Cellular respiration

Introduction to cellular respiration and redox reactions

- Catabolic reaction
Breakdown of glucose: C6H12O6 + 6O2  6CO2 + 6H2O
ΔG = -686 kcal/mol, meaning it releases energy
The energy is used to make ATP molecules
 Direct: substrate-level phosphorylation of ADP
 Indirect: oxidative phosphorylation (requires oxygen) in the
mitochondria
The electron transport chain
 Responsible for oxidative phosphorylation
 Electrons from glucose are used by electron transport chain
 Electrons move to a lower energy level and release energy
 Energy is used to pump protons out of the matrix
 Protons flow back down the gradient and drives the ATP
synthase to make ATP
- Cellular respiration is the breakdown of organic fuels (e.g. glucose) to
maintain the metabolism of the cell
- Electron carriers
Small molecules that carry electrons from glucose breakdown
reaction to the electron transport chain
NAD+ (nicotinamide adenine dinucleotide)
FAD (flavin adenine dinucleotide)
When NAD and FAD pick up electrons, they also gain hydrogen atoms
 NAD+ + 2e- + 2H+  NADH + H+
 FAD + 2e- + 2H+  FADH2
When they drop off electron, they return to their original form
 NADH  NAD+ + 2e- + H+
 FADH2  FAD + 2e- + 2H+
- Redox reactions
Reactions involving electron transfers
Oxidation is where a molecule “loses” electrons; reduction is where a
molecule “gains” electrons, due to difference in electronegativity
Examples:
 Mg + Cl2  Mg2+ + 2Cl- (ionic, where magnesium is oxidised and
chlorine is reduced)
 2H2 + O2  2H2O + energy (covalent, where hydrogen is oxidised
and oxygen is reduced)
In a biological context
 If a carbon-containing molecule loses H atom or gains O atoms,
it is likely to be oxidised
  If a carbon-containing molecule gains H atom or loses O atoms,
it is likely to be reduced
Breakdown of glucose: C6H12O6 + 6O2  6CO2 + 6H2O
 Glucose is oxidised because it lost hydrogen atoms, therefore
carbon has “lost” electrons to oxygen
 Oxygen is reduced because it gained hydrogen atoms, therefore
it has “gained” electrons from hydrogen
- The purpose of the redox
to get the energy out of the glucose molecule
Use the energy to synthesize ATPs as electrons move through the
electron transport chain to oxygen, passing to lower and lower energy
states and releasing energy at each step

Steps of cellular respiration

- Breakdown of glucose provides the electrons for oxidative phosphorylation
- Glycolysis
Glucose is broken down into two molecules of pyruvate, a three-
carbon molecule
2 ATPs are made and 2NAD+ is reduced
- Pyruvate oxidation
each pyruvate enters the matrix of mitochondria
they are converted into a two-carbon molecule bound to Coenzyme A,
known as acetyl CoA
Carbon dioxide is released and NADH is generated
- Krebs cycle
Acetyl CoA combine with oxaloacetate and becomes citric acid
Goes through a cycle of reactions and returns back to oxaloacetate
ATP, NADH and FADH2 are produced, carbon dioxide is released
- Oxidative phosphorylation
NADH and FADH2 carry electrons to the electron transport chain and
get oxidised (back to NAD and FAD)
The energy released by electrons moving down the chain is used to
pump protons out of the matrix
The ATP synthase use the gradient of protons to generate ATP
- Glycolysis can take place without oxygen in a process called fermentation,
but the other three steps all require the presence of oxygen
Glycolysis
- Glycolysis is a series of reactions that
extract energy from glucose by splitting it
into two three-carbon molecules called
pyruvates
- Has ten steps in total, which can be broken
down into two main phases
Energy-requiring phase
 Glucose is phosphorylated and
modified
 Split it in half and form two
phosphorylated three-carbon
sugars (glyceraldehyde-3-
phosphate)
 Two ATPs are used
Energy-releasing state
 Each three-carbon sugar is dephosphorylated and converted
 into pyruvate through a series of reactions
 Four ATP and two NADH are produced from a single glucose
- All the steps are carried out by enzymes
Hexokinase: phosphorylate the glucose
Phosphofructokinase: speeds up or slows down glycolysis in response
to the energy need within the cell
- Detailed steps of energy-requiring phase
Step1: A phosphate group is transferred to glucose, forming glucose-6-
phosphate
Step 2: glucose-6-phosphate is converted into fructose-6-phosphate
Step 3: another phosphate group is transferred and making it fructose-
1,6-biphosphate (catalysed by phosphofructokinase)
Step 4: the unable fructose-1,6-bisphosphate splits to form two three
carbon sugars: DHAP and glyceraldehyde-3-phosphate
Step 5: DHAP is converted into glyceraldehyde-3-phosphate
- Detailed steps of energy-releasing phase
Step 6: each glyceraldehyde-3-phosphate is oxidised and NAD is
reduced to NADH and H+, releasing energy to phosphorylate the
molecule to 1,3-bisphosphoglycerate
Step 7: each 1,3-bisphosphoglycerate is dephosphorylated and
produce one ATP
Step 8: each 3-phosphoglycerate is converted into its isomer, 2-
phosphoglycerate
Step 9: 2-phosphoglycerate loses one water molecule and becomes
PEP
Step 10: unstable PEP loses one phosphate to ADP, making a second
ATP, and is converted to pyruvate
 - What happens next?
With oxygen, two pyruvate molecules can be broken down (oxidised)
all the way to carbon dioxide, making many more ATPs
Two NADH carry electrons to the electron transport chain if there is
oxygen, or pass the electrons to an acceptor and allow fermentation if
there is no oxygen
Fermentation is a primary metabolic strategy for bacteria and red
blood cells

