RESPIRATION

UNIT 2
RESPIRATION is the process by which energy in the form of ATP is made
available to cells by the metabolic breakdown of organic compounds, primarily
glucose.

PHOTOSYNTHESIS is the process by which green plants are able to use

chlorophyll to harness the energy from light and use it to build up organic
molecules, such as glucose which are high in energy.

ATP – is considered the energy currency of cells which means it’s the form of
energy which is readily available to all cells to carry out all activities.

PHOSPHORYLATION – adding a phosphate group to an existing molecule,

primarily the addition of Pi to ADP to make ATP.
• Oxidative phosphorylation – phosphorylation by chemiosmosis, with the uptake of oxygen
• Substrate Level Phosphorylation – does not involve chemiosmosis but is linked to a high energy substrate
• Photophosphorylation – also involves chemiosmosis and uses light energy
CAPTURING AND RELEASING ENERGY
Energy is temporarily held as ATP by:
ADP + Pi --------- ATP
This is phosphorylation and Pi refers
to inorganic phosphate

ATP acts as an energy transfer

molecule and in order to release the
energy the process is reversed:
ATP --------- ADP + Pi

ATP cannot be transferred from one

cell to another, this is why Respiration
must take place in ALL cells and why
very active cells have many
mitochondria
 CO-ENZYMES

Co-Enzymes are derived from vitamins, and are available in very small amounts, so they have to be recycled,
that is, changed from the oxidized to the reduced form repeatedly.
They become reduced when they accept electrons- this takes place at different stages of respiration - then they
take these electrons to the Electron Transport Chain (ETC) on the inner membrane of mitochondria, where
they deliver the electrons to electron carriers, becoming oxidized themselves, so they can be recycled.

• NAD is derived from niacin and is the main co-enzyme of respiration, becoming reduced to NADH. Each
molecule of NADH yields 3 ATP in the Electron Transport Chain (ETC). In anaerobic respiration,
respiration does not proceed beyond glycolysis, so the electrons in NADH have to be accepted by other
molecules- ethanal or Pyruvate itself.

• FAD is derived from riboflavin and is reduced to FADH2. Each molecule of FADH2 yields 2 ATP in the
ETC.

[You should remember NADP is the co-enzyme used in Photosynthesis, and is reduced to NADPH2 or
NADPH +H]
THE MITOCHONDRION

Glycolysis takes place in the cytosol, but the other stages take place in the mitochondria:
• The matrix – the liquid part inside the mitochondrion – this is where enzymes are present and
both the Link Reaction and the Citric Acid Cycle take place
• The Inner membrane including infoldings called cristae – this is where the electron carriers of
the ETC and also ATPase (stalked particles) are located, so t is the site of oxidative
phosphorylation
• Inter-membrane space – the space between the inner and outer membranes, where hydrogen
ions accumulate to produce the proton motive force to drive chemiosmosis.
 STAGES OF RESPIRATION

The process of Respiration is generally considered to have 3 stages although there is a fourth stage in-
between stages 1 and 2. the stages are:

• GLYCOLYSIS – literally the splitting of sugar, it takes place in the cytosol and involves splitting
glucose into 2 molecules of Triose Phosphate and converting each TP to Pyruvate - this is the only
stage in anaerobic respiration and 2 molecules of ATP are produced as well as reduced carriers.

• THE LINK/TRANSITION STAGE – This takes place in the matrix of the mitochondria and converts
the 3C pyruvate to a 2C compound to enter the Kreb’s Cycle –electron carriers are reduced

• THE CITRIC ACID/KREB’S CYCLE –also in the matrix- the 2C compound from the link reaction is
added to a 4C compound producing a 6C compound (Citrate) and in a series of steps, electron carriers
are reduced and 2 ATP made

• ELECTRON TRANSPORT CHAIN (OXIDATIVE PHOSPHORYLATION) All reduced carriers take

their electrons to the inner membrane of the mitochondria, where chemiosmosis takes place,
producing most of the ATP in Respiration
OVERVIEW OF RESPIRATION
GLYCOLYSIS
1. Phosphorylation – ATP is used to
add a phosphate group to glucose
(C6), forming glucose-6-
phosphate to energize it and make
it more reactive.

2. Isomerisation – the molecule is

rearranged to form fructose-6-
phosphate

3. Phosphorylation – a second
phosphate group is now added to
form fructose -1,6-bisphosphate.

4. Lysis – Fructose bisphosphate is

now split to produce two
molecules of Triose Phosphate
(TP)

(TRIOSE PHOSPHATE) This is the energy investment phase,

as 2 molecules of ATP have been used
and none have been produced.
As glucose has now been split into 2 TP
molecules, from this stage onwards ,
everything is multiplied by 2

6. Each Triose phosphate is oxidized, coupled

to the reduction of a NADH , to produce the
high energy NADH, and forming an
intermediate compound.

7. Each molecule of the intermediate

compound undergoes substrate level
phosphorylation to generate a molecule of ATP
and more intermediate molecules.

8, 9, 10. intermediate molecules undergo

another substrate level phosphorylation to
generate another ATP, and Pyruvate is now
formed

At the end of Glycolysis, there are

• 4-2 = 2 ATP
• 2 NADH
• 2 Pyruvate
SUMMARY OF GLYCOLYSIS

Note that glycolysis takes place in the cytosol of the cell and not
mitochondria.

For each Glucose molecule:

• 2 ATP molecules have been used, while 4 ATP (2x2) have been
produced, for a net gain of 2 ATP

• 2 NADH (2x1) molecules have also been produced

• 2 Pyruvate molecules are the main molecules remaining.

