Cambridge International AS & A Level

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Biology
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 Maximise exam potential and prepare lear

With Cambridge International AS & A Level Complete Biology

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syllabus (9700). The chapters, topics and learning outcomes are all closely
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e learners for future study

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 Contents
Each chapter, along with
its topics and learning
outcomes, corresponds
with the latest Cambridge
syllabus (9700)

AS Level
01 Cell structure 07 Transport in plants

1.1 The microscope in cell studies 7.1 Structure of transport tissues

1.2 Cells as the basic units of living organisms 7.2 Transport mechanisms
Exam-style and practice questions Exam-style and practice questions

02 Biological molecules 08 Transport in mammals

2.1 Testing for biological molecules 8.1 The circulatory system

2.2 Carbohydrates and lipids 8.2 Transport of oxygen and carbon dioxide
2.3 Proteins 8.3 The heart
2.4 Water Exam-style and practice questions
Exam-style and practice questions

09 Gas exchange
A selection of pages
03 Enzymes
from this chapter are 9.1 The gas exchange system
included for you to
3.1 Mode of action of enzymes Exam-style and practice questions
evaluate
3.2 Factors that affect enzyme action
Exam-style and practice questions
10 Infectious diseases

10.1 Infectious diseases

04 Cell membranes and transport
10.2 Antibiotics
4.1 Fluid mosaic membranes Exam-style and practice questions
4.2 Movement into and out of cells
Exam-style and practice questions
11 Immunity

11.1 The immune system

05 The mitotic cell cycle
11.2 Antibodies and vaccination
5.1 Replication and division of nuclei and cells Exam-style and practice questions
5.2 Chromosome behaviour in mitosis
Exam-style and practice questions

06 Nucleic acids and protein synthesis

6.1 Structure of nucleic acids and replication of DNA

6.2 Protein synthesis
Exam-style and practice questions

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 Contents

A Level
12 Energy and respiration 18 Classification, biodiversity and conservation

12.1 Energy 18.1 Classification

12.2 Respiration 18.2 Biodiversity
Exam-style and practice questions 18.3 Conservation
Exam-style and practice questions

13 Photosynthesis

13.1 Photosynthesis as an energy transfer process 19 Genetic technology

13.2 Investigation of limiting factors
19.1 Principles of genetic technology
Exam-style and practice questions 19.2 Genetic technology applied to medicine
19.3 Genetically modified organisms in agriculture
Exam-style and practice questions
14 Homeostasis

14.2 Homeostasis in plants

Exam-style and practice questions
Glossary

Index
15 Control and coordination

15.1 Control and coordination in mammals

15.2 Control and coordination in plants
Exam-style and practice questions

16 Inheritance

16.1 Passage of information from parents to offspring

16.2 The roles of genes in determining the phenotype
16.3 Gene control
Exam-style and practice questions

17 Selection and evolution

17.1 Variation
17.2 Natural and artificial selection
17.3 Evolution
Exam-style and practice questions

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 3 Enzymes
3.1a
These pages help you to:
• describe enzymes as globular
proteins (3.1.1)
• understand that enzymes
catalyse reactions (3.1.1)
• explain the difference between
intracellular and extracellular
enzymes (3.1.1)
• understand the mode of action
of enzymes by considering the
role of the active site, enzyme–
substrate complex and lowering
activation energy (3.1.2)
• understand the difference
between the lock and key
hypothesis of enzyme action and
the induced fit hypothesis (3.1.2)
You will also:
• begin to appreciate the
importance of enzymes for
living organisms
• be able to explain terms
associated with enzyme structure
and mechanism of action
Figure 1 The ribonuclease A enzyme and its
substrate close to the enzyme’s active site
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Mode of action of enzymes: enzyme
structure and mechanism of action
Enzymes are globular proteins that catalyse metabolic reactions. A catalyst
alters the rate of a chemical reaction, but at the end of the reaction is
unchanged and can be used again. This means that enzymes are effective in
tiny quantities. Enzymes do not make a reaction happen; they simply speed up
ones that already occur, sometimes by a factor of many millions. All enzymes
are synthesised within the cell. Intracellular enzymes remain within the cell to
catalyse metabolic reactions. Extracellular enzymes are secreted from, and act
outside, the cell.
Enzyme structure
As globular proteins, enzymes have an overall spherical shape (Figure 1),
determined by their sequence of amino acids (Topic 2.3c). Despite their large
size, enzyme molecules only have a small region that is functional. This is known
as the active site. Only a few amino acids make up this active site. The active
site forms a hollow depression within the much larger enzyme molecule. The
substrate molecule is held within the active site by bonds, such as hydrogen
bonds, that temporarily form between the R groups of the amino acids of the
active site and groups on the substrate molecule. This structure is known as
the enzyme–substrate complex. Figure 2 shows part of an enzyme molecule
during enzyme–substrate complex formation:
• Amino acids 20 and 21 form temporary hydrogen bonds with the substrate
molecule at the binding site of the active site.
• The area of the active site that is directly involved in the reaction is composed
of amino acids 7, 8, 36 and 37 (the catalytic site).
• Amino acids 2 and 42 are not part of the active site but play an essential
role in helping to maintain the shape of the active site. There are R-group
interactions between these and amino acids 6 and 38.
Substrate molecule held within the
enzymne’s active site
Rest of
enzyme Substrate
molecule 42
38 6
One of the four (*) amino acids involved in
37 * * 7 2 catalysis at the active site
* 8
36 *
20 21 Temporary H-bonds that form to bind substratea
Bonds formed by interactions between R-groups
of amino acids (e.g. hydrogen bond, ionic bond,
disulfide bridge) to maintain enzyme tertiary
structure
Figure 2 Section of an enzyme molecule during the formation of an enzyme–substrate complex
Changes to amino acids in the active site or in amino acids maintaining the
shape of the active site can lead to loss of the specific three-dimensional shape
of the active site and reduce or prevent catalysis.
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 Enzymes

Enzymes and activation energy Remember

Consider a typical chemical reaction: The structure of an enzyme
2H2O2 → 2H2O + O2 molecule is suited to its function.
hydrogen peroxide water oxygen The active site has a specific shape.
Only a particular shape of substrate
(substrate) (products)
molecule can fit into this active
Catalase is an enzyme found in living tissue. It decomposes hydrogen peroxide, a site. There are amino acids with
toxic waste product of metabolic reactions. For such a reaction to occur naturally, hydrophilic R groups facing to the
the energy of the products must be less than that of the substrates. Such reactions, outside of the molecule. This makes
however, need an initial boost of energy to get them kick-started. This is known the enzyme soluble in the aqueous
internal environment of the cell or in
as the activation energy. In other words, there is an energy hill, or barrier,
extracellular fluids such as blood and
which must be overcome before the reaction can proceed. Enzymes lower this tissue fluid and allows it to catalyse
activation energy level so that the reaction can happen more easily (Figure 3). metabolic reactions.

