Cambridge International A Level Biology
BOX 12.1: Oxygen uptake
Oxygen uptake during respiration can be measured using
a respirometer. A respirometer suitable for measuring the
rate of oxygen consumption of seeds or small terrestrial
invertebrates at different temperatures is shown in Figure
12.19.
Carbon dioxide produced in respiration is absorbed by
a suitable chemical such as soda-lime or a concentrated
solution of potassium hydroxide (KOH) or sodium hydroxide
(NaOH). Any decrease in the volume of air surrounding the
organisms results from their oxygen consumption. Oxygen
consumption in unit time can be measured by reading the
level of the manometer fluid against the scale.
Changes in temperature and pressure alter the volume
of air in the apparatus, and so the temperature of the
surroundings must be kept constant while readings are
taken for example, by using a thermostatically controlled
water bath. The presence of a control tube containing
an equal volume of inert material to the volume of the
organisms used helps to compensate for changes in
atmospheric pressure.
Once measurements have been taken at a series
of temperatures, a graph can be plotted of oxygen
consumption against temperature.
The same apparatus can be used to measure the RQ
280
of an organism. First, oxygen consumption at a particular
temperature is found (xcm3min1). Then the respirometer is
set up with the same organism at the same temperature, but
screw-clip
non-vertebrates
to be studied
gauze platform
soda-lime
or KOH
or NaOH
experimental tube
Figure 12.19 A respirometer.
with no chemical to absorb carbon dioxide. The manometer
scale will show whether the volumes of oxygen absorbed and
carbon dioxide produced are the same. When the volumes
are the same, the level of the manometer fluid will not change
and the RQ = 1. When more carbon dioxide is produced than
oxygen absorbed, the scale will show an increase in the
volume of air in the respirometer (byycm3min1). The RQ can
then be calculated:
CO2 x + y
RQ = x
O2
Conversely, when less carbon dioxide is produced than
oxygen absorbed, the volume of air in the respirometer will
decrease (by zcm3min1) and the calculation will be:
CO2 x z
RQ =
O2 x
Another way of investigating the rate of respiration
of yeast is to use a redox dye such as a solution of
dichlorophenolindophenol (DCPIP) or of methylene blue.
These dyes do not damage cells and so can be added to a
suspension of yeast cells. When reduced, these blue dyes
become colourless. The rate of change from blue to
colourless is a measure of the rate of respiration of the yeast.
This technique can be used to investigate the effect of
various factors on yeast respiration, such as temperature,
substrate concentration or different substrates.
1 cm3 syringe
three-way tap
glass beads
soda-lime
or KOH
or NaOH
capillary U-tube
control tube containing
manometer fluid
 Cambridge International A Level Biology
QUESTION
Mitochondrial structure and
function
In eukaryotic organisms, the mitochondrion is the site
of the Krebs cycle and the electron transport chain.
Mitochondria are rod-shaped or filamentous organelles
about 0.51.0m in diameter. Time-lapse photography
shows that they are not rigid, but can change their shape.
The number of mitochondria in a cell depends on its
activity. For example, highly active mammalian liver cells
contain between 1000 and 2000 mitochondria, occupying
20% of the cell volume.
The structure of a mitochondrion is shown in Figure 12.13. Like
a chloroplast, each mitochondrion is surrounded by an envelope of
two phospholipid membranes (Chapter 4). The outer membrane
is smooth, but the inner is much folded inwards to form cristae
(singular: crista). These cristae give the inner membrane a large
276
total surface area. Cristae in mitochondria from different types of
cell show considerable variation, but, in general, mitochondria from
active cells have longer, more densely packed cristae than
mitochondria from less active cells.
Figure 12.13 Transmission electron micrograph of a
mitochondrion from a pancreas cell (15000).
The two membranes have different compositions and
properties. The outer membrane is relatively permeable to
small molecules, whereas the inner membrane is
less permeable.
The inner membrane is studded with tiny spheres, about
9nm in diameter, which are attached to the inner membrane
by stalks (Figure12.14). The spheres are the enzyme ATP
synthase. The inner membrane is the site of the electron
transport chain and contains the proteins necessary
for this. The space between the two membranes of the
envelope usually has a lower pH than the matrix of the
mitochondrion as a result of the protons that are released
into the intermembrane space by the activity of the
electron transport chain.
The matrix of the mitochondrion is the site of the
link reaction and the Krebs cycle, and contains the
enzymes needed for these reactions. It also contains small
(70S) ribosomes and several identical copies of looped
mitochondrial DNA.
ATP is formed in the matrix by the activity of ATP
synthase on the cristae. The energy for the production
of ATP comes from the proton gradient between the
intermembrane space and the matrix. The ATP can be
used for all the energy-requiring reactions of the cell, both
inside and outside the mitochondrion.
Figure 12.14 Transmission electron micrograph of ATP
synthase particles on the inner membrane of a mitochondrion
(400000).