2.

5 enzymes

1. Consider the metabolic pathway shown below.

If there is end-product inhibition, which product (B to E) would inhibit which enzyme (1 to 4)?

Product Enzyme
A. C 4
B. B 3
C. B 4
D. E 1

2. What can reduce the effect of a competitive inhibitor of an enzyme?

A. Decrease the temperature at which the reaction takes place.

B. Increase the temperature at which the reaction takes place.

C. Increase the substrate concentration.

D. Add a non-competitive inhibitor.

3. What effect do enzymes have on the activation energy of exergonic and endergonic reactions?

Activation energy of Activation energy of

exergonic reactions endergonic reactions
A. increases increases
B. decreases decreases
C. increases decreases
D. decreases increases

4. In the enzyme controlled pathway shown below, which compound is most likely to inhibit
enzyme (w)?

A. I

B. II

C. III

D. IV

5. The graph below shows the effect of changing the substrate concentration on an enzyme

1
 controlled reaction.

What is the correct interpretation of these data?

A. The rate of reaction increases continuously with increase in substrate concentration.

B. The rate of reaction decreases continuously with increase in substrate concentration.

C. The rate of reaction increases up to a point and then remains constant.

D. The rate of reaction is not affected by any change in the substrate concentration.

6. Which of the following could cause denaturation of an enzyme?

A. Substrate concentration

B. A competitive inhibitor

C. High temperature

D. Low salt concentration

7. What determines the specificity of an enzyme for its substrate?

A. The temperature at which it is operating

B. The optimum pH of the enzymes

C. The concentration of the substrate

D. The structure of the enzyme molecule

8. The graphs below show the energy changes during endergonic and exergonic reactions, with and
without enzymes.

Which line represents an endergonic reaction, without an enzyme?

A. I

B. II

C. III

D. IV

2
9. The reaction below shows the energy changes in a chemical reaction.

What would happen to the changes in energy if this reaction was controlled by an enzyme?

A. I would increase.

B. II would decrease.

C. I and IV would decrease.

D. II and III would decrease.

10. According to the induced fit model of enzyme function, which of the following statements is
correct?

A. Active sites on enzymes are specific to a single substrate.

B. The shape of the active site can be changed by the binding of an allosteric inhibitor.

C. The binding of the substrate changes the shape of the active site slightly.

D. Competitive inhibitors can change the shape of enzymes.

11. What is an active site?

A. The part of an enzyme that binds only to the product molecules.

B. The sequence of amino acids responsible for the catalytic activity of enzymes.

C. The sequence of amino acids responsible for the structure of an enzyme.

D. The specific area responsible for the activity of all proteins.

12. Which statement describes how allosteric enzymes work?

Competitive End-product Active and

Reversible
inhibition inhibition inactive forms
A.
B.
C.
D.

Key: = yes = no

13. The graph below shows enzyme activity plotted against temperature. What is happening at point
I?

A. The enzyme is being denatured.

3
 B. pH changes are slowing the reaction.

C. The concentration of the substrate remains constant.

D. The reaction is increasing in speed.

14. What occurs in the induced fit model for enzyme catalysed reactions?

A. There is an exact fit between a specific substrate and a specific enzyme.

B. The enzyme can change shape to accommodate the substrate.

C. The substrate can change its shape to fit a number of enzymes.

D. Other substrates can bind away from the active site.

15. The diagram below shows the energy levels of a reaction in the presence or absence of an
enzyme. What is the best explanation of the different energy levels labelled I, II and III?

I II III
A. Absence of an enzyme Presence of an enzyme Endergonic reaction
B. Presence of an enzyme Absence of an enzyme Exergonic reaction
C. Absence of an enzyme Presence of an enzyme Exergonic reaction
D. Presence of an enzyme Absence of an enzyme Endergonic reaction

16. Consider the metabolic pathway shown below.

If there is end-product inhibition, which product would inhibit which enzyme?

