EXPERIMENT NO.
Before you proceed, read carefully through the whole of Question 1 and Question 2.
Plan the use of the two hours to make sure that you finish the whole of Question 1 and Question 2.
1 During the manufacture of a fruit juice, an unwanted colour can sometimes appear in the juice. An
enzyme can be used to remove this colour.
You will carry out an investigation to determine the concentration of enzyme that is most effective
at removing the colour in mock fruit juice, J. Solution J is not real fruit juice, so is not safe to drink.
You are provided with the materials shown in Table 1.1.
Table 1.1
labelled contents hazard volume / cm3
J mock fruit juice harmful 60
E 2.0% enzyme solution harmful irritant 25
W distilled water none 100
If any solution comes into contact with your skin, wash off immediately under cold water.
It is recommended that you wear suitable eye protection.
(a) You will need to carry out a serial dilution of the 2.0% enzyme solution, E, to reduce the
concentration by half between each successive dilution.
You will need to prepare four concentrations of enzyme solution in addition to the 2.0%
enzyme solution, E.
After the serial dilution is completed you need to have 10 cm3 of each concentration available
to use.
(i) Complete Fig. 1.1 to show how you will prepare your serial dilution.
Fig. 1.1 shows the first two beakers you will use to make your serial dilution. You will
need to draw three additional beakers.
For each beaker add labelled arrows to show:
• the volume of enzyme solution transferred.
• the volume of distilled water, W, added.
Under each beaker, state the concentration of enzyme solution.
9700/33/O/N/21
0 cm3 of W
............................
20 cm3 of
2.0 % enzyme ............................
10am of&tim, o
solution, E
............................
of e
............................
commis ofsolution
1%
of
10cms w
.
1
10 cm3 of 2.0 %
enzyme
r
solution to use Y
of 1 0%
10cms
............................
.
solution
............................
engyme -----
to use
- -
............................ -
Y
-
-
............................
of
10cm3 0 .
5%
solution--
-----
ge se
-
-
0
10m 0 25
%
.
---
=
enzyme ----
Solution to
of
20cms 0 125
.
%
use
solution
enzyme
to use
Fig. 1.1
[3]
Carry out steps 1 to 10.
1. Prepare the concentrations of enzyme solution, as decided in (a)(i), in the beakers
provided.
2. Label the test-tubes with the concentrations you prepared in step 1.
3. Put 5 cm3 of J into each test-tube.
4. Using the beakers labelled hot water and cold water, set up a water-bath with water
at approximately 40 °C. Maintain the water-bath at approximately 40 °C during step 5 to
step 8.
5. Put the test-tubes from step 3 into the water-bath. Leave the test-tubes for 3 minutes.
6. Put 5 cm3 of the 2.0% enzyme solution into the appropriately labelled test-tube. Shake
gently to mix.
7. Repeat step 6 with the other concentrations of enzyme solution you prepared in step 1.
8. Start timing and leave the test-tubes in the water-bath for 10 minutes.
While you are waiting carry on with Question 1.
9. After 10 minutes (step 8) remove the test-tubes from the water-bath. Observe the colour
of the solution in each test-tube.
To see the colour more clearly, it may help to hold a piece of white paper behind the
test-tube.
You may see the same colour in more than one test-tube.
10. Record your results in (a)(ii) using the symbols shown in Table 1.2.
Table 1.2
intensity of colour symbol
dark blue +++++
++++
decreasing
intensity of blue +++
colour
++
no colour +
(ii) Record your results in an appropriate table.
You may use the same symbols for more than one test-tube.
Concentration
Solution
of enzyme Intensity of colour
(% )
+ + +
28
. + +
1 0
.
+ + + +
0 5 .
+ ++
0 25
.
+ +
0 125
.
[5]
(iii) Using your results in (a)(ii), state which concentration of enzyme removed the colour
most effectively.
2 0 %
.....................................................................................................................................
.
[1]
(iv) Using your knowledge of enzymes, explain the trend in your results.
More substrate
complexes form
...........................................................................................................................................
enzyme concentration
at
high of
...........................................................................................................................................
enzyme
.
..................................................................................................................................... [1]
(v) State one variable, other than temperature, that needs to be controlled in this investigation.
PH
..................................................................................................................................... [1]
(vi) The procedure used in this investigation has several sources of error. Table 1.3 shows
one of these sources of error.
Complete Table 1.3 by:
• stating two other sources of error
• describing an improvement to the procedure for each of the three sources of error.
Table 1.3
source of error how to improve the procedure
out tests
the test-tube with 2.0% enzyme was left for carry individually
longer than the other test-tubes
temperature difficult to Use thermos
tatically
controlled
control water-bath
.
the
difficult judge colour Useedorimeter.
to
[5]
(b) Grapes are a type of fruit that can be eaten freshly picked or dried.
Table 1.4 shows the sugar content of fresh grapes and dried grapes.
Table 1.4
sugar content / g per 100 g of grapes
type of sugar
fresh dried
glucose 6.5 27.0
fructose 7.5 29.5
sucrose 0.5 1.0
(i) Plot a bar chart of the data in Table 1.4 on the grid in Fig. 1.2.
Use a sharp pencil for drawing graphs.
30-
·
25
*
20-
gas
-
*
15 -
o
-
5 -
⑧
dried
fresh dried fresh fresh dried
Glucose
Fructose Sucrose
Type of Sugar
Fig. 1.2
[4]
(ii) The concentration of all sugars in dried grapes is higher than in fresh grapes.
Calculate the percentage increase in the concentration of glucose in dried grapes.
⑫65 x10
Show your working.
-
315
answer = ......................................................% [2]
(iii) Suggest why the glucose concentration is higher in the dried grapes than in the fresh
grapes.
Dried have less water
grapes
.
...........................................................................................................................................
...........................................................................................................................................
..................................................................................................................................... [1]
[Total: 23]
