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International A Level Biology Edexcel Revision Notes 7. Respiration, Muscles & the Internal Environment Respiration
7.7 Core Practical 15: Investigation of Respiration in Yeast

7.7 Core Practical 15: Investigation 1. Molecules, Transport & Health

of Respiration in Yeast
2. Membranes, Proteins, DNA &…
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3. Cell Structure, Reproduction …

Investigation of Respiration in Yeast

4. Plant Structure & Function, B…
A redox indicator is a substance that changes colour when it is reduced or oxidised
DCPIP and methylene blue are redox indicators
They are used to investigate the e"ects of temperature and substrate
concentration on the rate of anaerobic respiration in yeast 5. Energy Flow, Ecosystems & t…

These dyes can be added to a suspension of living yeast cells as they don’t damage
cells
Yeast can respire both aerobically and anaerobically, in this experiment it is their rate 6. Microbiology, Immunity & Fo…
of anaerobic respiration that is being investigated

7. Respiration, Muscles & the In…

Mechanism
Dehydrogenation happens regularly throughout the di"erent stages of aerobic
Respiration
respiration
The hydrogens that are removed from substrate molecules are transferred to the
final stage of aerobic respiration, oxidative phosphorylation, via the hydrogen 7.1 Overview of Respiration
carriers NAD and FAD
The enzyme dehydrogenase catalyses the production of reduced NAD in glycolysis 7.2 Glycolysis

When DCPIP or methylene blue are present they take up hydrogens from the organic
compounds and get reduced instead of NAD 7.3 Link Reaction & Krebs Cycle

Both redox indicators undergo the same colour change when they are reduced
7.4 The Electron Transport Chain
Blue → colourless
The faster the rate of respiration, the faster the rate of hydrogen release and the
7.5 Anaerobic Respiration
faster the dyes get reduced and change colour
This means that the rate of colour change can correspond to the
rate dehydrogenase would be working at and therefore, the rate of 7.6 Respiratory Quotient
respiration in yeast
The rate of respiration is inversely proportional to the time taken 7.7 Core Practical 15: Investigation of
Respiration in Yeast
Rate of respiration (sec-1) = 1 ÷ time (sec)
7.8 Core Practical 16: Respirometer to
Calculate RQ

Apparatus
Yeast Muscular Movement

Glucose solution
Test tubes
The Internal Environment
Stopwatch
DCPIP

8. Coordination, Response & G…

Method - Temperature
1. Add a set volume of yeast suspension to test tubes containing a certain
concentration of glucose
2. Put the test tube in a temperature-controlled water bath and leave for 5 minutes to
ensure the water temperature is correct and not continuing to increase or decrease
3. Add a set volume of DCPIP to the test tube and start the stopwatch immediately
4. Stop the stopwatch when the solution becomes colourless or lose all blue colour
This is subjective and therefore the same person should be assigned this task for
all repeat experiments
5. Record the time taken for a colour change to occur once the dye is added
Repeat across a range of temperatures. For example, 30oC, 35oC, 40oC, 45oC
6. The e"ect of substrate concentration can be investigated by adding di"erent
concentrations of a substrate to the suspension of yeast cells and recording the
time taken for a colour change to occur once the dye is added
For example, 0.1% glucose, 0.5% glucose, 1.0% glucose

Methylene blue or DCPIP is added to a solution of anaerobically respiring yeast cells in

a glucose solution. The rate at which the solution turns from blue to colourless gives
the rate of dehydrogenase activity

Controlling other variables

It is important when investigating one variable to ensure that the other variables in
the experiment are being controlled
Volume of dye added: if there is more dye molecules present then the time
taken for the colour change to occur will be longer
Volume of yeast suspension: when more yeast cells are present the rate of
respiration will be inflated
Feedback

Type of substrate: yeast cells will respire di"erent substrates at di"erent rates
Concentration of substrate: if there is limited substrate in one tube then the
respiration of those yeast cells will be limited
Temperature: an increase or decrease in temperature can a"ect the rate of
respiration due to energy demands and kinetic energy changes. The
temperature of the dye being added also needs to be considered
pH: a bu"er solution can be used to control the pH level to ensure that no
enzymes are denatured

Interpretation of results
A graph should be plotted of temperature against time
As the temperature increases, the rate of respiration also increases so the time
taken for the solution to go colourless reduces
This means hydrogens are released by the reactions more quickly, hence the
DCPIP accepts electrons/hydrogens more quickly until all molecules of DCPIP
are reduced. This means that it will take less time to turn from blue to colourless
At extreme high temperatures, the enzyme may denature and meaning the colour
change may not occur

Exam Tip
Although the DCPIP and methylene blue undergo a colour change from blue to
colourless it is important to remember that the yeast suspension in the test tube may
have a slight colour (usually yellow). That means when the dye changes to colourless
there may still be an overall yellow colour in the test tube. If this is the case it can be
useful to have a control tube containing the same yeast suspension but with no dye
added, then you can tell when the dye has completely changed colour.

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Author: Marlene
Marlene graduated from Stellenbosch University, South Africa, in 2002 with a degree in Biodiversity and Ecology. After
completing a PGCE (Postgraduate certificate in education) in 2003 she taught high school Biology for over 10 years at
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