Topic 5.1-5.

5 Respiration AM

Topic 5: Energy for biological processes

Respiration
Organisms and energy
 All organisms need energy to make and break bonds during chemical reactions which bring
about growth, reproduction and the maintenance of life

 Autotrophic organisms make their own food, mostly by photosynthesis

 Heterotrophic organisms eat and digest other organisms to obtain food

 The energy in the chemical bonds of food are transferred to the bonds of adenosine
triphosphate (ATP) during respiration
 ATP provides the energy for all other metabolic reactions
 Cellular respiration = the process by which energy from food molecules is transferred to ATP
 Glucose is almost always the respiratory substrate
 Glucose + oxygen water + carbon dioxide + energy
 C6H12O6 + 6O2 6H2O + 6CO2 + 36 ATP
 Energy yield (ΔH) of 1 mole (6 x 1023 molecules) of glucose is -2880kJ

(revision) 4.2.vi Know that phosphorylation of ADP requires energy and that hydrolysis of ATP
(adenosine triphosphate) provides an accessible supply of energy for biological processes.
 ATP is a mononucleotide
 ATP is formed from ADP + Pi by transfer of energy
from other reactions

 ATP is used for energy-requiring reactions – hence

‘universal energy currency’ in cells

 Reactions yielding ATP include photosynthesis and

respiration
 Reactions requiring energy include the synthesis of
proteins or cellulose and the contraction of muscles

5.1.i Know that cellular respiration yields ATP which is used as a source of energy for metabolic
reactions, and the process also generates heat.

 When the phosphoanhydride bonds are hydrolysed, large amounts of energy are released
 i.e 30.5 kJmol-1 of energy is released when ATP is broken down to ADP (it is the reaction of
phosphate with water that releases the energy)
 Some of this energy is lost as heat, but much free energy is available to do useful work
 ATP can move easily between and within cells by facilitated diffusion
 Unlike burning, the energy is
released from sugar in a series of
small enzyme-catalysed steps.
This means the energy can be
trapped in ATP, although some is
wasted as heat

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Topic 5.1-5.5 Respiration AM

5.1.ii Know the different stages in aerobic respiration, including:

● glycolysis in the cytoplasm
● link reaction
● Krebs cycle
● oxidative phosphorylation in mitochondria

 Glycolysis does NOT occur in the

mitochondria
 The enzymes controlling glycolysis are in
the cytoplasm

(revision) 2.1.v. Know the ultrastructure of

eukaryotic cells and the functions of
organelles, including: …..mitochondria, ….
Structure
 Large rod-shaped organelle, up to 1mm x 10mm
 Double membrane, inner one folded into cristae
 Fluid matrix inside of metabolites and enzymes of
the Krebs cycle
 Cristae carry stalked particles associated with ATP
sythesis
 Small circular piece of DNA in matrix
 Divide themselves for cell replication
 Number of mitochondria in a cell is related to the
energy requirements
Investigating respiration
 In the early 1930s this involved looking at whole organisms and the factors that affected their
rate of respiration
 We will be doing something similar in :-

CORE PRACTICAL 9: Investigate factors affecting

the rate of aerobic or anaerobic respiration using a
respirometer, taking into account the safe and
ethical use of organisms.

 Modern techniques have included:

 Centrifuging cell fragments to isolate
mitochondria which will produce ATP in the
presence of oxygen and glucose
 The high resolving power of electron microscopes
have enabled the structure of the cristae on the
inner membrane and stalked particles to be
discovered
 Stalked particles and associated membrane have been isolated and shown to synthesise ATP

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Topic 5.1-5.5 Respiration AM

5.2 Glycolysis
i Understand the conversion of monosaccharides to pyruvate during glycolysis in the cytoplasm,
including:
● the phosphorylation of hexose molecules by ATP
● breakdown to glycerate 3-phosphate (GP)
● production of reduced coenzyme (NADH) and ATP (details of
intermediate compounds and other reactions are not required).

