1.

Beetroot cells contain a red pigment.

Cut cubes of beetroot were washed to remove the red pigment from damaged surface cells.
The cubes were placed in test tubes containing 10cm 3 water, in waterbaths at a range of temperatures.
The tubes were left at each temperature for 10 minutes. The beetroot was removed from the tubes and the
colour of the solution in the tube measured on a colorimeter. The darker the red colour of the solution, the
greater the colorimeter reading.
The following results were obtained.

(i)

Describe the pattern of results from the above table.

[2]

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(ii)

Explain the results as fully as you can. Refer to cell membrane structure in your answer.

[3]

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(iii)
Explain the difference in the results if you were investigating the effect of ethanol on the
permeability of the membrane.

[2]

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(Total 7 marks)

2.

In an experiment the enzyme catalase, obtained from liver tissue, was used to break down
hydrogen peroxide releasing oxygen. The experiment was carried out at selected pH values and
the volume of oxygen given off was measured at intervals. The results obtained were plotted as
shown below.

(a)
Which one of the four pH values would be nearest to the optimum for
catalase?
[1]
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(b)

Select the letter which indicates:

(i)

The fastest rate of reaction.

[2]

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Give a reason for your
choice. ...................................................................................................................

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(ii)

The least concentration of hydrogen peroxide.

[2]

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Give a reason for your
choice......................................................................................................................
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(c)
[3]

Explain fully the results obtained at pH 4.4.

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(d)

A control was set up at optimum pH conditions using boiled and cooled liver. Suggest an
explanation for the shape of the control curve shown on the above graph.

[1]

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(e)

Outline how the experiment could be modified in order to investigate the effect of the
concentration of catalase on the rate of the reaction.

[2]

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(Total 11 marks)

3.
When a potato tuber is cut open, the surfaces exposed to the air gradually
turn brown due to
the production of dark brown coloured pigments from phenolic compounds (e.g.
catechol). The enzyme, polyphenol oxidase, catalyses this process.
The graph below shows the results of an experiment in which different
concentrations of the enzyme were added to tubes containing catechol solution.
The tubes were kept at a constant temperature and shaken periodically during the
experiment. The time taken for a standard brown colour to develop was recorded
and from this the rate of reaction was calculated, using time in minutes 1 for the
colour to develop.
A graph was then plotted of rate against enzyme concentration.

(a)
What general conclusion concerning enzyme action can be drawn from these
results? [1]
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(b)

At what enzyme concentration would the standard colour be obtained after

18 minutes? Show your working.
[2]

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(c)
Explain why the addition of lemon juice (citric acid) to the tubes stops any
colour change. [1]

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(d)

Explain what is likely to happen to the rate of colour change if the

temperature of a tube containing the enzyme polyphenol oxidase and
catechol was gradually raised from 10C to 100C.
[2]
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(e)

Suggest what might happen to the rate of reaction if a competitive inhibitor

was added. Explain your answer.
[2]

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(f) '

All enzymes are proteins', describe how you would test to show that
polyphenol oxidase is a protein.
[2]
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(Total 10 marks)

1.

(i) {very little increase/0-7 units} up to {40C/first 4 temperatures;

rapid rise at 50C/rise from 7 to 80 at 50C;
small rise from 80 to 100 units from 50 to 70C [2]
higher temperature, darker solution/higher reading
(ii) Membrane (proteins) are stable between 10 and 40C/at low
temperatures;
(Appearance of the red pigment means that) the (tonoplast and) cell
membrane has
been damaged between 40 and 50C/at high temperatures;
(meaning) pigment has leaked out of cells/sap vacuole/change in
membrane
permeability/damaged membrane;
proteins change shape/denature and come out of bilayer; [3]
(iii) Dissolves phospholipids/destroys cell membrane structure; [1]
(So) pigment leaks out. [1]
(not: ref. to higher or lower readings)
(Total 7 Marks)
2.
(a) 6.5 [1]
(b) (i) A [1]
the greatest volume of oxygen produced in unit time / reference to
minutes [1]
(ii) E [1]
the greatest number of substrate / hydrogen peroxide molecules have been
broken down to produce the oxygen / all the enzyme active sites are
occupied. [1]
(c) A change in pH in the environment of the active site alters the ionic
charge of the
acidic and basic groups of the amino acids and causes the shape of the
protein to change.
This may prevent the substrate molecules from making a close fit to
the active site. [3]
(d) The slight reaction is due to the decomposition of hydrogen peroxide in
air. [1]
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(e) (The substrate concentration is constant)

Use the optimum pH value [1]
Vary the concentration of catalase by dilution of minced liver / changing
the mass of
the minced liver added. [1]
(Total 11 Marks)
3.

(a) As the concentration increased the rate of reaction increased. [1]

(b) 5.5; (allow : 5.6) [1]
Fig. from graph (1.1 to 1.2) [1]
(c) Enzyme has a pH optimum / does not work in acid pH caused by lemon
juice [1]
(allow: denatured not: destroyed / inhibited)
(d) Faster (colour change) at start as temperature increases; [1]
No further colour change after enzyme denatured. [1]
(e) rate drops / stops if all active sites blocked; [1]
as it competes with normal substrate for active sites. [1]
(f) Biuret test / NaOH or any alkali plus CuSO 4; [1]
Purple / violet ring / colour develops if protein. [1]
(linked marks) (Total 10 Marks)