9

Pathways That Harvest

Chemical Energy

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Chapter 9 Key Concepts

Metabolic pathways:
• Cells get energy from glucose in a series of metabolic
pathways.
• Complex transformations occur in a series of separate
reactions.
• Each reaction is catalyzed by a specific enzyme.
• Many metabolic pathways are similar in all organisms.
• In eukaryotes, metabolic pathways are compartmentalized in
specific organelles.
• Key enzymes can be inhibited or activated to alter the rate of
the pathway.

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Energetics of living organisms

Second law of thermodynamics states that disorder or entropy

increases because of energy transformations during all chemical
and physical processes.
However, a living cell is in a low-entropy, nonequilibrium state
characterized by a high degree of structural organization.
It takes energy to impose order on a system. Unless energy is
applied to a system, it will be randomly arranged or disordered.
Living organisms preserve their internal order by taking free
energy from the surroundings in the form of nutrients and
returning to their surroundings an equal amount of energy as heat
and entropy.

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ATP
The most important molecule for capturing and transferring free
energy in biological systems is adenosine triphosphate, or
ATP.

Formation of Hydrolysis
ATP is of ATP is
endergonic: exergonic

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Figure 8.6 Coupling of Reactions
Through coupling reactions, ATP drives endergonic reactions
using energy derived from exergonic reactions.

ATP can be hydrolyzed to ADP and Pi , releasing a lot of energy

for endergonic reactions.
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ATP
The chemical basis for the relatively large, negative,
standard free energy of hydrolysis of ATP.

Electrostatic repulsion: The hydrolytic cleavage of

the terminal phosphoric acid anhydride
(phosphoanhydride) bond in ATP separates one of
the three negatively charged phosphates and thus
relieves some of the electrostatic repulsion in ATP.

Resonance stabilization: The Pi (HPO42-) released

is stabilized by the formation of several electron
resonance forms not possible in ATP.

ADP and Pi ionization: ADP2-, the other direct

product of hydrolysis, immediately ionizes, releasing
H+ into a medium of very low [pH] (~10-7 M).

Interactions with water: Improved solvation of free Pi

and ADP2- by surrounding water
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Energetics of living organisms

Energy relationships between catabolic and anabolic pathways

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9.1 Cells Harvest Chemical Energy from Glucose Oxidation

Burning or metabolism of glucose:

C6H12O6 + 6 O2 → 6 CO2 + 6 H2O + free energy

ΔG = - 686 kcal/mol
• Highly exergonic; drives endergonic formation of
many ATP molecules

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Figure 9.1 Energy for Life

Three catabolic processes harvest energy

from glucose
• Glycolysis (anaerobic)
• Cellular Respiration (aerobic)
• Fermentation (anaerobic)

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9.3 Oxidative Phosphorylation Forms ATP
Why does glucose utilization have so many steps?
A single reaction would release too much free energy all at
once.
In a series of reactions, each reaction releases a small amount
of energy that can be captured by an endergonic reaction.

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9.1 Cells Harvest Chemical Energy from Glucose Oxidation
Oxidation–Reduction (Redox) reactions: One substance transfers
electrons to another substance.

• Reduction: Gain of electrons.

• Oxidation: Loss of electrons.

Oxidation and reduction always occur together.

Transfer of electrons is
often associated with
transfer of hydrogen ions:
H = H+ + e –

• When a molecule loses

H atoms it becomes
oxidized.
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9.1 Cells Harvest Chemical Energy from Glucose Oxidation

In glucose metabolism, glucose is the reducing agent.

In a redox reaction some energy is transferred from the
reducing agent (glucose) to the reduced product.