Product Enzyme
A. X 4
B. W 3
C. W 2
D. Z 1

17. What happens during end product inhibition of the pathway shown below?

A. Enzyme I is inhibited by substance X.

B. Enzyme I is inhibited by substance Z.

4
 C. Enzyme III is inhibited by substance W.

D. Enzyme III is inhibited by substance Y.

18. Which graph shows the relationship between the substrate concentration and the rate of an
enzyme controlled reaction?

19. (a) Define the term active site of an enzyme.[1]

(b) Outline how enzymes catalyze biochemical reactions. (2)

(c) Explain the effect of pH on enzyme activity. [3]

(d) The rate of cellular respiration is controlled by the allosteric inhibition of

phosphofructokinase by ATP. Phosphofructokinase is the first enzyme in the respiration
pathway. Explain the meaning of allosteric inhibition using this example. [4]

20. Explain the effect of substrate concentration on enzyme activity. [3]

21. (a) Outline the induced fit model of enzyme activity. [3]

22. (a) State why each step in a biochemical pathway often requires a separate enzyme. (2)

(b) Explain the effects of either changing temperature or pH on enzyme activity. (3)

(c) Discuss the statement, “Enzyme inhibitors function by binding to the active site of the
enzyme”. [3]

23. Explain the effects of temperature, pH and substrate concentration on enzyme activity. [8]

24. Outline enzyme-substrate specificity. [5]

25. Explain how allosteric control of metabolic pathways by end-product inhibition includes negative
feedback and non-competitive inhibition. [8]

26. Outline two examples of the commercial application of enzymes in biotechnology. (6)

27. Explain competitive and non-competitive inhibition, including allostery. (8)

28. (a) (i) The graph below shows the energy changes in a reaction.

On the above graph draw the result you would obtain in this same reaction if an
enzyme that catalyses this reaction were added.
(1)

(ii) Explain how the enzyme produces this effect. [3]

(b) Annotate the graph representing an exergonic enzyme catalysed reaction.

(2)

(c) Explain how end-product inhibition controls metabolic pathways. [3]

5
29. At the start of glycolysis, glucose is phosphorylated to produce glucose 6-phosphate, which is
converted into fructose 6-phosphate. A second phosphorylation reaction is then carried out, in
which fructose 6-phosphate is converted into fructose 1,6-bisphosphate. This reaction is catalyzed
by the enzyme phosphofructokinase. Biochemists measured the enzyme activity of
phosphofructokinase (the rate at which it catalyzed the reaction) at different concentrations of
fructose 6-phosphate. The enzyme activity was measured with a low concentration of ATP and a
high concentration of ATP in the reaction mixture. The graph below shows the results.

(a) (i) Using only the data in the above graph, outline the effect of increasing fructose
6-phosphate concentration on the activity of phosphofructokinase, at a low ATP
concentration.

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................
(2)

(ii) Explain how increases in fructose 6-phosphate concentration affect the activity of the
enzyme.

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................
(2)

(b) (i) Outline the effect of increasing the ATP concentration on the activity of
phosphofructokinase.

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................

...........................................................................................................................
(2)

(ii) Suggest an advantage to living organisms of the effect of ATP on

phosphofructokinase.

...........................................................................................................................

...........................................................................................................................
(1)
(Total 7 marks)

30. Alcohol dehydrogenase is an enzyme that catalyses the reversible reaction of ethanol and ethanal

6
according to the equation below.

NAD+ + CH3CH2OH CH3CHO + NADH + H+

ethanol ethanal

The initial rate of reaction can be measured according to the time taken for NADH to be
produced.

In an experiment, the initial rate at different concentrations of ethanol was recorded (no
inhibition). The experiment was then repeated with the addition of
l mmol dm–3 2,2,2-trifluoroethanol, a competitive inhibitor of the enzyme. A third experiment
using a greater concentration of the same inhibitor (3 mmol dm–3) was performed. The results for
each experiment are shown in the graph below.

