Biology A2 Unit 4

Respiration
Mr. Woodward

What is Respiration?
• Respiration is the process by which organisms extract the energy stored in
complex molecules and use it to generate adenosine triphosphate (ATP).
• In this way they obtain energy to fuel their metabolic pathways.
• ATP provides the immediate source of energy for biological processes such as
active transport, movement, and metabolism.

Types of Respiration
• Aerobic Respiration
◦ During aerobic respiration, a respiratory substrate, for example, glucose, is
split in the presence of oxygen to release carbon dioxide and water. A large
number of ATP molecules are produced, releasing the energy from the glucose.
▪ C6H12O6 + 6O2 → 6CO2 + 6H2O + 36ATP
• Anaerobic Respiration

Where does respiration occur?

• Respiration occurs in all living cells. In eukaryotes, the early stages of respiration
occur in the cytoplasm, where the later stages are restricted to the
mitochondria.
• Mitochondria contain highly folded inner membranes that hold key respiratory
proteins (including the enzyme that makes ATP) over a large surface area.
• Mitochondria provide an isolated environment to maintain optimum conditions
for respiration.
• Mitochondria have their own DNA and ribosomes, so can manufacture their own
respiratory enzymes.

The Stages of Respiration

• Glycolysis is the first stage of both aerobic and anaerobic respiration. It occurs in
the cytoplasm, converting glucose to pyruvate and producing a small amount of
ATP. No oxygen is needed for glycolysis.
• Anaerobic respiration occurs without oxygen. The products of glycolysis are
broken down, allowing glycolysis to continue.
◦ It can produce either lactate, or ethanol and CO 2.
• Aerobic respiration uses oxygen. Pyruvate is broken down to regenerate ATP.
There are three stages to aerobic respiration after glycolysis: the link reaction,
Krebs cycle and the Electron Transport Chain (ETP).

Adenosine Triphosphate
• ATP contains a sugar (Ribose), a base (adenine) and three phosphate groups.
• When ATP is hydrolysed to form ADP, 37.5 kJ of energy are produced.
• Why ATP?
◦ ATP releases its energy instantly in a single reaction, and requires a small
amount of energy to hydrolyse.

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Phosphorylation of ADP
• The addition of an inorganic phosphate group, (Pi) to a molecule like ADP is
called phosphorylation. ADP is phosphorylated during respiration.
• Two types of phosphorylation occur during respiration:
◦ Substrate level: Glycolysis and Krebs cycle: A single reaction involving the
direct transfer of a phosphate group from a donor molecule to ADP.
◦ Oxidative: Electron transport chain: A series of oxidation reactions that
produce sufficient energy to form ATP from ADP and phosphate.

Coenzymes
• Coenzymes are molecules that bind with a specific enzyme or substrate, helping
to catalyze a reaction.
• Breaking the bonds between coenzyme and product after a reaction is crucial,
otherwise coenzyme concentration will drop, limiting respiratory rate.
• Three major enzymes are used in respiration:
◦ NAD
◦ CoA
◦ FAD
• NAD can accept a hydrogen molecule (Two hydrogen atoms), forming reduced
NAD (NADH)
◦ This is used to regenerate ADP in the Electron Transport Chain (ETC)
◦ Coenzyme A aids the transition between glycolysis and the Krebs cycle, by
converting pyruvate to acetyl coenzyme A.
◦ FAD, like NAD, can accept hydrogen to form reduced FAD (FADH2)

The stages of Respiration

• Glycolysis
◦ Glucose is initially phosphorylated by one ATP molecule. This creates a
molecule of glucose-6-phosphate.
◦ Glucose-6-phosphate is converted to fructose-1-phosphate. A second ATP
molecule phosphorylates fructose-1-phosphate, forming hexose-1,6-
bisphosphate.
◦ Hexose-1,6-biphosphate is split into two triose phosphate molecules. This is a
three-carbon sugar, with one phosphate group attached.
◦ Dehydrogenase enzymes remove two hydrogen atoms from each sugar. NAD
is the H acceptor, forming one NADH for each triose phosphate. Each sugar
also produces one ATP by substrate-level phosphorylation.
◦ The three-carbon intermediate is now converted to pyruvate by enzyme
action. This also regenerates a molecule of ATP.
▪ Glycolysis uses one molecule of glucose to produce two molevules of
pyruvate, 2 ATPs and 2 NADH. The entire process is dependent on the
supply of NAD. This is a limiting factor in the rate of glycolysis.

