CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: WORKBOOK
Exam-style questions and sample answers have been written by the authors. In examinations, the way marks are awarded
may be different.
Workbook answers
Chapter 3
Exercise 3.1 Answering questions
about graphs
1 a As time increases, the total volume of
oxygen collected also increases. The rate
of increase is fastest at the start of the
reaction, gradually, decreasing, until from
about 330 s onwards there is no further
increase and the graph levels off at a value
of 11.6 cm3.
b The concentration of the substrate,
hydrogen peroxide, is greatest at the start.
This is when collisions between enzyme
and substrate will be most frequent, and
therefore when the rate of reaction is
greatest. This explains the high rate of
production of oxygen during this stage.
As time increases, the concentration
of substrate decreases, because it is
being changed into product. The rate of
reaction therefore gradually slows, until it
becomes zero at 330 s onwards, because
all of the substrate has been converted to
product.
2  he curve for catalase from apple is similar
T increased frequency of collision between
to that for carrot, in that it shows a rate of
reaction that decreases with time. However,
the rate (that is, the slope of the curve) is
always less than that for carrot. The maximum
volume of oxygen given off in 360 s is only 6.8
cm3, which is 4.8 cm3 less than the maximum
volume for carrot. Unlike the curve for carrot,
the apple curve has still not levelled off at
this time, showing that the reaction is not
yet complete and there is still more substrate
present.
3  he value that you obtain will depend on
T molecules is complete and no reaction
exactly how you draw the tangent, which is
tricky to do precisely. Your answer should be
somewhere close to 0.13 cm3 s–1.
1 Cambridge International AS & A Level Biology – Jones & Parkin © Cambridge University Press 2020
Exercise 3.2 Effects of temperature
and pH on enzyme activity
1 a At temperatures from 20 to 44 °C, the rate
of activity remains constant at about 15
arbitrary units. As temperature increases
from 44 °C to just over 60 °C, the rate of
activity increases steeply, reaching a peak
of 140 a.u. at about 61 °C. At temperatures
higher than this optimum value, activity
decreases very steeply, reaching 0 at 75 °C.
b At low temperatures, the kinetic energy
of enzyme and substrate molecules is
relatively low, and collisions are infrequent,
resulting in a relatively low rate of reaction.
However, we would normally expect the
rate of activity to increase as temperature
increases, but this does not happen until
the temperature reaches 44 °C. Perhaps
the activation energy for the reaction is not
reached at temperatures below this so, even
when enzyme and substrate collide, the
reaction only rarely takes place.
As temperature increases above 44 °C,
enzyme and substrate, which happens
with more energy, result in an increased
rate of reaction.
At temperatures above 61 °C, the high
kinetic energy of the molecules causes
vibrations in the enzyme molecules,
breaking hydrogen bonds and causing the
enzymes to lose their three-dimensional
shape. The active site no longer fits the
substrate molecule and, at a temperature
of 75 °C, denaturation of the enzyme
occurs.
2 a We would normally expect activity to
be greatest at an optimum pH and to
decrease at pHs lower or higher than that.
 CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: WORKBOOK

Here, however, we have a ‘two-humped’ temperature. (Note: you cannot keep pH

curve, with two pHs at which rate is the same, because the dependent variable
higher than either side of them. There is a change in pH.)
appear to be two optimums, one at pH 5,
2 I t is possible that the inhibitor might be
and the other over the range 8–9.
harmful, so care should be taken not to get it
b The two peaks on the pH curve suggest onto the skin, or to breathe in the powder.
there are two forms of the enzyme present,
each one with its own optimum pH value.  he solution produced when ammonia
T
dissolves in water may be very alkaline, so
3  emon juice contains citric acid, which will
L this should not be allowed to touch skin or
lower the pH. The graph for pH shows that the clothing. You should wear goggles and a
enzymes are relatively inactive at low pH values. protective laboratory coat; you could also
wear gloves.
Exercise 3.3 Finding Vmax and Km 3 a Use a top pan balance to measure out
1  he concentration of the lipase and the
T 10 g of urea.
temperature. Put this into a 100 cm3 volumetric flask.
2 35
Add a small volume of distilled water.

