Edexcel GCSE Biology Your notes
Enzymes
Contents
The Action of Enzymes
Factors Affecting Enzymes
Practical: Enzymes & pH
Rate Calculations for Enzyme Activity
Enzymes as Biological Catalysts
Practical: Food Tests
Practical: Energy Content in Food
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The Action of Enzymes
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The Action of Enzymes
Enzymes
Enzymes are proteins that act as biological catalysts to speed up the rate of a chemical reaction
without being changed or used up in the reaction
They are biological because they are made in living cells
Enzymes are necessary to all living organisms as they allow all metabolic reactions to occur 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 mechanism of enzyme action
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
When the substrate moves into the enzyme’s active site, the enzyme-substrate complex is formed
After the reaction has occurred, the products leave the enzyme’s active site, which is then free to take
up another substrate
The steps of an enzyme catalysed reaction are shown in the diagram below and can be summarised as
follows:
Step One: Enzymes and substrates randomly move about in solution
Step Two: When an enzyme and its complementary substrate randomly collide, an enzyme-
substrate complex forms and the reaction occurs
Step Three: A product (or products) forms (from the substrate) and is then released from the
active site. The enzyme is unchanged and will go on to catalyse further reactions
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Your notes
How enzymes work
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Denaturation of enzymes
Enzymes are proteins and have a specific shape, held in place by bonds Your notes
This is extremely important around the active site, as the specific shape of this area of the enzyme is
what ensures the substrate will fit into the active site and enable the reaction to proceed
If the bonds that hold the enzyme together are disrupted or broken the active site it will lose its shape
- this is known as denaturation
The enzyme is said to be denatured
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 the reaction they are catalysing will stop
Denaturation can occur due to high temperatures or extremes of pH
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Factors Affecting Enzymes
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Factors Affecting Enzyme Action: Temperature
Enzymes work fastest at their ‘optimum temperature’
In the human body, this optimum temperature is about 37⁰C
Heating to high temperatures (beyond the optimum) will break the bonds that hold the enzyme
together and the active site will lose its shape
The enzyme has been denatured
The effect of temperature on enzyme activity
As temperature increases (towards the optimum) the activity of enzymes increases
This is because the molecules have more kinetic energy, move faster and have more successful
collisions with the substrate molecules. This leads to a faster rate of reaction
This means that low temperatures do not denature enzymes, they just make them work more slowly
due to a lack of kinetic energy
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Your notes
Graph showing the effect of temperature on the rate of enzyme activity
Factors Affecting Enzyme Action: pH
The optimum pH for most human enzymes is pH 7
Some enzymes that are produced in acidic conditions, such as the stomach, have a lower
optimum pH (pH 2)
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 far above or too far below the optimum, the bonds that hold the amino acid chain
together to make up the protein can be disrupted or broken
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 the reaction it is
catalysing will stop
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Your notes
Effect of pH on enzyme activity
Graph showing the effect of pH on the rate of activity for an enzyme from the duodenum
Factors Affecting Enzyme Action: Substrate
Concentration
The greater the substrate concentration, the greater the enzyme activity and the higher the rate of
reaction:
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As the number of substrate molecules increases, the likelihood of enzyme-substrate complex
formation increases
Your notes
If the enzyme concentration remains fixed but the amount of substrate is increased past a certain
point, however, all available active sites eventually become saturated and any further increase in
substrate concentration will not increase the reaction rate
When the active sites of the enzymes are all full, any substrate molecules that are added have
nowhere to bind in order to form an enzyme-substrate complex
For this reason, in the graph below there is a linear increase in reaction rate as substrate is added,
which then plateaus when all active sites become occupied
At this point (known as the saturation point), the substrate molecules are effectively ‘queuing up’
for an active site to become available
The effect of substrate concentration on the rate of an enzyme-catalysed reaction
Examiner Tips and Tricks
Remember the terminology when writing about enzymes is very important. Make sure you refer to an
enzyme becoming 'denatured' not 'dying'.Being able to describe AND explain the effect of each
environmental condition on enzyme action is [Link] describing and explaining using the
graphs and then check your descriptions against your notes.
