Chapter 12

A2 Level
Energy and Respiration
Chapter Outline
Part 1: Aerobic and Anaerobic Respiration
• The need for energy • Mitochondria structure and
• ATP function

• Aerobic respiration • Anaerobic respiration

1. Glycolysis • Ethanol Pathway
• NAD, FAD, coenzyme A • Lactate Pathway
2. Link reaction • Oxygen debt
3. Krebs cycle
4. Oxidative phosphorylation
• Electron transport chain
• ATP Synthase
Chapter Outline
Part 2: Respiration using other biomolecules
• Respiration using carbs, lipids and protein
• Energy values of carbs, lipid and protein
• Respiratory quotients (RQ) of carbs, lipid and protein

• P5: respirometers
• P5: redox indicators (e.g. DCPIP, methylene blue)

• Adaptation of rice with submerged roots in water

 Net 2 2 28

Chapter 12 (Part 1)

Aerobic and Anaerobic Respiration

Chapter Outline
Part 1: Aerobic and Anaerobic Respiration
• The need for energy • Mitochondria structure and
• ATP function

• Aerobic respiration • Anaerobic respiration

1. Glycolysis • Ethanol Pathway
• NAD, FAD, coenzyme A • Lactate Pathway
2. Link reaction • Oxygen debt
3. Krebs cycle
4. Oxidative phosphorylation
• Electron transport chain
• ATP Synthase
Why do living organisms need energy? To work!
Examples of Work:
• Anabolic reactions
Eg: protein synthesis, synthesis
of glycogen (glycogenesis), DNA
replication, polymerisation

• Active transport
Eg: Na+- K+ pump, movement of
vesicles in exo/endocytosis • Maintenance of a constant body
temperature
in “warm-blooded animals”
i.e. endotherms
• Movement
Eg: Muscle contraction,
cilia/flagella, movement of
chromosomes • Bioluminescence / electrical
discharge
E.g. in jellyfish, electric eels
 ATP
Structure and Characteristics
• Adenosine triphosphate
• It is a phosphorylated nucleotide

Its structure has 3 components:

1) Adenine (organic, nitrogenous base)
Adenosine (nucleoside)
2) Ribose sugar (pentose sugar)
3) Three phosphate groups
ATP
Structure and Characteristics
Characteristics of ATP:
• Small
• Water-soluble
→ Easily transported around the cell
• Readily hydrolysed / lose phosphate to release energy
• Small packets of energy released at one time
• ATP can be synthesised and broken down quickly
→ High turnover rate

This makes it ideal as an energy currency

in all organisms
ATP
Roles of ATP
Roles/Functions of ATP:
a) The universal link/intermediate energy molecule
• Between energy-giving reactions and energy-requiring reactions

• Example of energy-giving reactions: aerobic respiration aka

complete oxidation of glucose
→ Gives a large quantity of energy of 2870 kJ per mole of glucose

C6H12O6 + 6O2 6CO2 + 6H2O + 2870kJ

P/S: Oxidation of glucose is also possible in anaerobic conditions….but we will talk about that later!
 ATP
Hydrolysis of ATP → Energy
• But energy transfers are inefficient
→Excess energy is lost at different stages in the multi-step reaction
→As thermal/heat energy

heat energy heat energy

ATP
Roles of ATP

b) ‘Energy currency’ of the cell

• Energy giving/yielding-reactions are linked to production of ATP
first, then energy-requiring reactions

Energy currency molecule Energy storage molecule

• Energy storage molecules store energy in the form of chemical

potential energy
E.g.
• Short term – glucose, sucrose
• Long term – glycogen, starch,
triglyceride
ATP
Hydrolysis of ATP → Energy
c) Immediate donor of energy
• To reactions requiring energy

Chemical equation of ATP hydrolysis:

ATP ADP + Pi + 30.5kJ
where Pi is inorganic phosphate
or
ATP + H2O ADP + H3PO4 + 30.5kJ

• Reactions are all reversible

• ATP can be synthesised and broken down quickly
• Rate of turnover/interconversion of ATP is high
E.g. At rest, we use about 40kg of ATP in 24hour
ATP
Hydrolysis of ATP → Energy
• Removal of 1st phosphate group from ATP → ADP
→ 30.5 kJmol-1 energy released

• Removal of 2nd phosphate group from ADP → AMP

→ 30.5 kJmol-1 energy released

• Removal of last phosphate group from AMP → Adenosine

→ 14.2 kJmol-1 energy released
ATP
ATP Synthesis
• ATP is synthesized from energy-yielding reactions
• E.g. oxidation of glucose in cellular respiration OR light-
dependent stage in photosynthesis
• In a series of reactions
What is respiration?
• Process where….
• Organic molecules
(such as glucose, amino acids, glycerol, fatty acids)
• Are broken down in a series of stages
• To release energy
• Which is used to synthesise ATP

