HKALE BIOLOGY | 4 Enzyme and Metabolism | P.

1. 1995/IB/8
(a) An experiment was conducted to investigate the effect of temperature on the activities of amylases I
and II. These enzymes were obtained from the same type of tissue of vertebrate animals I and II
respectively. Similar pH, enzyme concentration and substrate concentration were used. The
experimental data obtained are recorded in the following table:

Time for complete digestion of 1 unit

Temperature/℃ of substrate/minute
Amylase I Amylase II
10 2.0 5.0
15 1.6 3.5
20 1.5 2.5
25 1.6 2.0
30 2.0 1.6
35 4.0 1.4
40 10.0 1.5

(i) Plot a graph to demonstrate the effect of temperature on the activities of amylases I and II.
(5 marks)

(ii) Explain the results for amylase I.

(2 marks)

(iii) What are the optimum temperatures for amylase I and amylase II? What thermoregulatory
ability would the data for amylase suggest for vertebrate animal I?
(2 marks)

(b) The graph below shows the effects of two chemicals, A and B, on the activity of amylase I.
Similar pH, temperature, enzyme concentration and chemical concentration were used in the experiment.

(i) Compare and contrast the effects of chemical A and chemical B on the activity of amylase I.
(3 marks)
(ii) Explain how each chemical, A and B, exerts its effect on amylase I.
(3 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.2

2. 1996/IB/10
(a) A student performed an experiment to monitor the progress of a simple enzyme-catalysed reaction
involving one substrate and one product. He prepared replicate reaction tubes. In each tube, the
concentration of only one reaction component, either the substrate or product, was measured. The
results of three selected tubes are shown as curves A, B and C below:

(i) Identify which measured component, substrate or product, is represented by each curve.
(1.5 marks)

(ii) It was realized that the reaction mixture in one of the three tubes had been wrongly prepared.
This resulted in a different reaction condition.

(1) State which curve represents the result of an error in the preparation. Give a reason for your
choice. Suggest two possible mistakes in the student’s preparation of this reaction mixture.
(3.5 marks)

(2) What evidence shows that the other two curves represent identical reaction conditions?
(2.5 marks)

(iii) With reference to curve C,

(1) calculate the rate of enzyme reaction at the 3rd minute, given that 100% concentration is
equivalent to 100 mmoles of the measured component (Show the readings you take from the
graph in your calculation);
(2 marks)

(2) compare and explain the difference in enzyme reaction rate at the 3rd and 15th minute.
(2.5 marks)

(iv) Curve A will finally level off and will not reach 100 %. Explain this phenomenon.
(1.5 marks)

(b) Explain what is meant by the ‘active site’ of an enzyme.

(1.5 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.3

3. 1997/IA/2
Compare and contrast the characteristics of competitive and non-competitive inhibitions on enzyme
activity. (4 marks)

4. 1998/I/6
Give one application of enzyme and explain its effect. (2.5 marks)

5. 1999/IB/12
The following reaction is catalysed by the enzyme succinate dehydrogenase:

(a) State three parameters that can be used to determine the rate of this reaction. (3 marks)
(b) The graph below shows the effect of succinate concentration on the rate of reaction under
optimum pH and temperature:

The curve flattens out at X. Give two explanations for this observation based on the
mechanism of enzymatic reaction. (2 marks)
(c) Malate and succinate are metabolic intermediates in the Krebs cycle. They have the following
structural formulae:

(i) Suggest what effect malate might have on the rate of reaction catalysed by succinate dehydrogenase.
(1 mark)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.4

(ii)The positions of malate, succinate and fumarate in the Krebs cycle are shown in the following flowchart:

Malate can play a role in regulating the rate of reactions in the Krebs cycle. Explain the possible
mechanism by which malate can achieve such a control.
(41/2 marks)

6. 2000/IA/1
Yeast produces an extracellular invertase to break down sucrose into glucose and fructose. Design an
experiment to show the presence of extracellular invertase activities in a yeast culture and state two
appropriate controls. (7 marks)

7. 2000/IB/9
(a) Figure 1 shows a standard curve obtained by using a colorimetric method for determining the
concentration of protein solutions.

In an experiment to study the effect of heat treatment on the digestibility of a protein substrate and
the effect of raw bean extract (taken from bean X) on protease activity, various reaction mixtures
were prepared and were incubated for 30 minutes. The protein concentration of each reaction
mixture at the beginning and at the end of the incubation period was determined by the colorimetric
method. Intensities of colour developed are shown in Table 1.
Table 1
Colour intensity of the reaction mixtures / arbitrary unit
Incubation Heated protein Unheated protein Unheated protein
period / min substrate + substrate + substrate + raw bean
protease protease extract + protease
0 10 10 10
30 4 7 9

(N.B. The protein content of the raw bean extract added is negligible.)

(i) Use the curve shown in Figure 1 to determine

(1) the protein concentration of the reaction mixtures at the beginning of the incubation period.
HKALE BIOLOGY | 4 Enzyme and Metabolism | P.5

(2) the protein concentration of the following reaction mixtures after digestion by the protease for
30 minutes:

mixture with the heated protein substrate

mixture with the unheated protein substrate
(1 mark)

(ii) Which is more digestible by the protease, the heated or the unheated substrate? Justify your
choice by interpreting the experimental data.
(1.5 marks)

(iii) Based on the structure of protein molecules, account for the effect of heat treatment on the
digestibility of this protein substrate.
(3 marks)
(iv) What is the effect of the raw bean extract on protease activity?
(0.5 mark)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.6

8. 2002/I/9
The following flowchart summarizes an experiment to determine whether compound X is an inhibitor of
amylase :

(a) Suggest a dependent variable to indicate the result of the experiment and suggest a test to
measure the dependent variable.
(2 marks)
(b) Predict experimental result B is X is an inhibitor of amylase.
(1 mark)
(c) How will you modify the experiment to show that X is a competitive inhibitor of amylase ?
(2 marks)

9. 2003/IIA/2
(a) In the human body, the metabolism of ethanol (the alcohol in beer and wine) is catalysed by two
enzymes E1 and E2, in the liver as shown in the following reactions:

(i) After alcohol consumption, acetaldehyde causes facial reddening, dizziness and vomiting (i.e. alcohol
sensitivity) in some people. About 50% of Chinese show alcohol sensitivity whereas Westerners
normally do not show alcohol sensitivity. This was found to be caused by different activities of E1 and
E2 between the two groups of people.