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Practical: Enzymes & pH
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Practical: Enzymes & pH
Amylase is an enzyme that digests starch (a polysaccharide of glucose) into maltose (a disaccharide
of glucose)
The effect of different pH levels on the activity of amylase can be investigated
Apparatus
Spotting tile
Measuring cylinder
Test Tube
Syringe
Pipette
Stopwatch
Buffer solutions
Iodine
Starch solution
Amylase solution
Method
Add a drop of iodine to each of the wells of a spotting tile
Use a syringe to place 2 cm3 of amylase into a test tube
Add 1cm3 of buffer solution (at pH 2) to the test tube using a syringe
Use another test tube to add 2 cm3 of starch solution to the amylase and buffer solution, start the
stopwatch whilst mixing using a pipette
Every 10 seconds, transfer a droplet of the solution to a new well of iodine solution (which should turn
blue-black)
Repeat this transfer process every 10 seconds until the iodine solution stops turning blue-black (this
means the amylase has broken down all the starch)
Record the time taken for the reaction to be completed
Repeat the investigation with buffers at different pH values (ranging from pH 3.0 to pH 7.0)
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Your notes
Investigating the effect of pH on enzyme activity
Results and Analysis
Amylase is an enzyme which breaks down starch
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When the iodine solution remains orange-brown, all the starch has been digested
This investigation shows: Your notes
At the optimum pH, the iodine stopped turning blue-black and remained orange-brown within the
shortest amount of time
This is because the enzyme is working at its fastest rate and has digested all the starch
At higher or lower pH's (above or below the optimum) the iodine took a longer time to stop turning
blue-black or continued to turn blue-black for the entire investigation
This is because on either side of the optimum pH, the enzymes are starting to become denatured
and as a result are unable to bind with the starch or break it down
Limitations
The starch and amylase solutions that need to be used should be placed in a water bath at optimum
temperature before being used
A colorimeter can be used to measure the progress of the reaction more accurately by measuring the
absorbance/transmission of light through the coloured solution
A control of iodine solution would be used for comparison
A graph showing the optimum pH for an enzyme from a region of the small intestine
Applying CORMS to practical work
When working with practical investigations, remember to consider your CORMS evaluation
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Your notes
CORMS Evaluation
In this investigation, your evaluation should look something like this:
C - We are changing the pH of the environment
O - This is not relevant to this investigation as we aren't using an organism
R - We will repeat the investigation several times to ensure reliability
M1 - We will measure the time taken for
M2 - the iodine to stop turning black
S - We will control the concentration and volume of the amylase, iodine and starch solution used in
the investigation
Examiner Tips and Tricks
When describing the effect of pH on enzyme activity, it is important to remember that any pH
outside of the optimum can lead to the enzyme becoming permanently denatured.
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Rate Calculations for Enzyme Activity
Your notes
Rate Calculations for Enzyme Activity
Rate calculations are important in determining how fast an enzyme is working (i.e. the rate of reaction)
To perform a rate calculation, use the following formula:
rate = change ÷ time
Change = the change in the substance being measured
E.g. the amount of substrate used up in the reaction or the amount of product formed
Time = the time taken for that change to occur
Worked Example
Amylase catalyses the breakdown of starch into maltose. 15 grams of starch were added to a
solution containing amylase. It took 2 hours for all the starch to be broken down.
Calculate the rate of reaction.
Answer:
Step 1: write out the equation for calculating the rate of enzyme activity
rate = change ÷ time
In this case: rate = amount of substrate used ÷ time
Step 2: substitute in the known values and calculate the rate
rate = 15 g ÷ 2 hours
rate = 7.5 g / hr or 7.5 g hr⁻¹
Worked Example
The enzyme catalase catalyses the breakdown of hydrogen peroxide into water and oxygen. In one
experiment, a student found that 45 cm³ of oxygen was released in 5 minutes.
Calculate the rate of reaction.
Answer:
Step 1: write out the equation for calculating the rate of enzyme activity
rate = change ÷ time
In this case: rate = amount of product formed ÷ time
Step 2: substitute in the known values and calculate the rate
rate = 45 cm³ ÷ 5 minutes
rate = 9 cm³ / min or 9 cm³ min⁻¹
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In some situations you may not be told how much something has changed during a reaction; instead,
you may only be told the time taken for the reaction to occur
Your notes
In this case you can still calculate the rate of reaction by using the following (slightly different) formula:
rate = 1 ÷ time
Worked Example
A student adds a set volume of starch solution to a set volume of amylase solution at a range of
different pH values. At each pH, the student times how long it takes for the amylase to break down
all of the starch. At pH 6 the time taken for amylase to break down all of the starch was 50 seconds.
Calculate the rate of reaction at pH 6.