The Fate of Pyruvate

• Without oxygen, pyruvate cannot enter the aerobic pathway. Glycolysis becomes
the main source of ATP, but this needs a constant source of of NAD+.
• The NADH produced in glycolysis is reoxidised to NAD+. Pyruvate accepts hydrogen
ions from NADH and the cytoplasm to form lactate.

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 ◦ The NAD can now be used to produce more pyruvate.

Anaerobic respiration outside the animal kingdom

• Outside the animal kingdom, pyruvate is converted to ethanol in order to
replenish the level of NAD needed for glycolysis.
◦ Firstly enzymes cause pyruvate to release a molecule of carbon dioxide to form
ethanal.
• NADH then donates an H+ ion to the ethanal molecule. This, along with a H + from
the cytoplasm, forms ethanol and NAD+.
◦ The NAD+ is re-used in glycolysis, making more pyruvate and ATP.

The Link Reaction

• The link reaction allows pyruvate to enter the aerobic pathway by converting it to
acetate.
◦ The NAD+ needed for glycolysis is regenerated later in the process.
• The link reaction occurs in the mitochondrial matrix. This means that pyruvate
produced in the cytoplasm must be actively transported into the mitochondria
before the reaction can begin.
• In the mitochondrial matrix, pyruvate reacts with coenzyme A to form acetyl CoA.
CO2 is released in the process and another molecule of NAD + is reduced.
◦ The sugar molecule now contains only two carbon atoms.
• Each molecule of glucose entering glycolysis produces two molecules of pyruvate.
This means that the link reaction yields two NADH and two molecules of CO2 per
molecule of glucose.

The stages of the Krebs Cycle

• Acetyl CoA is produced in the link reaction. Acetate is removed from CoA,
combining with oxaloacetate to form a six-carbon sugar, citrate. CoA is reused in
the link reaction.
• Citrate is decarboxylated, releasing a CO 2. Is is also dehydrogenated, releasing two
hydrogen atoms that are used to reduce NAD +. This leaves a five-carbon
compound.
• This five-carbon compound is again decarboxylated and dehydrogenated,
releasing CO2 and reducing another NAD+. A four-carbon compound is produced.
• The first four-carbon compound is converted into another four-carbon compound.
This regenerates a moleculeof ATP by substrate-level phosphorylation.
• The conversion of the next four-carbon compound involves dehydrogenation. Two
hydrogen atoms are released, and these reduce FAD +, producing FADH2 and a
different four-carbon compound.
• The final four-carbon intermediate is converted to oxaloacetate by
dehydrogenation. This again produces a molecule of NADH.
• Acetyl CoA is produced in the link reaction. Acetate is removed from CoA,
combining with oxaloacetate to form a six-carbon sugar, citrate. CoA is reused in
the link reaction.

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Keeping track of the products
• For each molecule of glucose, glycolysis produces:
◦ 2 x ATP
◦ 2 x Pyruvate
◦ 2 x NADH
• For each molecule of glucose, the link reaction produces:
◦ 2 x Acetyl CoA
◦ 2 x CO2
◦ 2 x NADH
• For each molecule of glucose, the Krebs cycle generates:
◦ 4 x CO2 produced by decarboxylation.
◦ 6 x NADH produced by redox reactions.
◦ 2 x FADH2 produced by redox reactions.
◦ 2 x ATP produced by substrate-level phosphorylation.
• The NADH and FADH2 contain the potential energy originally locked in glucose. This
energy is now transferred to ATP by oxidative phosphorylation in the electron
transport chain.

