NANYANG JUNIOR COLLEGE H2 Biology

Practical 8: Enzymes & Starch Grains

Name: ANSWERS Class: Date:

PRACTICAL 8: ENZYMES & STARCH GRAINS

QUESTION 1: Modified JUNE 1999 Q1 [55min]

You have been supplied with an extract from a potato tuber, some squares of filter paper and four
solutions of hydrogen peroxide of different concentrations. Hydrogen peroxide is easily decomposed
as follows:

2H₂O₂[l] ⟶ 2H₂O[l] + O₂[g]

Note: Hydrogen peroxide is corrosive. If any should come into contact with your skin wash

immediately under cold water.

Put sufficient of the 1.5 mol dm-3 hydrogen peroxide solution into a specimen tube, or small beaker,
to a depth of about 4 cm.

Using a pair of forceps, dip one of the squares of filter paper into the potato extract. Shake once to
remove the excess liquid. Hold the filter paper about 1 cm above the surface of the hydrogen peroxide
solution and drop it into the solution.

Observe the filter paper over the next minute.

a) Record your observations, then remove the square of filter paper.

Sinks, bubbles/effervescence form around paper, floats back up to surface
Observations ……………………………………………………………………………………………………………. [2]

b) State two factors, apart from enzyme concentration, that should be kept constant if investigating
the effect of substrate concentration. [2] [Additional qns to ask students: Describe how you would
keep each of the variable constant.

1. Enzyme concentration (blending ensures that a homogenous liquid potato solution is

obtained, hence conc. of catalase is constant throughout the liquid potato solution.)
2. Volume of H2O2 solution
3. Size of beaker (same size and length to ensure distance the filter paper discs have to travel is
the same for each run)
4. Filter paper disc (same diameter and same thickness. So same amt of O2 is required to lift it
to the surface for each run)
5. Temperature (Enzyme activity affected by temp. Carry out expt in water bath whose
temperature is assumed to remain constant throughout duration of expt)

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Practical 8: Enzymes & Starch Grains

c) Devise and carry out a procedure to determine if your observations in (a) were due to the presence
of an enzyme in the potato extract.
Describe briefly your procedure. State your conclusions about the presence of an enzyme and the
reasons for making your conclusions. [6]
Boil and cool potato extract; reject distilled water.
dip one square of filter paper into the boiled and cooled extract, shake to remove excess;
repeat experiment by dropping the filter paper into a fresh tube of 1.5moldm-3 hydrogen
peroxide solution;
enzyme present in potato extract because the filter paper dipped in boiled enzyme did not float
up;
boiling extract denatures the enzymes within it;
enzymes are protein in nature, high temperature provides energy for thermal agitation;
weak hydrogen bonds/ ionic bonds between R groups are broken, specific 3D structure of enzyme
is distorted;

Teaching Points on Control:

To ensure that any effects observed are due to the changes in the values of the independent
variable and not some unidentified variable.

A negative control group is a control group that is not exposed to the experimental treatment or
to any other treatment that is expected to have an effect.
E.g. (i) replacing a solution with the same volume of water, (ii) denaturing an enzyme by boiling

d) Account for your observations in (a) in the light of your conclusions in (c). [3]
Filter paper initially sinks as it is denser than the solution;
hydrogen peroxide is broken down by catalase found in the potato extract to give oxygen gas
and water;
oxygen gas forms bubbles around the filter paper giving it buoyancy/ allowing it to float

e) Investigate the behaviour of squares of filter paper, soaked with potato extract, in different
concentrations of hydrogen peroxide solution. Start by dropping a square of filter paper, dipped
into potato extract, into the sample of 1.5 mol dm-3 hydrogen peroxide solution used previously.

Start a stopwatch or clock as soon as the filter paper touches the bottom of the container. Stop
timing as soon as the square rises from the bottom of the container. Remove the square of filter
paper.

i) Record this time period in seconds:

Repeat this procedure twice more using the 1.5 mol dm-3 solution and three times for each of
the four other hydrogen peroxide solutions provided, using a fresh square of filter paper each
time. Draw up a table for your results before you begin and include in it the value you obtained
in (d)(i).