 Glycolysis iterally means ‘splitting of sugar’

 Occurs in cytoplasm
 Glucose is main respiratory substrate
 Glycogen must first be converted to glucose
 There are 4 stages to glycolysis:

1) Phosphorylation
 Glucose can potentially yield 2880kJ mol-1
 But is stable and unreactive
 First needs an input of energy from ATP
 Two phosphate groups are added to glucose from 2 ATP molecules (phosphorylation)
 Forms fructose bisphosphate
 Glucose is now more reactive and cannot leave the cell

2) Lysis
 Fructose bisphosphate is now split into 2 x 3C compounds -
glycerate-3 phosphate (GP)

3) Oxidation

 A dehydrogenase enzyme removes 2 H atoms from the 3-

carbon molecule
 A coenzyme and hydrogen acceptor, nicotinamide adenine
dinucleotide (NAD) accepts the H+ ions and electrons and
becomes reduced NAD
 NAD + + 2H + + 2e- NADH + H +
(or NADH2)

4) ATP formation

 Two ATP molecules are formed by substrate level phosphorylation as the 3-carbon molecule is
converted to pyruvate
 As there are 2 x 3-carbon compounds there are:
 2 molecules of pyruvate
 2 ATPs
Summary of glycolysis - there has been a net gain of :-
 Two ATPs
 Two pairs of H atoms which have formed two reduced NAD (involved in ATP formation later)
 Two molecules of the 3C pyruvate
 No oxygen has been used so far

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Topic 5.1-5.5 Respiration AM

What happens next depends on whether oxygen is present or not

 If plenty of oxygen is present the pyruvate will enter the mitochondria and be used in aerobic
respiration
 If oxygen is in short supply the pyruvate remains in the cytoplasm and is used in anaerobic
respiration

Aerobic respiration
5.3 Link reaction and Krebs cycle
i Know that the link reaction and Krebs cycle take place in the mitochondrial matrix.
ii Understand that during the complete oxidation of pyruvate the events of the link reaction and
the Krebs cycle result in the removal of carbon atoms to produce:
● carbon dioxide
● reduced coenzyme (NADH)
● ATP (detailed knowledge of the intermediate compounds in the Krebs cycle is not required).

 Fate of pyruvate if oxygen is present

 Pyruvate enters the mitochondria
 The link reaction occurs, pyruvate is:
 decarboxylated (CO2 released)
 Dehydrogenated (2H are taken up by coenzyme NAD)
 Resulting 2C molecule combines with coenzyme A to form acetyl coenzyme A (acetyl CoA) which
enters Krebs cycle

 The reduced NAD (with 2 H atoms) will be involved in ATP formation later

Understanding the chemistry of respiration

 There are four types of reaction, all catalysed by enzymes:-

 Phosphorylation
o E.g. ADP + Pi ATP

 Decarboxylation
o Break off CO2

 Redox reactions (OIL RIG (of electrons or H))

o Oxidised NAD + electrons/H reduced NAD

 Pyruvate acetyl CoA + 2H (dehydrogenation)

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Topic 5.1-5.5 Respiration AM

Krebs cycle
 Takes place in mitochondrial matrix where the necessary enzymes are located
 Named after Sir Hans Krebs (also known as the citric acid cycle or TCA cycle)

 The two-carbon acetyl co-enzyme A reacts with a four-carbon organic acid to form a 6-carbon
molecule
 The co-enzyme A is reused in the link reaction

 There are two decarboxylations with the formation of CO2

 There are four dehydrogenations with the removal of hydrogen atoms to H acceptors - 3 NAD
and 1 FAD (flavine adenine dinucleotide)

 There is one substrate-level phosphorylation with the direct formation of ATP

Summary of Krebs cycle

 For each 2-carbon molecule entering:-
 Two CO2 molecules are made
 One ATP is made (substrate-level phosphorylation)
 Three pairs of H atoms are accepted by NAD
 and one by FAD

 However glucose was converted to two pyruvate in glycolysis so Krebs cycle turns twice for
every molecule of glucose

