12 Energy and respiration

12.1 Energy Image: Cambridge Intl A-Level Coursebook

a) The need for energy in living organisms

All living organisms need a continuous supply of energy
to maintain their metabolism. They either absorb light
energy (autotrophs) or chemical potential energy
(heterotrophs). This is to do the work necessary to live.
Work includes: 6) rapid turnover rate
7) readily available in cells upon demand as stores are
1) anabolic reactions
released in manageable amounts, meaning no
2) active transport (transport of substances against
energy is wasted
concentration gradient)
3) movement e.g., muscle contraction, cilia movement, c) ATP synthesis
locomotion
4) bioluminescence, electrical discharge, temperature ATP is synthesised in substrate-linked reactions in
regulation, etc. glycolysis and in the Krebs cycle.

Ø anabolic reactions - synthesis of complex high H+ concentration

∴ positive
substances from simpler ones

Examples of anabolic reactions

1) DNA replication
2) protein synthesis
3) active transport low H+ concentration ∴
4) movement negative
5) the maintenance of body temperature
making polysaccharides, triglycerides, polynucleotides
(DNA/RNA)
6) phosphorylation
Image: bio.libretexts.org

b) ATP 1) the process by which ATP is synthesised is called

Features of ATP that make it suitable as the universal chemiosmosis
energy currency 2) it occurs in the inner mitochondrial membrane (or
the thylakoid membrane of a chloroplast)
Image: ‘Structure of Adenosine Triphosphate’ by Phil Schatz.
3) ATP is generated using electrical potential energy
(this is energy from the transfer of electrons by
electron carriers like NADH and FADH in the electron
transport chain)
4) the electrons cause conformation changes in the
shapes of the proteins and cause them to pump H+
across a selectively permeable cell membrane
5) the uneven distribution of H+ ions across the
membrane establishes both concentration and
electrical gradients (thus, an electrochemical
gradient) due to their positive charge as well as them
collecting at one side of the membrane
6) H+ are allowed to flow down their concentration
gradient via facilitated diffusion through ATP
synthase
1) small
2) water-soluble 7) the transfer of 3 H+ allows the production of 1 ATP
3) easily transported around the cell molecule provided that ADP and Pi are available
4) easily hydrolysed to release energy inside the mitochondrion
5) relatively large quantity of energy released 8) as H+ travel through the ATP synthase complex, it
drives the down part by a rotation force provided by
ATP + H2O ⇌ ADP + Pi (inorganic phosphate) the electrochemical gradient that produces ATP

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 Image: Cambridge Intl A-Level Coursebook

Structure of ATP synthase Why more ATP can be synthesised in aerobic respiration
Has a total of 6 binding sites; from one gram of lipid
three of them are 1) lipids contain relatively more hydrogen atoms
catalytically inactive and they 2) more reduced NAD and FAD are produced
bind ADP. The three other 3) more electrons are passed along ETC
subunits catalyze the ATP 4) more hydrogens pumped across inner mitochondrial
synthesis. membrane
Transverse section of ATP synthase

d) Roles of coenzymes in respiration g) Respiratory quotient (RQ)

1) NAD (nicotinamide adenine dinucleotide) • the ratio of oxygen taken in and carbon dioxide
reduction
released is 1:1, however when other substrates are
NAD+ + H+ + 2e- ⇌ NADH being respired, this ratio differs
oxidation • measuring this ratio therefore can show what
substrate is being in respiration
• a hydrogen carrier molecule, accepts a hydrogen from
one reaction (getting reduced) and donates it to • change in RQ indicates that the substrate being
another respired has changed
• NADP (NAD + phosphate group) is used a hydrogen • the ratio can also show whether or not anaerobic
carrier molecule in photosynthesis respiration is occurring

2) FAD (flavin adenine dinucleotide) Ø respiratory quotient - the ratio of carbon dioxide
reduction
molecules produced to oxygen molecules taken in
during respiration
FAD + 2H+ + 2e- ⇌ FADH2
oxidation
RQ = CO2 produced
• similar in function to NAD
• used in respiration in the Krebs cycle O2 consumed