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Figure 9.3 Oxidation, Reduction, and Energy

Most reduced state

Highest free energy

Most oxidized state

Lowest free energy

The more reduced a molecule is, the more energy it

has.
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9.1 Cells Harvest Chemical Energy from Glucose Oxidation
Coenzyme NAD+ is a key electron carrier in redox
reactions.
Reduction:
NAD+ + H+ + 2e– → NADH

Glucose NAD+

NADH
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9.1 Cells Harvest Chemical Energy from Glucose Oxidation
The final reaction: oxidation
O2 accepts electrons from NADH:
NADH + H+ + ½ O2 → NAD+ + H2O

The reaction is exergonic:

NADH O2
ΔG = –52.4 kcal/mol
O2 is the oxidizing agent, it is the

ENERGY
final electron acceptor.
NAD+ H2O

ATP
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9.1 Cells Harvest Chemical Energy from Glucose Oxidation

Cells harvest energy from glucose

using different combinations of
metabolic pathways.
Some species can harvest energy by
anaerobic respiration.

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Figure 9.4 Energy-Yielding Metabolic Pathways (Part 2)

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Glycolysis
Glycolysis: *
Preparatory phase

Payoff phase

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*
Figure 9.6 Glycolysis Converts Glucose into Pyruvate (Part 1)

Glycolysis
– Takes place in the cytosol
– Converts glucose into 2 molecules of
pyruvate
– Produces 2 ATP and 2 NADH
– Occurs in 10 steps.

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Figure 9.5 Glycolysis Converts Glucose into Pyruvate (Part 1)

Steps 1–5 Glycolysis

require 2 – Takes place in the
ATP. cytosol
– Converts glucose into 2
molecules of pyruvate
– Produces 2 ATP and 2
NADH
– Occurs in 10 steps.

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Figure 9.5 Glycolysis Converts Glucose into Pyruvate (Part 2)

Steps 6–10 yield NADH

and ATP (energy-
harvesting reactions).

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Concept 9.2 In the Presence of Oxygen, Glucose Is Fully Oxidized
(3)

Two types of reactions occure repeatedly in many

metabolic pathways:
Oxidation–reduction: energy released by glucose
oxidation is trapped via the reduction of NAD+ to
NADH.
Substrate-level phosphorylation: energy released
transfers a phosphate from the substrate to ADP,
forming ATP.

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 Oxidation–reduction: The first reaction is exergonic—more than 50
kcal/mol of energy are released in the oxidation of glyceraldehyde 3-
phosphate. Some energy is trapped via the reduction of NAD+ to
NADH.
2. Substrate-level phosphorylation: The second reaction in this
series is also exergonic, but in this case less energy is released,
sufficient to transfer a phosphate directly from the substrate to ADP,
forming ATP.
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9.2 In the Presence of Oxygen, Glucose Is Fully Oxidized
Pyruvate Oxidation
• Occurs in the mitochondrial matrix
• Produces acetate and CO2
• Acetate binds to coenzyme A to form acetyl CoA
• A multistep reaction catalyzed by the pyruvate
dehydrogenase complex (3 enzymes)
• Exergonic; one NAD+ is reduced to NADH

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Figure 9.6 The Citric Acid Cycle
Citric acid cycle
Acetyl CoA is the starting point, it donates
its acetyl group to oxaloacetate, forming
citrate. This initiates the citric acid cycle.
Eight reactions completely oxidizes the
acetyl group to 2 molecules of CO2.
Energy released is captured by GTP,
NADH, and FADH2.

Oxaloacetate is regenerated in the last

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step.
Figure 9.9 The Citric Acid Cycle

GTP can transfer its high-energy

phosphate to form ATP.
Overall, the oxidation of one
glucose molecule yields:
• 6 CO2
• 10 NADH
• 2 FADH2
• 4 ATP

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Figure 9.9 The Citric Acid Cycle

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In-Text Art, Ch. 9, p. 178 (2)

For the citric acid cycle to continue the starting molecules (acetyl
CoA and oxidized electron carriers) must be replenished.
The electron carriers are reduced and they must be reoxidized.

Oxygen (O2) becomes reduced as NADH becomes oxidized,

so that another molecule of glucose can be catabolized.
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Figure 9.8 The Respiratory Chain and ATP Synthase Produce
ATP by a Chemiosmotic Mechanism (Part 1)
Oxidative phosphorylation: ATP is synthesized
by reoxidation of electron carriers in the
presence of O2.

