Cytology Year 05 Enzyme Paper I 5(4)

Questions:
5 (a) State one factor that causes the reduction in the activity of gastric enzymes
in the small intestine (1 mark)

(b) Explain how changes in the factor (a) affecter the activity of enzyme
(3 marks)
Answer:
5 (a) pH / alkaline medium (1)
(b) Changes in pH affect ionization of the amino acids of enzymes (1). This
may cause a change in the conformation of the active site / enzyme /
denaturation of enzyme (1), thus affects the formation of the
enzyme-substrate complex (1). As a result, the activity of enzymes will be
affected.

Cytology Year 04 Enzyme Paper II 1(20)

Questions:
1
Vitamin A and vitamin D are fat-soluble while vitamin C is water-soluble.They are essential for
normal growth and body functions.

(a) Explain why an excessive intake of fat-soluble vitamins in general is undesirable

whereas an excessive intake of water-soluble vitamins is not (2 marks)

(b) Give one major function in the human body of vitamin A and vitamin C (2 marks)

(c) By means of a flow chart, show the major vessels involved in the transport of vitamin D
from the small intestine to the heart (3 marks)

(d) Vitamin D undergoes the following enzymatic activation to form a highly active metabolite
Y:

(i) 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)
(ii) It is known that blood calcium level is kept relatively constant by a negative
 feedback mechanism regulated by hormone H. H acts to stimulate both
enzyme 1 and enzyme 2. Use a flow chart to show a possible mechanism
for this negative feedback. (4 marks)
(iii) Explain how vitamin D deficiency might lead to skeletal problems in adults.
(4 marks)
(iv) Why is vitamin D deficiency more damaging to children than to adults?
(3 marks)

Answers:
1
(a)
Main concept:
˙ whether the vitamins can be excreted due to their solubility in water
˙ whether the vitamins can accumulate and cause harm to the body

e.g.
˙ fat-soluble vitamins are not readily excreted in urine, but excessive water-soluble
vitamins can be excreted in urine
˙ fat-soluble vitamins accumulated in the body to reach toxic levels/cause harm to
the body

(b) ˙ vitamin A － retinal pigment / visual pigment / rhodopsin formation (also

accept pigment in rods, responsible for dim light vision) OR control normal
epithelial structure & growth
˙ vitamin C (any one function) :
> antioxidation / get rid of free radicals
> proper metabolism / formation of connective tissue
> necessary for the synthesis of collagen fibre
> healing of wound
(N.B. do not accept deficiency disease as answer)

(c) small intestine →

Lymphatic → thoracic duct → (left) → (anterior) → heart

vessels / lacteal subclavian vein
vena cava

(d) (i) Any two:

 ˙ indoor life, lack of sunlight for vitamin D synthesis
˙ lack of vitamin D in food
˙ poor vitamin D absorption
˙ lower enzyme 1 activity / absence of or not enough enzyme 1 production
(any other acceptable answer)

(ii)

Main concept:
˙ effect of deviation from normal blood Ca++ level (above or lower normal on
hormone H level
˙ effect of H on E1 activity
˙ effect of E1 and E2 activity on Y
˙ effect of activity or level of Y on Ca++ uptake in maintaining
normal Ca++ level
˙ path way for the deviation from normal Ca++ level in the
opposite way
(iii)
Main concept:
˙ effect of vitamin D deficiency on Ca++ uptake
˙ blood Ca++ level lower than norm
˙ Ca++ drawn from bone to restore norm
˙ adverse affects on bone / description of such effect on skeleton
e.g.
˙ deficient vitamin D → decreased dietary Ca++ absorption
˙ Ca++ level in blood decreased / lower than normal
˙ Ca++ drawn from bone to maintain homeostasis
˙ reduce bone mass / bone weakens / bone easily break

(iv)
Main concept:
˙ children need more calcium as the growth of children is more than that of adults e.g.
˙ effect of vitamin D deficiency on bone growth / teeth development ˙

children need more calcium than adults as children continue to grow whereas
adults stop growth / less growth
˙ insufficient vitamin D → poor bone formation / bone deformity / rickets / soft
bone / poor teeth development

Cytology Year 04 Enzyme Paper I 4(3)

Questions:

Answer:
Main concept:
˙ name of inhibitor
˙ evidence for deduction based on the graph / description of the enzymatic activity
observed
˙ explanation of inhibitory action

e.g. 1
˙ non-competitive inhibitor / allosteric inhibitor (Name)
˙ inhibitory action of inhibitor cannot be overcome by increased substrate
concentration (Deduction)
Or
lowers enzymatic activity irrespective of substrate concentration (Description)
˙ because inhibitor does not bind to the active site (Explanation)

e.g. 2
˙ irreversible inhibitor (Name)
˙ lowers enzymatic activity irrespective of substrate concentration (Description)
˙ binds to enzyme permanently / forms a covalent bond with enzyme (Explanation)

