Ch.
5 Enzymes
What Are Enzymes?
● Enzymes are:
○ Catalysts that speed up the rate of a chemical reaction without being
changed or used up in the reaction
○ Enzymes are 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
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 Action & 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
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.
Factors that affect Enzymes
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 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
Enzymes & pH
● 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
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
Investigating the effect of pH on enzyme activity