Respiration

Lesson 1:
Introduction:
Why do we need energy?
- All living organisms need energy to drive their biological process/metabolism
- A sequence of metabolic reactions form a metabolic pathway
- The individual molecules in the pathway are called metabolites or intermediates
- Anabolic pathways build larger molecules to smaller molecules (they use energy -
endergonic)
- Catabolic pathways break down larger molecules to smaller molecules (they release
energy - endergonic)
- The net result of anabolism and catabolism determines metabolism
- Aerobic happens in the mitochondria, anaerobic happens in the cytoplasm
- Regulated by enzymes
What biological process can you think of that requires energy?
- Active transport
- Endocytosis
- Exocytosis
- Synthesis of large molecules such as proteins e.g.
collagen, enzymes and antibodies.
- DNA replication
- Cell division
- Movement - such as movement of bacterial flagella od
eukaryotic cilia and undulipodia and motor proteins that
walk along cytoskeleton threads in cells, moving
organelles
- Activation of chemicals - glucose is phosphorylated at
the beginning of respiration so that it becomes more
reactive and able to be broken down to release more
energy.
What is energy and where does it come from?
- Energy is the ability to do work
- Energy is never created or destroyed but converted
from one form to another
- It is measured in kilojoules or joules
- There are many forms of energy: sound, light, heat, chemical, electrical and atomic

The structure and functions of ATP:

ATP is the universal energy currency of all living organisms
 Glycolysis Happens in cytoplasm of all cells and 6C sugar is broken down into two
molecules of pyruvate (3C compound) does not need oxygen

Link reaction Happens in the matrix of mitochondria. Pyruvate is dehydrogenated

and decarboxylated and converted to acetate

Krebs cycle Happens in the matrix of mitochondria. Acetate is decarboxylated and

dehydrogenated

Oxidative Happens on the folded inner membrane (cristae) of mitochondria this is

phosphorylation where ADP is phosphorylated of ATP

Lesson 2:
Glycolysis:
Starting compound - one molecule of glucose
1. Phosphorylation
2. Splitting of hexose 1,6-bisphosphate
3. Oxidation of triose phosphate to an intermediate
4. Conversion of the intermediate to pyruvate
End compound = two molecules of pyruvate
1. Substrate Level Phosphorylation:
Why phosphorylate?
Changes the glucose molecule so it is unable to leave the
cell through glucose transporters in the membrane
Maintains the glucose concentration for diffusion of glucose
out of the blood and into the cell.
Takes the glucose to a higher energy level. This makes it
unstable and therefore more reactive, so it is easier to split
2. Splitting of hexose 1-6bisphosphate:
3. Oxidation of triose phosphate:
At this stage 2 molecules of ATP are produced by substrate level phosphorylation
A dehydrogenase enzyme removes two hydrogen atoms from each triose phosphate
molecule
It gives these hydrogen atoms to NAD (a coenzyme)
The triose phosphate has been oxidised as it has lost hydrogen, when the NAD has been
reduced it has gained it
4. Conversion of the intermediate to pyruvate:
A series of enzyme controlled reactions produce
pyruvate
Again, 2 molecules of ATP are formed by
substrate level phosphorylation

Nicotinamide adenine dinucleotide:

Organic, non-protein coenzyme. Made of 2 linked
nucleotides. Made in the body from nicotinamide (vitamin B3),
2 ribose, adenine and 2 phosphate groups.
It is the nicotinamide ring that can accept hydrogen atoms.
When NAD transports the hydrogen to the inner mitochondrial
membrane, where it will be used to generate more ATP in the
electron transport chain.
Once it has dropped off the hydrogen in the mitochondrion
the NAD is in its oxidised form again and free to accept more
hydrogens.
Lesson 3:
Citric acid cycle:

Link reaction
Pyruvate moves through the cytoplasms to the mitochondria for the link reaction. Pyruvate is
the end product of glycolysis. Pyruvate dehydrogenase dehydrogenated (loses hydrogen)
pyruvate. Pyruvate decarboxylase decarboxylate (loses a carboxyl group) pyruvate.
These processed turn pyruvate into acetate

Coenzyme A (CoA)
 - Made from pantothenic acid (B-group vitamin), adenosine (ribose +adenine), 3
phosphate groups and cysteine.
- CoA carries the acetate (ethanoate) groups made in the link reaction into the Krebs
Cycle
Link and Krebs Summary
- Both take place in mitochondrial matrix
- Series of enzyme controlled reactions
- Remember one glucose molecule produces 2 pyruvate molecules during glycolysis

