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Ans. to let the co2 pass easily. Co2 is lighter than normal air. So it can pass through the cotton
wool easily. Otherwise excess co2 can kill the seeds.
An investigation into the effect of temperature on the activity of amylase
The digestive enzyme amylase breaks down starch intosugar maltose. If the speed at which the
starch disappears is recorded, this is a measure of the activity of the amylase.
The figure shown apparatus which can be used to record how quickly the starch is used up.
Spots of iodine are placed into the depressions on the spotting tile. 5 cm3 of starch suspension is
placed in one boiling tube, using a syringe, and 5 cm3 of amylase solution in another tube, using
different syringe. The beaker is filled with water at room temperature. Both boiling tubes are
placed in the beaker of water for 5 minutes, and the temperature recorded.
The amylase solution is then poured into the starch suspention, leaving the tube containing the
mixture in the water bath. Immediately , a small sample of the mixture is removed from the tube
with a pipette and added to the first drop of iodine on the spoting tile. The colour of the iodine is
recorded.
The indicator Janus Green B changes colour according to the amount of oxygen present.
Oxygen present
Indicator oxidised to blue colour
Oxygen absent
Indicator reduced to pink colour
Method
1
Place 20cm3 of glucose solution with some yeast added to in a test tube. Add two
drops of indicator solution. The colour produced will tell you whether oxygen is
in the mixture.
2
Pour enough liquid paraffin over the mixture to form a layer right over the
surface. This layer will prevent more oxygen from the air getting into the
mixture.
3
As soon as the colour of the mixture indicates that there is no oxygen present, fix
up a delivery tube leading to a small amount of bicarbonate/indicator in another
test tube.
4
Design and set up a suitable control. Remember that in this experiment you are
trying to find out whether the yeast is respiring in the absence of oxygen.
5
Leave the apparatus for 10-20 minutes and then record your observations.
Questions
1
2
3
(a)
the bicarbonate indicator solution?
(b)
The Janus Green Bb indicator solution?
What can you conclude from your two answers to question 1?
Why is a layer of liquid paraffin used in the experiment?
If yeast is left in a sugary liquid for some days, a characteristic smell is produced.
This is the result of ethanol accumulating in the mixture. Fermentation is the
name given to this process in which yeast turns sugar into ethanol:
Sugar
ethanol
+
carbon dioxide
Suggest a way of discovering whether any energy is released during this process.
experiments on respiration
An equation summarizing the process of aerobic respiration is given below.
C6H12O6
6O2
6CO2
6H2O
Energy
glucose
(decrease
in dry weight)
uptake of oxygen
Close the screw clips and note any change in the level of liquid in the
manometer over a period of 10 to 20 minutes.
Discussion
I What change in the liquid level did you observe in the two tubes ?
2 What does this change tell you about the volume of air in each boiling tube ?
3 What are the principal gases in the atmosphere and what is the percentage volume of
each ?
4 What is the function of the soda-lime in this experiment? What might happen if it were
not included?
5 Which atmospheric gases could be responsible for the change in the volume of air in
the boiling tube?
6 Say why you regard your results as reasonable evidence that respiration is taking place
in the organisms under investigation.
7 Why were the two tubes placed in a water bath?
8 Tube B is a 'control' to the experiment. What is its purpose?
5 Discussion answers
I In tube A the liquid should rise. In tube B there should be little or no change.
2 A rise in level indicates that there is a reduction in the volume of air in tube A.
3 Air contains approximately 20% oxygen, 79% nitrogen and 0.04% carbon dioxide.
4 The soda lime absorbs any carbon dioxide produced by the living material. If the living
organism produced as much carbon dioxide as it consumed oxygen, there would be no
change in volume unless the carbon dioxide were absorbed.
5 The absorption of the tiny amount of carbon dioxide from the air will make no
perceptible difference to the manometer. Thus the decrease in volume could be due to
removal of oxygen, nitrogen or both. Since nitrogen does not usually participate in living
reactions, the most likely cause of the diminution in volume is the removal of oxygen.
6 Aerobic respiration involves the production of carbon dioxide and the absorption of
oxygen. Since the soda lime absorbs any carbon dioxide given out, the reduction in
volume is consistent with the hypothesis that the organisms are using up oxygen.
7 Any change in temperature of the tubes will cause the air inside them to expand or
contract and so affect the manometer. The water is less susceptible to temperature
fluctuations and will maintain a fairly constant temperature throughout the experiment.
8 The control tube (a) shows up any change in volume due to temperature change,
(b) shows that removal of oxygen is the result of living activities rather than simple
physical absorption of oxygen by the material.
(f) Turn the tap downwards, push the syringe plunger to the bottom and close the tap
(horizontal).
(g) Take two more readings of the volume of gas produced in one minute and calculate
the average of the three.
(h) Check that the tap is open (upwards) and use a small Bunsen flame to raise the
temperature of the water bath by about 5 C. Leave the apparatus for two minutes after
heating has stopped.
Record the new temperature and take three more one-minute readings as before, but this
time
record the temperature at the end of the three readings to obtain the mean temperature
during
the experiment.
(i) Repeat the operation from (g) raising the temperature about 5 C each time until you
reach 45
or 50 C. At temperatures between 30-40 C the three readings should be made in rapid
succession to avoid volume changes due to cooling; at 40-50 c you should add 0.1 cm3
to all readings to compensate for the cooling effect.
Discussion
1 In general terms what was the effect of rising temperature on the rate of carbon dioxide
production?
2 Was the change in rate consistent over the whole of the temperature range which you
tried? If
not, suggest an explanation for any deviation.
3 As the water bath cooled down during each series of three readings, there would be a
contraction in the volume of gas. How could this affect the accuracy of your readings?
Suggest a control which could be employed to correct any errors of this kind.
4 The solubility of gases in water decreases with increasing temperature. How might this
property affect your results?
5 What factors, apart from temperature, might be limiting the rate of carbon dioxide
production
from yeast cells?
Discussion - answers
1 In general, a rise in temperature is accompanied by an increase in carbon dioxide
production
and the relationship is linear, at least up to 35 oC.
2 At temperatures beyond 40 oC the rate of carbon dioxide production will level out and
then start to decrease. This may be due at first to limiting factors, e.g. the rate of intake of
substrate, and finally to increasing numbers of yeast cells being killed by the high
temperature (thermal death point).
3 The effect of cooling would be to make the readings at the higher temperatures too low,
since the decrease in volume would counteract the increase due to gas production.
A suitable control would be an identical respirometer in the water bath but containing 10
cm3 of glucose solution only. The decrease in volume due to cooling would be measured
by the syringe,
and its volume added to the reading in the experimental tube.
4 The reduced solubility of carbon dioxide at higher temperatures would result in the
expulsion of some dissolved gas with each increase of 5 oC. This might make the first
reading of each group of three slightly higher than the others.
5 Limiting factors are likely to be the availability of the substrate in solution, the rate at
which it
can be taken up by the yeast cells, the availability of enzymes, co-enzymes etc. in the
cell, the rate at which the carbon dioxide can be expelled from the cell and possibly the
accumulation of inhibitory metabolites.