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A REPORT ON THE EFFECT OF TEMPERATURE ON MEMBRANES

PREPARED FOR BIOLOGY PRACTICAL 2.1

BY GAAJEEN A/L PARMAL @ PERUMAL

(NRIC: 920621-10-5801)

(SID NO: 2010646046)

IN GROUP 11M7

UNDER MADAM ILYANIE HJ. YAACOB

SUBMITTED ON 04 OCTOBER 2010

1 The Effect of Temperature on Membranes

TEAM MEMBERS

FROM LEFT:

MUHAMMAD HAQIMI BIN ZULKEFLI (920614 – 04 – 5273)

GAAJEEN A/L PARMAL @ PERUMAL (920512 – 04 – 5192)

THERESA NG LEK WEI (920512 – 04 – 5192)

2 The Effect of Temperature on Membranes

Title
The effect of pH value on enzyme activity

Objective
To investigate how different pH value can affect rate of enzyme activity

Problem Statement
What is the effect of different PH value on rate of enzyme activity?

Hypothesis
The rate of enzyme reaction is highest when the pH value is at optimum level. The rate of
enzyme activity is fastest at pH 6.5, the optimum pH value for potato catalase.

Introduction
Beetroot

Kingdom: Plantae

Phylum: Magnoliophyta

Class: Magnoliopsida

Order: Caryophyllales

Family: Amaranthaceae
1
Diagram 1: Picture of raw beetroot Table 1: Scientific Classification of
Beetroot2 Genus: Beta

Species:
Beetroot are biennial plants grown as annuals B. vulgaris
and harvested for their swollen root
tuber and leaves. It tastes like earthy caramel and its freshness is similar to earthy mint. The
usually deep-red roots of beetroot (refer to Diagram 1) are eaten boiled either as a cooked
vegetable, or cold as a salad after cooking and adding oil and vinegar, or raw and shredded,
either alone or combined with any salad vegetable. A large proportion of the commercial
production is processed into boiled and sterilised beets or into pickles.

1 Image source: www.greatgrubclub.com/?location_id=70

2 Information source: http://en.wikipedia.org/wiki/Beet

3 The Effect of Temperature on Membranes

 Diagram 2: Chemical structure of beetroot3

The bright red colour in beet roots is caused by betalains, or specifically betacyanin, a
water soluble plant pigment replacing anthocyanins in plants of the order Caryophyllales.
Betalains contain nitrogen while anthocyanins do not (refer to Diagram 2). Betalains are
found in the vacuole. The red-violet vesicles obtained from protoplasts are intact beet
vacuoles, so betalain pigments have to cross two membranes. In the living tissues of beet
slices, there can be a leakage by a heat shock, by acid treatment or by incubating them in
acidified 80% methanol, due to damage of the membrane structure and denatured plasma
proteins. Besides, putting beet slices in the deep-freeze will kill them, and the pigments will
leak out. On the other hand, a pH of about 3-4 stabilises the pigments and protects them
against oxidation. Pigment extracts must also be protected from direct sun light and should
be kept in a cool and dark place.

Diagram 3: Betalains present in flowers4

The function of betalains is not provably known, but it is assumed that when present
in the flowers (refer to Diagram 3), they serve to attract pollinating insects and when present
in seeds they may attract birds to disperse the seeds. Man favours betalains because they are
attractive to look at, and the colour may have co-segregated with another useful trait which
improves the flavour. This explains why betanin, a red glycoside food dye obtained from
beets, is commercially used. There is no indication that betalains protect plants against
herbivores and pathogens such as fungal, bacterial or viral, and they do not absorb UV light.

3 Image source: www.absoluteastronomy.com/topics/Betalain

4 Image source: www.webexhibits.org/causesofcolor/7H.html

4 The Effect of Temperature on Membranes

 As for the goodness for human, betalains are often claimed to have antioxidant
properties and are believed to be good for health. Beetroots are good sources of folic acid,
potassium and dietary fibre. They also contain sugars that contribute to its calorie content.
Beetroot also has stimulating effects on the liver's detoxification processes. The betacyanin
content gives beetroot its rich purple-crimson colour and is a potent cancer-fighting agent.
Beetroot's fibre promotes both healthy cholesterol levels and bowel function.5

Diagram 4: Structure of plasma membrane6

When beetroot is heated or cooked, there will be lots of red dye in the cooking
water. This is because leakage of betalains has occurred as a result of the disruption of the
cell membranes. A biological membrane is made of a phospholipid bilayer. A phospholipid
bilayer is formed because the phospholipids that line up side by side have a polar "water-
loving" (hydrophilic) head and a non-polar “water-hating” (hydrophobic) hydrocarbon tail.
The tails pack together and face inwards, exposing only the polar heads to the water. It
resembles two blankets one atop the other, with the fatty acid tails towards each other.