How enzymes work

Energy barrier
(a) The lock and key hypothesis. without enzyme
In one sense, enzymes work in the same way as a key operates a lock: each key
Energy barrier
has a very specific shape which, on the whole, fits and operates only one lock. with enzyme
In the same way, a substrate will only fit into the active site of one particular Energy level
enzyme. Enzymes are therefore specific in the reactions that they catalyse. The SUBSTRATE
of substrate

Free energy
shape of the substrate (key) is complementary to the shape of the active site of Lower activation
energy
the enzyme (lock). This is known as the lock and key hypothesis the specificity
of enzymes. Figure 4 shows how two different substrates have a complementary
shape to regions within the active site. The enzyme–substrate complex results in
one product molecule. Enzymes can catalyse a reverse reaction.
Substrate Energy level PRODUCTS
Substrate of the products
Product
Time during reaction
Figure 3 How enzymes lower activation
Active site free to energy
accept substrate
molecules
Extension
Active site Enzyme–substrate
complex Enzyme molecule How enzymes lower activation
Enzyme energy
molecule
Supplying heat energy will increase
Figure 4 The lock and key mechanism of enzyme action
the rate of a slow reaction because
(b) The induced fit hypothesis the proportion of molecules that
In practice, rather than being a rigid lock, the active site of the enzyme actually have activation energy will increase.
changes its form slightly to fit the shape of the substrate. This means that the This is not possible in cells as protein
substrate fits better into the active site. This is the induced fit hypothesis of denaturation will occur. Using the
active site, enzymes help to lower
enzyme action.
the activation energy required.
Ways that this can be achieved are:
Summary test 3.1a holding the substrates close together
for easier bond formation; exposing
Enzymes act as biological (1). They are (2) proteins that have a specific shape bonds; holding a substrate to slightly
within which there is a functional portion known as the (3). Enzymes lower strain bonds to be broken; providing
the (4) of a reaction, allowing it to proceed at a lower temperature than it a hydrophobic region for a reaction
would normally. In an enzyme-controlled reaction, the general term for the involving non-polar substrates;
substance on which the enzyme acts is (5) and the substances formed at transfer of electrons.
the end of the reaction are known as the (6). The enzyme molecule and the
substance it acts on fit together very precisely, giving rise to the name (7)
hypothesis of enzyme action. In practice, most enzymes have active sites that
change shape slightly and so mould around the shape of their substrates. This
is called the (8) hypothesis of enzyme action.
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 3.1b Mode of action of enzymes:
investigating the progress of
enzyme-catalysed reactions

These pages help you to:
• learn how to investigate the
progress of enzyme-catalysed
reactions (3.1.2)
• understand that the rate of an
enzyme-catalysed reaction can
be calculated by measuring rate
of product formation or rate of
substrate formation (3.1.2)
You will also:
• understand the difference
between independent and
dependent variables
• understand what is meant by
standardised variables
• understand what is meant by a
control
10
Volume of oxygen produced / cm3
Before considering how different factors affect enzymes, it is worth bearing in
mind that, for an enzyme to work, it must:
• come into physical contact with its substrate
• have an active site which fits the substrate.
Almost all factors that influence the rate at which an enzyme works do so by
affecting one or both of the above two features. In order to investigate how
enzymes are affected by various factors we need to be able to measure the
reactions they catalyse.
Measuring enzyme-catalysed reactions
To measure the progress of an enzyme-catalysed reaction we usually measure
its time-course, i.e. how long it takes for a particular event to run its course.
The two ‘events’ most frequently measured are:
• the formation of products of the reaction, e.g. the volume of oxygen
produced when catalase acts on hydrogen peroxide (Figure 1)
• the disappearance of the substrate, e.g. the reduction in concentration of
starch when it is acted upon by amylase (Figure 2).
Although the graphs in Figures 2 and 3 differ, the explanation for their shapes is

9 the same:
8 • At first there is a lot of substrate (hydrogen peroxide/starch) but no product
7 (water and oxygen/maltose).
6 • It is very easy for substrate molecules to come into contact with the empty
5 active sites on the enzyme molecules.
4 • All enzyme active sites are filled and the substrate is rapidly broken down into
3 its products.
2 • The quantity of substrate decreases as it is broken down, resulting in an
1
increase in the amount of product.
• As the reaction proceeds, there is less and less substrate and more and more
0
0 20 40 60 80 100 120 product.
Time / s • The product produced per unit time decreases as there are fewer substrate
Figure 1 Measurement of the formation of oxygen molecules and so some active sites may not be filled at any one moment.
due to the action of catalase on hydrogen peroxide • The rate of reaction continues to slow as the substrate concentration decreases.
• The graphs flatten out because all the substrate has been used up and so no
10 new product can be produced.
9
Practical skills: Following the progress of a catalase-controlled reaction
Concentration of starch /

8
7
Figure 3 shows one method to measure the volume of oxygen produced over
arbitrary units

6
time. Hydrogen peroxide solution, the substrate, and catalase solution, the
5
enzyme, are mixed together at time 0 and the volume of oxygen produced is
4 recorded at set time intervals.
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12
Time / min
Figure 2 Measurement of the disappearance of
starch due to the action of amylase

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 Enzymes

Syringe for adding

hydrogen peroxide
Remember
As an enzyme-catalysed reaction
Rubber connecting tubing proceeds, substrate molecules are
used up. The enzyme will only be
able to work at its maximum rate in
the initial stage of the reaction when
there are most substrate molecules.
For this reason, the initial rate of
Gas syringe for measuring
volume of oxygen produced
reaction for an enzyme is frequently
calculated. Figure 1 shows that the
curve is almost a straight line until
approximately 30 seconds, so the
Conical flask initial rate can be calculated using
the gradient of this straight line.
After this time, the rate slows down.
Catalase and hydrogen
peroxide solution