3) coenzyme A Typical RQs for the aerobic respiration of different

substrates:
pyruvate + CoA + NAD ⇌ acetyl COA + CO2 +
respiratory substrate energy density / kJ g-1
reduced NAD carbohydrate 1.0
• carries acetyl groups made from pyruvate during the lipid 0.7
link reaction to Krebs cycle protein 0.9
• can also carry acetyl groups made from fatty acids
RQ values when respiration is anaerobic
e) Explain that the synthesis of ATP is Example: yeast
associated with the electron transport chain on C6H12O6 → 2C2H5OH + 2CO2 + energy
the membranes of mitochondria and RQ = CO2 = 2 = ∞
chloroplasts (refer to 12.2g) O2 0
• in reality, some respiration in the yeast will be
f) Relative energy values of carbohydrates, aerobic so a small volume of oxygen will be taken up
lipids & proteins and the RQ will be less than infinity
• energy in aerobic respiration comes from the • high values of RQ indicate that alcoholic
oxidation of hydrogen to water when reduced NAD fermentation is occurring
and FAD are passed onto the electron transport chain
(ETC) • no RQ can be calculated for muscle cells using the
• therefore, greater the number of hydrogens, greater lactate pathway as no carbon dioxide is produced
the energy value per unit mass (energy density)
• typical energy values can be determined by burning a h) using respirometers to determine RQ
known mass of a substrate in a calorimeter
using a respirometer to determine the RQ of germinating
seeds or small invertebrates (e.g., blowfly larvae)
TYPICAL ENERGY VALUES
respiratory substrate energy density / kJ g-1 1) set up a respirometer
carbohydrate 15.8 • control tube must contain an equal amount of
lipid 39.4 inert material (e.g., glass beads) to the volume of
protein 17.0 organism being used
2) use the manometer reading to calculate change in gas
volume within a given time (x cm3 min-1)

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x – oxygen consumed (distance moved by fluid in 3) this raises the energy level of glycose / lowers the
experimental tube) activation energy of the reaction
y – distance moved by fluid in control tube 4) fructose bisphosphate is lysed to produce 2 triose
x - y – carbon dioxide given out phosphate molecules (3C)
5) hydrogen atoms and phosphate groups are removed
RQ = x-y from triose phosphate by coenzyme NAD to produce
x 2 molecules of pyruvate and reduced NAD
• since removal of hydrogen is oxidation:
12.2 Respiration TRIOSE PHOSPHATE
OXIDATION
2 PYRUVATE
a) The 4 stages of aerobic respiration • removed phosphate groups are added to ADP to 4
Ø respiration – the process whereby chemical ATP, however since 2 were used, there’s a net gain of
potential energy from the breakdown of organic 2 ATP
molecules is used to synthesise ATP

NET ATP
PROCESS LOCATION
PRODUCED
glycolysis cytoplasm 2
link reaction 0
mitochondrial 2 (one for
Krebs cycle matrix every acetyl
group)
oxidative mitochondrial
32
phosphorylation membrane

Image: bioninja.com.au

• overall, 2 molecules of ATP are used

• 4 are made during the glycolysis of one glucose
molecule
• ∴ net gain of 2 ATP per glucose

c) Link reaction
• when oxygen is available, pyruvate is converted into
acetyl (2C) coenzyme A in the link reaction
• pyruvate enters by active transport from the
cytoplasm into the mitochondrial matrix

1) pyruvate is decarboxylated (CO2 is removed)

2) then it is dehydrogenated (hydrogen is removed and
picked up by NAD)
• this converts pyruvate into a 2C compound
b) Glycolysis 3) then it’s combined with coenzyme A to produce
acetyl coenzyme A (ACoA)
• phosphorylation of glucose
• the subsequent splitting of fructose 1,6- CO2

bisphosphate (6C) into two triose phosphate pyruvate (3C) + CoA acetyl (2C) CoA
molecules NAD+ NADH
• these are further oxidised to pyruvate with a small
yield of ATP and reduced NAD 4) coenzyme A transfers the acetyl group with 2C to
the Krebs cycle
1) glucose is phosphorylated (substrate-level
phosphorylation) using 2 ATP molecules
2) this results in the formation of fructose
bisphosphate (6C) (also known as hexose
bisphosphate)

