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Beetroot Lab February 2022

This document provides instructions for an experiment to determine the thermal death point of membrane proteins in beetroot cells. Students are asked to design an experiment using beetroot tissue samples heated to varying temperatures to test the hypothesis that membrane proteins will denature and allow betanin pigment to leak out at a certain temperature. The experiment involves heating identical beetroot samples, collecting the water, and measuring betanin concentration to find the temperature where proteins denature and pigment leaks out.

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johnosborne
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298 views3 pages

Beetroot Lab February 2022

This document provides instructions for an experiment to determine the thermal death point of membrane proteins in beetroot cells. Students are asked to design an experiment using beetroot tissue samples heated to varying temperatures to test the hypothesis that membrane proteins will denature and allow betanin pigment to leak out at a certain temperature. The experiment involves heating identical beetroot samples, collecting the water, and measuring betanin concentration to find the temperature where proteins denature and pigment leaks out.

Uploaded by

johnosborne
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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THE THERMAL DEATH POINT OF MEMBRANE PROTEINS

Beetroot cells contain an intensely red, water-soluble pigment, betanin, which is found in the central vacuoles.
When a beetroot is sliced, the plasma membranes of some cells are damaged and the betanin escapes and
stains your hands. Heating beetroot tissue produces a similar effect - membrane proteins are irreversibly
denatured by heat. The proteins will no longer be able to control the flow of the red pigment, so the betanin
pigment will be found outside the tissue.

EXPLORATION
o GENERAL QUESTION, WHICH DRIVES THIS INVESTIGATION

At which temperature would you expect the proteins in the plasma membrane of beetroot cells to be denatured
by heat?

o PREPARATION

Research and read around the topic of protein denaturation, to give you some background from which you can
propose a hypothesis.

o HYPOTHESIS

Devise a hypothesis, based upon your knowledge of proteins, to suggest at which temperature the proteins will
be denatured and betanin will leak from the beetroot tissue in large quantities. (This hypothesis will be formally
written in the Planning document.)

EXPERIMENTATION
o TESTING THE HYPOTHESIS

A Read the information below and then complete the planning format, including your hypothesis and
its justification, determining the variables and deciding an appropriate procedure.

Carry out an investigation to determine the approximate temperature at which membrane proteins are denatured,
using replicate (identical) sticks or discs cut from well-washed beetroot tissue. Much of the information needed
to design the investigation is given below:

1 Identical sticks or discs of tissue, cut with a knife to a size which fit easily into the available tubes or
cups which contain water, should be subjected to a fixed period (10 minutes?) in water at a range of
temperatures between 0°c and boiling. Suggested temperatures might be 0°c, 20°c, 40°c, 60°c and 80°c
(ie boiling in Quito). Use the same number of equally-sized beetroot sticks or discs in, perhaps, 5
different temperatures. (At this stage, the volume of water is not important – the water simply should
surround the sticks, so that they are properly heated at the necessary temperature.)
2 After heating for 10 minutes, the beetroot pieces should be removed and placed into clean containers,
containing the same volume of (distilled) water in each container. They should be left for a fixed period
– say 15 minutes - at room temperature. The pieces of beetroot can then be thrown away. The remaining
water sample will now contain any betanin pigment that has leaked out of the tissue, through the
denatured proteins.
3 The intensity of the colour of the solutions reflects how much pigment is present. This can be measured
accurately (ie. quantitatively) with a colorimeter, using a filter which is complementary to red. But,
without the service of a colorimeter, a quantitative scale must be devised, probably from 0 to 4 (ie 5
levels of colour saturation).

ANALYSIS
o DATA PRESENTATION

B Make a table of the quantitative results obtained and include any qualitative results of importance.
The table should be in digital form. Make sure to show margins of error (+/-).

C Plot the results against temperature in a graph. The graph should be completed digitally and should
show whisker lines which indicate margins of error or degrees of uncertainty.

You should obtain a curve from which you can deduce the approximate temperature at which the escape of
betanin was accelerated. At the point where the proteins are denatured, a greater density of the pigment should
be apparent.

EVALUATION
o CONCLUSIONS

D Support or reject your hypothesis by commenting upon and discussing the results and the graph
which you obtained.

What does the graph show? What happened? What is the explanation? If the value for the denaturing of the
membrane proteins is different from the one proposed in your hypothesis, comment on the possible explanations
by analysing your experimental method, or reviewing the hypothesis.

o EVALUATION

E List uncertainties of measurement as well as degrees of precision (+/-) of measuring apparatus.


Comment about other errors which might have affected the validity of the results.

F Review the investigation, commenting upon possibilities for improvement and follow-up.
o HELP & GUIDANCE

Matrials and equipment


Beetroots (2 or 3 large) Scissors
Knife or scalpel or disc cutter Forceps or sharp point to life beetroot sticks
10 cups or containers Ruler
Ice & boiling water Some way to measure volumes of water
(Distilled) water Cutting surface
Paper towels
YouTube video

NB: Of course you cannot follow the exact process in this video but the general idea is presented. This scientist
completes the investigation in only one stage but this is unlikely to generate great results.

https://www.youtube.com/watch?v=Hc3Mg0Yc7kI

Doing this investigation at home presents several special problems! You will need to be creative to find ways to
overcome these problems.

 You probably will not have distilled water. Use tap water.
 Cutting pieces of beetroot: You could use lots of small discs, cut with a straw or similar, or use sticks.
Whatever you decide, make sure the pieces are identical in their dimensions. The bigger the surface area of
your sample, the better.
 You will probably not have a 0°c to 100°c thermometer. No problems! 0°c is easy – ice! 20°c is easy – room
temperature. 40°c – finger hot; or do you have a medical thermometer? 60°c – this is the hardest! 80°c –
more or less the temperature of boiling water in Quito. (Maybe you have an oven thermometer?)
 Measuring volumes of water – easy! The actual volumes are not so important; what is important is that the
volumes are the same.
 Quantitative colour scale – there are some free phone apps which provide excellent results. One I have used
which worked well is called ON Color Measure from Google Play Store.

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