CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: COURSEBOOK

Sample answers have been written by the authors.

Coursebook answers
Chapter 3
Self-assessment questions
1 Explanation of results: 3 a
• Catalase, liver and potato were much more

Amount of starch in mixture

efficient than the inorganic catalysts:
The catalase enzyme has evolved to
be the correct shape to catalyse the
decomposition of hydrogen peroxide
efficiently.
• Pure catalase was more efficient than
the liver and potato: The concentration
of the pure enzyme is higher than the
concentration in liver and potato. The
higher the concentration of an enzyme, Time
the faster it works.
• Liver was more efficient than potato: Liver b Calculate the slope of the curve right at
is an animal tissue. Animal tissues have a the beginning of the reaction.
higher metabolic rate than plant tissues.
Hydrogen peroxide therefore probably 4 Measure the volume of oxygen given off over
builds up faster in liver cells than potato regular time intervals for several hydrogen
cells and must therefore be got rid of peroxide–catalase reactions at different
faster. Liver cells therefore probably have temperatures. In each case, all conditions
a higher concentration of catalase than other than temperature must remain constant.
potato cells. In particular, the volume and concentration of
• Ground-up liver was more efficient than hydrogen peroxide solution, and the volume
pieces of liver: Grinding up the liver and concentration of catalase solution must
breaks open the cells and releases the be kept constant each time. Plot volume
contents, including catalase. The catalase of oxygen against time for each reaction.
therefore has easier access to the substrate Calculate the slope of the line at the beginning
(hydrogen peroxide). of the reaction in each case to give the initial
reaction rate. Then plot initial reaction rate
2 In case of inaccuracy of measurement at against temperature.
30 seconds. The overall shape of the curve is Alternatively, the volume of oxygen given
more likely to give an accurate value. off in a given time period, for example
1 or 2 minutes, could be found for each
temperature. The volume is proportional to
the rate, so a graph of volume (rate) against
temperature could be plotted. Another
alternative would be to time how long it takes
to collect a given volume of oxygen for each

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 CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: COURSEBOOK

temperature. In this case, the inverse of the 8 As soon as the reaction starts, the substrate
time taken would be a measure of rate. starts to be used up, so the substrate
concentration starts to fall. This results in a
5 a Haemoglobin is the coloured pigment corresponding fall in the rate of reaction. It is
that causes bloodstains. Haemoglobin the initial rate at the start of the experiment
is a protein. Protein-digesting enzymes that is the true rate.
(proteases) catalyse the hydrolysis of
haemoglobin to amino acids, which are 9 a carbonic anhydrase
colourless. They are also soluble, so will
wash away in water. b lysozyme, because it has the lowest Km

b Many protein-digesting enzymes have an 10 a

optimum temperature of around 40 °C,
which is a relatively low temperature for
a washing machine. Washing at lower
temperatures also helps to save money. enzyme B

Rate of reaction
c Other components of washing powders, enzyme A
such as the oil-removing detergents, work
best at high temperatures.

6 One possible answer is as follows; other answers

might be equally acceptable.
Set up two sets of five tubes containing equal Substrate concentration
volumes of the same concentration of milk
suspension. Make up five buffer solutions b i enzyme B
of varying pH. Add equal volumes of buffer
ii enzyme B
solution, one of each pH, to each set of five
tubes containing milk suspension. To one iii enzyme B  
set of tubes, add equal volumes of trypsin
solution. To the other set of tubes, add the iv enzyme B
same volume of water; these act as controls.
Time the disappearance of cloudiness in each 11 a Prepare a solution of lactase and a
tube containing trypsin. Cloudiness should separate sample of lactase immobilised
remain in the control tubes. Plot rate of in alginate beads. Suspend the beads in
reaction (1/time taken) against pH. water. Heat separate samples of lactase
in solution and immobilised lactase
7 beads suspended in water at a range of
temperatures (e.g. 40 °C to 100 °C) for
10 minutes each. Allow to cool to room
If substrate temperature.
becomes
limiting, the If the enzyme is still active, it will catalyse
Initial rate of reaction

addition of the hydrolysis of lactose to glucose

extra enzyme
and galactose: 1 mole of lactose will
cannot increase
the rate of produce 1 mole of glucose and 1 mole of
reaction. galactose. All are reducing sugars,
but the concentration of reducing sugar
will double as a result of the reaction.
A semi-quantitative Benedict’s test on a
sample before and after the reaction
Enzyme concentration can therefore be used to find out if the

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 CAMBRIDGE INTERNATIONAL AS & A LEVEL BIOLOGY: COURSEBOOK

enzyme is active. This can be done by

mixing the enzyme with lactose (or milk)
Reflection
at a suitable temperature (e.g. 37 °C), and A range of answers is possible, which may include
leaving it for a few minutes. In the case of the following:
the lactase beads, you would first have to
• amino acid sequence / primary structure
tip the beads into a sieve to remove the
surrounding water before adding them to • tertiary structure (to match any potential
a solution of lactose. substrate)

b Suspend the lactase beads in water. Leave • temperature optimum

samples of the enzyme solution and the • range of temperatures over which effective
immobilised lactase beads for different
lengths of time at 90 °C. Then test for • pH optimum and range
enzyme activity as in a above. • affinity for substrate
c Prepare a series of lactase solutions of • Vmax
different pH using appropriate buffer • ability to stick to a substrate for
solutions. For each pH, use some of immobilisation
the solution to make alginate beads
containing immobilised enzyme. Then • longevity when immobilised / resistance to
carry out the reaction using milk or denaturation; disulfide bonds may increase
lactose as a substrate and test for enzyme stability
activity as in a above. • ability to add useful prosthetic groups.
12 Immobilised enzymes do not contaminate the There are many possible new uses for enzymes.
product. They are not lost, so they can be re- Being able to break down plastics using enzymes is
used. They are able to work at a wider range an obvious use. Scientists would like to be able to
of pHs than enzymes in free solution, and also carry out nitrogen fixation using enzymes.
at a wider range of temperatures. They are
more resistant to denaturation.

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