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d, e) Krebs cycle energy for ATP synthesis (details of ATP synthase
are not required)
• oxaloacetate (a 4C compound) acts as an acceptor
of the 2C fragment from acetyl coenzyme A to form
• energy for phosphorylation of ADP to ATP comes
citrate (a 6C compound), which is reconverted to
from the activity of the electron transport chain
oxaloacetate in a series of small steps
(ETC)
• reactions in the Krebs cycle involve decarboxylation
and dehydrogenation and the reduction of NAD and 1) reduced NAD and FAD are passed onto the ETC in
FAD the inner mitochondrial membrane (cristae)
2) hydrogen released from reduced NAD and FAD splits
1) acetyl coenzyme A combines with a 4C compound into electrons and protons
called oxaloacetate to form citrate (6C) 3) as these electrons pass along electron carriers from
• CoA is reformed, the cycle turns twice for each a higher to lower energy level, energy is released
original glucose molecule 4) energy released pumps H+ into the intermembrane
space creating a proton gradient across the crista
2) citrate is decarboxylated and dehydrogenated in a 5) protons diffuse back through channel proteins which
series of steps have ATP synthase associated with them into the
• this produces CO2 (waste gas) and hydrogens matrix
(which are accepted by NAD and FAD) 6) 1 ATP molecule is produced from ADP and Pi when 3
• 1 FAD and 3 NAD molecules are reduced each H+ pass down via the electrochemical gradient
turn of the cycle (hydrogen in reduced NAD/FAD 7) oxygen is the final electron acceptor at the end of the
will be released in oxidative phosphorylation) ETC in the matrix
• main role of the Krebs cycle in respiration is to 8) protons combine with electrons and oxygen atoms
generate many reduced hydrogen carriers to to form water
pass onto the next stage
h) Effect of factors such as temperature and
3) oxaloacetate is regenerated and can recombine
with ACoA
substrate concentration on the rate of
Image: bioninja.com.au
respiration of yeast using a redox indicator
(e.g., DCPIP or methylene blue)
Mechanism of redox indicators when determining
respiration rates
1) dehydrogenation happens regularly throughout the
different stages of aerobic respiration
2) the hydrogens that are removed from substrate
molecules are used in oxidative phosphorylation and
are transferred by NAD and FAD
3) when DCPIP and methylene blue are present, they
can also take up hydrogens and get reduced (blue →
colourless)
4) faster the rate of respiration, the faster the rate of
hydrogen release and faster the dyes get reduced
and turn colourless
5) therefore the rate of colour change can correspond
to the rate of respiration in yeast
Summary 6) rate of respiration (sec-1) = 1 / time (sec)
• for each turn of the cycle, 2CO2 and 1 ATP produced,
1 FAD and 3 NAD reduced Effect of temperature on the rate of respiration of yeast
• for every glucose molecule, 4CO2 and 2 ATP using a redox indicator
produced, 2 FAD and 6 NAD reduced 1) add a redox dye such as DCPIP or methylene blue to
a suspension of yeast cells
f, g) Oxidative phosphorylation 2) add the test tubes to a temperature-controlled water
bath
• energetic electrons release energy as they pass
3) record the time taken for a colour change to occur
through the electron transport system
once the dye is added (& repeat across a range of
• the released energy is used to transfer protons
temperatures)
across the inner mitochondrial membrane
4) when reduced, the blue dyes become colourless
• protons return to the mitochondrial matrix by (rate of change from blue to colourless is a measure
facilitated diffusion through ATP synthase providing of the rate of respiration of the yeast)

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Effect of substrate concentration on the rate of 6) matrix contains enzymes for the link reaction and
respiration of yeast using a redox indicator Krebs cycle, 70S ribosomes are circular DNA are
1) add different concentrations of a substrate to the also present
suspension of yeast cells (e.g. 0.1%, 0.5% glucose)
2) record the time taken for a colour change to occur Function
once the dye is added (& repeat across a range of 1) ATP synthesis
temperatures) 2) aerobic respiration
3) when reduced, the blue dyes become colourless
(rate of change from blue to colourless is a measure j, k) anaerobic respiration
of the rate of respiration of the yeast)
*BOTH OCCUR IN THE
Variables to be controlled in redox indicator experiment ALCOHOLIC CYTOPLASM LACTIC
to investigate respiration rate in yeast FERMENTATION FERMENTATION
1) volume of dye added PURVATE → ETHANOL PURVATE → LACTATE
2) volume of yeast suspension (by yeast) (by mammalian
muscle)
3) type of substrate
4) concentration of substrate
5) temperature • when free oxygen is not present, hydrogen cannot be
removed by combining with O2 so reduced NAD is
i) structure and function of the mitochondrion not recycled
Image: https://teachmephysiology.com/
• the ETC stops working
• oxidative phosphorylation cannot take place as
there’s nothing to accept e- (& H+)
• the mitochondrion then runs out of NAD or FAD that
can accept hydrogen from the Krebs cycle
• Krebs cycle and link reaction pause
• glycolysis can still continue as long as pyruvate can
be removed and reduced NAD → NAD