In a cell, the phospholipids form sacks. The biggest sack goes all around the whole
cell to form the plasma membrane; others may form the tonoplast of vacuoles. In these lipid
seas, there will be a number of proteins in various degrees of submersion (refer to Diagram 4).
Some span all the bilayer, thus being exposed on both sides, is known as integral, trans-
membrane or intrinsic proteins. Others just drift on either of its surfaces, which are called
peripheral or extrinsic proteins. Typically, 70% of a cell membrane is protein, mostly for
transport. The water in the cytoplasm around and within the compartments formed by the
phospholipid bilayers is also crammed with protein.

5 Information source:
http://blogs.abc.net.au/nsw/2009/09/kitchen-gardener-episode-7-beetroot.html?
program=702_drive

6 Image source: www.tutorvista.com/.../plasma-membrane.php

5 The Effect of Temperature on Membranes

Variables
Manipulated variable (Independent variable): Different pH values

Responding variable (Dependent variable): Rate of enzyme reaction

Fixed variables: the time for the action of enzyme potato catalase,
the volume of buffer solution used
the volume of hydrogen peroxide used.

Apparatus
Stopwatch, calibrated tube, test tube, boiling tube, boiling tube rack, 250cm3 beakers
,delivery tube, cork, syringe, spatula, measuring cylinders.

Diagram 5: cuvettes7

Materials
Citric acid and sodium phosphate buffer solutions, hydrogen peroxide solution 6% w/v,
Blended potato, water baths.

7 Image source: www.labdepotinc.com/c-489-spectrophotometer.php

6 The Effect of Temperature on Membranes

Procedures
Preparation of the materials and apparatus

1. 1 spatula of blended potato is added into a boiling tube.

2. 5cm3 of buffer solution of pH 4.4 is transferred into the boiling tube.
3. It is swirled to mix.
4. Graduated tube is filled with water and is inverted carefully into the beaker.
5. 2.5 cm3 of hydrogen peroxide is measured into the syringe and added to the boiling
tube.
6. The flask is connected.
7. The graduated tube is immediately placed into the end of the delivery tube.
8. Stopwatch is started. The volume of oxygen collected is measured every 30 second
for 5 minutes.
9. Steps 1 to 8 are repeated using different buffer solution of pH 5.2, 6.5, and 7.5.
10. The experiment is repeated twice to obtain the average value.
11. All the results obtained are tabulated.
12. A graph of rate of catalase activity against pH value is plotted.