Summary test 3.1b

Figure 3 Measuring the volume of oxygen give off over time in a catalase-controlled reaction
We can measure the progress of
an enzyme-catalysed reaction by
Variables
measuring its (1). This is usually
In this experiment:
done by measuring either the (2)
The independent variable is the variable that will be changed, time. On a graph, of the substrate or the formation
this is on the x-axis. The volume of oxygen given off every 10 or 20 seconds is of the (3). For example, in the
recorded for 120 seconds. Note that there should be at least 5 readings. case of the enzyme amylase, we
could either measure the rate
The dependent variable, the volume of oxygen given off, is the variable that is
at which (4) is produced or the
affected by the independent variable. It is on the y-axis of a graph. Figure 3 shows
rate at which (5) is used up. To
that a gas syringe can be used to collect oxygen. Another method is to collect the
follow the progress of a reaction
gas under water, with the oxygen displacing water in a graduated cylinder.
catalysed by the enzyme catalase,
Standardised variables are other variables that may have an effect on the the volume of oxygen produced
results and need to be controlled. For example, in this experiment it is important by the breakdown of (6) can be
to standardise temperature as this has a large effect on enzyme action (see 3.1d). measured. This is the (7) variable.
The hydrogen peroxide solution and catalase solution should be at the standardised A graph can be drawn of total
temperature before they are mixed, and the conical flask should be kept at the volume of oxygen produced on
same temperature throughout the experiment. It is also important when repeating the y-axis and (8) on the x-axis.
experiments to use the same temperature and the same volume and concentration The steepest part of the graph
of enzyme solution and substrate solution as the first experiment. is in the first 30 seconds and
by using the change in volume
Controls over time, an (9) rate of reaction
A control run should also be conducted and the results compared to the can be calculated. In a reaction
experimental results. This allows verifies that it is only the action of catalase involving the hydrolysis of starch
that allows oxygen to be released and that no other variable is acting. by the enzyme (10), the colour
change that occurs when testing
Here, the same quantity of boiled (denatured) enzyme should confirm that no
samples at regular time intervals
oxygen is produced. In reality, adding the hydrogen peroxide solution at time 0
with (11) solution can be
will cause some displacement of air in the conical flask. The control will show
observed and the time to reach
how much air is displaced and this can be taken into account when the results
an (12) can be recorded.
are analysed.

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 3.1c Mode of action of enzymes: using a
colorimeter

These pages help you to: Understanding the colorimeter

• learn how to use a colorimeter A colorimeter is a light-sensitive piece of equipment that gives a quantitative
to measure the progress of measurement for a coloured solution.
enzyme-catalysed reactions
(3.1.4) The solution is placed into a small container, known as a cuvette, that typically
holds 3 or 4 cm3 of solution. The cuvette is clear on two opposite sides and
You will also: frosted (translucent) on the other two sides (Figure 1).
• have an understanding of how
colorimeters work
• see how to apply the main
ideas to the example of starch
hydrolysis by amylase
• understand how colorimeter
measurements can be converted
to concentrations

Figure 1 A colorimeter and cuvettes

The cuvette is placed into the sample well of a colorimeter and a beam of light
is aimed through the clear sides of the cuvette. Some light is absorbed by the
solution and the rest of the light is transmitted through the solution. The light
reaches a sensor and a numerical reading is obtained:
• a transmission value is given as a percentage transmission or
• an absorbance value is given in absorbance units (au).
Different colour intensities, or colour shades, will give different readings
(Figure 2) that can be compared.
Light source Sample Sensor Reading

Transmission or Absorbance

Lower Higher
percentage absorbance
transmission units (au)

Darker shade of colour

Higher Lower
percentage absorbance
transmission units (au)

Lighter shade of colour

Figure 2 Using a colorimeter to compare sample solutions of two different intensities of colour

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 Enzymes

A particular wavelength of light is used. This is achieved by using LEDs or a

Remember
coloured filter. Some colorimeters have a slot to add a filter, others incorporate
a rotating wheel of coloured filters. To produce the most accurate results, the Carry out a trial to determine an
wavelength that is absorbed best by the coloured solution is chosen as this makes appropriate concentration of I in KI
it easier to detect small differences in colour change. solution to use in the cuvettes. If the
concentration is too high, a reading
The colorimeter is calibrated (standardised) using a reference, or blank, cannot be obtained as the blue-black
solution in a cuvette. This may be distilled water. Using this reference solution, colour will be too intense.
the transmission is set as 100% (and the absorbance is set as 0 au) as the
baseline reading.
The transmittance or absorbance readings of the sample solution can be taken
visually. Alternatively, readings can be recorded by downloading to a computer,
or the data can be sent to a phone, tablet or laptop.
Extension

Using the colorimeter to measure an enzyme-catalysed reaction Preparing a calibration curve to

determine concentrations
involving a colour change
It is possible to determine the
A simple way to follow the progress of the breakdown of starch by amylase is concentration of starch in a
to use iodine in potassium iodide (I in KI) solution. Colour changes over time sample cuvette by preparing a
reflect the decrease in starch (substrate) that is occurring owing to its hydrolysis calibration curve.
by the enzyme. The reaction mixture will change from a blue-black colour at You can use the colorimeter to obtain
time 0 to the orange colour of I in KI solution (when all starch has been used absorbance readings for known
up). This is the end point. concentrations of starch solution. Plot
a graph of absorbance (y-axis) against
A sample of reaction mixture removed at time 0, when added to a cuvette
concentration of starch (x-axis) and
containing I in KI solution, will give the darkest blue-black colour. This will
draw in the calibration curve. This
produce the highest absorbance reading. means that the absorbance readings
At the end of the reaction, all of the starch will be hydrolysed to reducing obtained during the experiment
sugars. A sample taken at the end of the reaction will produce a pale orange can be converted directly to starch
concentration.
colour (the colour of the I in KI in the cuvette) and will have the lowest
absorbance reading. Samples taken at set time intervals during the reaction will
show a decrease in the absorbance readings.
A graph can be plotted of absorbance (y-axis) against time (x-axis) to show the Remember
progress of the reaction.
A thermostatically controlled
water bath can be used to allow
Summary test 3.1c the amylase and starch solutions
to equilibrate to the reaction
A (1) is a piece of equipment that is sensitive to light and can be used to temperature (e.g. 30°C) before they
provide a (2) measurement for a coloured solution. The sample solution are mixed together.
to be tested is added to a small container known as a (3). This is placed in
the sample well and the quantity of light that passes through the sample
is detected by a (4). A reading of percentage (5) is obtained. The result can
also be obtained as the quantity of light that is (6) by the sample.