Pyruvate oxidation
- Occurs in the matrix of mitochondria, therefore it is aerobic
- Converts the pyruvate into acetyl CoA, a two-carbon molecule attached to
Coenzyme A
- Carbon dioxide is released and one NAD is reduced from this process
- Detailed steps
Step 1: a carboxyl group is snipped off of pyruvate and released as a
carbon dioxide
Step 2: the two-carbon molecule is oxidised, thus reducing one NAD
Step 3: the oxidised two-carbon molecule is attached to Coenzyme A, a
molecule derived from Vitamin B5, to form acetyl CoA
Catalysed by an enzyme called pyruvate dehydrogenase complex,
which regulates the amount of acetyl CoA entering into the Krebs cycle
- Two NADHs and two acetyl CoA are produced, two carbon dioxides are
released from one single glucose

The citric acid cycle (Krebs cycle)

- Occurs in the matrix of the mitochondria, therefore requires oxygen
- A closed loop, oxaloacetate is being reused after each cycle
- An overview of the process
Acetyl CoA combines with a four-carbon molecule called oxaloacetate,
to form citrate or citric acid
Citrate releases two carbons as CO2 and producing one NADH each
time
The remaining four-carbon molecule undergoes a series of reactions to
make ATP or GTP, then reducing FAD to FADH2, and finally generate
another NADH
Overall, four carbon dioxides are released, six NADH and two FADH 2,
and two GTPs or ATPs are produced from a single glucose
- Detailed steps
Step 1:
 acetyl CoA combines with oxaloacetate
 releasing the CoA group and forming citrate (6C)
Step 2:
 Citrate is converted into its isomer, isocitrate (6C)
 Involves the removal and addition of a water molecule
Step 3:
 isocitrate is oxidised and releasing one carbon dioxide, leaving
behind α-ketoglutarate (5C)
 one NAD is reduced
 catalysed by isocitrate dehydrogenase, which regulates the rate
of the cycle
Step 4:
 α-ketoglutarate is oxidised and one CO2 is released
 one NADH is produced
 the remaining four-carbon molecule combines with Coenzyme A
again and forms the unstable succinyl CoA (4C)
 catalysed by α-ketoglutarate dehydrogenase, also regulates the
rate of the cycle
Step 5:
 Succinyl CoA replaces the Coenzyme A by a phosphate group
 The phosphate group is then passed on to GDP/ADP and make
GTP/ATP
 Succinate (4C) is produced in this step
Step 6:
 Succinate is oxidised, reducing one FAD molecule to FADH2
 FAD is used because succinate is not a great electron donor, and
FAD is a better electron acceptor than NAD
 Fumarate (4C) is made
Step 7:
 water is added to fumarate, converting it into malate (4C)
Step 8:
 malate is oxidised and becomes oxaloacetate (4C) again
  another molecule of NAD is reduced

Oxidative phosphorylation
- made up of two closely connected components: the electron transport chain
and chemiosmosis
- role of oxygen
oxygen is used at the end of ETC, where it accepts electrons and picks
up protons to form water
if oxygen is absent, the ETC would stop running, and ATP will no
longer be produced by chemiosmosis
- Overview of oxidative phosphorylation
Electrons are delivered by NADH and FADH2 to the ETC
As electrons are passed on, they release energy to pump H+ out of the
matrix
Electrons are transferred to O2, which splits in half and takes up H + to
form water (H2O)
As protons flow down their gradient, it drives ATP synthase to
produce ATP molecules
- The electron transport chain
Four large membrane-embedded proteins complexes (I to IV)
Located in the inner mitochondrial membrane in eukaryotes, and in
the plasma membrane in prokaryotes
 Regenerates electron carriers and makes a proton gradient
Electrons travel to a lower energy level (from less electronegative to
more electronegative molecules) and release energy