(1) With reference to the given reactions, deduce how the activities of E1 and E2 in alcohol-sensitive
Chinese differ from those in Westerners, Explain your deduction. (5 marks)

(2) Enzyme inhibitors can be used as drugs to discourage people from excessive alcohol drinking. For
this purpose, should me inhibitor of E1 or E2 be used?

Explain your choice. What would be die harmful effect to the body if the inhibitor of the other
enzyme were used?
(4 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.7

(ii) Methanol and ethanol have similar structural characteristics. It is illegal to add methanol to alcoholic
drinks as it is convened to formaldehyde by the reaction catalyzed by E1, and formaldehyde damages
many tissues including the eye.

Explain why administering a large quantity of ethanol can be used as a medical treatment to reduce
the toxic effect of methanol.
(2 marks)
(b) Enzymes are used in some industrial processes. Based on die nature and properties of enzymes,
give
(i) two advantages, and
(2 marks)
(ii) two limitations
(2 marks)
(c) Some types of biological washing powder contain protease and lipase.
(i) Explain the hazard to the body when such biological washing powder is used to handwash clothes.
(1 mark)
(ii) Propose a safety precaution that should be taken when using such biological washing powder for
handwashing clothes.
(1 mark)
10. 2004/I/3
The graph below shows the effect of substrate concentration on the rate of an enzymatic reaction in
the presence and absence of an enzyme inhibitor:

Deduce the possible type of inhibitor used. Explain your deduction. (3 marks)

11. 2004/IIA/1di
Vitamin A and vitamin D are fat-soluble while vitamin C is water-soluble. They are essential for
normal growth and body functions.Vitamin D undergoes the following enzymatic activation to form a
highly active metabolite Y:

Elderly people usually have problems in calcium absorption due to a decreased concentration of X.
Suggest two possible causes for this decrease of X. (2 marks)

12. 2005/IA/5
(a) State one factor that causes the reduction in the activity of gastric enzymes in the small intestine.
(1 marks)

(b) Explain how changes in the factor in (a) affect the activity of enzymes. (3 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.8

13.2006/IIA/1ciii
The uptake of cholesterol into liver cells requires a membrane protein(receptor) and the abundance of
this receptor is determined by a gene.People heterozygous for this gene have half the normal number of
the receptors on the liver cell membrane. Their blood cholesterol level tends to be higher than normal, but
it can be reduced by a combination of drug administration and lifestyle modification.

(1) One group of drugs has a similar chemical structure as intermediate X. Explain how these drugs
could reduce the blood cholesterol level. (2 marks)

14. 2007/I/3
A student carried out an experiment to study the effect of chemical P on the rate of an enzymatic reaction.
The same temperature and enzyme concentration were used in the experiment. The results are shown in
the graph below:

(a) Referring to the graph, what interpretations can be made about chemical P?
(4 marks)
(b) Besides temperature and enzyme concentration, state one other variable that needs to be kept constant
for the results to be comparable. Give reason(s) for your answer. (3 marks)

15. 2007/IIA/1
Benzopyrene is a harmful compound commonly found in coal tar, automobile exhaust fumes, tobacco
smoke, burnt toast, and the crust of barbecued meat. When taken into the body, benzopyrene can be
metabolized in the liver by two enzyme systems, known as phase 1 and phase 2, as shown in the pathway
below:

phase 1 product A
phase 1 enzyme system (mutagenic)
benzopyrene enzyme system intermediate X
product B
(fat soluble) Phase2 enzyme (non-toxic and non-
system mutagenic; excreted
glutathione by kidney)

(b) In order to protect the body against the mutagenic consequence of benzopyrene metabolism,
which enzyme system (phase 1 or phase 2) should be more active? Explain your answer.
(4 marks)

(c) Glutathione is manufactured in the mitochondria. However, the number of mitochondria in the
liver decreases when one ages. Explain the effect of this phenomenon on the detoxification of
benzopyrene in the liver. (3 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.9

16. 2008/IIA/2bi
Fluoride is an inhibitor of another enzyme in the glycolytic pathway.
Although fluoride does not structurally resemble the substrate of this enzyme, its mode of inhibition is
found to be competitive. Briefly outline the principle of an experiment that you would carry out to test this
mode of inhibition of fluoride.
(2 marks)
17. 2010/IIA/2b
Some cancers are treated with anticancer drugs that interfere with cell division in various ways. The figure
below shows the biochemical pathway in which anticancer drugs A and B exert their suppressive effects on
the biosynthesis of thymine nucleotide within body cells at specific stages of the pathway:

(ii) Anticancer drug A is an irreversible inhibitor of enzyme 1. Describe how drug A affects the
biosynthesis of thymine nucleotide. (2 marks)
(iii) Anticancer drug B is structurally similar to compound X. Describe how drug B affects the biosynthesis
of thymine nucleotide. (4 marks)
 HKALE BIOLOGY | 4 Enzyme and Metabolism | P.10

15. 2009/I/4