Ethanol pathway
1) pyruvate and reduced NAD formed from glycolysis
2) pyruvate is decarboxylated by pyruvate
decarboxylase producing ethanal
3) ethanal is the hydrogen acceptor from reduced NAD
4) ethanol is formed – this reduction is catalysed by
alcohol dehydrogenase
Image: http://kolibri.teacherinabox.org.au/ 5) irreversible (so chemical potential energy of ethanol
is wasted)
6) NAD is regenerated so it can accept more hydrogen
atoms
7) therefore, glycolysis can continue

Lactate pathway
Pyruvate itself accepts the hydrogen and is converted to
lactate by lactate dehydrogenase. Again, NAD is released
and allows glycolysis to continue.
Image: https://en.wikipedia.org/wiki/Cori_cycle

Structure
1) 0.5-1.0 µm
2) double membrane: inner one is folded inwards to
form cristae
3) cristae increase surface area for reactions
(mitochondria from more active cells have more
densely packed cristae)
4) outer membrane is more permeable than the inner
one
5) inner membrane has a lower pH than the matrix due
to H+ released due to the ETC The lactate pathway is reversible (in the liver).

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Why less ATP can be synthesised from the same mass 2) rate of respiration is represented by gradient of the
of glucose in anaerobic respiration than aerobic graph
respiration
1) glucose is not completely broken down and only

distance moved by
manometer fluid
glycolysis occurs
2) pyruvate still contains energy
3) ETC stops as there’s no oxygen to act as the final m = rate
electron acceptor
4) so oxidative phosphorylation stops, Krebs cycle (no
NAD or FAD to accept hydrogen) and link reaction
pause 0
time
Both reactions ‘buy time’ by allowing some (2 molecules)
ATP to be produced although oxygen is not available as a
hydrogen acceptor. They, however, cannot continue as Using a respirometer to investigate the rate of uptake of
the products are toxic. oxygen
1) organisms to be investigated are placed in one tube
Oxygen debt and non-living material (glass beads) of the same
• during exercise, it takes time for the heart and lungs mass in the other tube
to meet demand, so anaerobic respiration occurs 2) soda lime is placed in both tubes to absorb carbon
• after exercise, the person continues deep breathing dioxide
to absorb more oxygen 3) coloured fluid is poured into the manometer
• the extra oxygen is needed for: reservoir and allowed to flow into the capillary tube
1) converting lactate into glycogen (ensure that there are no air bubbles and volume of
2) reoxygenation of haemoglobin liquid is the same in both tubes)
3) high metabolic rate 4) rubber bungs are fitted on both tubes; spring clips
are closed, and the manometer is then attached to
the bent glass tubing (ensure that it's airtight)
l) adaptations of rice for wet environments 5) open spring clips (to allow pressure throughout the
apparatus to equilibrate with atmospheric pressure)
How it helps the plant to
6) as organisms respire, oxygen is taken from air in the
Feature survive when roots are
tube, reducing the volume and pressure, causing the
submerged
manometer fluid to flow towards the organisms
When roots are 7) carbon dioxide is removed by the soda lime which
submerged in water, less ensures that distance moved by the fluid is only
oxygen is available than affected by oxygen uptake
cells are tolerant of high
when the soil contains air 8) distance moved by the manometer fluid can be
concentrations of ethanol
spaces. Cells therefore calculated using πr2h
respire anaerobically, 9) volume of oxygen taken up can be calculated if the
producing ethanol. diameter of the tube is known
stems have tissues called Aerenchyma allows
aerenchyma, containing oxygen from the air to
large air spaces diffuse down to the roots

some types of rice are The leaves remain

able to grow elongated exposed to air, which
stems to keep their leaves facilitates gas exchange
above the water as its for photosynthesis and
level rises respiration.

m) Using simple respirometers to measure the

effect of temperature on the respiration rate of
germinating seeds or small invertebrates
1) respirometer is placed in water baths at different
temperatures

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