Two types:
1) Aerobic respiration
2) Anaerobic respiration

Yeast/
 Aerobic Respiration
• Breakdown of organic molecules
(i.e. glucose, but fatty acids, glycerol and
amino acids too!)
• To release energy
• Which is used to synthesise ATP
• In the presence of oxygen

• 4 stages in aerobic respiration of glucose:

Stage Location
1) Glycolysis Cytoplasm
2) Link Reaction Mitochondrial matrix
3) Krebs Cycle Mitochondrial matrix
4) Oxidative Phosphorylation Inner mitochondrial membrane / cristae
 Aerobic Respiration
Oxidation of Glucose
C6H12O6 + 6O2 6CO2 + 6H2O + 2870kJ

• IRL oxidation of glucose in cellular respiration is a

multi-step reaction
• Each step: releasing a small quantity of energy
(ATP)

Why use small, multiple steps?

1) Allow precise control
2) Cells could not fully harness total energy released
if all were made available at one instant

• Also, reactions do not happen easily! Glycolysis, the 1st stage of glucose
Why ah? oxidation in aerobic respiration
Aerobic Respiration
Oxidation of Glucose
Why doesn’t the reaction happen easily?
Because….glucose is quite a stable substance
→ It requires a high activation energy for
reaction to take place

SO…how do organisms overcome this?

a) Usage of enzymes to lower activation energy

b) Raising energy level of glucose by

phosphorylation
→ More reactive
Stage 1: Glycolysis aka Glycolytic Pathway
The Steps
glucose (6C)

1) Glucose (6C) is phosphorylated C C C C C C

• By 2 ATP ATP
• Form hexose / fructose bisphosphate (6C)
fructose phosphate (6C)
• This raises chemical potential energy of glucose
• Provide activation energy for split C C C C C C P
ATP
2) Fructose bisphosphate breaks down to
2 triose phosphate (3C) fructose bisphosphate (6C)
• 6C → 2 × 3C P C C C C C C P

3) 2 hydrogen atoms are removed

• 2 reduced NAD formed 2 x triose phosphate (3C)
• This is a dehydrogenation / oxidation reaction P C C C C C C P
2 ATP
4) 4 ATP produced 2 NAD
2H
• 4 ATP – 2 ATP = net gain of 2 ATP
2 NADH
• Chemical potential energy is released from intermediates
intermediate steps 2 ATP
2 x pyruvate (3C)
5) 2 pyruvate (3C) produced
C C C C C C
Stage 1: Glycolysis aka Glycolytic Pathway
The Steps

Phosphorylation

Lysis

Oxidation /
Dehydrogenation

ATP is synthesized through

substrate-level phosphorylation
Stage 1: Glycolysis
Summary
@ Cytoplasm
• Lysis of glucose
• Multi-step process
• Involves many enzymes at each step

Initial reactants:
• 1 Glucose (6C)
• 2 ATP
• 4 ADP, 2 NAD

Final products per molecule of glucose:

• 2 Pyruvate (3C)
• 2 NADH
• 4 ATP → But 4 - 2 = net gain of 2 ATP only!
Okay let’s pause and talk about….
Hydrogen Carrier Molecules
• Also called hydrogen acceptor molecules

You will learn 3 types:

1. NAD – nicotinamide adenine
dinucleotide
(used in respiration)
2. NADP – nicotinamide adenine
dinucleotide phosphate
(used in photosynthesis)
3. FAD – flavin adenine dinucleotide
(used in respiration)

• They are all coenzymes = a non-protein

complex organic substance that is
required for an enzyme's activity.
Hydrogen Carrier Molecules
1. NAD
• Nicotinamide Adenine Dinucleotide
• Coenzyme
• H carrier molecule in respiration

Structure:
• Two linked nucleotides
• Both have ribose sugar and a phosphate group each
• 1 has adenine base, the other nicotinamide ring
• Nicotinamide ring – accepts H
Hydrogen Carrier Molecules
1. NAD
Function:
• H carrier molecule in respiration
• Carry hydrogens from all stages of respiration (Stage 1 2 3)
• To take part in oxidative phosphorylation (Stage 4) where
most ATP is synthesised
Hydrogen Carrier Molecules
2. NADP
• Nicotinamide Adenine Dinucleotide Phosphate
• Coenzyme
• Hydrogen carrier molecule used in
photosynthesis (Chapter 13)
• Different form of NAD

Structure:
• Similar to NAD NAD
• But has a phosphate group instead of H on NADP
carbon 2 on ribose ring with adenine
Hydrogen Carrier Molecules
3. FAD
• Flavin Adenine Dinucleotide
• Coenzyme
• H carrier molecule in respiration
• Used to carry H produced in
Krebs Cycle (Stage 3) only
→ Then used in oxidative
phosphorylation (Stage 4)