Results
Time(second) 30 60

pH value Volume of oxygen Volume of oxygen released

released (cm3) (cm3)
Trial
1 2 3 Avera 1 2 3 Average
ge

4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

5.2 1.0 1.0 0.0 0.67 1.0 1.0 1.0 1.0

6.5 16.0 5.0 7.8 9.6 23.6 7.6 12. 14.6

6

7.5 10.0 10.6 12. 10.9 16.0 17.8 20. 18.0

0 0

7 The Effect of Temperature on Membranes

Table 1: Volume of oxygen collected in respective to different pH values

Time(second) 90 120

pH value Volume of oxygen Volume of oxygen released

released (cm3) (cm3)
Trial
1 2 3 Avera 1 2 3 Average
ge

4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

5.2 2.0 1.8 2.0 1.9 2.6 2.0 2.0 2.2

6.5 27.0 11.4 16. 18.1 28.0 13.0 19. 20.2

0 6

7.5 21.0 23.0 25. 23.0 25.0 26.0 29. 26.7

0 0

Table 2: Volume of oxygen collected in respective to different pH values

Time(second) 150 180

pH value Volume of oxygen Volume of oxygen released

released (cm3) (cm3)
Trial
1 2 3 Avera 1 2 3 Average
ge

4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

5.2 2.7 2.2 2.0 2.3 2.8 2.2 2.4 2.5

6.5 28.4 15.4 21. 21.7 28.4 16.0 23. 22.5

4 0

7.5 27.0 28.8 31. 29.1 29.8 30.8 33. 31.3

6 4

8 The Effect of Temperature on Membranes

Table 3: Volume of oxygen collected in respective to different pH values

Time(second) 210 240

pH value Volume of oxygen Volume of oxygen released

released (cm3) (cm3)
Trial
1 2 3 Avera 1 2 3 Average
ge

4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

5.2 2.8 2.2 2.6 2.5 2.8 2.4 2.8 2.7

6.5 28.4 16.0 23. 22.7 28.6 16.0 25. 23.2

8 0

7.5 31.4 32.6 34. 32.9 32.4 32.9 35. 33.6

6 6

Table 4: Volume of oxygen collected in respective to different pH values

Time(second) 270 300

pH value Volume of oxygen Volume of oxygen released

released (cm3) (cm3)
Trial
1 2 3 Avera 1 2 3 Average
ge

4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

5.2 2.8 2.6 2.8 2.7 2.8 2.8 2.8 2.8

6.5 28.6 16.0 26. 23.5 28.6 16.0 26. 23.5

9 The Effect of Temperature on Membranes

 0 0

7.5 33.4 33.6 36. 34.3 34.0 35.4 36. 35.3

0 4

Table 5: Volume of oxygen collected in respective to different pH values

Graph 1: Graph of volume of oxygen released against pH value of buffer solution

Graph 2: Graph of rate of catalase reaction against pH value of buffer solution

Discussion
10 The Effect of Temperature on Membranes
Analysis and Interpretation of Results

In this experiment to examine the effect of temperature on cell membranes, the

surrounding temperature of the cell membrane of beetroot is modified to see the effect it has
on the membrane structure. Graph 1 above which data is plotted using the average value of
colorimeter reading in Table 2 shows the general trend of data with increasing temperature.
From the graph, we are able to see that the colorimeter reading decreases from 0°C to 27°C
and increases from 27°C to 65°C. To understand this, we ought to know the functioning of
the colorimeter in this experiment. The colorimeter is used to measure the percentage
absorbance of light by the solutions where the beetroot cylinders are immersed into after the
heat treatment. The higher the colorimeter reading brings about a higher percentage
absorbance and so the higher is the amount of red dye present in the solution because there is
more red dye in the solution to absorb the light transmitted from the colorimeter. The
increase in amount of red dye in the solution is related to the increase of the permeability of
beetroot membrane. Hence, an increase in the reading of colorimeter indicates that the
permeability of the beetroot membrane increases.

The colorimeter shows a reading of 0.207 arbitrary units when the beetroot cylinder is
immersed into water bath at 0°C. It is accurately drawn in the graph that when the beetroot
cylinder is immersed into distilled water at room temperature, that is 27°C, the colorimeter
shows a reading of 0.144 arbitrary units. This colorimeter reading is lower compared to
reading taken at 0°C. Presumably we understand that the leakage of betalains located within
the cell vacuole is due to the damaged membrane structure and denatured plasma proteins at
high temperatures. But why is there any pigment released from what should be intact cell
membranes at room temperature? This may be due to when washing the beetroot, all we are
washing off is the pigment which has leaked out of the damaged cells at that time. The
pigment does not necessarily all leak out at once, however, and the leakage is probably
continuing when we start the experiment.

While for higher temperatures, from the distilled water which the beetroot cylinder
stands in after it is immersed in water bath at 35°C, the colorimeter reading is 0.197 arbitrary
units. Similarly when the temperature of water bath is at 45°C, the colorimeter reads a value
of 0.442 arbitrary units. The colorimeter shows a reading of 0.969 arbitrary units for the
condition where the beetroot cylinder which is being immersed in water bath at 55°C. Lastly
from the distilled water which the beetroot cylinder stands in after it has been heat treated at
65°C, we obtain a colorimeter reading of 1.142 arbitrary units.