Cytology Year 03 Enzyme Paper II 2(20)

2
(a)
(i)
(1) ˙ Westerners (alcohol insensitive): E1 and E2 activities are the same (1) / E2
activity is greater than that of E1
˙ alcohol sensitive Chinese have higher / faster / more active E1 activity than
that of E2 (1), leading to the production of more acetaldehyde (1)
˙ the acetaldehyde formed cannot be quickly metabolized (1), acetaldehyde
accumulates (1) (5 marks)

(2) ˙ an inhibitor of E2 (1) to cause acetaldehyde accumulation (1)

˙ acetaldehyde induces dizziness and vomiting / alcohol sensitivity which will
discourage drinking (1)
˙ if the inhibitor of E1 was used, ethanol will accumulate in blood / liver (1),
this will cause damage to liver / other body organs (1) (max. 4)

(ii) ˙ concept of competitive inhibition, e.g. ethanol competes with methanol for
the binding site on E1 (1)
˙ rate of formation of formaldehyde will be slowed down (1) / less
formaldehyde will be formed, ∴ toxic effect reduced (2 marks)

(iii) ˙ ethanol cannot be completely metabolized at once (1) / alcohol needs time
to break down
˙ stay in the blood (1) / alcohol transported by blood to lung,
˙ ethanol diffuses from blood into alveoli air (1) and exhaled stay in the blood
 (Bonus : alcohol is volatile ∴ easily vaporize (1)) (3 marks)
+bonus =1
(poss. max. = 4)

(b) (i) advantages (any two) :

˙ speed up chemical reactions in industrial process for mass production
of products (1)
˙ enzymes are specific in action ∴ can catalyze specific processes (1)/
less likely to generate undesirable products
˙ enable artificial manipulation of rate of industrial process by
controlling reaction temperature or pH (1)
(accept other correct / reasonable alternatives) (2 marks)
(ii) limitations (any two) :
˙ the industrial processes will be sensitive to changes in temperature as
enzyme works in specific temperature range (1) (accept similar
concept such as enzyme denatured by high temperature)
˙ the industrial processes will be sensitive to pH changes as enzyme
works within specific pH range (1)
˙ vessels / utensils used in the industrial processes have to be clean so as
to avoid degradation of the enzyme (1) / easily affected by inhibitors
(accept other correct / reasonable alternatives, but do not accept en-product
inhibiton) (2 marks)
(c) (i) ˙ protease and lipase may damage the skin / eye / mucous membrane as these body
parts consist of protein and lipid (1)
(ii) ˙ wear gloves (1) (accept other correct / reasonable alternatives)
(2 marks)

Cytology- enzyme
0219(5)
Q: 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 if 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)
(Total: 5 marks)

A:
(a)- Amount of reducing sugar/ maltose formed/ at the end of the experiment (1)
- Benedict’s test (1) OR
- Rate of disappearance of starch (1)/ the concentration of starch/ the amount of
remaining starch at the end of the experiment
- Iodine test (1)
(b) No/ less reducing sugar/ no maltose is formed compared with result A (1) OR
No red precipitate is formed/ no/ little color change (1) OR
Starch disappeared more slowly in B compared with A (1)/ starch did not disappear/
the mixture remains blue black in B
(c) Increase the concentration of starch (1), the inhibitory effect of X will be reduced (1)/
removed

0119(6)
Q: A pharmaceutical company has produced a new drug, X, which it claims can lower
plasma cholesterol level. To verify this claim, an in vitro experiment can be performed
using the following:
- drug X
- precursor A
- an enzyme which converts precursor A to cholesterol
- an indicator which combines with cholesterol to form a blue
complex. (it is known that the intensity of the blue color is
proportional to the cholesterol concentration)
(a) Outline the principal steps in the procedure of this experiment. (5 marks)
(b) Various tests have to be carried out to ensure that X is safe for human consumption
before putting X onto the market. Suggest one of these tests. (1 mark)
A: (a) Incubate X with the precursor A and the enzyme for some time (1), a control
where X was not added (1), add the color indicator and compare the color intensity (1)
with a colorimeter. If the color intensity in the control tube is higher than that in the
treatment tube, X can lower cholesterol (2). (accept reverse description)
(b) Any one of the following:
- test for toxicity (1)
- test for allergy (1)
(Accept correct alternatives)

0019a(6.5)
Q: 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 incubation
period was determined by the colorimetric method. Intensities of color developed are
shown in table 1.