Structure:
• Two linked nucleotides
• One nucleotide with phosphate, ribose and adenine
• Another nucleotide with phosphate, ribitol and flavin
Stage 2: Link Reaction
The Steps
@ Mitochondrial matrix
• Pyruvate (3C) is transported from the cytoplasm
• Into the mitochondria matrix
• When oxygen is available
• By active transport coenzyme A
pyruvate (3C)
C C C + CoA
• When oxygen is available, pyruvate (3C) is:
1) Decarboxylated CO2
NAD
→ to form carbon dioxide
NADH
2) Dehydrogenated
→ NADH produced
3) Combined with coenzyme A (CoA) acetyl coenzyme A (2C)
→ To form acetyl coenzyme A (2C) CoA
C C
Stage 2: Link Reaction
The Steps

decarboxylation

dehydrogenation

acetyl CoA
(simplified chemical structure)
Stage 2: Link Reaction
What is coenzyme A?
• Complex molecule
acetyl CoA
• Made of a nucleoside (adenine + ribose) and (simplified chemical structure)

a vitamin (pantothenic acid)

Function:
• Carry acetyl groups (2C) to Krebs Cycle (Stage 3)
Stage 2: Link Reaction
Summary
@ Mitochondrial matrix

Initial reactants:
• 1 Pyruvate (3C)
• 1 NAD
• 1 CoA But since 1 molecule of glucose (6C) is
oxidized into 2 pyruvate…
Final products per molecule of pyruvate: Final products per molecule of glucose:
• 1 Acetyl CoA (2C) • 2 Acetyl CoA (2C)
• 1 NADH • 2 NADH
• 1 CO2 → waste gas, released • 2 CO2 → waste gas, released
Stage 3: Krebs Cycle
The Steps
• Aka citric acid cycle / tricarboxylic acid cycle
@ Mitochondrial matrix
• Enzyme-controlled pathway CoA
C C
CoA
1) Acetyl coenzyme A (2C) C C C C
→ combines with oxaloacetate (4C) C C C C C C
→ To form citrate (6C)
→ CoA removed and can be used again in
Link Reaction (Stage 2)

CO2

CO2
 Stage 3: Krebs Cycle
The Steps
2) Citrate (6C) goes through series of dehydrogenation and decarboxylation
→ 7 steps, by products of each step are
1. Nothing
2. Reduced NAD and CO2
3. Reduced NAD and CO2
4. ATP
7
5. Reduced FAD
6. Nothing 1
7. Reduced NAD 6 C C C C C C

5
2
3) Oxaloacetate (4C) regenerated C C C C C
→Can combine with another acetyl CoA
→ Kreb cycle continues
C C C C 3
4
 Stage 3: Krebs Cycle
The Steps

dehydrogenation

dehydrogenation
decarboxylation
dehydrogenation

substrate-level dehydrogenation
phosphorylation decarboxylation
Stage 3: Krebs Cycle
The REAL Steps
(which you fortunately do not need to memorise!)
Can I Keep Selling Substances for Money, Officer?

Take away messages:

• You need to remember the names
of 3 compounds: oxaloacetate (4C),
acetyl CoA (2C), citrate (6C)
• You need to know the byproducts
formed at each step
• You need to know the no. of C of
each intermediate compound
• Remember that each step requires
an enzyme
(ATP)
Stage 3: Krebs Cycle
Summary
@ Mitochondrial matrix
• Enyzme-controlled pathway

Initial reactants:
• 1 Acetyl CoA (2C)
• 1 Oxaloacetate (4C)
But since 1 molecule of glucose (6C) is
• 1 ADP, 3 NAD and 1 FAD oxidized into 2 pyruvate, which is converted to
2 acetyl CoA in the Link Reaction…
Final products per molecule of acetyl CoA: Final products per molecule of glucose:
(This is 1 turn of the Krebs Cycle) (This is 2 turns of the Krebs Cycle)
• 3 NADH • 6 NADH
• 1 FADH2 • 2 FADH2
• 2 CO2 → waste gas, released • 4 CO2 → waste gas, released
• 1 ATP • 2 ATP
• Oxaloacetate (4C) → regenerated • Oxaloacetate (4C) → regenerated twice
Stages 1, 2, 3:
Glycolysis, Link Reaction, Krebs Cycle
Products so far per molecule of glucose – Fill this up!
Stage ATP CO2 NADH FADH2