[Source: R Taber, Biochemical Education, (1998) 26, pages 239-242]

(a) Outline the effect of increasing the substrate concentration on the control reaction (no
inhibition).

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................
(2)

(b) (i) State the initial rate of reaction at an ethanol concentration of 50 mmol dm–3
in the presence of the inhibitor at the following concentrations.

1 mmol dm–3: .....................................................................................................

3 mmol dm–3: .....................................................................................................

(1)

(ii) State the effect of increasing the concentration of inhibitor on the initial rate of
reaction.

...........................................................................................................................

...........................................................................................................................
(1)

(c) Explain how a competitive inhibitor works.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

7
 .....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................
(3)
(Total 7 marks)

31. The enzyme aspartate carbomyltransferase (ACTase) is a key regulatory enzyme in nucleotide
metabolism in bacteria. The activity of this enzyme was studied in the bacterium Helicobacter
pylori, an important human pathogen. ACTase activity and the growth of H. pylori were
measured at different concentrations of carbomoyl aspartate (CAA), the end product of the
reaction catalysed by ACTase.

[Source: Burns, et al, Biological Procedures Online, (1998), http://www.biologicalprocedures.com]

(a) (i) State the growth of H. pylori at a CAA concentration of 30 mmol dm–3.

...........................................................................................................................
(1)

(ii) Calculate the change in ACTase activity between CAA concentrations of 20 and 40
mmol dm–3.

...........................................................................................................................

...........................................................................................................................
(1)

(b) Compare the effect of increasing CAA concentration on the growth of H. pylori and
ACTase activity.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................
(2)

(c) Explain the effect of CAA on ACTase activity.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

8
 .....................................................................................................................................
(2)

(d) Suggest a direct medical application of this information.

.....................................................................................................................................

.....................................................................................................................................
(1)
(Total 7 marks)

32. The hydrolysis of inorganic phosphate (PPi) by phosphatase enzyme provides energy for a wide
range of reactions. A phosphatase (PPase) occurs bound to thylakoid membranes. This enzyme
was purified from the thylakoid membranes of spinach leaves using chromatography. The activity
of the membrane bound enzyme and the purified enzyme was measured.

The effect of the concentration of magnesium ions (Mg2+) on the relative activity of these
enzymes was determined using different concentrations of magnesium chloride. The
concentration of inorganic phosphate used in both cases was of 1 mmol dm–3.

Activity of phosphatase (arbitary units)

Membrane bound Purified
12 618 1215

[Reprinted from Po-Yin Cheung et al., “Thiols Protect the Inhibition of Myocardial Aconitase by Peroxynitrite”, Archives of
Biochemistry and Biophysics, vol. 350, issue 1, pp. 104-108 © 1998, with permission from Elsevier.]

(a) State the percentage of relative activity of the purified enzyme when the concentration of
magnesium chloride is

(i) 1 mmol dm–3 ...................................%

(ii) 2 mmol dm–3 ...................................%

(2)

(b) Outline the effect of magnesium chloride on the relative activity of the membrane bound
enzyme.

....................................................................................................................................

....................................................................................................................................

....................................................................................................................................

....................................................................................................................................
(2)

(c) Calculate the approximate ratio of inorganic phosphate to magnesium chloride

concentration needed to achieve maximum activity in membrane bound enzymes.

....................................................................................................................................

9
 (1)

(d) (i) State the difference in phosphatase activity when membrane bound and when
purified.

..........................................................................................................................

..........................................................................................................................
(1)

(ii) Suggest a reason for this difference.

..........................................................................................................................

..........................................................................................................................
(1)
(Total 7 marks)

33. Inflammation of human tissues often causes pain. Cyclooxygenases (COX) are a group of
enzymes that play a role in causing inflammation. Analgesics are drugs that can reduce pain. The
graph below shows how increasing concentrations of the analgesic drug dipyrone, affects the
activity of three different cyclooxygenases, COX-1, COX-2 and COX-3.