– Two stages:
• Electron transport
• Chemiosmosis

– Chemiosmotic ATP synthesis:

Chemiosmosis converts the potential
energy of a proton concentration gradient
to chemical energy in ATP.

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Figure 9.7 The Oxidation of NADH and FADH2 in the Respiratory
Chain

The electron transport chain is located in

the folded inner mitochondrial membrane.

Electron transport:
Electrons from NADH and FADH2
pass through the respiratory
chain of membrane-associated
complexes.
Energy is released as electrons
are passed between carriers.
Electron flow results in a proton
concentration gradient across
the inner mitochondrial membrane.
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 Redox forms of coenzyme Q. CoQ oxidized form (ubiquinone)
can be reduced to ubiquinol (CoQH2) by two steps of one
electron each through semiquinone form, or by one reaction of
two electrons, without the semiquinone intermediate.
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 The electron transport chain

Is the movement of
electrons through the
electron transport chain
spontaneous?

What is the driving force?

Redox potential is a measure of the tendency of a chemical

species to acquire electrons from or lose electrons to an
electrode and thereby be reduced or oxidized respectively.
A species with a higher reduction potential possesses a
higher tendency to acquire electrons and be reduced.

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9.3 Oxidative Phosphorylation Forms ATP
The Respiratory Chain and ATP Synthase Produce ATP by a
Chemiosmotic Mechanism
Protons are actively transported into the intermembrane space
during electron transport, where H+s accumulate and create:
- a proton concentration gradient
- charge difference, an electrical membrane potential
difference.
Δp= Δ ψ−Z ΔpH
• This potential energy is called the proton-motive force, wich
is an electrochemical gradient generated by the
movement of protons across a membrane, which drives
ATP synthesis in biological systems.
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Figure 9.8 The Respiratory Chain and ATP Synthase Produce
ATP by a Chemiosmotic Mechanism
Cytoplasm Outer
mitochondrial
membrane

Intermembrane
space (high H+
Cytochrome c
concentration and Cytochrome c oxidase
Cytochrome c
postive charge) NADH-Q Ubiquinone reductase
reductase

IV
III
I

Inner
mitochondrial
membrane I
I

Matrix of mitochondrion
(low H+ concentration and
negative charge)

This
whichcreates
Electrons an protons
(carried
transport imbalance
(H+of
by NADH H+—and
) and
out the thus
FADH
of a charge
2) from
matrix difference—between
glycolysis
to and the citric
the intermembrane acid the
space. cycle “feed” the electron
intermembrane space and
carriers…
the
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University Press
9.3 Oxidative Phosphorylation Forms ATP
ATP synthase is conserved in all living
organisms.
It is a molecular motor with two parts:
• F0 unit— it has a transmembrane
H+ channel and has an attached
central stalk. Both parts rotate as
protons flow through the H+
channel.
• F1 unit—projects into the matrix;
has 3 active sites for ATP
synthesis.
• The movement of protons is
coupled to the formation of ATP in
the F1 unit.

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Figure 9.8 The Respiratory Chain and ATP Synthase Produce
ATP by a Chemiosmotic Mechanism

Because of the proton-motive force,

protons return to the matrix by
passing through the H+ channel of High
ATP synthase (the F0 unit). concentration of
H+

When H+ diffuses through the ATP

synthase
channel, the potential energy is
converted to kinetic energy, causing
the central polypeptide to rotate. F0 unit

The energy is transmitted to the

catalytic subunits of F1, resulting in
Low
ATP synthesis. F1 unit
concentration of
H+

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 In chemiosmosis, the proton-motive force - a proton (H+)
concentration gradient and an electric potential (voltage gradient)
across the membrane - drives an energy-requiring process such
as ATP synthesis

e e
- -

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 A human ATP synthase consists of 29
polypeptide chains of 18 different subunits.