Glycolysis 4-2=2 0 2 0

Link Reaction 0 2 2 0

Krebs Cycle 2 4 6 2

TOTAL 4 6 10 2
Yay! Waste gas,
released
Used in oxidative phosphorylation
(Stage 4)
Stage 4: Oxidative Phosphorylation
Introduction
@ Inner membranes/cristae of mitochondria
• High release of electrical potential energy here
→For the production of ATP ( ADP + Pi → ATP )
• Involves a chain of electron carrier molecules in the inner membranes
→Electron transport chain (ETC)
• ATP synthesis is catalyzed by ATP Synthase

Outer mitochondrial membrane

Intermembrane space

Inner mitochondrial
membrane

Mitochondrial matrix
Stage 4: Oxidative Phosphorylation
The Steps
1) Hydrogen atoms removed from reduced NAD and FAD
• NADH and FADH2 are from glycolysis, link reaction, Krebs cycle
• At the inner mitochondrial membrane
• Through dehydrogenation / oxidation reaction
• Catalysed by dehydrogenase enzymes
• NAD and FAD regenerated
→ Can be reduced again in glycolysis, link reaction, Krebs cycle

Reduced NAD → NAD + H

Reduced FAD → FAD + 2H

• Hydrogen atoms split into electrons and protons (H+ ions)

H → H+ + e-
Stage 4: Oxidative Phosphorylation
The Steps
2) Electrons are passed along ETC
• Passed along a series of electron carriers in inner membranes
• Electron carriers are associated with 4 types of membrane proteins
→ forms a functional unit called a respiratory complex
• Energetic electrons release energy as they pass through the ETC
Stage 4: Oxidative Phosphorylation
The Steps
3) Energy released is used to pump protons
• Across inner mitochondrial membrane
• From mitochondrial matrix to intermembrane space
• Since inner membrane is impermeable to protons
• And there is high [H+] in intermembrane space
→ proton gradient formed / electrochemical gradient formed
Stage 4: Oxidative Phosphorylation
The Steps
4) Protons move down electrochemical gradient
• By facilitated diffusion through ATP synthase
• Back into the mitochondrial matrix
• This provides energy for ATP synthesis
• This process is called chemiosmosis

• ATP synthase enzyme rotates

ADP+ Pi → ATP
• ATP is synthesised
• Movement of 3 H+ ions back
into matrix = 1 ATP molecule
Stage 4: Oxidative Phosphorylation
ATP Synthase
• aka ATP synthetase/stalked particles
• transmembrane channel protein + enzyme

Found in:
• Inner membranes of mitochondria
https://www.youtube.com/watch?v=W3KxU63gcF4
• Thylakoid membranes of chloroplasts

Structure:
• Enzyme is connected to a part of the channel protein
– able to rotate as H+ ions pass thru
• 3 binding sites for ADP +Pi
• As it rotates, enzyme’s binding site changes structurally to
allow 3 different processes to occur:
a)Binding of ADP and Pi
b)Forming tightly bound ATP
c)Releasing ATP
Stage 4: Oxidative Phosphorylation
The Steps
5) Oxygen is the final hydrogen acceptor and final electron acceptor
O2 + 4H+ + 4e- → 2 H2O
• Forms water
• So previous electron carrier in ETC can be reduced again
• Ensures electrons can keep flowing along ETC
• Oxygen is why it’s called oxidative phosphorylation and aerobic
respiration
Stage 4: Oxidative Phosphorylation
Animated
Stage 4: Oxidative Phosphorylation
Summary
@ Mitochondrial inner membrane / cristae
• Requires the electron transport chain and ATP Synthase
• ATP is synthesized from electrical potential energy

Initial reactants per molecule of glucose:

• 10 NADH
• 2 FADH2
• O2 and ADP + Pi

• Theorectically: 1 NADH → 3 ATP; 1 FADH2 → 2 ATP

• But bcs some energy may be needed to transport ADP and Pi into
mitochondria and new ATP into cytoplasm
• So on average: 1 NADH → 2.5 ATP; 1 FADH2 → 1.5 ATP
Final products per molecule of glucose:
• 10*2.5 + 2*1.5 = 28 ATP
• Water
Stage 1 to 4: Balance Sheet of ATP synthesized for
1 molecule of Glucose in respiration – Fill this up!
Stages of Respiration ATP used ATP made Net gain in ATP

Glycolysis -2 4 +2

Link reaction 0 0 0

Krebs cycle 0 2 +2

28
Oxidative phosphorylation 0 (From 10 NADH and +28
2 FADH2)

Total -2 34 +32
Stage 1 to 4: Summary of reactants and products
of each stage per molecule of glucose – Fill this up!
Stage Reactants Products
1 glucose 4 ATP (but net gain 2 ATP)
Glycolysis 2 ATP, 4 ADP + Pi 2 pyruvate
2 NAD 2 reduced NAD