Answer:
Step 1: write out the equation for calculating the rate of enzyme activity
rate = 1 ÷ time
Step 2: substitute in the known values and calculate the rate
rate = 1 ÷ 50 seconds
rate = 0.02 s⁻¹
The units for the calculation above are in s⁻¹ because rate is given per unit time.
Examiner Tips and Tricks
In an exam you could be asked to plot the reaction rates (from an enzyme catalysed reaction) on a
graph. However, using the equation 'rate = 1 ÷ time' often gives small numbers that are difficult to
plot on a graph. In these cases, you can also use the equation:
rate = 1000 ÷ time
This equation give you bigger numbers that are easier to plot on a graph. So, for the calculation in
the worked example above, you would get:
rate = 1000 ÷ 50 seconds
rate = 20 s⁻¹
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Enzymes as Biological Catalysts
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Enzymes as Biological Catalysts
The purpose of digestion is to break down large, insoluble molecules into smaller, soluble molecules
that can be absorbed into the bloodstream
Food is partially digested mechanically (by chewing, churning and emulsification) in order to break
large pieces of food into smaller pieces of food
This increases the surface area for enzymes to work on
Digestion mainly takes place chemically, where bonds holding the large molecules together are
broken to make smaller and smaller molecules
Chemical digestion is controlled by enzymes that are produced in different areas of the digestive
system
Enzymes are biological catalysts – they speed up chemical reactions without themselves being used
up or changed in the reaction
There are three main types of digestive enzymes: carbohydrases, proteases and lipases
Carbohydrases
Carbohydrases are enzymes that break down carbohydrates into simple sugars such as glucose
Amylase is a carbohydrase that is made in the salivary glands, the pancreas and the small intestine
Amylase breaks down starch into maltose
Maltase then breaks down maltose into glucose
Starch is broken down into glucose using two enzymes: amylase and maltase
Proteases
Proteases are a group of enzymes that break down proteins into amino acids
Pepsin is an enzyme made in the stomach that breaks down proteins into smaller polypeptide
chains
Proteases made in the pancreas and small intestine break the polypeptides into amino acids
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Your notes
Proteins are broken down using pepsin and other proteases
Lipases
Lipases are enzymes that break down lipids (fats) to glycerol and fatty acids
Lipase enzymes are produced in the pancreas and secreted into the small intestine
Lipids are broken down by lipase enzymes
Synthesis of carbohydrates, proteins and lipids
Enzymes are not just important in breaking down larger molecules into smaller ones
They are also required for the synthesis of larger molecules (building small molecules back up into
bigger ones)
Enzymes are required by organisms to synthesise carbohydrates, proteins and lipids
Carbohydrates are synthesised by joining simple sugars together
For example, glycogen synthase is an enzyme that joins together many chains of glucose
molecules to form glycogen (an energy-storage molecule in animals)
Proteins are synthesised by joining amino acids together
Again, enzymes catalyse the reactions required to do this
Many enzymes are involved in the synthesis of lipids from fatty acids and glycerol
Examiner Tips and Tricks
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The pancreas is an accessory organ in the digestive system. Food does not pass directly through
it, but it has a key role in producing digestive enzymes, as well as the hormones that regulate blood
sugar (insulin and glucagon). Your notes
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Practical: Food Tests
Your notes
Practical: Food Tests
Preparing a sample
Before you can carry out any of the food tests described below, you may need to prepare a food
sample first (especially for solid foods to be tested)
To do this:
Break up the food using a pestle and mortar
Transfer to a test tube and add distilled water
Mix the food with the water by stirring with a glass rod
Filter the mixture using a funnel and filter paper, collecting the solution
Proceed with the food tests
Test for glucose (a reducing sugar)
Add Benedict's solution to the sample solution in a test tube
Heat in a boiling water bath for 5 minutes
Take the test tube out of the water bath and observe the colour
A positive test will show a colour change from blue to orange / brick red
The Benedict's test for glucose
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Test for starch using iodine
We can use iodine to test for the presence or absence of starch in a food sample Your notes
Add drops of iodine solution to the food sample
A positive test will show a colour change from orange-brown to blue-black
In the presence of starch, iodine will turn from brown to blue-black
Test for protein
Add drops of Biuret solution to the food sample
A positive test will show a colour change from blue to violet / purple
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Your notes
The Biuret test for protein
Test for lipids
Mix the food sample with 4cm3 of ethanol and shake
Allow time for the sample to dissolve in the ethanol
Strain the ethanol solution into another test tube
Add the ethanol solution to an equal volume of cold distilled water (4cm3)
A positive test will show a cloudy emulsion forming
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Your notes
The ethanol test for lipids
Food Test Results Table
Important hazards
Whilst carrying out this practical you should try to identify the main hazards and be thinking of ways to
reduce harm
Biuret solution contains copper (II) sulfate which is dangerous particularly if it gets in the eyes, so
always wear goggles
Iodine is also an irritant to the eyes
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Sodium hydroxide in biuret solution is corrosive, if any chemicals get onto your skin wash your hands
immediately
Your notes
Ethanol is highly flammable; keep it away from any Bunsen burner
The Bunsen burner itself is a hazard due to the open flame
Worked Example
Food tests: analysis
Write a conclusion to state which food groups are present one of the food samples you tested and
an explanation of how you know this.