11 The Effect of Temperature on Membranes

 Diagram 6: Effect of temperature on the packing of the hydrocarbons8

To function correctly a cell needs to be able to control transport across the partially
permeable cell surface membrane, so normally the betalains in the cell vacuole cannot pass
through it. In addition, in order for the betalain to leave the cell it needs to pass through two
different membranes, which are the membrane bounding the vacuole and the membrane
enclosing the cell. At normal room temperature, the phospholipid bilayer is in a gel state and
tightly packed (refer to Diagram 6). At higher temperatures, the bilayer actually ‘melts’ and
the interior is fluid allowing the lipid molecules to move around, rotate, exchange places. As
more phospholipids move around, more betalains are able to leak out. The cellulose cell wall
on the outside of the cell is fully permeable and so no barrier to the egress of the pigment.
Consequently at higher temperature, the red dye in the distilled water where the beetroot
cylinder is placed is more concentrated.

Not only that, when we increase the temperature of the water baths, the transport
proteins embedded on the plasma membrane is denatured. Heat can be used to disrupt
hydrogen bonds and non-polar hydrophobic interactions of the polypeptide chain. This occurs
because heat increases the kinetic energy and causes the molecules to vibrate so rapidly and
violently that the bonds are disrupted. This damages the vacuole and makes holes in the cell
membrane, inducing leakage of cytoplasm and other substances contained within the
membrane, which is proven by the presence of red dye in the distilled water.

From the temperature of 27°C to 35°C, the colorimeter reading increases gradually
and steadily, meaning that the percentage absorbance rises slowly. This shows there is only a
little increase in the amount of betalains present in the distilled water which the beetroot
cylinder stands in after it is immersed in water bath. The rise may not seem to be significant

8 Image source: Wolfe S.L., Molecular and Cellular Biology, (1993). Wadsworth Publishing
Company.

12 The Effect of Temperature on Membranes

because 35°C is not high enough to denature the plasma proteins. The increase in heat
energy only increases the kinetic energy of the phospholipid molecules to move around,
rotate and exchange places which at the same time increases the rate of random movement of
betalains out through the cell membrane. Still, more leakage of betalains has occurred at
35°C as compared to that at 27°C. This indicates that the permeability of cell membrane has
increased even by an increase in room temperature.

From the temperature of 35°C to 45°C, there is a rapid rise in the colorimeter reading,
showing that the percentage absorbance increases in a sudden manner due to a big increase in
the amount of betalains present in the distilled water which the beetroot cylinder stands in
after it is immersed in water bath. As most proteins denature by which its tertiary structure
unravels as a result of the destroy of the strong covalent bond between the R groups of amino
acids in the polypeptide chain at a temperature over 40°C, there is less control of transport of
substances across the partially permeable cell surface membrane. Hence much more
betalains is able to pass out of the membrane, causing the permeability of cell membrane
increases so much as shown by the gradient of the line from the temperature of 35°C to 45°C
on the graph.

From the temperature of 45°C to 55°C, the gradient of the line increases again. This
shows the permeability of the cell membrane is even higher due to more and more plasma
proteins have been denatured, in addition to the ‘melting’ of the bilayer because of the violent
movement of the phospholipid molecules.

As the temperature goes on to 65°C, the curve starts to flatten out, meaning that the
rate of leakage of betalains has decreased. This is probably due to most of the plasma
proteins have been denatured by the heat, and the leakage is already at its maximum rate. In
other words, the amount of the plasma proteins has become the limiting factor for this
experiment. However, there is also another suggestion that says that this happens because
although the denaturing of the plasma proteins causes a rapid rise in the amount of betalains
released, when the temperature begin to get higher still, the tertiary structure of the protein
blocks parts of the holes in the cell membrane 9. Therefore, this slows down the leakage of
betalains. On the other hand, it could also be the effect of a very high temperature which
causes the red dye from the beetroot to change colour, and so influences the colorimeter
reading. The red pigment is heat labile, which means it is denatured by heat and would turn
into a yellow chemical. Since the colorimeter is set to measure absorbance at one
wavelength, a change in colour will result in a lower absorbance at that wavelength and so
apparently the amount of betalains decreases.