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These pages help you to:
• understand how to investigate
the effects of temperature
and pH on enzyme-catalysed
reactions (3.2.1)
• explain why changing
temperature or pH will affect an
enzyme catalysed reaction (3.2.1)
You will also:
• appreciate the importance of
standardising variables
• be able to make decisions
relating to measurement and
observations
• be able to interpret and explain
results
Rate at which reaction
increases due to increased
kinetic energy of substrate
and enzyme molecules
Rate of a reaction, i.e. Velocity [V]
Rate at which reaction
decreases due to
denaturation of
enzyme molecules
Optimum
temperature
0 10 20 30 40
Temperature / °C
Figure 1 Effect of temperature on the rate of
an enzyme-controlled reaction
Figure 2 Bacteria (red) growing in this hot
spring in New Zealand are not killed, and their
enzymes are not denatured, at temperatures in
excess of 80 °C
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3.2a Factors that affect enzyme action:
temperature and pH
Effect of temperature on enzymes
A rise in temperature increases the kinetic energy of molecules, which
therefore move around more rapidly and collide with one another more
often. In an enzyme-catalysed reaction, this means that the enzyme and
substrate molecules come together more often in a given time, so that the rate
of reaction is increased. Shown on a graph, this is a rising curve. However,
the temperature rise also increases the energy of the atoms that make up the
enzyme molecule. Its atoms begin to vibrate and cause bonds to break, with
weaker bonds such as hydrogen bonds, breaking first. Gradually, the shape of
the active sites is changed. At first, the substrate fits less easily into the active
site slowing the rate of reaction. For many human enzymes, this may begin at
temperatures of around 45 °C. At some point, usually around 60 °C, the tertiary
structure of the enzyme and shape of the active site is so changed that it stops
working altogether. It is said to be denatured. Shown on a graph, the rate of
this reaction follows a falling curve. The actual effect of temperature on the rate
of an enzyme reaction is a combination of these two factors, increased kinetic
energy of molecules and denaturation of the enzyme (Figure 1). The optimum
working temperature differs from enzyme to enzyme. Some work best at
around 10 °C, while others continue to work well at 80 °C (Figure 2). Each
enzyme in the human body has a different optimum working temperature. Our
Actual rate of
reaction as a
body temperatures have, however, evolved to be 37 °C because:
result of the • Although higher body temperatures would increase the metabolic rate slightly,
combined
the advantages are offset by the additional energy (food) that would be needed
effect of these
two influences to maintain the higher temperature.
• Proteins, other than enzymes, may be denatured at higher temperatures.
• At higher temperatures, any further rise in temperature, e.g. during illness,
might denature the enzymes.
Denaturing enzymes using high temperatures prevents spoilage (breakdown)
of food materials. This is the basis for heating food before canning or bottling it
and for blanching vegetables before freezing.
Effect of pH on enzymes
The pH of a solution is a measure of its hydrogen ion concentration. Each
50 60 enzyme has an optimum pH, i.e. a pH at which it works fastest (Figure 3).
At a pH lower or higher than this optimum, the activity of the enzyme will
decrease. Large changes in the pH, or hydrogen ion concentration, can cause
hydrogen bonds and ionic bonds to become disrupted and break. This can lead
to a change in the shape of the active site because these bonds are important
in maintaining the tertiary structure of the enzyme, including its active site. A
slight change in the active site shape may allow the enzyme to function, but
less effectively. At extremes of pH, denaturation occurs and as a result, the
substrate can no longer become attached to the active site and the enzyme–
substrate complex cannot be formed. This is why foods can be preserved in
vinegar: the low pH denatures the enzymes that would otherwise cause the
food to break down. Decreasing or increasing the pH from the optimum can
also affect the transfer of electrons that may occur during catalysis. Changes
to the substrate may also occur. Solutions, known as buffer solutions, can be
used to prevent fluctuations in pH.
16/12/2019 12:35
 Enzymes

Making decisions in investigations Remember

The effect of temperature on the action of catalase Enzymes are not alive and so cannot
You have been asked to determine the optimum temperature of the enzyme catalase be ‘killed’. Use the correct term
at its optimum pH of 7. The optimum is stated to be between 30°C and 40°C. which is denatured.

The initial rate of reaction can be determined by the volume of oxygen given off in
the first 30 seconds collected in a gas syringe.
Salivary
The apparatus is set up for you and you have been instructed to transfer 2 cm3 of Pepsin Sucrase amylase Arginase

Rate of a reaction, i.e. Velocity [V]

buffered enzyme solution to the test tube containing the substrate. You have a choice
of a 50 cm3 beaker, a 10 cm3 measuring cylinder and a 5 cm3 syringe to use for this.
You are supplied with buffer solutions from pH 4 to 8.

Example of a decision you may need Decision made, with reasons

to make
Choosing the appropriate chemicals
to use
Choosing the correct apparatus
to use
Identifying the independent variable
Identifying the dependent variable
How to change the independent
variable
Range and spacing to be used for
the independent variable and the
number of values at which the
dependent variable is to be recorded
How frequently the dependent
variable should be measured
Whether the experiment should be
repeated
Which variables it is possible to
standardise
Whether a control should be
carried out
The effect of pH on the action of catalase
In this experiment, many of the decisions made for investigating the effect of
temperature will be the same. However, the independent variable is pH. Temperature
must be constant, but pH needs to be altered using buffers. You can check the pH of
a solution by using a pH probe and meter or by using universal indicator paper.
Use buffer solution at pH 7, as this is the optimum pH
for catalase.
Use the 5 cm3 syringe, as this has the smallest
graduations and so is the most precise apparatus.
Temperature, as this is the variable that is to be 2 4.5 7 10
changed pH
Volume of oxygen given off, as this is being affected
Figure 3 Effect of pH on the rate of an
by the temperature change
enzyme-controlled reaction
Use a thermostatically controlled water bath, as it can
be set at the desired temperatures
• Rangebetween30°Cand40°C,asthiscoversthe
given range and you are limited for time (ideally Summary test 3.2a
28°Cand42°Ciftimeallows)
If the temperature is increased,
• Takemeasurements,at2°Cintervalstopinpoint
more accurately the optimum temperature (a
the rate of enzyme action will
minimumoffivemeasurements) (1) up to a point at which its
molecular structure is disrupted
Takereadingsofoxygenvolumeevery5seconds
for30secondstoallowagraphtobedrawnfor and the shape of its (2) is altered
calculation of initial rate of reaction so that the substrate no longer
fits it. At this point the enzyme
Repeat each temperature three times to obtain means
and to detect any anomalous results is said to be (3). Many human
enzymes have an optimum
At each temperature use: same volumes of substrate;
same volumes of enzyme solution; same volumes of
working temperature of (4).
buffer solution to add to the enzyme to maintain pH, Enzymes also have an optimum
as these are factors that affect enzyme action pH at which they operate. Some,
Usethesamequantityofboiled(denatured)enzyme
like pepsin, work fastest at a
(andwithallothervariableskeptconstant)toensure pH of (5) while others, such as
that it is only the enzyme action that allows oxygen to (6), function fastest in neutral
be released and no other variable is acting conditions.
Changing the pH changes the
concentration of (7). This will
affect hydrogen and (8) bonds
between amino acids and can
affect the (9) of the active site
so that the enzyme is prevented
from operating at its maximum
rate. The ability to transfer (10) to
catalyse the reaction may also be
hindered.