 All the H atoms carried by NAD and FAD from glycolysis, the link reaction and Krebs cycle are
now involved in the production to ATP in the electron transport chain

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Topic 5.1-5.5 Respiration AM

5.4 Oxidative phosphorylation

i Know that the electron transport chain takes place in the inner mitochondrial membrane.

ii Understand the role of the electron transport chain in generating ATP

(oxidative phosphorylation).
iii Understand the role of oxygen as a terminal electron acceptor forming water.
iv Understand how ATP is synthesised by chemiosmosis.
v Understand the importance of mitochondrial membranes in this process.

 Fate of hydrogen – the electron transport chain

 The reduced coenzymes NAD and FAD shuttle the
hydrogen to the electron transport chain
 This is on the inner mitochondrial membrane

 Each H atom’s electron (e-) and proton (H+)

separate
 Electrons pass along a chain of electron carriers in the inner
mitochondrial membrane = electron transport chain
 The H ions are used in the process of chemiosmosis to
produce ATP

 Each electron carrier is reduced when it accepts an electron, and oxidised when it passes it on –
a series of redox reactions

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Topic 5.1-5.5 Respiration AM

 Chemiosmosis won a Nobel prize for Peter Mitchell in 1978

 Energy is released as electrons pass along the electron transport chain
 This energy is used to pump hydrogen ions from the matrix into the intermembranal space
 The inner mitochondrial membrane is impermeable to H ions

 This forms a steep electrochemical gradient across the inner membrane

 The high H+ concentration means the intermembranal space is more positive than the matrix

 The hydrogen ions diffuse back through hollow protein channels in stalked particles
 ATPase located in each stalked particle catalyses
ATP synthesis

 Electron carriers are:

 NAD and FAD

 Cytochromes are protein pigments with an iron

group rather like haemoglobin

 Cytochrome oxidase is an enzyme that is

reduced when it receives the electrons and
oxidised when it passes it on

 Oxygen is the final electron acceptor

 Cyanide acts on cytochrome oxidase preventing the

production of ATP, causing death swiftly!

ATPase
 Hydrogen ions cause a conformational change in
the enzymes’ active site so ADP and Pi can bind

 In the matrix the H+ and electrons combine to form

hydrogen atoms
 These combine with oxygen to form water

 Oxygen can therefore be said to be the final carrier in the

electron transport chain so is known as the terminal
electron acceptor

O2 + 4H+ + 4e- 2H2O

 This method of synthesising ATP is known as oxidative phosphorylation

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Topic 5.1-5.5 Respiration AM

Summary of respiration
 Respiration has many steps – in A level we study a simplified version

 In respiration the energy follows this sequence:

 glucose NADH electron transport chain proton-motive force ATP

How much ATP is produced?

 Total number of ATP produced by one glucose varies according to efficiency of cell
 Scientists are continually revising their estimates in the light of new knowledge
 38 ATP are produced if we
assume:
 Reduced NAD results in 3
ATP
 Reduced FAD results in 2 ATP

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Topic 5.1-5.5 Respiration AM

However this is too simplified:

 It takes 2 ATP to transport reduced NAD from glycolysis across the mitochondrial membrane –
hence 36 ATP is often quoted
 Between 3-4 H ions need to flow through ATPase to generate 1 ATP
 However ATP yields are now thought not to be in whole numbers:
 2 reduced NAD appear to yield 5 ATP, and 2 FAD yield 3 ATP
 The electrochemical gradient across the inner membrane is also involved in the transport of
other ions and molecules
 The exchange of ADP and ATP between matrix and cytoplasm uses up some H ions
 Pyruvate needs to be moved into the mitochondrion which takes energy
 Therefore not all H ions are available for ATP generation
 Actual yield is probably about 30 ATP

Energy released
 Complete oxidation by combustion of 1 mole glucose (180g) releases 2880kJ
 However only 1163kJ is released from the ATP made from 1 mole of glucose (assuming 38 ATPs
are produced from each glucose molecule)
 = 40% of potential chemical energy