The monomeric F1Fo-ATPas from mammalian

mitochondria consist of:
- a soluble F1 head domain (subunits α3β3),
the site of ATP synthesis
- a membrane embedded Fo domain (several
c subunits forming a ring structure),
Fo being responsible for proton
translocation.

These two domains are linked by a central

stalk (subunits γ, δ, and ε) rotating inside the
F1 region and a stationary peripheral stalk
(subunits ab2).

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 As the central stalk (shown as a green subunit in the
figure) rotates inside the F1 region it induces
conformational changes in the three (3 and 3 β
subunits) F1 subunits resulting in ATP synthesis.

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Number of protons pumped by 2 electrons as they move
through the electron transport chain

1 NADH generates: 10 H+, and 10/4 H+ per ATP = 2.5 ATP per NADH
For every 4 H+ ions
1 FADH2 generates: 6 H+, and 6/4 H+ per ATP = 1.5 ATP per FADH2
1 ATP is produced
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9.3 Oxidative Phosphorylation Forms ATP

In principle enzyme activity is reversible!

ATP synthase can also act as ATPase, hydrolyzing ATP to
ADP and Pi.
Why is ATP synthesis favored?
• ATP leaves the matrix as soon as it is made, keeping
ATP concentration in the matrix low.
• H+ gradient is maintained by active transport.

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9.3 Oxidative Phosphorylation Forms ATP
In brown fat cells, a protein called UPC1 inserts into the
mitochondrial membrane and makes it permeable to protons.
This uncouples electron transport and chemiosmosis, and
energy is released as heat rather than being trapped in ATP.

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9.3 Oxidative Phosphorylation Forms ATP
Many bacteria and archaea have evolved pathways that allow
them to exist where O2 is scarce or absent, by using other
electron acceptors— anaerobic respiration.

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9.4 In the Absence of Oxygen, Some Energy Is Harvested from
Glucose
Absence of O2:
- some ATP can be made by
glycolysis and fermentation.
Fermentation:
- occurs in the cytoplasm.
- glucose is only partially oxidized
- 2 ATP per glucose are produced
by substrate-level phosphorylation
in glycolysis.
- NAD+ is regenerated to keep
glycolysis going.

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9.4 In the Absence of Oxygen, Some Energy Is Harvested from
Glucose
• Lactic acid fermentation
• Lactic acid fermentation uses NADH as a
reducing agent to reduce pyruvate to
lactate, thus regenerating NAD+ to keep
glycolysis operating.
• Lactate dehydrogenase catalyzes
fermentation
! • Pyruvate is the electron acceptor and
lactate is the product.
• Occurs in microorganisms and in
vertebrate muscle cells.

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Concept 9.4 In the Absence of Oxygen, Some Energy Is
Harvested from Glucose (3)

In large vertebrate muscle cells during intense exercise,

O2 cannot be delivered fast enough for aerobic
respiration.
Muscle cells then break down glycogen and carry out
lactic acid fermentation.
When lactate builds up, the increase in H+ lowers pH and
causes muscle pain.

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9.4 In the Absence of Oxygen, Some Energy Is Harvested from
Glucose
Alcoholic fermentation
• Requires two enzymes to metabolize
pyruvate to ethanol.
• In alcoholic fermentation, pyruvate from
glycolysis is converted first into
acetaldehyde, and CO2 is released.
• NADH from glycolysis is used to reduce
acetaldehyde to ethanol, thus regenerating
NAD+ to keep glycolysis operating.
!
• Yeasts

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Figure 9.12 Cellular Respiration Yields More Energy Than
Fermentation
Cellular respiration yields more
energy than fermentation.
– Glycolysis plus fermentation =
2 ATP/glucose
– Glycolysis plus cellular
respiration = 32 ATP/glucose
Glucose is only partially oxidized in
fermentation, more energy remains
in the products.

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Figure 9.13 Relationships among the Major Metabolic Pathways
of the Cell

Metabolic pathways do not

operate in isolation.
Many pathways share
intermediate molecules.
Pathways are regulated by
enzyme inhibitors.