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 3.2b Factors that affect enzyme
action: enzyme and substrate
concentration

These pages help you to: In addition to external factors, such as temperature and pH, substrate and
enzyme concentrations affect the rate of enzyme-catalysed reactions. An
• understand how to investigate the
enzyme reaction is always most rapid at first because the enzyme and substrate
effects of enzyme concentration
molecules can freely collide with one another. As the reaction proceeds,
and substrate concentration on
substrate molecules decrease in concentration and there are fewer successful
enzyme-catalysed reactions (3.2.1)
collisions between the molecules. The rate of reaction therefore slows.
• explain why changing enzyme
concentration or substrate
Effect of enzyme concentration on the rate of reaction
concentration will affect an
enzyme-catalysed reaction (3.2.1) Once an active site on an enzyme has acted on its substrate, it is free to
repeat the procedure on another substrate molecule. This means that enzymes
You will also:
are not used up in the reaction and therefore work efficiently at very low
• be able to interpret and explain concentrations. In some cases, a single enzyme molecule can act on millions of
results substrate molecules in one minute.
• identify sources of error
As long as there is an excess of substrate, an increase in the quantity of enzyme
• suggest improvements to your
leads to a proportionate increase in the rate of reaction. A graph of the rate
investigations
of reaction against enzyme concentration will initially show a proportionate
increase (straight line). This is because there is more substrate
Extension than active sites. If we increase the enzyme concentration,
more substrate will be acted upon and the rate of reaction
Non-protein biological catalysts
will increase. If, however, the substrate is limiting, i.e. there
It used to be thought that all biological catalysts were is not sufficient to supply all the enzyme’s active sites at one
enzymes and were therefore made of protein. We now know time, then any increase in enzyme concentration will have
that some reactions in cells are catalysed by RNA molecules, no effect on the rate of reaction. The rate of reaction will
also known as ribozymes. Some ribozymes work on therefore stabilise at a constant level, i.e. the graph will level
other RNA molecules, such as those that cut out unwanted off. This is because the available substrate is already being
sections from messenger RNA. This could answer the used as rapidly as it can be by the existing enzyme molecules.
‘chicken or egg’ question – which came first, the enzyme These events are summarised in Figure 1.
(protein) needed to make nucleic acids, or the nucleic acids,
needed to make enzymes? The answer could be RNA which,
in this sense at least, is both nucleic acid and ‘enzyme’.

Excess Substrate molecules

substrate in active sites of Empty active sites
molecules enzyme molecules
Enzymes
molecules

Low enzyme concentration Intermediate enzyme concentration High enzyme concentration

There are too few enzyme molecules to allow all
substrate molecules to find an active site at one
time. In this example the rate of reaction is
therefore only half the maximum possible for the
number of substrate molecules available.
Rate of reaction
With twice as many enzyme molecules
available, all the substrate molecules can
occupy an active site at the same time.
The rate of reaction has doubled to its
maximum because all active site are filled.
Rate of reaction
An increase in the number of enzyme
molecules present has no effect as there will
not be enough substrate to fill all the available
active sites. There is no increase in the rate of
reaction.

Rate of reaction

Substrate
concentration
Enzyme limiting
concentration
limiting

Enzyme concentration Enzyme concentration Enzyme concentration

Figure 1 Effect of enzyme concentration on the rate of enzyme action
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 Enzymes

Effects of substrate concentration on the rate of enzyme action Remember

If the concentration of enzyme is fixed at a constant level and substrate The active site and the substrate are
concentration is increased, the rate of reaction increases in proportion to the not the same, any more than a lock
increase in substrate concentration. If a higher concentration of substrate is and key are the same. The correct
used the active sites become fully occupied. They are said to be fully saturated term is complementary.
at the point where they are all working as fast as they can. The rate of reaction
is at its maximum (Vmax). After that, the addition of more substrate will have
no effect on the rate of reaction. In other words, when the substrate is in
excess the rate of reaction levels off. A summary of the effect of substrate
concentration on the rate of enzyme action is given in Figure 2.
Excess substrate molecules unable
Substrate molecules
Empty active sites to find any free active sites
in active sites of
Enzyme
Substrate molecules enzyme molecules
molecules

Low substrate concentration Intermediate substrate concentration High substrate concentration

There are too few substrate
molecules to occupy all the available
active sites in this example. The rate
of reaction is therefore only half the
maximum possible for the number of
enzyme molecules available.
Rate of reaction
With twice as many substrate molecules
available in this example, all the active
sites are occupied at one time. The rate
of reaction has doubled to its maximum
because all the active sites are filled.
A higher concentration of substrate has
no effect as all active sites are already
occupied at one time. There
is no increase in the rate of reaction
Remember
The Michaelis–

Rate of reaction

Rate of reaction Enzyme

concentration Menten
limiting constant is
Substrate a substrate
concentration
concentration.
limiting

Substrate concentration Substrate concentration Substrate concentration

Figure 2 Effect of substrate concentration on the rate of enzyme action Maximum velocity [ Vmax ]
Rate of reaction, i.e. velocity [ V ]

Maximum rate of reaction and the Michaelis–Menten constant

The Michaelis–Menten constant (Km) is the substrate concentration needed
for an enzyme reaction to proceed at half of its maximum rate (Figure 3). The Half maximum
constant is the same for any one enzyme but varies for different enzymes. It velocity
gives a measure of how easily an enzyme reacts with its substrate. In other
words, it measures the affinity of an enzyme for its substrate.
Compare enzyme X with enzyme Y:
• Enzyme X has a higher Km than enzyme Y.
• So, enzyme X has a lower affinity for its substrate than enzyme Y. Substrate concentration
• This means that enzyme X needs a higher concentration of substrate to reach Vmax.
The Michaelis–Menten constant can therefore be used to compare the Michaelis–Menten constant [ Km]
efficiency of different enzymes for their substrates. Figure 3 