2 pyruvate 2 acetyl coenzyme A

Link reaction 2 CoA 2 reduced NAD
2 NAD 2 carbon dioxide
2 ATP
2 acetyl CoA
6 reduced NAD
1 oxaloacetate (used twice)
2 reduced FAD
Krebs cycle 6 NAD
4 carbon dioxide
2 FAD
2 CoA (regenerated)
2 ADP + Pi
1 oxaloacetate (regenerated twice)
10 reduced NAD 28 ATP
Oxidative 2 reduced FAD 10 NAD (regenerated)
phosphorylation 28 ADP + Pi 2 FAD (regenerated)
6 oxygen 6 water
 ATP can be synthesized in 2 ways in respiration:
ATP 1) Substrate-level phosphorylation
ATP Synthesis 2) Oxidative phosphorylation

Link
reaction

2 2 28
ATP
ATP Synthesis
ATP can be synthesized in 2 ways in respiration:
1) Substrate-level phosphorylation
• During glycolysis @ cytoplasm → 2 ATP
• During Krebs cycle @ matrix → 2 ATP
• 4 ATP produced by substrate-level phosphorylation in total
• Transfer of phosphate group from one molecule to another
• Chemical potential energy released
ATP
ATP Synthesis
2) Oxidative phosphorylation
• @ inner mitochondrial membrane / cristae
• Requires proton/electrochemical gradient, ATP synthase, ETC
• Electric potential energy released by chemiosmosis is used by ATP
synthase to catalyse formation of ATP
• 28 ATP produced by oxidative phosphorylation in total
• Also happens in the chloroplast during photosynthesis! (Chap 13)
Mitochondria
Structure and Function
• The mitochondria is the site for:
- Link reaction (matrix)
- Krebs cycle (matrix)
- Oxidative phosphorylation
(inner membrane)

• Typically rod-shaped
• Able to change shape and move in the cell
• 0.5 - 1.0 μm in diameter
• Double membrane
• No. of mitochondria in a cell, no. of cristae and length of crista in
mitochondrion – depends on cell
Mitochondria
Structure and Function
1) Matrix
• Has small, circular mitochondrial DNA
• Has 70S ribosomes
→ Synthesise mitochondrial proteins

• Have many enzymes in matrix for link reaction and Krebs cycle
Mitochondria
Structure and Function
2) Inner membrane / cristae
• Inner membrane is the site of
ETC and oxidative phosphorylation

• Inner membrane holds many special proteins and electron carriers

→i.e. enzyme ATP synthase, channels for H+ ions
• Linear arrangement of ETC on inner membrane
→ Greater efficiency

• Inner membrane / cristae is folded

→ Increase total surface area for ATP synthase and membrane proteins
→ Active cells have more foldings / dense cristae

• Inner membrane impermeable to H+ ions

→ Maintains proton gradient
→H+ only go through channels i.e. ATP Synthase
→Energy released by chemiosmosis used to synthesise ATP
Mitochondria
Structure and Function
3) Outer membrane
• Different in composition from inner membrane
• Smooth, not folded
• More permeable to small molecules than inner membrane
• Contains transport proteins to transport pyruvate into the
mitochondria for link reaction and Krebs cycle

4) Intermembrane space
• Space between the two membranes
• Allows accumulation of H+
• Lower pH than mitochondria matrix
• More acidic
→Due to pumping of H+ ions from matrix by the activity of ETC
Mitochondria
Structure and Function
Anaerobic Respiration
• Synthesis of ATP in the absence of oxygen

When free oxygen is NOT available:

• No final electron and H acceptor
• H from reduced NAD and FAD cannot be removed
→ ETC stops working
→ No ATP from oxidative phosphorylation
• NAD and FAD not regenerated
→ No Krebs cycle and link reaction
→ No ATP from Krebs cycle
 Anaerobic Respiration
However,
• Glycolysis still occurs @ cytoplasm
• Only net 2 ATP made per glucose molecule
by substrate level phosphorylation
• So glucose not completely broken down without oxygen
• Pyruvate still contains energy

• 2 NADH are formed too!

