Enzymes are biological catalysts
This is because they are:
o Catalysts that speed up the rate of a chemical reaction without being changed or used up in
the reaction
o Made from the biological molecule protein
They 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
o 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?
The enzyme is made from protein that is folded into a very specific 3D shape
Part of this 3D shape is a section of the enzyme called the active site
The active site is the section of the enzyme that binds to the substrate (the reactant)
When the enzyme binds to the substrate it is called the enzyme-substrate complex
The formation of the enzyme-substrate complex diagram
The enzyme-substrate complex forms when the substrate binds to the active site of the enzyme
The product is made from the substrate(s) and is released
After the product is released the enzyme is free to bind to a new substrate and repeat the reaction
again
Diagram showing how enzymes work
Enzymes act as biological catalysts
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 3D shape
o Different types of enzymes have a different combination of amino acids in the
protein, which results in a different shape
This is known as the lock and key hypothesis
o The active site is the lock and the substrate is the key
o Only specific shaped keys can fit into each lock
The lock and key model of enzyme action diagram
The lock and key model of enzyme action results in enzyme specificity
Measuring Reactants & Products
The progress of enzyme-catalysed reactions can be investigated by:
o Measuring the rate of formation of a product
o Measuring the rate of disappearance of a substrate
For example, the enzyme catalase releases oxygen gas as a product as it breaks down the substrate
hydrogen peroxide
o The oxygen gas can be collected and the volume is measured using a measuring cylinder or a
gas syringe
Measuring the rate of product formation of an enzyme-catalysed reaction diagram
The rate of oxygen production is measured in order to determine the rate of the catalase enzyme activity
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 amylase diagram
The rate of amylase activity can be determined by how quickly the wells of iodine stop turning blue-black
when the reaction solution is added
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 amylase diagram
The rate of amylase activity can be determined by how quickly the wells of iodine stop turning blue-black
when the reaction solution is added
Examiner 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 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.
Enzymes: Temperature & pH
The effect of temperature
The specific shape of an enzyme is determined by the amino acids that make the enzyme
The three-dimensional shape of an enzyme is especially important around the active site area;
this ensures that the enzyme’s substrate will fit into the active site enabling the reaction to proceed
Enzymes work fastest at their ‘optimum temperature’ – in the human body, the optimum
temperature is around 37°C
Heating to high temperatures (beyond the optimum) will start to break the bonds that hold the
enzyme together – the enzyme will start to distort and lose its shape – this reduces the
effectiveness of substrate binding to the active site reducing the activity of the enzyme
Eventually, the shape of the active site is lost completely and the enzyme is described as
being ‘denatured’
Substrates cannot fit into denatured enzymes as the specific shape of their active site has been lost
Enzyme denaturation diagram
Denaturation is largely irreversible – once enzymes are denatured they cannot regain their
proper shape and activity will stop
Increasing temperature from 0°C 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, but at lower temperatures with less
kinetic energy both enzymes and their substrates collide at a lower rate
The effect of temperature on enzyme activity diagram
This graph shows the effect of temperature on the rate of activity of an enzyme
The effect of 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
Enzyme denaturing due to pH changes diagram
If pH is increased or decreased away from the optimum, then the shape of the enzyme is
altered
The effect of pH on enzyme activity diagram
This graph shows the effect of pH on the rate of activity of an enzyme from the duodenum