Summary test 3.2b

Enzymes work fastest at the start of a process and this is called the (1). As the enzyme concentration increases, the rate
of reaction (2), provided there is excess substrate. The graph may later ‘tail off’ if the concentration of substrate is
limited because not all the (3) of the enzyme molecules are filled. If the substrate concentration of an enzyme-controlled
reaction is halved then the rate of reaction will be (4), but when the substrate is in excess the rate of reaction will (5).
A higher (6) constant for enzyme X compared to enzyme Y tells us that enzyme Y has a higher (7) for its substrate.
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 3.2c
These pages help you to:
• understand the difference
between competitive and
non-competitive inhibitors
(3.2.1)
• explain the effects of reversible
inhibitors on enzyme
activity (3.2.3)
You will also:
• appreciate the importance of
inhibitors in allowing the cell to
function efficiently
• understand how to investigate
how inhibitors affect the rate of
reaction of an enzyme
Extension
Inhibitors, helpful or harmful?
Many inhibitors regulate metabolic
reactions within the cell. Allosteric
regulators are reversible non-
competitive inhibitors that bind to
the allosteric site of the enzyme. They
control the activity of the enzyme
when enough product has been made.
Some inhibitors are irreversible and
bind to the enzyme active site or
allosteric site permanently. These can
damage the cell and may lead to cell
death. Heavy metal poisoning is due
to irreversible inhibition. Irreversible
inhibitors have been used as
therapeutic drugs: for example,
penicillin.
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Factors that affect enzyme action:
inhibitors
Enzyme inhibitors are substances that directly or indirectly interfere with the
functioning of the active site of an enzyme and so reduce enzyme activity.
Most inhibitors only make temporary attachments to the active site. These are
called reversible inhibitors and are of two types:
• competitive (active site directed) – inhibitor binds to the active site of the
enzyme
• non-competitive (non-active site directed) – inhibitor binds to the enzyme at
a position other than the active site.
Competitive (active site directed) inhibitors
Competitive inhibitors have a molecular shape that is complementary to that of
the substrate, which allows them to occupy the active site of an enzyme. They
therefore compete with the substrate for the available active sites (Figure 1).
It is the difference between the concentration of the inhibitor and the
concentration of the substrate that determines the effect this has on enzyme
activity: if the substrate concentration is increased, the effect of the inhibitor
is reduced. The inhibitor is not permanently bound to the active site and so,
when it leaves, another molecule can take its place. This could be a substrate
or inhibitor molecule, depending on how much of each type is present. Sooner
or later, all the substrate molecules will find an active site, but the greater the
concentration of inhibitor, the longer this will take.
Substrate molecule
occupying the active site
of the enzyme Substrate
molecule
unable to
enter the
active site
Inhibitor
molecule
occupying the
active site of
Enzyme the enzyme
molecule
Figure 1 Competitive inhibition
Non-competitive (non-active site directed) inhibitors
Non-competitive inhibitors attach themselves to the enzyme at a binding site
which is not the active site. This is known as the allosteric site (allosteric = ‘at
another place’). Upon attaching to the enzyme, the inhibitor alters the shape of
the enzyme’s active site in such a way that substrate molecules can no longer
occupy it, and so the enzyme cannot function (Figure 2). As the substrate
and the inhibitor are not competing for the same site, an increase in substrate
concentration does not decrease the effect of the inhibitor (Figure 3).
16/12/2019 12:35
 Enzymes

In this example, the substrate

molecule cannot enter the active
site because the the active site
has changed shape. This means
Substrate molecule that it is no longer complementary
occupying the active site to the shape of the substrate.
of the enzyme In some cases, the substrate may
be able to enter the active site but
1. Inhibitor absent – 2. Inhibitor present – the changes are too great to allow
The substrate attaches The inhibitor prevents binding or catalysis to occur.
to the active site of the the normal enzyme–
enzyme in the normal substrate complex Enzyme molecule shape is
way. An enzyme-substrate being formed. The changed due to presence of
complex forms and the reaction rate is reduced. the inhibitor molecule, changing
reaction takes place as the shape of the active site
normal.
Inhibitor molecule attached
Enzyme molecule to enzyme molecule

Figure 2 Non-competitive inhibition

Investigating the effect of inhibitors

The rate of reaction of the enzyme needs to be determined over a range of
Maximum rate of reaction

Rate of reaction, i.e. Velocity [v]

substrates, and without the presence of an inhibitor at each substrate concentration.
The main variables that need to be standardised are: temperature, enzyme Absence of
inhibitor Fixed quantity
concentration, volume of enzyme solution, volume of substrate solution and pH. of competitive
The quantity of inhibitor (concentration and volume) also needs to be standardised. inhibitor

How the presence of inhibitors affects Vmax and Km Fixed quantity of

non-competitive
Competitive inhibition inhibitor
• At higher substrate concentrations, the effect of a competitive inhibitor
decreases and Vmax is reached.
• Km increases (Figure 4). Substrate concentration
• The enzyme has a lower affinity for its substrate. Figure 3 Comparison of competitive and
Vmax non-competitive inhibition on the rate of
Absence of an enzyme-controlled reaction at different
inhibitor
substrate concentrations
Rate of reaction

Presence of
1/2 Vmax
competitive inhibitor

Summary test 3.2c

Figure 4 The effect of
Km Km competitive inhibition In competitive inhibition, the
Substrate concentration on Vmax and Km
inhibitor has a similar shape to
Non-competitive inhibition the (1). This allows it to fit into
• The effect of a non-competitive inhibitor remains the same and Vmax is not the active site of the enzyme,
reached. preventing the formation of an
• Km remains the same (Figure 5). (2). Increasing the substrate
• The enzyme has the same affinity for its substrate. concentration decreases the
inhibition and (3) is reached.
Vmax
Km is increased, showing that
Absence of the enzyme has less (4) for its
inhibitor
Rate of reaction

substrate. In non-competitive
Vmax

1/2 Vmax Presence of non-

inhibition, the inhibitor does not
competitive inhibitor bind to the active site but to the
1/2 Vmax (5), causing the (6) of the active
site to change. The substrate is no
Figure 5 The effect longer (7) in shape to the active
Km of non-competitive site and catalysis cannot occur.
Substrate concentration inhibition on Vmax and Km