 If only 30 ATPs are made then the energy released would be 918kJ – 32%

 The remaining energy raises the cell’s temperature

Anaerobic respiration
5.5 Anaerobic respiration
i Know that anaerobic respiration is the partial breakdown of hexoses to produce a limited yield
of ATP in the absence of oxygen.
ii Understand the difference in ATP yields from one molecule of hexose sugar in aerobic
conditions compared with anaerobic conditions.
iii Understand how lactate as a by-product of anaerobic respiration affects mammalian muscle
contraction.
iv Understand how anaerobic respiration in plants results in ethanol formation.

 In the absence of oxygen pyruvate does not enter the link reaction
 Anaerobic respiration occurs in the cytoplasm

 Living organisms have developed two types of anaerobic respiration

 Plants and yeast produce ethanol and carbon dioxide
 Animals and some bacteria produce lactic acid

 Glycolysis still occurs yielding 2 ATP

 Glucose (hexose) is only partially broken down to 2 pyruvate molecules

 Without oxygen the electron carriers cannot function and eventually all the NAD and FAD is
reduced
 Metabolism would stop = death
 This cannot be allowed to happen

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Topic 5.1-5.5 Respiration AM

Anaerobic respiration (also known as fermentation)

 H ions from reduced NAD are taken up by pyruvate to produce either lactate or ethanol
 Glycolysis can therefore continue

Anaerobic respiration in plants and yeast

 Pyruvate is decarboxylated
 The 2C ethanol (acetaldehyde) is the H acceptor
 NAD is oxidised
 Ethanol and carbon dioxide are the products and are excreted
from the cell

Anaerobic respiration in animals and some bacteria

 Lactate is the H acceptor
 NAD is oxidised
 Bacteria excrete lactate

Anaerobic respiration in mammalian muscles

 As levels of lactate increase in muscles the pH
falls and the tissue becomes acidic
 Muscles feel ‘stiff’
 It used to be thought that this acidity directly
affected muscle contraction

 Modern research suggested that the

movement of lactic acid and H ions into the
blood lowers its pH
 This affects the CNS, reducing nervous
stimulation to the muscles and eventually
causing them to stop contracting

 This is a protective mechanism allowing the

muscle cells to recover, resume aerobic
respiration and raise the blood pH again

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Topic 5.1-5.5 Respiration AM

Repaying the oxygen debt

 At the end of exercise the lactate levels are
raised
 As lactate is toxic some is converted in the
muscle cells to pyruvate so it can enter the
Krebs cycle
 Most is transported to the liver where it is
oxidised back to pyruvate and then to
glucose (gluconeogenesis)
 Then blood takes it to replenish glycogen
supplies in muscles
 It takes oxygen to oxidise the lactate,
hence you need to breathe deeply even after exercise has stopped

 It appears that more oxygen is needed than the oxidation of lactate requires
 This is know as the EPOC – excess post oxygen consumption and appears to be due to:

 Myoglobin supplies in muscles need reoxidising

 Raised temperature and adrenaline increase oxygen usage
 Breathing deeply means intercostal and diaphragm muscles use more oxygen
 ATP and phosphocreatine supplies need replenishing in muscle cells which takes oxygen

Anaerobic respiration and athletes

 100m athletes run up to 95% of their race relying on anaerobic respiration
 Long-distance runners must maintain aerobic respiration for their muscles to continue to work

 Training involves the ability to:

 Get more oxygen to the muscles
 Tolerate higher levels of lactate
 Repeated exposure to high lactate levels increases the lactate transporter proteins in the
mitochondrial membranes so lactate can be processed more quickly once oxygen is available

Efficiency of anaerobic respiration

 Yield of ATP from both forms of anaerobic respiration is 2 ATP per glucose molecule (in
glycolysis)
 Yield = 2 x 100 = 5 – 6 % efficiency of aerobic respiration
30 – 38

 All early forms of life respired this way

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