.2 Enzyme Investigations
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 temperature on enzyme activity
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 pH
Investiga
ting the effect of pH on enzyme activity
Exam Tip
Describing and explaining experimental results for enzyme experiments is a common
type of exam question so make sure you understand what is happening and, for a 7, 8
or 9, can relate this to changes in the active site of the enzyme when it has denatured,
or if it is a low temperature, relate it to the amount of kinetic energy the molecules
have.
5.1.3 Enzyme Action & Specificity: Extended
Enzyme Action & Specificity: Extended
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
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.
5.1.4 Enzymes & Temperature: Extended
Enzymes & Temperature: Extended
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 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
Graph showing the effect of temperature on the rate of enzyme activity
5.1.5 Enzymes & pH: Extended
Enzymes & pH: Extended
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
Effect of pH on enzyme activity
Graph showing the effect of pH on rate of activity for an enzyme from the
duodenum