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 3.2d Factors that affect enzyme action:
immobilised enzymes

These pages help you to: One method of immobilising enzymes is to trap them in small alginate beads.
This is carried out in a laboratory as follows.
• understand how to investigate
the difference in activity • The enzyme to be immobilised is mixed with a solution of sodium alginate.
between an enzyme that is • Tiny droplets of this mixture are added to a solution of calcium chloride, one
immobilised in alginate and at a time using a syringe.
the same enzyme free in • A reaction takes place between the sodium alginate–enzyme mixture and the
solution (3.2.4) calcium, causing calcium ions to replace sodium ions.
• state the advantages of using • As a result of this reaction, jelly-like beads are formed in which the enzyme is
immobilised enzymes rather trapped.
than enzymes free in
To catalyse reactions, the beads are packed into a long column in a vessel and the
solution (3.2.4)
substrate is poured in at the top. As the substrate enters the beads, it is converted
You will also: to the product by the enzymes immobilised in the beads. Although the process
cannot continue indefinitely, as impurities accumulate, it can proceed for a
• improve your ability to describe
considerable time without renewing the enzyme (Figure 1).
and interpret graphical results
Enzymes are used in a wide range of industrial processes, including the
Substrate in
production of foods, agrochemicals and drugs. In many cases, the enzyme and
(e.g. milk containing lactose) substrate are mixed together to form a product. This is then extracted and the
rest of the mixture, including the enzyme, is discarded. Enzymes are costly to
produce so this is both wasteful and expensive. As enzymes are not used up
Tap to control in reactions, keeping them for future use clearly has a cost benefit. One way
flow of substrate of doing this is to immobilise the enzymes so that they can be retained rather
into the column
than discarded.

Alginate beads
containing Advantages of using immobilised enzymes or cells
immobilised enzyme
e.g. β-galactosidase
There are a number of advantages to immobilising enzymes or cells.
• With enzyme immobilisation, the enzyme can be used repeatedly as it is not
Long column
containing lost in the process making it more economic, especially where the enzyme is
the beads expensive.
• Enzymes are vulnerable to changes in temperature and pH. The beads in
which they are trapped can buffer them against these changes.
• The immobilised enzymes and cells, being held in place, cannot contaminate
the substance being made leading to a purer product.
• With whole cell immobilisation, a number of enzymes can act together at the
same time in a single process.
• Downstream processing, the steps needed to obtain a purified, high quality
Tap to control product, is easier. This is because very little, or no enzyme will be lost from the
flow of substrate reaction vessel.
out of the column
• The shelf-life of the enzyme is longer because it is less vulnerable to extremes
Product out of pH and temperature.
(e.g. milk with glucose and galactose,
but no lactose)
• The protection given by the matrix to higher temperatures means that the
reaction can proceed at higher temperatures to give greater productivity.
Figure 1 Using immobilised enzymes to • The process can be carried out on a continuous basis as enzyme does not need
produce lactose-free milk to be continually recovered from the product mixture.

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 Enzymes

Extension
(a) Improving the productivity of an immobilised enzyme
The experimental set-up shown in Figure 1 can be used to determine the An example of the commercial use
conditions that will give the highest rate of product formation. One factor should be of immobilised enzymes is in the
investigated at a time, with all other conditions kept constant. Examples of factors manufacture of lactose-free milk for
that can be varied: temperature, substrate concentration, enzyme concentration, people with lactose intolerance. In
pH, size of bead, rate of flow of substrate through the column. this case the immobilised enzyme is
β-galactosidase. Milk contains the
(b) Investigating the difference in activity between an immobilised enzyme and an sugar lactose which is the cause
enzyme free in solution. of the discomfort experienced by
The optimum temperature or the optimum pH for an enzyme that has been lactose-intolerant individuals. To
immobilised is not always the same as those for the enzyme when it is free in solution. produce lactose-free milk, the milk
is passed over the immobilised
When comparing the rate of reactions of an immobilised enzyme and the enzyme
β-galactosidase which catalyses the
free in solution at different temperatures, pH or substrate concentrations, remember
hydrolysis of lactose to glucose and
that all factors other than the one being investigated should be kept constant
galactose. Figure 1 illustrates the
(standardised variables).
process.
Once you have obtained your raw data, it is helpful to construct one graph that
You can check specifically for the
contains the results for both, for easier comparison. Remember that comparing
presence of glucose using test strips
means that you need to look for similarities as well as differences.
known as glucose dipsticks:
For temperature or pH, compare:
• Test the lactose at the start of the
• the optimum for the immobilised enzyme with the enzyme free in solution experiment to confirm that there is
• the pattern that is shown as temperature (or pH) increases (look at the overall no glucose
shape of the curves) • Test the product mixture to
• any differences in the level of activity shown at each temperature (or pH) tested confirm that hydrolysis has taken
• any differences in the steepness of the curve ‘before’ the optimum and ‘after the place.
optimum’.
You can check if any enzyme has
For substrate concentration compare: ‘leaked’ into the product mixture by
taking a small sample of the mixture
• the maximum velocity, Vmax and the Michaelis–Menten constant, Km
and carrying out a test for the
• the steepness of the increase up to the Vmax.
presence of proteins (using Biuret
solution).

Summary test 3.2d

To immobilise an enzyme in beads, the enzyme solution is mixed with

sodium (1) and the mixture is placed into a (2) to make small drops that
can be added to a solution of (3). Advantages of using immobilised enzymes
include: the product is not (4) by enzyme; the enzyme can be recovered
and (5); the enzyme is more tolerant to changes in (6) and temperature; (7)
processing is easier.