If there is no Oxygen present, Respiration cannot proceed any

further, giving a net production of ATP, but in the presence of
oxygen, the other stages take place with a potential production of
38 molecules of ATP per glucose molecule
THE LINK REACTION

Coenzyme
A This is an oxidative decarboxylation,
in which a carbon is removed and
pyruvate oxidized, coupled to the
reduction of NAD to NADH.
Coenzyme A is added, forming a 2C
acetyl CoA
 In the Citric Acid Cycle, 2C Acetyl CoA is added
to 4C Oxaloacetate, forming 6C Citrate.
2CC
In a series of Oxidative decarboxylation,
Oxidation and Reduction Reactions, Citrate is
6C reconverted to Oxaloacetate, with the formation
of :
4C • 2x1 = 2ATP
• 2x3 = 6 NADH
• 2x1 = 2 FADH2

5C Total ATP molecules formed up to now, is just 4,

however there 10 NADH and 2 FADH2 molecules.
NADH and FADH2 will go to the ETC, where
they will be used to generate ATP.
4C4 1 NADH – 3ATP
4 1FADH2 - 2 ATP

Please note that the names of intermediate stages

do not have to be known
 OXIDATIVE PHOSPHORYLATION- ELECTRON
TRANSPORT CHAIN

In a process very similar to photosynthesis, electrons are brought by NADH and FADH2, to the inner
membrane of the mitochondria. As electrons pass down an ETC, energy given off is used to pump H+ ions
into the inter-membrane space, creating an electro-chemical gradient which establishes a proton motive
force. When this has built up, H+ are released through ATP ase, by chemiosmosis fuelling the making of ATP.
Note – the final electron acceptor is oxygen, which is reduced to water, and the enzyme responsible is
cytochrome oxidase which can be inhibited by cyanide.
CHEMIOSMOSIS

• Reduced coenzymes NADH and FADH2 from the Citric Acid/Krebs cycle, bring electrons to the
inner membrane (including cristae) of mitochondria.
• Electrons pass along a series of electron carriers in an Electron Transport Chain.
• Energy is given off at each stage and this energy is used to pump H+ into the inter-membrane
space
• This builds up an electro-chemical gradient, which provides a proton motive force, which
drives the H+ out through the ATP ase located on the inner membrane.
• The release of the H+ fuels the making of ATP
• This is oxidative phosphorylation, as the making of ATP is linked to the uptake of oxygen, which
also serves as the final electron acceptor in the ETC
ATP PRODUCTION AT DIFFERENT STAGES OF
RESPIRATION

Note that 38 is the

theoretical
maximum yield,
but in reality, since
ATP is used in the
process, the real
yield is likely to be
closer to 34 ATP
SITES OF RESPIRATION
GLYCOLYSIS takes place in the cytosol.
As anaerobic respiration does not go beyond
glycolysis, the entire process takes place in the
cytosol and does not involve mitochondria.

THE KREBS CYCLE (and the Link Reaction)

takes place in the matrix of the mitochondria,
which is a liquid medium, in which enzymes and
other compounds are present.

THE ELECTRON TRANSPORT CHAIN takes

place on the inner-membrane of the mitochondria,
which is a membrane with proteins which are
electron carriers as well as the membrane-bound
enzyme, ATP-ase, which is responsible for the
production of most ATP molecules
 ANAEROBIC RESPIRATION

At the end of glycolysis, if there is no Oxygen present, respiration cannot go any further – as Oxygen is the
final electron acceptor which completes the process, so NADH molecules need electron acceptors so they
can be oxidized back to NAD – to be reused.
There are two possible pathways : Ethanol pathway in yeast and plant cells – pyruvate is decarboxylated
to ethanal /acetaldehyde, which accepts electrons and is reduced to ethanol
Lactate pathway – in bacteria and animal cells – the pyruvate itself
accepts electrons and is reduced to lactate.
ALCOHOL FERMENTATION
 COMMERCIAL USES OF ANAEROBIC RESPIRATION -
BIOTECHNOLOGY

ALCOHOLIC FERMENTATION produces

alcohol and carbon dioxide.
The respiring organism is yeast, and this is the
basis of production of alcoholic beverages, as well
as a rising agent in bread making – as the CO2
causes the dough to rise,
LACTATE FERMENTATION is used in the
making of dairy products – the respiring
organisms are bacteria
When energy demands are very high,
oxygen cannot be delivered to muscle cells
quickly enough, so cells respire
anaerobically.
The build-up of lactate is toxic and causes
fatigue, cramps and nausea, and must be
broken down very quickly, by taking in
additional oxygen.
REPAYING OXYGEN DEBT
LACTATE VS
ALCOHOL
FERMENTATION
INVESTIGATING RESPIRATION – A simple respirometer

A Respirometer is used to measure the rate of respiration of small organisms, by oxygen uptake.
A respirometer must have the following features:
• A tube where the living organisms are placed
• A tube leading from this to a capillary tube
• A means of keeping the apparatus airtight and watertight
• A chemical to remove CO2 given off, so pressure changes are due to oxygen uptake only
• Gauze or mesh to keep the organisms away from the corrosive chemical
• A drop of coloured liquid which can be seen moving as oxygen is used up
• A ruler to measure the movement
• An even simpler respirometer can be made using a syringe attached to a capillary tube
This respirometer has
respiring organisms in one
tube and glass beads in the
other.

What is the purpose of the

glass beads?
 CALCULATING VOLUME OF OXYGEN USED

• A capillary tube is a very narrow tube (cylinder)of approximately 0.6mm diameter.

• It is usually housed within an outer glass tube as it is very delicate.
• The distance travelled by the fluid inside a respirometer, can be used to calculate the volume of O2 used.
• Consider the following : in 10 minutes the fluid moved from the 25mm to the 12mm position on the ruler
• Calculate the rate of respiration of the respiring organism