• Special pathways are used to regenerate NAD
• But this is not sustainable / cannot go on indefinitely
• Due to toxic byproducts

Yeast/
Anaerobic Respiration
Occurs in:
• In yeast cells / plant tissues
→ ethanol pathway / alcoholic fermentation
• In animals / mammalian muscles / some bacteria
→ Especially when muscle activity is high and cells do not
have enough oxygen to perform aerobic respiration
→ lactate pathway / lactic fermentation

• These pathways are used to regenerate NAD

• But this is not sustainable / cannot go on indefinitely
• Due to toxic byproducts
 Anaerobic Respiration in Yeast
Ethanol Pathway / Alcoholic Fermentation
The Steps
• 2-step reaction: Pyruvate → Ethanal → Ethanol
• Irreversible

1) Glycolysis: Glucose → Pyruvate (3C)

• Net gain of 2 ATP
• 2 NADH produced glucose (6C)
C C C C C C
2) Pyruvate (3C) undergoes
decarboxylation → Ethanal (2C)
• CO2 produced 2 ADP 2H 2 NADH 2 NAD

2 ATP

2 x pyruvate (3C) 2 x ethanal (2C)

C C C C C C C C C C
2 CO2
Anaerobic Respiration in Yeast
Ethanol Pathway / Alcoholic Fermentation
The Steps
3) Ethanal (2C) / acetaldehyde acts as a H acceptor
• Reduced by receiving hydrogen from reduced NAD
• Ethanal → Ethanol (2C)
• Catalysed by alcohol dehydrogenase
→This prevents H+ from lowering pH in yeast cell

glucose (6C)
4) NAD is regenerated C C C C C C
→Allows glycolysis
to continue
→To produce ATP
2 ADP 2H 2 NADH 2 NAD

2 ATP

2H

2 x pyruvate (3C) 2 x ethanal (2C) 2 x ethanol (2C)

alcohol
C C C C C C C C C C dehydrogenase C C C C
2 CO2
Anaerobic Respiration in Yeast
Ethanol Pathway / Alcoholic Fermentation

• Ethanol is toxic
• Reaction is irreversible!
• Remaining chemical potential energy in
ethanol is wasted
Anaerobic Respiration in Mammals
Lactate Pathway / Lactic Fermentation
The Steps
• 1-step reaction: Pyruvate → Lactate
• Reversible

1) Glycolysis: Glucose → Pyruvate (3C)

• Net gain of 2 ATP
• 2 NADH produced
glucose (6C)

2) Pyruvate acts as the H acceptor C C C C C C

• Receive H from NADH
• Pyruvate reduced to lactate /
lactic acid 2 ADP 2H 2 NADH 2 NAD
• Catalysed by 2 ATP
lactate dehydrogenase
2H
3) NAD is regenerated
2 x pyruvate (3C) 2 x lactate (3C)
→ Allows glycolysis to continue lactate
C C C C C C dehydrogenase C C C C C C
→ To produce ATP
Anaerobic Respiration in Mammals
Lactate Pathway / Lactic Fermentation
• Lactate is toxic
• Causes drop in pH / acidic
• But the reaction is reversible!

• Lactate is transported by blood plasma

• From muscles to be broken down in liver

In liver:
• Lactate converted back to pyruvate
• By lactate dehydrogenase
When oxygen is present again:
• Pyruvate is further broken down / oxidized in aerobic respiration
• i.e. link reaction → Krebs cycle → oxidative phosphorylation
• Produce carbon dioxide + water

• Excess lactate converted to glycogen

Anaerobic Respiration in Mammals
Oxygen Uptake Before, During and After Strenuous Exercise

Oxygen is a measure of metabolic rate

At rest:
• Rate of oxygen uptake at resting
levels is low
• Based on this graph… 0.2dm3 min-1

During exercise:
• Oxygen demand and uptake by cells increases
• But heart and lungs are unable to meet demand immediately
• Oxygen deficit = the volume diff between ideal and real O2 uptake
• So anaerobic respiration occurs
• Heart rate and breathing rate takes time to increase to meet O2 demand
Anaerobic Respiration in Mammals
Oxygen Uptake Before, During and After Strenuous Exercise

Extra oxygen is also needed for:

1. For the conversion of lactate
→ glycogen
2. For the reoxygenation of
haemoglobin in the blood
3. To meet the high metabolic
rate in organs

During recovery:
• Breathing rate / oxygen uptake is still higher than resting levels
• To “pay back” the oxygen debt
• Oxygen debt = volume of oxygen required to metabolise lactate
accumulated during anaerobic respiration to CO2 and water after exercise
Anaerobic Respiration in Mammals
Oxygen Debt
Oxygen debt = volume of oxygen required to metabolise lactate
accumulated during anaerobic respiration to CO2 and water after exercise

Calculating oxygen debt:

• oxygen consumed = oxygen inhaled – oxygen exhaled;
• measure oxygen consumption at rest (x) and after exercise stops (y)
• extra oxygen consumed = oxygen debt = y – x
• Divide by mass of organism
Similarities Between Ethanol and Lactate
Pathway
1. Both occur when oxygen is absent/low in concentration
2. Both occur in the cytoplasm
3. Both involve glycolysis
4. Both produces only 2 ATP net per glucose molecule
5. Both involve usage and regeneration of NAD
Differences between Ethanol and Lactate Pathwa
Fill this up!
Features Ethanol Pathway Lactate Pathway