Mix until the urea has all dissolved.
30
Initial rate of activity of lipase/µmol fatty acid

Add more distilled water to make up the

solution to the 100 cm3 mark on the flask.
25
Mix thoroughly.
produced min-1

20 b You could use a more concentrated

3  his is the value at which the graph levels off.
T
Vmax is 33 µmol of fatty acid per minute.
4 You should find the Km is just above 10.0%
concentration of substrate.
5  he second enzyme has a greater affinity for its
T b You could try 1%, 0.1%, 0.01%, 0.001%
substrate than the one from B. cepacia.
Exercise 3.4 Planning an
investigation into the effect of
inhibitor concentration on urease
activity
1 a
b the rate of activity of urease, measured as
the rate of change of pH
c The concentration of the enzyme; the
concentration of the substrate; the
solution of urease.
15 4 a You could use serial dilution. Take 1 cm3
of the 1% solution and add it to 9 cm3 of
10 water, to make a 0.1% solution. Then take
1 cm3 of the 0.1% solution and add it to
9 cm3 of water, to make a 0.01% solution.
5
Repeat to make a range of solutions, each
10 times as dilute as the previous one.
0
0 10 20 30 40 50 Alternatively, you could take 8 cm3 of the
Concentration of substrate / % 1% solution and add it to 2 cm3 of water,
to make a 0.8% solution. Then take 6 cm3
of the 1% solution and add it to 4 cm3 of
water to make a 0.6% solution. Continue
until you have a sufficient range of
concentrations.
and 0. Alternatively, you could try 1%,
0.8%, 0.6%, 0.4%, 0.02% and 0. The
sample with no enzyme is a control.
c It would be good to do ten replicates at
each concentration of inhibitor.
5  easure a known volume of urease solution
M
– say 5 cm3 – into several test tubes. Place the
the concentration of the inhibitor
test tubes in a water bath at a temperature
known to be close to the optimum for this
enzyme.
 easure a known volume of each concentration
M
of inhibitor (including 0) – say 5 cm3 – and add

2 Cambridge International AS & A Level Biology – Jones & Parkin © Cambridge University Press 2020
 CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: WORKBOOK

to each of the test tubes containing the enzyme, Exercise 3.5 Calculating actual
in the water bath.
and percentage error
 easure a known volume of urea solution – say
M
5 cm3 – into several test tubes. Place these test 1 ±5 cm3
tubes in the water bath. 2  he uncertainty or error for each
T
 eave all the tubes for at least ten minutes,
L measurement is ± 1 cm3, so the total error
to allow their contents to come to the when calculating the difference between two
temperature of the water bath. measurements is ± 2 cm3.

 lace a pH probe into one of the test tubes

P 3  he error in the measurement is 0.05 cm3. The
T
containing the enzyme and inhibitor. Note the percentage error is therefore (0.05 ÷ 10) × 100 =
pH. Pour the contents of one of the urea test 0.5%.
tubes into the enzyme and inhibitor. Note the 4  he error in each measurement is 0.5 mm, so
T
pH at suitable intervals of time, or after a set the error in the measurement of the change in
period of time. Record. length is 1.0 mm.
 epeat with each of the concentrations of
R The change in length is 3.35 cm.
inhibitor.
 he percentage error is therefore
T
Repeat the whole experiment three times. (1.0 ÷ 33.5) × 100 = 2.99%.
6  he precise design of the results chart will
T 5  he error in each measurement is 0.005 g, so
T
depend on the method for recording the the error in the measurement of the change in
results. This could either be the pH reached at mass is 0.01 g.
a particular time, or the time taken to reach a
particular pH. The change in mass is –0.07 g.

One possible design could be:  he percentage error is therefore (0.01 ÷ 0.07)
T
× 100 = 14.3%.
Concentration pH after 5 minutes
of inhibitor / % Trial 1 Trial 2 Trial 3 Mean  otice that we ignore the minus sign when
N
calculating actual error or percentage error –
it makes no difference whether the mass has
gone up or down.

7  he curve should be drawn on axes labelled

T
with the concentration of the inhibitor (x-axis)
and the dependent variable (e.g. the pH after
five minutes) on the y-axis. The curve should
show that the rate of reaction decreases as
the concentration of the inhibitor increases,
plateauing at high concentrations of inhibitor.
So, if the dependent variable is the pH after
five minutes, the curve should start high when
the concentration of inhibitor is 0, then fall to
a constant value.

3 Cambridge International AS & A Level Biology – Jones & Parkin © Cambridge University Press 2020