The Electron Transport Chain

• The electron transport chain performs oxidative phosphorylation, a process
that regenerates large amounts of ATP.
• The ETC is formed from a series of proteins, bound in the inner mitochondrial
membrane.
• The cristae maximise the surface area of this membrane, and this increases the
rate of ATP production.
• NADH, produced in the matrix during Krebs cycle, is oxidized by the first protein in
the ETC, NADH dehydrogenase.
◦ This produces a proton (H+) and NAD+, along with two electrons (2e -) that bind
to the protein.
• The electrons are now passed between the ETC proteins in a series of redox
reactions. As they move, they lose energy. Some of this is used to pump H + ions
from the matrix into the intermembrane space; the rest is lost as heat.
• As the inner mitochondrial membrane is impermeable to H + ions, a concentration
gradient forms.
• H+ ions move down their concentration gradient into the matrix using protein
channels. These are associated with the enzyme ATP synthase, which
phosphorylates one ADP for each H+ ion passing through it.
• The use of energy in a chemical gradient to generate ATP by the flow of hydrogen
ions through ATP synthase is called chemiosmosis.
• The final protein in the ETC (cytochrome oxidase) donates the electron pair to an
oxygen atom. This is the final proton and electron acceptor, as it binds with the H +
ions in the matrix to form water.
◦ ½O2 + 2H+ + 2e- → H2O
• The donation of the electrons to the oxygen molecule releases enough energy to
pump another H+ ion across the membrane. This can be used to regenerate
another molecule of ATP.
• FADH2 is also oxidised by the ETC; however, it interacts with the second protein in
the chain. This means that it causes less H + to be pumped into the intermembrane
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 space than NADH, so regenerates less ATP.

How much ATP is produced?

• Via the electron transport chain and chemiosmosis, each NADH can yield 2.5 ATP
and each FADH2 1.5 ATP.
• From one molecule of glucose, glycolysis yields 2 NADH, the link reaction yields 2
NADH and the Krebs cycle yields 6 NADH and 2 FADH2.
◦ 10 x 2.5 = 25 ATP from NADH
◦ 2 x 1.5 = 3 ATP from FADH2
◦ Total = 2 + 2 + 25 + 3
◦ 32 ATP overall

Efficiency of aerobic respiration

• The theoretical yield of 32 ATPs for each glucose molecule is rarely achieved. In
fact respiration is only about 32% efficient.
◦ Some protons leak across the mitochondrial membrane, so not all are available
to generate ATP via chemiosmosis.
◦ Some ATP is used up moving pyruvate into the mitochondria by active
transport.
◦ Some ATP is used up moving pyruvate into the mitochondria by active
transport.
◦ Some ATP is used up moving hydrogen from reduced NAD made during
glycolysis into the mitochondria.
◦ Some energy is lost as heat. This heat helps to maintain a suitable body
temperature for enzyme-controlled reactions.

Evidence for chemiosmosis

• The theory of chemiosmosis states that the energy in a chemical gradient
established by electron movement is used to generate ATP.
• Evidence includes:
◦ The protein gradient across the inner membrane can be measured as it
corresponds to a pH gradient.
◦ Isolated ATP synthase enzymes can produce ATP using a proton gradient even
if no electron transport is occurring.
◦ Chemicals that block the ETC inhibit the formation of a proton gradient and
prevent ATP synthesis.

Respiratory Rate
• The respiratory rate is the rate at which an organism converts glucose to CO 2 and
water. It can be calculated by measuring an organism's rate of oxygen
consumption.
• Studies on simple animals often use a respirometer.
• Respirometers measure the change in gas volume in a closed system Any change
is due to the respiratory activity of the study organisms. Potassium hydroxide or
soda lime is used to absorb the carbon dioxide produced, meaning any changes in
volume are due to oxygen consumption.

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Respiratory substrates
• Other substances as well as glucose can be respired. Different respiratory
substrates release different amounts of energy.

Respiratory substrate Mean energy value (kJg-1)

Carbohydrate 15.8
Lipid 39.4
Protein 17

Respiratory Quotient
• Respiratory Quotient is the ratio of the volume of carbon dioxide produced to
the volume of oxygen used in the same period of time.
• RQ = volume of CO2 given out / volume of O2 taken in
• RQ gives an indication of the respiratory substrate being respired and whether
respiration is aerobic or anaerobic.
• If the amount of oxygen in is the same as the amount of carbon dioxide given out,
the RQ will be 1.

Type of respiration Substrate RQ

Anaerobic Glucose >1
Aerobic Carbohydrate 1
Protein approx. 0.9
Lipid approx. 0.7

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