[Source: Adapted from N V Chandrasekharan, et al, (2002), Proceedings of the National Academy of Sciences, USA, 99, (21),
pages 13926−13931]

(a) Outline the relationship between dipyrone concentration and COX-3 activity.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................
(2)

(b) Deduce whether dipyrone is an inhibitor of COX-2.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................
(2)

(c) Evaluate the potential of dipyrone as an analgesic using the data in the graph.

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

.....................................................................................................................................

10
 (2)
(Total 6 marks)

END OF PAPER

11
Suggested answer:

1. D 2. C 3. B 4. D 5. C
6.C 7. C 8. C 9. D 10. C
11.B 12. B 13. A 14. B 15. C 16. D 17. B 18. B

19. (a) the site (on the surface of an enzyme) to which substrate(s) bind / the site (on the
enzyme) where it catalyzes a chemical reaction; 1

(b) bring substrates close together in active site / in correct orientation;

forms enzyme-substrate complex / substrate(s) bind to active site;
lowers the activation energy for the reaction;
weakens bonds in the substrate; 2 max

(c) enzymes have an optimal pH;

lower activity above and below optimum pH / graph showing this;
too acidic / basic pH can denature enzyme;
change shape of active site / tertiary structure altered;
substrate cannot bind to active site / enzyme-substrate complex cannot form;
hydrogen / ionic bonds in the enzyme / active site are broken / altered; 3 max

(d) ATP inhibits phosphofructokinase at (allosteric) site away from the active site;
inhibition alters the enzyme’s conformation / structure;
the active site does not accept the substrate molecule;
when respiration increases ATP levels phosphofructokinase is inhibited;
respiration slows down;
phosphofructokinase is the first enzyme in the respiration pathway so
there is no build up of metabolic intermediates;
as ATP is used up by the cell the inhibition of phosphofructokinase is reduced;
respiration speeds up again;
this is an example of negative feedback; 4 max

20. as substrate concentration increases enzyme activity increases;

at high substrate concentration enzyme reaches maximum activity;
active sites saturated;
labelled sketch-graph showing above relationship;
[3]

21. (a) substrate binds / approaches active site;

shape of active site changes;
bonds in substrate weaken;
activation energy decreases;
explains broad specificity of some enzymes;
eg proteases; 3 max

22. (a)enzymes are specific for their substrate / lock and key model / energy requirements
for reactions with substrates vary;
each step of the pathway is unique / different substrate at each step;
finer control of metabolic pathways; 2

12
 (b) Either, temperature: [3 max]
each enzyme has an optimal temperature for its maximum activity;
(small) temperature increases result in increased enzyme activity to a point / optimum;
increase activity due to increased movement of molecules / increased kinetic energy or
conversely stated;
temperature increases above the optimum causes (progressive) loss of activity due to
denaturation / shape changes

or, pH: [3 max]

each enzyme has an optimal pH for its maximum activity;
as pH varies from optimal pH, enzyme activity diminishes / becomes inhibited;
loss of activity is due to denaturation / shape changes;
gain or loss of hydrogen ions distorts tertiary shape of enzyme;
homeostatic mechanisms maintain optimal conditions for enzyme activity; 3 max
Credit marking points above if illustrated by a suitably annotated graph.
[5]
(c) competitive inhibitors are structurally similar to the substrate;
competitive inhibitors bind / block active site;
non-competitive inhibitors do not bind to the active site / bind somewhere else on enzyme;
they function by changing the shape of the enzyme so the substrate cannot fit in to the
active site; 3 max

23. enzymes have an active site;

that fits the substrate precisely;
changes in the chemical environment of the enzyme can lead to a shape / conformational change
in the protein;
leading to a change in the shape of the active site;
may interfere with the binding of the substrate with the active site;
altering pH can alter intermolecular interactions within the protein;
or within the active site;
enzymes have an optimum pH;
increase in temperature can increase molecular motion leading to disruption of intermolecular
interactions;
increases chance of enzyme substrate collisions so enzyme activity increases;
optimal temperature;
temperature changes / pH changes can denature the protein;
the more substrate, the more product / more enzyme-substrate complex forms;
after a point, all active sites are bound to substrate / all active sites occupied;
additional substrate will not lead to a greater rate of product formation at this point; 8 max
For full marks all three conditions must be included, otherwise award [6 max].