Table 1
Incubation period/ Color intensity if the reaction mixtures/ arbitrary unit
min Heated protein Unheated protein Unheated protein
substrate substrate substrate
 +
+ + raw bean extract
Protease protease +
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 incubation
period (½ mark)
(2) the protein concentration of the following reaction mixtures after digestion by the
protease for 30 minutes: (1 mark)
mixture with the heated protein substrate
mixture with the unheated protein substrate
(ii) Which is more digestible by the protease, the heated or the unheated substrate?
Justify your choice by interpreting the experimental data. (1½marks)
(iii) Based on the structure of protein molecules, account for the affect 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? (½ mark)

A: (i) [No unit, deduct 1/2 mark, denote as U-1/2; no decimal place, deduct 1/2 mark,
denote as D-1/2; deduct max. 1/2 mark for whole of (i)]
(1) 5.0% (1/2)
(2) mixture with heated protein substrate: 2.0%(1/2)
mixture with unheated protein substrate: 3.5% ( 1/2)
(ii) heated (1/2)
less protein remained after incubation with the protease(1)
(iii) Heat destroyed/ denatured/ changed the 3D structure/ conformation/ folding of the
protein substrate (1), it broke the hydrogen bonds ( 1/2) and ionic bonds (1/2).
This exposed the various parts/ increased the surface area of the molecule to the
action of the enzyme (1).
(iv) The raw bean extract inhibits/ lowers the protease activity ( 1/2)

0016(7)
Q: Yeast produces an extracellular invertase to break down sucrose into glucose and
fructose. Design an experiment to show the presence of extracellular invertase acticities
in a yeast culture and state two appropriate controls. (4 marks)
A: Incubate yeast culture with sucrose solution for some time (1), centrifuge/ dialyse/
filter to separate cell from supernatant (1), test supernatant (1) with Benedict’s solution
(1), red ppt (1) indicates presence of invertase activity
Control (any two): --yeast without sucrose (1),
--boiled yeast with sucrose (1),
--sucrose without yeast (1)
99112(10.5)
Q: 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)

(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. (4 marks)

A: (a) (Any 3 of the following, 1 mark each)

The concentration / amount of:
﹣fumarate
﹣FADH2
﹣succinate
﹣FAD
( b ) ﹣all enzyme molecules are saturated with substrate molecules ( 1 ) / All active sites
are saturated with substrate molecules ( 1 )/ enzyme is limiting
﹣FAD is limiting ( 1 ) / FAD is not sufficient to accept the H from succinate ( 1 )
Page total=5
(c) (I) Malate slows down the reaction (1).
(II) As reactions in the krebs cycle proceeds (1/2), malate concentration will increase (1/2).
Malate is structurally similar to succinate (1/2), hence will compete with succinate (1/2) for
binding to the active site on succinate dehydrogenase (1/2)/ malate is a competitive
inhibitor (1). This slows down the reactions in the krebs cycle (1/2). Malate concentration
is in turn lowered (1/2). Inhibition is removed (1/2), reaction rate in the Krebs cycle
increases again (1/2).
[Bonus: negative feedback (1/2)]
Page total=5.5
(poss. Page total=6 )

9816(2.5)
Q: Give one application of enzyme and explain its effect. (21/2 marks)
A: Meat tenderizer (1)
(Accept correct alternatives)
contains protease (1/2) which breaks down protein in the meat (1/2), renders the meat
more tender (1/2)

9712(4)
Q: Compare and contrast the characteristics of competitive and non-competitive
inhibitors on enzyme activity. (4 marks)

A: Both lower the activity of enzymes (1/2) and their actions are reversible (1/2)/ no
permanent damage to the enzyme.
Competitive inhibitor – inhibitor molecules have similar structure as the substrate
molecule (1/2), compete with the substrate for the active sites on the enzyme molecule
(1/2), inhibition effect overcome by increased substrate concentration ( 1/2).
Non-competitive inhibitor molecules attack the enzyme molecule not at the active site
(1/2), but changes the conformation of the enzyme ( 1/2) prevents the active site from
catalyzing reactions, inhibition effect not overcome by increased substrate concentration
(1/2).

96110(15)
Q: (a) A student performed an experiment to monitor the progress of a simple
enzyme-catalyzed 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. (11/2 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 1/2 marks)

(2) What evidence shows that the other two curves represent identical
reaction conditions? (2 1/2 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 moles 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 1/2 marks)

(iv) Curve A will finally level off and will not reach 100 %. Explain this phenomenon.
1
(1 /2 marks)
(11/2 marks)
(b) Explain what is meant by the 'active site' of an enzyme. (1 1/2 marks)

Total: 15 marks
A: (a) (i) Curve A = product (0.5)
Curve B = product (0.5)
Curve C = substrate (0.5)
(11/2)

(ii) (1) Curve B (0.5) because it indicates a slower rate of reaction

(0.5) when compared to the reaction rates indicated by
curves A and C (0.5)
Possible errors are: (any two, 1mark each)
－ lower enzyme concentration was used / less enzyme was added
－ lower substrate concentration was used / less substrate was added
－ possible contaminant(s) / inhibitor(s) was introduced into
the reaction mixture (any other acceptable answer, do
not accept temperature difference as an answer) (2)
(31/2)
(2) The remaining two curves / curves A and C show a corresponding
conversion to the product from the substrate (1) according to
(with respect to) their % concentration changes (1) at various time
points (0.5). (2.5)
OR Rate of formation of the product as shown in curve A is the same as
the rate of disappearance of the substrate as shown in curve C (2)
throughout the time points (0.5).