Conclusion:
The apple contained both starch and sugar as it tested positive for both the iodine test (orange →
blue - black) and the benedict's test (blue → orange).
The apple did not contain protein or lipid (fat) as the biuret and emulsion tests were both negative.
Applying CORMS to practical work
When working with practical investigations, remember to consider your CORMS evaluation.
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Your notes
CORMS evaluation
In this investigation, your evaluation should look something like this:
C - We are changing the type of food in the sample
O - This is not relevant to this investigation as we aren't using an organism
R - We will repeat the investigation several times for each food sample to ensure a reliable result
M1 - The presence of the specific biological molecule in each food type by noting the colour
change
M2 - ....after testing with each specific testing agent
S - We will control the volume of each testing agent used, the quantity of the food sample, the
concentration of the testing agents, the temperature of the water bath for the Benedicts test.
There may be other examples that you can think of
Examiner Tips and Tricks
When describing food tests in exam answers, make sure you give the starting colour of the solution
and the colour it changes to for a positive result.
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Practical: Energy Content in Food
Your notes
Practical: Energy Content of a Food Sample
We can investigate the energy content of food in a simple calorimetry experiment
Apparatus
Boiling tube
Boiling tube holder
Bunsen burner
Mounted needle
Measuring cylinder
Balance/scales
Thermometer
Water
Food samples
Method
Use the measuring cylinder to measure out 25 cm3 of water and pour it into the boiling tube
Record the starting temperature of the water using the thermometer
Record the mass of the food sample
Set fire to the sample of food using the bunsen burner and hold the sample 2 cm from the boiling tube
until it has completely burned
Record the final temperature of the water
Repeat the process with different food samples
E.g. popcorn, nuts, crisps
Investigating the energy content of food samples diagram
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Your notes
Different food samples can be burned in a simple calorimetry experiment to compare the energy
contents of the samples
Results
The larger the increase in water temperature, the more energy is stored in the sample
We can calculate the energy in each food sample using the following equation:
g × temperature increase
energy transferred per gram of food J = mass of water mass ( ) (
oC ) × 4.2
( )
of food sample g ( )
4.2 kJ is the specific heat capacity of water, meaning that it is the energy required to raise 1 kg of water
by 1 °C
1 cm3 of water has a mass of 1 g
The energy content of food samples table
Food Mass of Mass Initial water Final water Change in Energy
sample water / of temperature / temperature / water transferred per
g food / °C °C temperature / gram of food
g °C (J)
Popcorn 25 8.5 20.5 31.2 10.7 132.2
Walnut 25 8.1 20.4 34.1 13.7 177.6
Limitations
Incomplete burning of the food sample
Solution: Relight the food sample until it no longer lights up
Heat energy is lost to the surroundings
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Solution: Whilst heat lost means that the energy calculation is not very accurate, so long as the
procedure is carried out in exactly the same way each time (with the same distance between food
sample and boiling tube), we can still compare the results Your notes
Applying CORMS evaluation to practical work
When writing about practical investigations the CORMS evaluation can be used:
CORMS provides a framework for writing about practical investigations
In this investigation CORMS can be applied as follows:
Change
We are changing the type of food in the sample
Organisms
This is not relevant to this investigation as we aren't using an organism
Repeat
We will repeat the investigation several times for each food sample
Measurement 1
We will measure the change in temperature of the water
Measurement 2
The mass of the food will be measured after the food sample has burned out
Same
We will control the volume of water used and the distance between the food sample and the
boiling tube during burning
The food will also be relit every time it goes out until it no longer relights
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