24. active site of enzyme binds to specific substrate;

shape of the active site and substrate fit / complement each other;

lock and key model;

chemical properties of substrate and enzyme attract / opposite charges;
enzyme / active site is not rigid and substrate can induce slight changes in shape;
allows substrates of similar structure to bind with same enzyme;

induced fit;
causes weakening of bonds in substrate to lower activation energy;
[5]

13
25. allosteric enzyme has binding site(s) away from / other than the active site;
(shape of an) allosteric enzyme alternates between active and inactive (form);
non-competitive inhibitor binds to allosteric site / away from active site;
non-competitive inhibitor changes shape of active site;
non-competitive inhibitors do not compete with substrate for the active site;
end-product can inhibit enzyme needed for early / first step in metabolic pathway;
negative feedback since increased level of product decreases rate of its own production;
metabolic pathway regulated according to the requirement for its end-product;
idea that inhibition is reversible;
Award [1] for named enzyme and [1] for its non-competitive / end-product inhibitor. [8]

26. the name of the enzyme and the substrate;

the name(s) of the product(s);
a statement as to why the application is useful commercially;
Award [3 max] per example,
eg pectinase acts on (soluble) pectin;
produces smaller, more soluble carbohydrates;
used in fruit juice clarification / to increase yield;
eg endonuclease DNA acts on DNA;
produces DNA fragments;
used in genetic engineering;
eg protease acts on (insoluble) proteins;
produces amino acids;
washing powders - stain removal;
[6]
Accept any other suitable examples.
27. competitive:
a molecule structurally similar to the substrate binds to the active site;
preventing substrate binding;
eg inhibition of butanedioic acid (succinate) dehydrogenase by propanedioic acid (malonate) in
the Krebs cycle / other valid example;
competitive inhibition is reversible;

non-competitive:
an inhibitor molecule binds to an enzyme;
not at the active site;
causes a conformational change in the active site;
preventing substrate binding;
eg CN inhibition of cytochrome oxidase by binding to SH groups / other valid example;
Award [6 max] for explanation of competitive and non-competitive inhibition.

most allosteric enzymes have multiple allosteric sites;

allosteric inhibition is a form of non-competitive inhibition;
metabolites can act as allosteric inhibitors of enzymes earlier in a metabolic pathway to regulate
metabolism;
binding (of end product) to an allosteric site changes shape of enzyme;
[8]

14
 Characteristic Competitive inhibition Non-competitive
structurally / chemically very
inhibitor: different from substrate;
similar to substrate
binds to different site / not active site /
site of binding: active site
allosteric site;
effect: blocks active site changes 3° structure of enzyme /
Conformational change of active site;
substrate cannot bind / reaction
competes with substrate /
effect: not catalyzed / decreased enzyme
prevents substrate binding
activity;
example: Butanedioic acid (succinate) metal ions / Hg+ / Ag+ / Cu2+ /
dehydrogenase by CN– inhibit enzymes (cytocrome
propanedioic (malonate) acid oxidase) by breaking disulfide
in the Krebs cycle / any valid linkages / any valid example;
example eg
Folic acid synthesis in bacteria
by sulfonamide Prontosil;
effect of substrate can be reduced by increasing increasing substrate concentration
concentration: substrate concentration does not reduce effect of inhibitor;