(iii) (1) 8 mmoles min -1(1) N.B. No mark for no unit

Y2  Y1
Readings (1) (2)
X 2  X1
 (2) At 3rd minute: faster rate of enzyme reaction / or at 15th
minute: slower rate of enzyme reaction (1)
At 3rd minute, substrate concentration is higher (0.5) / at 15th minute,
substrate concentration is lower, at 3rd minute chance of collision
between enzyme and substrate molecules is higher (0.5) / at 15th
minute chance of collision between enzyme and substrate molecules
is less, at 15th minute rate is limited by product inhibition (0.5). (1.5)
(2.5)
(iv) The reaction is reversible (0.5). End product effect pushes the
reaction backwards and equilibrium is reached (1) (11/2)

(b) It is a special region on the enzyme molecule with a specific

configuration (0.5) that can bind with the substrate to form an enzyme
－ substrate complex (0.5). Reaction occurs at the active site (0.5) (1
1
/2)
(Total: 15 marks)

9518(15)
Q: (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:

Temperature/ Time for complete digestion of 1

℃ unit of substrate / minute

Amylas Amylase II
e

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 I 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 concentrations
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)
(Total: 15 marks)
A: (a) (i) Refer to the graph shown below
－ title of graph (0.5)
－ properly scaled and fully labelled X and Y axes with correct units (1)
－ 1 curve labelled or keyed (0.5)
－ correctly plotted points (correct conversion for 1/t) for the 2
curves (1.5 marks for each curve) (if students plot time VS
temp : title (0.5) properly scaled and fully labelled  axis with
correct unit (0.5).)

Deduct 0.5 mark if 1-2 points are plotted wrongly,

and if extra points are shown. No mark if > 2 points
are plotted wrongly.
 (Bonus: 0.5 mark if student can distinguish 'reaction rate' and 'enzyme
activity'. Activities of enzyme can be expressed as arbitrary unit- of
substrate over time, to be indicated on Y axis.)

Effect of temperature on the activities of amylase I and II

5
Reaction Rate / min

Amylase I
3
Amylase II
-1

10 15 20 25 30 35 40
temperature

(correct conversion for 1/t. For marker's reference in checking accuracy of points)

Temperature / Rate of react on 1/t / min-1

C Amylase I Amylase II
10 0.5 0.2
15 0.625 0.29
20 0.67 0.4
25 0.625 0.5
30 0.5 0.625
35 0.25 0.71
40 0.1 0.67
(Possible Max. 5.5)
(ii) As temperature increases from 10C to 20C, the kinetic energy
(0.5) of the molecules increases and there is increased collision
 (0.5) between enzyme and
1
substrate molecules (to form the enzyme substrate complex
which subsequently breaks down to form the products). As
temperature increases further, the active site of the enzyme is
progressively destroyed / the enzyme undergoes thermal
denaturation (1), and the reaction rate declines.
1
(2)
(iii) The optimum temperature for amylase I is around 20C (0.5)
while that for amylase II is the value shown on the candidates
graph (0.5).
1
It is possible that vertebrate animal I is poikilothermic / cold blooded
animal.
1
(2)
OR
The thermoregulatory ability cannot be deduced owing to
insufficient data.

Deduct 0.5 mark for spelling mistake.

(b) (i) Both chemicals A and B inhibit the activity of amylase I.

1

The inhibitory activity of A is competitive / can be overcome by

increasing the substrate concentration but the inhibitory activity of
B is non-competitive / cannot be overcome by increasing
substrate concentration.
2
(3)
OR
Contrast the effect at high substrate concentration (1) and at low substrate
concentration (1).

(ii) Chemical A competes with the substrate for the active site (0.5)
on amylase I and produced inhibitory effect at low substrate
concentration (0.5). Chemical A is structurally similar to the
substrate (0.5).
max. 1
 However, when substrate concentration increases, more
substrate molecules are available to compete for the active site
on amylase I.
1
Thus inhibitory effect is overcome at high substrate concentration.
1
Chemical B binds with enzyme in such a way that it reduces the
catalytic properties of the enzyme.
(Total: 15 marks)