Occurs in Yeast, Plants Animals, mammalian muscles

Hydrogen acceptor Ethanal Pyruvate

Decarboxylation
Yes No
reaction

Production of CO2 Yes No

Reversibility of
Irreversible Reversible
reaction
Final product
Ethanol Lactate / lactic acid
formed

Enzymes involved Alcohol dehydrogenase Lactate dehydrogenase

Chapter 12 (Part 2)

Respiration
using Other
Biomolecules
Chapter Outline
Part 2: Respiration using other biomolecules
• Respiration using carbs, lipids and protein
• Energy values of carbs, lipid and protein
• Respiratory quotients (RQ) of carbs, lipid and protein

• P5: respirometers
• P5: redox indicators (e.g. DCPIP, methylene blue)

• Adaptation of rice with submerged roots in water

What is respiration?
• Process where….
• Organic molecules
(such as glucose, amino acids, glycerol, fatty acids)
• Are broken down in a series of stages
• To release energy
• Which is used to synthesise ATP

Two types:
1) Aerobic respiration
2) Anaerobic respiration

Yeast/
Respiratory Substrates
1) Glucose
• Essential respiratory substrate for neurones in the brain, RBC, lymphocytes
• Oxidised in glycolysis, link reaction, Krebs cycle

2) Lipids
• Converted to acetyl coA (2C)
• Oxidised in Krebs cycle

3) Protein
• Amino acid converted to pyruvate (3C) or acetyl coA (2C)
• Oxidised in link reaction or/and Krebs cycle
---
• NADH and FADH2 are produced
• Passed to the ETC in oxidative phosphorylation
• To produce ATP
But which molecule contains the most energy?
Respiratory Substrates
Respiratory Substrates
Energy Values
How do we know which molecule contains the most energy?

• Most ATP comes from oxidative phosphorylation

→ Oxidation of hydrogen from NADH and FADH2 to water
→ Electrons from NADH and FADH2 pass through ETC
→ Hydrogens from NADH and FADH2 are pumped into intermembrane space
→H ions move back into matrix via ATP synthase
→ combine with oxygen to form water
→ ATP synthase rotates
→ produce ATP

• H needed for ATP production /chemiosmosis

Respiratory Substrates
Energy Values
Hence,
• The higher the number of C-H bonds / H atoms in a respiratory substrate
→ The more hydrogens can be carried by hydrogen carriers (NAD, FAD)
→ More reduced NAD produced
→ More oxidative phosphorylation / ETC
→ More hydrogen ions pumped across inner mitochondrial membrane /
steeper proton gradient
→ The more ATP made per gram of respiratory substrate
→ The greater the energy value
 Respiratory Substrates
Energy Values of Lipids
• Has most C-H bonds / no. of H atoms per unit mass
• Compared to carbs and protein
• So releases most energy per unit mass
• High energy density

• Many more H available to reduce oxygen to water

• More water produced from metabolism of lipid

• Oxidation of lipids can only happen in the presence of oxygen

 Respiratory Substrates
Energy Values of Proteins
• Slightly more C-H bonds per unit mass
• Than carbohydrates
• Used during starvation / lack of fat or carbs

• Oxidation of amino acids can only happen in the

presence of oxygen
Respiratory Substrates Memorise these values!

Energy Values

Respiratory
Energy Values (kJ g-1)
Substrate

Less C-H bonds,

Carbohydrate 15.8 more oxygen

Lipid 39.4 Most C-H bonds

Protein 17.0
Respiratory Substrates
How to determine energy values?
• Using calorimeter
• Burning a known mass of substance
• With oxygen
• Measure rise in temperature
• Of a known mass of water
Respiratory Substrates
Respiratory Quotient (RQ)
• RQ = Ratio of the volume of carbon dioxide produced to oxygen used
• Per unit time
• Can use no. of molecule / moles also

• Diff respiratory substrate, diff ratio of the volumes of oxygen used and
carbon dioxide produced

• Function of RQ:
1) Shows the type of substrate being used in respiration
2) Shows if anaerobic respiration is occurring
Respiratory Substrates
Respiratory Quotient (RQ)
In aerobic respiration of glucose:

RQ = 6/6
= 1.0

In aerobic respiration of fatty acids (in this case oleic acid, from olive oil):

C18H34O2 + 25.5O2 → 18CO2 + 17H2O + energy

RQ = 18/25.5
= 0.7
Respiratory Substrates
Respiratory Quotient (RQ)
In anaerobic respiration of glucose in yeast: (ethanol pathway)
C6H12O6 → 2C2H5OH + 2CO2 + energy

RQ = 2/0
=∞

In reality, some respiration in yeast cells will the aerobic as well, so wont reach infinity.
High values of RQ = Anaerobic respiration is occurring

But in anaerobic respiration of glucose in muscle cells: (lactate pathway)

C6H12O6 → 2C3H6O3 + energy

• No carbon dioxide produced

• No RQ can be calculated
Respiratory Substrates Memorise these values!