Product per molecule of Link reaction Krebs cycle

glucose

Reduced NAD 2 6

Reduced FAD 0 2

Carbon Dioxide 2 4

ATP 0 2

Lesson 4:
The ETC, Oxidative Phosphorylation and Chemiosmosis:
ETC:
where - occurs in the inner mitochondrial membrane where the electron carriers are
embedded in protein complexes. the folded cristae of this membrane increase the surface
area for housing the protein complexes
why - results in net synthesis of ATP and water
what - it is a series of coupled redox reactions catalysed by oxidoreductase enzymes, and
requires oxygen as the final electron acceptor for the process to work.
1. reduced NADP and reduced FAD donate hydrogen atoms, which are split into
protons and electrons, to the electron carriers in the inner mitochondrial membrane.
2. the electrons are passed along the chain of electron carriers, otherwise known as the
electron transport chain
3. as this happens, protons are pumped across the membrane to the inter membrane
space
4. This builds up a proton gradient. this is also known as an electrochemical gradient
5. The hydrogen ions cannot diffuse through the lipid part of the inner membrane. They
diffuse back into the matrix of the mitochondria, down a concentration gradient,
through channels containing the ATP synthase enzyme. Another name for these
channels containing the ATP synthase enzyme. Another name for these channels is
stalked particles. the ATP synthase produces ATP as this happens. the process is
chemiosmosis

Chemiosmosis:
The pumping of H+ ions into the inter membrane space generates an electrochemical
gradient and a proton (pH) gradient. This generates a proton motive force that should lead to
the diffusion of H+ ions back into the matrix.
however, the membrane is relatively impermeable to H+ ions, and the only way they can
return to the matrix is through the ATPsynthase enzyme complex
as the H+ moves through this complex it catalyses the formation of ATP
It is this stage that is known as oxidative phosphorylation. you are phosphorylation the ADP
to form ATP by using oxygen as the final electron acceptor if the electron transport chain/
How much ATP is synthesised from 1 molecule of glucose?
number of reduced coenzymes formed during respiration

Glycolysis Link reaction Krebs cycle

Reduced NAD 2 2 6

Reduced FAD 0 0 2
The protons and electrons from 10 molecules of reduced NAD and can theoretically produce
25 molecules of ATP and those 2 molecules of reduced FAD can theoretically produce 3
molecules of ATP
stage of respiration net theoretical gain of ATP per molecule of
glucose

glycolysis 2

link reaction 0

the krebs cycle 2

oxidative phosphorylation 28

total 32
BUT
- some of the ATP produced is used to actively transport pyruvate from the cytoplasm
of the cell into the matrix of the mitochondrion
- some ATP is used to bring reduced NAD from glycolysis in the cytoplasm into the
mitochondrion
- Some H+ ions leak back across the mitochondrial membrane reducing the proton
motive force that generates ATP
The theoretical yield of ATP is rarely achieved
The actual yield is about 30 ATP molecules per molecule of glucose
Notes on page 144-145:

Lesson 5:
Anaerobic respiration:
Without oxygen to act as the final electron acceptor in oxidative phosphorylation:
1. water is not formed by the protons diffusing into the mitochondrial matrix combining
with electrons and oxygen
2. the concentration of protons in the matrix therefore increases, reducing the
concentration gradient of protons between the inner membrane space and the matrix
3. oxidative phosphorylation stops
4. reduced NAD and FAD cannot unload their hydrogen atoms so there is less NAD and
FAD
5. the Krebs Cycle and link reaction cease
In order to allow glycolysis to continue, NAD must be regenerated from reduced NAD.
mammals and heat achieve this in different ways
Reoxidised ATP so glycolysis can continue to produce ATP
In mammals:
The lactate formed is carried away in the blood to the liver.
when more oxygen is available the lactate is converted back until pyruvate to enter the krebs
cycle, or it is recycled back into glucose and glycogen
muscle fatigue is not caused by a buildup of lactic acid but it is specifically the reduction in
pH that reduced enzyme activity in the muscle cells.
In yeast and plants:
Yeast is a facultative anaerobe - it can live without oxygen.
If ethanol builds up beyond 15% then it will kill the yeast. Yeast grows faster under aerobic
conditions, so in the brewing industry the yeast is first grown with oxygen, then grown under
anaerobic conditions for alcoholic fermentation to take place.

Yield of ATP:
The yield of ATP through anaerobic respiration is approximately 1/15 that for aerobic
respiration as glycolysis is the only method by which it is generated.