28. (a) (i) 1

Award [1] if drawn line is same shape as original, starting and

finishing in same place and activation energy lower.
(ii) enzyme binds to substrate;
lowers activation energy;
by weakening bonds;
making substrate more likely to react; 3 max
(b)

One mark for each correct annotation. The marks for a+b / reactants or substrates or c+d
/ products can only be awarded if the label is on the horizontal parts of the line.2 max
(c) example of negative feedback
eg ATP inhibition of phosphofructokinase, in glycolysis;
controls rate of product synthesis / amount of product produced / inhibits earlier stage /
switches off pathway when too much product made;
binds to enzyme but not active site / allostery / allosteric site;
enzyme changes shape / substrate cannot bind / enzyme cannot catalyse; 3 max

29. (a) (i) increasin

in activity;
activity levels out / remains constant as (substrate) concentration continues
to rise; 2
(ii) more collisions with active site as concentration rises;
at high substrate levels all active sites are occupied so no further increase
in rate / enzyme working at maximum rate; 2

15
 (b) (i) decreases activity;
at all fructose 6-phosphate concentrations;
most effect at intermediate fructose 6-phosphate concentrations / little difference
at high fructose 6-phosphate concentrations;
ATP acts as an inhibitor; 2 max

(ii) end-product inhibition;

respiration rate decreased if ATP already available; 1 max
[7]

30. (a) directly proportional / greater concentration, greater rate of reaction;

at high concentrations the increase is smaller / plateau / levels-off (at
approximately 70 mmol dm–3); 2

(b) (i) 1 mmol dm–3 : 0.70 (± 0.02)

3 mmol dm–3 : 0.55 (± 0.02) 1 max
Both needed for [1]. For 1 mmol dm–3 accept 0.7.
(ii) lower reaction rate at inhibitor concentration of 3 mmol dm–3 / the greater the
inhibitor concentration the slower the rate of reaction;
trend / overall shape are the same / increases but then levels-off;
but lower at greater concentration of inhibitor; 1 max

(c) substrate and inhibitor (structurally) similar;

inhibitor binds to active site;
prevents substrate from binding;
activity of enzyme prevented;
named example (eg malonate inhibits succinate dehydrogenase as it is similar to succinate); 3 max
[7]
31. (a) (i) 72.5 (.1.
(ii) 40% (decrease) 1

(b) both are inversely proportional /

as CAA concentration increases both activity and growth decrease;
growth is more irregular at low concentrations;
ACTase activity decreases more at higher
concentration / after 30 mmol dm–3 than growth;
both show linear decreases between 10 and 30 mmol dm–3; 2 max
Any other valid comparisons.

(c) end-product is slowing enzyme activity;

allostery / non-competitive inhibition; 2

(d) use CAA to control / inhibit bacterial growth / antibiotic functions;

use CAA to treat H. pylori infection; 1 max
[7]
32. (a) (i)
27 (±1)

16
 (ii) 90 (±1) 2

(b) as the concentration of Mg increases the rate increases;

most rapid increase between 1 and 2 mmol dm−3;
peaks at 4 mmol dm−3;
until it plateaus (at 5 mmol dm−3 ) / no more increase / drops slightly; 2 max

(c) 1:4 1

(d) (i) membrane bound is 10 times more efficient (12000 to 1200);

difference is (12618-1215) 11403 arbitrary units greater in membrane
bound;
about 1000% greater / 938.5% greater; 1 max
(ii) purification could have affected structure of protein;
bound to membrane allows interactions / other molecules in
membrane may help it / be acting as coenzymes; 1 max
[7]

33. (a) at low dipyrone concentrations, there is no effect on enzyme function;

as concentration of dipyrone increases, increased inhibition of enzyme activity; 2

(b) over range of concentrations in the experiment appears to have no effect on

enzyme function (of COX-2);
may be inhibited at concentrations higher than those used in the experiment; 2

(c) it would limit inflammation due to COX-1 and COX-3;

but not COX-2;
effective only at high doses; 2
[6]

17