9 Information source: http://www.123helpme.com/view.asp?id=123054

13 The Effect of Temperature on Membranes

Evaluation
Limitations and Improvements

Laboratory coats were worn during the investigation to prevent

chemicals from spoiling clothes. Care was also taken whilst handling
the chemicals as hydrogen peroxide is corrosive and the manometer
fluid is permanently staining.Assumingly that we have done the experiment under
full attention, there are still some techniques themselves which are faulty. One of the
limitations of the experiment is that the red dye of betalains may not be uniformly dispersed
throughout the distilled water even though the solution has been shaken. In other words, the
solution being measured using the colorimeter may not be a homogenous solution. Thus, the
reading of the colorimeter obtained may not accurately show the actual percentage
absorbance of the red dye in the solution. This might explain why the colorimeter reading at
27°C is lower than that of 0°C. To eliminate this limitation we can possibly stir the solution
using a stirrer or glass rod.

Besides that, a little amount of the red dye in the solution may be lost when we
transfer the solution into the cuvette. To test for the percentage absorbance we placed 2 cm3
of the dye solution into the colorimeter cuvette by using a dropper. There is a high chance
which a small amount of the solution is still left in the dropper, resulting in the volume of
solution in the cuvette is less than 2 cm3 and is not constant for the testing of all the dye
solution after the beetroot is being heat treated under different temperatures. Consequently,
the accuracy of results is affected. So to get rid of this limitation, we could use a pipette for
measuring 2 cm3 instead of a dropper.

In addition to that, the reading of colorimeter may not necessarily be perfectly

accurate. This is due to another limitation in this experiment using colorimeter that lies in the
imperfections in the glass or plastic cuvette. We need to understand that colorimeter
functions by simply directing a light through a cell cuvette containing the dye solution to be
analyzed. A portion of the light beam is absorbed by the sample and the remaining portion
passes through the cuvette to the photodetector means for measurement. Very minor
variations in a cuvette, for example any foreign particles or fingerprints on the cuvette will
substantially affect measurements made by the photodetector means. So to reduce the effect
of this limitation, we shall clean the surface of the cuvette properly by wiping using a clean
cloth. We should also make sure the cuvette surface is smooth on all sides.

As explained in the discussion for the flattening of the curve from the temperature of
55°C to 70°C, a very high temperature can cause the red dye from the beetroot to change
colour to yellow. Since the filter dial of the colorimeter is set to the blue/green filter, the
colorimeter can only measure absorbance at one wavelength; a change in colour will result in
a lower absorbance at that wavelength and so apparently the amount of betalains decreases.
This limitation is hard to eliminate.

Other than that, when we measure and cut 1 cm length slices from sections of
beetroot, we assumed that each of the length slices has roughly the same surface area to
volume ratio, but in actual they are not exactly the same. This would lead to slight

14 The Effect of Temperature on Membranes

inaccuracy of the results because we know the rate of diffusion would increase if the total
surface area to volume ratio increases. There might be more betalains diffused into the
distilled water for thinner and smaller beetroot cylinders. The best way to eliminate this
limitation is to repeat the experiment and get the average of the results.

Besides, as mentioned before why is there any pigment released from what should be
intact cell membranes at room temperature, this is may be due to one of the limitations in this
experiment too. When we cut the beetroot cylinders and place them in a beaker of distilled
water, the limited time may not be able to wash away all the red pigments on the outside of
the beetroot cylinder which comes from the damaged cells. What we can possibly do is to
leave them in the distilled water for a longer time, perhaps 15 minutes if not left overnight.

Systematic and Random Errors

When we are measuring the length of beetroot cylinders, parallax error may occur
when our eye level is not perpendicular to the scale of the ruler. This error would contribute
to the limitation of inconstant length of beetroot cylinders as mentioned above.

Parallax error may also occur when we are measuring the volume of distilled water
needed to be placed into each boiling tube if our eye level is not perpendicular to the bottom
meniscus of the water. When the volume of distilled water is inconstant, the concentration of
the betalains that leak out of the beetroot will vary too. To avoid these two parallax errors,
we might as well pay total attention and concentration by making sure our eyes is
perpendicular to the scale of the measuring tools.

Systematic error may occur when zero error is present on the colorimeter. It is
therefore very important when we calibrate the colorimeter. The colorimeter is adjusted to
read zero absorbance for clear water. We also made sure no one alter the setting again during
the experiment especially when we share the colorimeter with other groups.

In this experiment, we used electric water baths and not prepare our individual water
baths. Hence thermometer used in this experiment is just to confirm the temperature of the
water bath, as there is minimum fluctuation of temperature of water baths.