NADH is good at donating electrons, so it can transfer electrons

directly to complex I
FADH2 is not as good at donating electrons, so it transfers electrons to
complex II, causing fewer protons to be pumped
Four electrons are required to reduce each molecule of O2, therefore
two water molecules are made
- Chemiosmosis
Complexes I, III and IV are proton pumps, creating a proton gradient
or proton-motive force
As protons flow through the ATP synthase, it turns the turbine that
catalyses the phosphorylation of ADP

Some organisms utilise the proton gradient to generate heat by letting

 protons pass through proteins other than ATP synthase
- ATP yield
In theory, the maximum ATP yield from a single glucose is 38; however,
the actual figure is about 30-32 ATPs
2 ATP from glycolysis; 2 from pyruvate oxidation; 2 from the citric acid
cycle; and 24-26 from oxidative phosphorylation (each NADH yields
about 2-2.5 ATP and each FADH2 yields 1.5 ATP)

Fermentation
- A method to get energy from fuels anaerobically (not the only anaerobic
way)
- Other anaerobic cellular respiration ways
Using sulfate as the electron acceptor, producing hydrogen sulfide as a
waste product
Using carbon dioxide as the electron acceptor, producing methane
- Consists of glycolysis with some extra reactions
Glycolysis occurs normally
Pyruvate molecule does not continue through oxidation and the Krebs
cycle, and the ETC
NADH needs to return back to NAD+ (because ETC is not functional),
which is accomplished by two ways
- Lactic acid fermentation
NADH directly transfer their electrons to pyruvate
Pyruvate is reduced to lactate
Carried out by RBC and bacteria which do not have mitochondria; or
by muscle cells when there is not enough oxygen
- Alcohol fermentation
The carboxyl group (COO) is removed from pyruvate and released as
CO2
Two acetaldehyde molecules are produced, and reduced by NADH to
ethanol
Alcohol is toxic to cells in large quantities
- Most bacteria and archaea are facultative anaerobes, meaning they can
switch between aerobic respiration and anaerobic pathways
- Other bacteria and archaea are obligate anaerobes, meaning they can only
live in the absence of oxygen

Connection between cellular respiration and other pathways

- Amino acids, lipids and other carbohydrates can be converted to
intermediates and undergo cellular respiration
- Some intermediates may be removed to complete other work
- How
carbohydrates enter the pathway
Most enter through glycolysis (e.g. breakdown of glycogen to glucose)
Sucrose (fructose and glucose) can also enter easily
- How proteins enter the pathway
Amino acids are used to mostly construct proteins, but can enter
cellular respiration if there is a need
Amino acids first have the amino group removed, producing ammonia
Ammonia is converted to urea and excreted by kidney
Deaminated amino acids enter at various stages depending on their
chemical properties (e.g. glutamate is converted into α-ketoglutarate)

- How lipids enter the pathway

Triglyceride is broken down to glycerol and three fatty acid tails
Glycerol is converted to glyceraldehyde-3-phosphate
Fatty acid tails undergo beta-oxidation in the mitochondria to become
two-carbon units, and form acetyl CoA
- Many molecules can be pulled out at many stages for other bodily functions
Regulation of cellular respiration
- ATP is unstable therefore controlling the rate of production is important
- Allosteric enzymes and pathway control
Regulation of the activity of enzymes in the pathway
Often by controlling the enzyme that catalyses the first committed
step (the step is not readily reversible)
Controlled by the binding of regulatory molecules at allosteric sites
Feedback inhibition
 Mostly promoted by ADP and AMP
 Mostly inhibited by NADH and the products
- Regulation of glycolysis
Phosphofructokinase (PFK), which is the enzyme converts fructose-6-
phosphate to fructose-1,6-biphosphate
Regulation of PFK
 ATP and citrate: inhibit PFK activity
 AMP: promotes PFK activity
- Pyruvate oxidation
Pyruvate dehydrogenase is made less active by ATP and NADH; and
more active by ADP
Activated by pyruvate (substrate), and stopped by acetyl CoA
(product)
- Citric acid cycle
Mostly regulated by the pyruvate dehydrogenase in the previous
phase, but can also be regulated by the enzymes involved in CO2
release step
Isocitrate dehydrogenase
 Promoted by ADP
 Inhibited by ATP and NADH
α-Ketoglutarate dehydrogenase
 inhibited by ATP, NADH and succinyl CoA (product)