Calculate, to the nearest whole number, the mean of the three readings for each
concentration of hydrogen peroxide solution and add these to the table. [6]

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Practical 8: Enzymes & Starch Grains

Tube Concentration of H2O2/ Time taken for filter paper to float to surface/s
moldm-3
1 2 3 Mean

[2] Correct headings with units

[1] 3 readings
[1] Correct trend in values
[1] calculation of mean
[1] precision – nearest whole numbers
Rej average; average can be mean, median or mode.

Recap teaching points on table format:

1. Each column in the table should be headed with the quantity and units (when
applicable).
2. The first column in the table should be independent variable, i.e. the variable that you
manipulated in the investigation.
3. The subsequent columns record the values for dependent variable which are the
measurements/observations you made.
4. Observations recorded should be phrased in this manner “solution turned from _____
to _______”
5. Recorded values should be according to the precision of the colorimeter.
6. When there are two or more sets of readings, there should be an additional column to
process the mean data.
7. Note that the slash is here used to separate what is measured from the unit in which it
is measured. Bracket is acceptable but slash is preferred for standardization across
cohort. Energy content / J kg-1
8. For rate, write m s-1 instead of m/s. Although either is acceptable, the use of the slash,
e.g. m/s, in tables can be misleading and is best avoided.
9. Use mean instead of average

ii) Plot a graph of the mean readings against concentration of the hydrogen peroxide solution.
[4] students are provided with a 12 x 10 grid
1. correct axes;
2. correct axes labels with units
3. correctly plotted points with best fit line (points equidistant on each side of line);
4. axes scaled appropriately, graph takes up 50% of grid;
🔹 Guideline for drawing scales of graphs -
Each axis should be scaled using multiples of 1, 2, 5 or 10 for each 20mm square on the grid.
This makes it easy for you to plot and extract data. Never use multiples of 3.

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Practical 8: Enzymes & Starch Grains

A – axes labels with units

P – Plot points
BF – best fit line

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Practical 8: Enzymes & Starch Grains

iii) Account for the shape of the graph as fully as possible, assuming that the reaction is influenced
by the presence of an enzyme in the potato extract. [3]

Describe general trend and quote figures: As concentration of hydrogen peroxide increases
from XXX to XXX, mean time taken decreases until [x], where it slows down/ plateaus;
At low concentration of hydrogen peroxide, substrate concentration is the limiting factor,
Past [x], [enz] becomes the limiting factor;
At high H2O2 conc: increase substrate concentration, increase frequency of effective collisions
between substrate and enzyme;
All available active sites are saturated; maximum rate of E-S complex formation; maximum rate
of pdt formation;
Avoid commenting on the gradient!

f) Identify one significant source of error in measuring the dependent variable in this investigation.
[1]

(Dependent variable) Time taken for the filter paper disc to reach the surface + idea of delay in
recording time

g) One way to increase confidence in the conclusions of this investigations would be to repeat the
experiment several times.
Describe a modification to the method that would increase confidence in the conclusions, and
explain how these modifications would achieve this. [2]

Using a water bath; to control temperature fluctuations;

Standardise soaking time in enzyme for each pc of blotting paper; to ensure the same volume
and concentration of enzyme Reject: amount in each blotting paper;
Measure the amount of hydrogen peroxide used in volume (e.g. 20cm3) instead of measuring
4cm in height; to ensure that the volume of substrate used across all the tubes are the same;
Increase number of hydrogen peroxide concentrations within the range to obtain more plot
points for a better best fit line;
[Total: 29]

Teaching point:
Improvements to a procedure that will increase the accuracy of the procedure for the
investigation or reliability of the observations / results:
- Standardise other relevant variables which may affect the dependent variable
- use a measurement method for the dependent variable which is more accurate
- collect more data by taking replicate measurements to obtain a mean

Suggest an alternative experimental protocol for the same practical.

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Practical 8: Enzymes & Starch Grains

QUESTION 2: Modified December 72/2

(a) Prepare a wet mount of the potato extract provided, using the potato pulp given. Observe
under the microscope, draw and annotate the structures observed.

(b) Irrigate the mount with iodine. Suggest what the structures are and state the changes seen
in the structures after irrigating with iodine.
starch grains;
concentric rings cannot be seen any more; starch grains appear blue-black;

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