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9.5 Metabolic Pathways Are Interrelated and Regulated
Catabolic interconversions:
• Polysaccharides are hydrolyzed to
glucose → enters glycolysis
• Lipids are broken down to

 glycerol → DHAP → glycolysis

 fatty acids → acetyl CoA → citric
acid cycle
• Proteins are hydrolyzed to amino
acids, which feed into glycolysis or
the citric acid cycle.

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Beta oxidation of fatty acids

Cytric acid cycle

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In-Text Art, Ch. 9, p. 190

Example: Glutamate is converted into α-ketoglutarate, an

intermediate in the citric acid cycle.

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9.5 Metabolic Pathways Are Interrelated and Regulated
• Anabolic interconversions
• Most catabolic reactions are reversible.

• Gluconeogenesis: Glucose formed from citric acid cycle

and glycolysis intermediates.
• Acetyl CoA can be used to form fatty acids.

• Citric acid cycle intermediates can be used to synthesize

nucleic acid components:
• α-ketoglutarate → purines
• Oxaloacetate → pyrimidines

• α-ketoglutarate is also a starting point for synthesis of the

amino acid glutamate.
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Glyconeogenesis

Gluconeogenesis is the pathway by

which glucose is formed from non-hexose
precursors such as glycerol, lactate,
pyruvate, and glucogenic amino acids.

Gluconeogenesis is essentially the

reversal of glycolysis.

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 Anabolic interconversions

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 Regulation

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9.5 Metabolic Pathways Are Interrelated and Regulated

How do cells “decide” which pathways to use?

Levels of substances in the metabolic pool are quite
constant.
Organisms regulate enzymes to maintain balance
between catabolism and anabolism.
Several mechanisms regulate the rate of each step in
a biochemical pathway.

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9.5 Metabolic Pathways Are Interrelated and Regulated

• Change the amount of active

enzyme by regulating gene
expression
• Change enzyme activity by
covalent modifications, such as
phosphorylation
• Feedback inhibition by allosteric
enzymes
• Substrate availability: if the
substrate of a particular enzyme is
used up by another pathway, the
first enzyme can no longer function
and the pathway shuts down.

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 Entry of pyruvate to fuel the citric acid cycle
leading to the aerobic production of ATP and intermediates for
biosynthesis is a key metabolic step.

Hence both the pyruvate dehydrogenase

complex and key enzymes in the cycle are targets for
regulation.

This occurs through:

- substrate availability,
- product inhibition,
- allosteric effectors,
- post-translational modifications of key enzymes in the
pathway.

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Concept 9.5 Metabolic Pathways Are Interrelated and Regulated (8)

Glycolysis and the citric acid cycle are subject to allosteric

regulation of key enzymes.
A high concentration of the final product can inhibit an enzyme
that catalyzes an earlier reaction.
An excess of product of one pathway can activate an enzyme
in another pathway.

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Figure 9.16 Allosteric Regulation of Glycolysis and the Citric Acid
Cycle (Part 1)

• The main control point in glycolysis

is phosphofructokinase enzyme,
which is inhibited by ATP.
• In fermentation, phosphofructo-
kinase operates at a high rate to
produce ATP.
• If O2 is present, more ATP is
produced, which inhibits the
enzyme and slows down
glycolysis.

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Figure 9.16 Allosteric Regulation of Glycolysis and the Citric Acid
Cycle (Part 2)

• The main control point in the citric

acid cycle is isocitrate
dehydrogenase enzyme
(step 3).
• It is inhibited by NADH and ATP;
if too much of either accumulates,
the citric acid cycle shuts down.

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9.5 Metabolic Pathways Are Interrelated and Regulated

Acetyl CoA is another control

point.

• If ATP levels are high and the

citric acid cycle shuts down,
accumulation of citrate
activates fatty acid synthesis
from acetyl CoA, diverting it to
storage.

• Fatty acids may be metabolized

later to produce more acetyl
CoA.

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Lecture

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Seminar

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