Respiratory Quotient (RQ)

Respiratory Substrate Respiratory Quotient

Carbohydrate 1.0

Lipid 0.7

Protein 0.9

x/0 = ∞
Anaerobic in yeast
Greater than 1.0

How do we determine Respiratory Quotient (RQ)

 P5
Respirometer
• Read manometer scale
• To measure rate of oxygen uptake during respiration
• Of small terrestial invertebrates/germinating seeds
 P5

Respirometer
1) Pressure and temperature must be kept constant while reading is taken
→Use thermostatic water bath
2) Use control tube with equal volume of inert material (e.g. glass beads)
→ compensate for changes in atmospheric pressure
3) Soda lime/ KOH / NaOH
→ Absorb carbon dioxide produced
4) Volume of air in test tube with organisms decrease = oxygen consumption
→Level of manometer fluid nearer to experimental tube increase

before after
Respirometer P5

Using the respirometer to measure RQ

• Use same apparatus twice

1) Find volume of oxygen (x cm3 min-1) consumed at a fixed temperature with

soda lime

2) Repeat by setting up with the same organism at the same fixed temperature
but remove chemical that absorbs carbon dioxide
→ Manometer fluid is now affected by both oxygen consumption
AND carbon dioxide released
→ Check changes to manometer scale fluid
 P5
Respirometer
Using the respirometer to measure RQ
1) If manometer fluid (nearer to experimental tube) level does not change:
→oxygen absorbed = carbon dioxide produced

RQ = 1

2) If manometer fluid lowers by y cm:

→carbon dioxide produced > oxygen absorbed
→Volume of air in respirometer increase
x
y
RQ = (x + y) / x
 P5
Respirometer
Using the respirometer to measure RQ
3) If manometer fluid increase by z cm:
→Oxygen absorbed > carbon dioxide produced
→Volume of air in respirometer decrease

RQ = (x - z) / x

x z
Respirometer P5

Using the respirometer to measure the effect of

temperature on oxygen consumption
• Take measurements at various temperatures
• Plot graph of oxygen consumption against temp

• Expected result:
• Higher temperature, higher rate of respiration
• Therefore faster absorption of oxygen
 P5
Redox indicators
(e.g. DCPIP, methylene blue)
• Can be used to investigate the rate of respiration
of yeast

• Redox indicator / dye does not damage cells

• So can add dye to suspension of yeast cells
• When reduced, the blue dye become colourless

• Measure time taken for dye to change from blue to colourless

= rate of respiration of yeast

• Can measure time taken at diff temp, with diff [S] or diff substrates
• To measure effect on rate of respiration
Adaptation of rice with
submerged roots in water
• Oryza sativa (rice)
• 20% of human population’s energy source
• Paddy fields – intentionally flooded
• Tolerate growing in water but competitor weeds cannot
Adaptation of rice with
submerged roots in water
Challenges:
• Low oxygen in water, roots do not get
enough oxygen
• Gas diffuse much slowly through water
than in air
• Anaerobic respiration occurs
• Toxic alcohol produced

Adaptations:
1) rice tolerant to higher levels of alcohol
• rice produces high levels of alcohol dehydrogenase to break down alcohol
• can respire anaerobically for longer periods
Adaptation of rice with
submerged roots in water
2) Presence of aerenchyma
• air spaces between cells in mesophyll /
cortex of stems
• oxygen able to diffuse through
aerenchyma from aerial tissues to roots
• this provides oxygen for aerobic
respiration
• Also allows for:
• escape of ethane
• buoyancy
• enables active transport in roots bcs
aerobic respiration produces energy
Adaptation of rice with
submerged roots in water
3) Grow taller in response to flooding
• So leaves, flower and spikes are above water
• O2 and CO2 can diffuse in/out through stomata on leaves
Adaptation of rice with
submerged roots in water
4) Leaves underwater have hydrophobic, corrugated surface
• Air trapped in between ridges
• Holds thin layer of air in contact with leaf surface
Videos
Cellular Respiration Glycolysis, Krebs cycle, Electron Transport 3D
Animation
https://www.youtube.com/watch?v=7J4LXs-oDCU

Glycolysis (a rap lecture) – for fun lol

https://www.youtube.com/watch?v=EfGlznwfu9U

ATP and Respiration: Crash Course Biology #7

(DO NOT follow the calculations for no. of ATP per glucose molecule)
https://www.youtube.com/watch?v=00jbG_cfGuQ