Lactate fermentation Ethanol fermentation

Hydrogen acceptor pyruvate ethanol

Is carbon dioxide produced no yes

Is ATP produced? no no

End products lactate and NAD ethanol, carbon dioxide and

NAD

Enzymes involved lactate dehydrogenase pyruvate decarboxylase and

ethanol dehydrogenase

In NAD reoxidised yes yes

 Site of pathway cytoplasm of cells cytoplasms of cells

Lesson 6:
Respiratory substrates
A respiratory substance is an organic substance that can be used for respiration
Different respiratory substrates:
- It takes 30.6kJ of energy to produce 1 mole of ATP
- The more hydrogen atoms in a molecule of respiratory substrates, the more ATP it
can can generate as there are more protons produced for chemiosmosis
- The more hydrogen atoms per mole of respiratory substrate, the more oxygen
needed to respire that substrate
- Lipids have far more hydrogen molecules than glucose or proteins, and hence
release about twice as much energy from their respiration than carbohydrates or
proteins
- Respiration is not an efficient process, and much of the energy is lost as heat rather
than generating ATP

Substrate Mean energy value (kJg-1)

Carbohydrates 15.8

Proteins 17.0

Lipids 39.4

Carbohydrates:
Some cells such as those in your brain can only respire glucose
Monosaccharides such as fructose and galactose can be changed to glucose for respiration
Glycogen stores can also be broken down to produce glucose
1 mole of glucose will produce about 32 moles of ATP. This is about one third of the
theoretical maximum. The rest of the energy is transformed into heat energy which is used
for maintaining a constant body temperature
Proteins:
Deaminated amino acids may be converted into glycogen and fat for storage
Some amino acids may be respired as they can enter the kreb’s cycle
Some amino acids can be converted into pyruvate or acetate and so can be passes to the
krebs cycle
Amino acids reduce slightly more molecules of NAD per mole than glucose to produce
slightly more ATP than the same mass of carbohydrates
When an organism is starving, proteins may be hydrolysed to amino acids for respiration
Lipids:
Lipids such as triglycerides can be hydrolysed to produce fatty acids and glycerol
Glycerol can be converted to glucose for respiration
Fatty acids contain many carbon atoms and so are a good source of protons for oxidative
phosphorylation
Fatty acids are combined with CoA using energy from the hydrolysis of ATP
The fatty acid-CoA complex is transported into the mitochondrial matrix where it is broken
down into 2 carbon acetyl groups. This is known as the beta oxidation pathway and reduced
NAD and FAD are formed
The acetyl CoA groups enter the Krebs cycle
The large amounts of reduced NAD and FAD produced are reoxidised via the electron
transport chain.

Lesson 7:
Calculating respiratory quotients:
Working out RQ values - respiratory quotient is a measure of the ratio of carbon dioxide
produced by an organism to the oxygen consumed over a given period. Quantities of gas
may be measured in moles or by volume.
RQ = carbon dioxide produced / oxygen used
RQ can be used to determine the respiratory substrate used. An RQ of 1 suggests
carbohydrates were respired, 0.9 suggests protein and 0.7 suggests fats. If the value is in
between two of the values it is a mixture of two.
What does it mean?
- The experiment using potassium hydroxide gives us the rate at which oxygen is
being used.
- The experiment using glass beads gives us the rate at which oxygen is being used -
the rate at which carbon dioxide is produced.
- The difference between the two is the volume of the carbon dioxide produced.

Homework - p152 Q1-7 - Due 16/11/22

Q1. explain why a diet high in fat is also high in energy
Fats have lots of hydrogen atoms, to be used in oxidative phosphorylation so more ATP can
be produced

Q2. explain why palmitic acid, a large molecule, can pass into the matrix of the mitochondria
It is non-polar so it can be diffused through the phospholipid bilayer.
Q3. explain why children whose diet does not contain sufficient fat or carbohydrate nay
sudder muscle wastage
Carbohydrates and fats are respiratory substrates, so with insufficient levels, there muscle
proteins will be broken down so the amino acids can be used in respiration to make ATP.

Q4. camel humps contain stored fat. One kilogram of fat, when respired produces 1 kg
metabolic water. Thus these humps provide energy and water for the camels. Explain why,
when fat is respired, it produces more metabolic water than respiration of an equivalent
quantity of stored carbohydrate
The fat from camel humps will contain many hydrogen atoms, which will react with oxygen to
give water. So because the fats have more hydrogen atoms than carbohydrates, more
metabolic water can be produced

Q5. is the lipid in camel humps respired aerobically or anaerobically? Explain your answer
They can be respire both aerobically and anaerobically, as for the krebs cycle to take place
oxygen must be present, and glycolysis is anaerobic

Q6. in an investigation into respiration by woodlive, the RQ value was found to be 0.8.
Suggest which type(s) of respiratory substrate were being used
A mixture between proteins and lipids and carbohydrates

Q7. in an investigation into respiration by germinating seeds, the RQ value was found to be
2.3. What may be deduced from this finding?
As the RQ value is higher than 1.0 there must have been some anaerobic respiration as well
as aerobic respiration