Validity and Reliability of Results

There are some apparent anomalies in our results. This can be seen in the colorimeter
reading of beetroot placed in water bath of 0°C and 27°C. There are some limitations and
sources of error in the experiment that can deem our results unreliable. In order to make
sure that the results are reliable and valid, besides adhering to the other
constant variables and follow all the necessary precautionary steps as
mentioned in the beginning of discussion, replication of experiment is very
important. Replication of studies helps to support findings and eliminate random errors.
If repeated experiments confirm unexpected results in this experiment, then the hypothesis
should probably be revised if there is any hope of it becoming a theory. In this case, since the
colorimeter readings we obtain are about the same for each temperature, and the average
values we calculated does not exactly follow the expected trend, we could conclude that our
hypothesis should not be accepted.

15 The Effect of Temperature on Membranes

Safety Precautions
When handling a cork borer to cut sections from a single beetroot, it is necessary to
hold the beetroot securely and ensure that our hand is not in the path of the borer. It is also
very important to screw the borer away from our body.

These sections had to be then cut into 1 cm length slices using a sharp knife to cause
less damage to the membranes. However it may be dangerous if a person do not apply his
full concentration when using the knife. Users must be careful concerning how the knife is
held and used.

When we place or remove the boiling tubes into and out of the electric water bath, it is
essential to use a pair of boiling tube holder, if not, a wet cloth to protect our hand from the
heat. We must not attempt to hold the boiling tubes with bare hands even after they are left
over some time to prevent any accidents.

Besides, we should also handle the colorimeter with care as colorimeter is an

expensive instrument.

We ought to wear a laboratory coat and a pair of gloves as spillage of beetroot juice
will stain our skin and clothing very badly, even though it is harmless to us.

Extension
This experiment can be modified to investigate effect of alcohol concentration on
membrane permeability. Solutions with different concentration of alcohol are prepared by
mixing the same amount of alcohol with variable amount of distilled water. The beetroot
cylinders are placed in solutions with different concentration of alcohol instead of different
temperature of water baths. The amount of red dye which leaked out is measured using a
colorimeter.

Conclusion
An increase in temperature does not exactly increase the permeability of the cell membrane.
The hypothesis is not accepted.

References
1. Image on website (The Great Grub Club)
The Great Grub Club – Being Healthy is Fun. Retrieved September 29th, 2010 from
www.greatgrubclub.com/?location_id=70

16 The Effect of Temperature on Membranes

2. Image on website (Absolute Astronomy)
Absolute Astronomy – Betalains. Retrieved September 29th, 2010 from
www.absoluteastronomy.com/topics/Betalain

3. Image on website (Causes of Color)

Causes of Color – Plants and Flowers. Retrieved September 29th, 2010 from
www.webexhibits.org/causesofcolor/7H.html

4. Image on website (TutorVista)

TutorVista – Plasma Membrane. Retrieved September 29th, 2010 from
www.tutorvista.com/.../plasma-membrane.php

5. Image on website (The Lab Depot)

The Lab Depot – Spectrophotometers. Retrieved September 29th, 2010 from
www.labdepotinc.com/c-489-spectrophotometer.php

6. Article on website (Wikipedia)

Wikipedia – Beet. Retrieved September 29th, 2010 from
http://en.wikipedia.org/wiki/Beet

7. Article on website (Kitchen Gardener)

Kitchen Gardener – Episode 7 – Beetroot. Retrieved September 29th, 2010 from
http://blogs.abc.net.au/nsw/2009/09/kitchen-gardener-episode-7-beetroot.html?
program=702_drive

8. Article on website (123HelpMe)

123HelpMe - The Effect of Temperature. Retrieved September 29th, 2010 from
http://www.123helpme.com/view.asp?id=123054

9. Wolfe S.L., Molecular and Cellular Biology, (1993). Wadsworth Publishing

Company.

10. C.J.Clegg, Edexcel Biology for AS, (2008). Hodder Education.

11. Campbell, Reece, Urry, Cain, Wasserman, Minorsky, Jackson, Biology Eighth
Edition, (2008). Pearson.
(NUMBER OF WORDS: 4242)

17 The Effect of Temperature on Membranes