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 3 Exam-style and Practice Questions

1 Phosphatases are a group of enzymes that catalyse the A 2 cm3 volume of lipase solution was incubated at the
removal of phosphate groups from organic compounds. same temperature in a separate test tube before being
added to the vegetable oil.
A group of students investigated the effect of substrate
concentration on the initial rate of the reaction The initial pH of the reaction mixture was measured
catalysed by phosphatase 1. The results are shown in using a pH meter. The pH was recorded at 5-minute
Figure 1. intervals for 60 minutes.
Figure 1 a State why the lipase solution was incubated
separately before being added to the
4
vegetable oil. (1 mark)
b Suggest why the vegetable oil was adjusted
× × × to pH 8.0 before the lipase was added. (1 mark)
3
Rate of reaction / umol min–1

×
Figure 2 shows the results of the investigation.
Figure 2

2 8
×

1 7
pH

0 × 6
0 1 2 3 4 5
0 5 10 15 20 25 30 35 40 45 50 55 60
Substrate concentration / mmol
Time / min
a Explain the results shown in Figure 1. (5 marks) c With reference to Figure 2, describe and
b i State the lowest substrate concentration to explain the results of the investigation. (5 marks)
give the maximum rate of reaction, Vmax. (1 mark) d Lysozyme A is an enzyme found in human tears
ii Determine the Michaelis–Menten and saliva. It hydrolyses the β-1,4 glycosidic bonds
constant, Km. (1 mark) present in compounds found in bacterial cell walls.
c The students repeated the investigation, but Lysozyme uses the induced fit mechanism.
added glycine, a competitive inhibitor of Explain the mode of action of an enzyme that
phosphatase 1, to each reaction mixture. uses the induced fit mechanism. (4 marks)
i Sketch a curve on Figure 1 to show the e Each molecule of lysozyme consists of a single
expected results. (2 marks) polypeptide. In a mutated form of lysozyme, variant
ii Explain the effect that glycine will have lysozyme, a mutation has caused a single amino
on the action of phosphatase 1. (2 marks) acid, phenylalanine, to be replaced by the amino
acid isoleucine. Variant lysozyme is less active that
(Total 11 marks) normal lysozyme and has also been linked to a
disease called renal amyloidosis, where protein fibrils
2 A student carried out an investigation into the digestion are deposited in the kidneys.
of triglycerides using lipase.
Suggest how the difference in one amino acid is
3
A volume of 20 cm of vegetable oil, adjusted to pH 8.0, responsible for the lower activity of variant
was added to a test tube, which was then put in a water lysozyme compared with normal lysozyme. (2 marks)
bath at 40 °C for 5 minutes. (Total 13 marks)

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 Exam-style and Practice Questions

3 Enzymes can function at a wide range of temperatures. 4 a Explain why enzymes function less well at lower
Shrimps that live in Arctic waters have enzymes that temperatures. (2 marks)
function best around 4 °C and are denatured at around
b Explain how high temperatures may prevent
15 °C. By contrast, bacteria that live in hot springs have
enzymes from functioning at all. (2 marks)
enzymes that function best at 95 °C and continue to
operate effectively above 100 °C. These bacteria are c Enzymes produced by microorganisms are
called thermophilic (heat-loving) bacteria. responsible for spoiling food. Using this fact and your
knowledge of enzymes suggest why the following
Enzyme X is produced by thermophilic bacteria and procedures are carried out:
hydrolyses many proteins, including haemoglobin and i Food is heated to a high temperature before being
egg albumin. canned. (1 mark)
ii Some foods, such as onions, are preserved in
Enzyme Y is found in the stomach of young mammals, vinegar. (1 mark)
where it acts on a single soluble protein found in milk,
d The figure below represents an enzyme and its
causing it to coagulate (clot).
substrate. Of the other four molecules shown, one is
a i From the descriptions, comment on the differences a competitive inhibitor and one is a non-competitive
in the specificity of the two enzymes. (2 marks) inhibitor of the enzyme.
ii Enzymes X and Y are each used for different
commercial purposes. Suggest what this might be 1
in each case. (2 marks)
iii Suggest a possible purpose of enzyme Y in the Enzyme
Active
molecule 2
mammalian stomach. (1 mark) site
iv Use the information about the two enzymes
to suggest a possible difference in the type of 3
bonding found in the tertiary structure of each.
Explain your reasoning. (3 marks) Substrate 4
b An experiment was carried out with enzyme X in molecule
which the time taken for it to fully hydolyse 5 g i State the number of the molecule that is a
of its protein substrate was measured at different competitive inhibitor. (1 mark)
temperatures. The following data were obtained: ii State the number of the molecule that is a non-
5 60
competitive inhibitor. (1 mark)
Temperature/°C Time for hydrolysis Rate of reaction
(Total 8 marks)
of protein/min 1
time

15 5.8
25 3.4
35 1.7
45 0.7
55 0.6
65 0.9
75 7.1

i Calculate the values for 1/time for each of the

temperatures. (2 marks)
ii Sketch a graph that shows the effect of temperature
on the rate of reaction of enzyme X. (3 marks)
iii State the optimum temperature for the action of
enzyme X. (1 mark)
iv Suggest how you might determine this optimum
temperature more precisely. (2 marks)
(Total 16 marks)

57

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 Notes
Student Book Contents Fully
matched to
the latest
syllabus
AS Level A Level
Introduction 6. Welcome to A Level
➞ Thinking about language
1. A toolbox for textual analysis
➞ Exploring meaning A selection of
➞ Genre and context
➞ Denotation and connotation pages from this
➞ Voice and point of view chapter are
➞ Structure, form, cohesion 7. Language change included for you
➞ Lexis and diction ➞ Attitudes to language change to evaluate.
➞ Register and tone ➞ Dealing with historical language data
➞ Grammar ➞ Language change 1500-present day
➞ Metaphorical language ➞ Language change reflecting society
and culture, politics and technology
➞ Spoken and written language
➞ Consequences of language change
2. Language issues
8. Child Language Acquisition (CLA)
➞ The language of advertising
➞ Ways of representing spoken language in
➞ Language – unconsciously conveying
written form
attitudes
and values, plus gender ➞ Stages of children’s language development
➞ Language change and variation ➞ Exploring the functions of children’s
language
➞ English, the Internet and electronic
communication ➞ Concepts, theories and research studies
in CLA
➞ Diaries, autobiographies, and biographies
9. English in the world
3. Paper 1: Reading
➞ Historical development of English as
➞ Phase 1 – Tackling the question
a ‘global’ language
➞ Phase 2 – Creating a text
➞ Roles and status of English language
➞ Phase 3 – Comparing texts
➞ Varieties of English
➞ Sample questions
➞ Issues and problems with English Language
➞ Analysing texts: Contextual and linguistic
analysis 10. Language and the self
➞ Construction and communication of
4. Paper 2: Writing a senseof self
➞ Shorter writing and reflective commentary ➞ Social identity: Individuals and groups
➞ Creating longer texts – extended writing: ➞ Relationship between language and thought
➞ Route 1: Imaginative writing
➞ Route 2: Review/critical writing 11. Papers 3 and 4: Exam technique
and preparation
➞ Route 3: Discursive / argumentative writing
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5. AS Level: A conclusion

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 Cambridge International AS & A Level
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Enhanced Online Book 978 138 200543 2

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