Cellular Respiration and

Fermentation

by Biological Science
Opening Roadmap
What Happens When Glucose Is Oxidized?

C6H12O6 + 6 O2 → 6 CO2 + 6 H2O + Heat and light

glucose oxygen carbon water
dioxide

§ In cells, glucose is oxidized through a long series of

carefully controlled redox reactions
§ The released free energy is used to synthesize
ATP
§ These reactions comprise cellular respiration
 Energy conversion
Photosynthesis
CO2 + H2O + sunlight

(CH2O)n

Energy storage

Glucose Starch, glycogen, fats

(synthesized from
glucose)

Energy use

Cellular Respiration Fermentation If there is no oxygen

Glucose + O2 + ADP + Pi Glucose + ADP + Pi
in the enviroment?
CO2 + H2O + ATP Small organic molecules + ATP © 2017 Pearson Education, Ltd.
What Happens When Glucose Is Oxidized?

Two fundamental requirements of cells:

1. Energy to generate ATP
2. A source of carbon to use as raw materials
for synthesizing macromolecules
 Cellular Respiration

§ Cellular respiration is a set of four processes:

1. Glycolysis
2. Pyruvate processing
3. Citric acid cycle
4. Electron transport and oxidative phosphorylation
 Cellular Respiration Oxidizes Glucose to Make ATP

NADH
NADH NADH
FADH2
Electron transport chain
CITRIC establishes proton gradient
Glucose Pyruvate Acetyl CoA ACID CO2 that is used to produce ATP
CYCLE
(two for (two for
every glucose) every glucose) O2 H2 O

ATP CO2 ATP ATP

1. Glycolysis 2. Pyruvate 3. Citric Acid Cycle 4. Electron Transport and
Processing Oxidative Phosphorylation
Occurs in: Inner membrane of
Cytosol of eukaryotes and Matrix of mitochondria or cytosol mitochondria or plasma
prokaryotes of prokaryotes membrane of prokaryotes

What goes in:

What comes out:

Homework

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Catabolic Pathways Break Down a
Variety of Molecules
§ For ATP production, cells first use!
§ Carbohydrates
§ Then fats
§ And finally proteins
 Cellular Respiration Interacts with Other
Catabolic and Anabolic Pathways
Carbohydrates Fats and phospholipids Proteins

Sugars Glycerol Fatty acids Amino acids

Catabolic
NH3
pathways

CITRIC
Glucose Pyruvate Acetyl CoA ACID
GLYCOLYSIS CYCLE

Anabolic Lactate Fatty acids

pathways (from fermentation)

Glycogen Substrates for Substrates for amino

nucleotide synthesis Phospholipids Fats
or starch acid synthesis
Homeostasis
§ Metabolism comprises thousands of different
chemical reactions
§ Organizing them into pathways allows them
to be regulated
§ Maintains a stable internal environment
even under different environmental
conditions
§ Referred to as homeostasis
 Lipid
Carbohydrate metabolism
metabolism

Nucleotide
metabolism
Amino acid
metabolism

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Four Steps of Cellular Respiration

§ What goes in?

§ What comes out?
§ What happens to the energy that is
released?
§ Where does each step occur?
§ How is it regulated?
1)Glycolysis
§ Glycolysis is a series of 10 chemical reactions that
occur in the cytosol
§ Three key points:
1. Glycolysis starts by using two ATP in the energy
investment phase (reactions 1–5)
2. During the energy payoff phase (reactions 6–10),
NADH is made and ATP is produced by
substrate-level phosphorylation
3. The net yield is two NADH, two ATP, and two
pyruvate
 Substrate-Level Phosphorylation Involves
an Enzyme and a Phosphorylated Substrate

ATP
ADP
Enzyme

Phosphorylated
substrate

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 All 10 reactions
of glycolysis occur
in the cytosol investment phase
Dihydroxyacetone phosphate
What goes in: ATP ATP

Enzyme
Glucose Glucose- Fructose- Fructose-
6-phosphate 6-phosphate 1,6-bisphosphate

Glycolysis begins with an

energy-investment phase: 2 ATP→ 2 ADP
What comes out: ADP ADP Glyceraldehyde-3-phosphate

The “2” indicates that fructose-1,6-

bisphosphate has been split into two
3-carbon sugars (only one is shown) payoff phase

2 NAD + 2 ADP 2 ADP

1,3-Bisphosphoglycerate 3-Phosphoglycerate 2-Phosphoglycerate Phosphoenolpyruvate Pyruvate

During the energy-payoff phase,

4 ATP are produced for a net gain of 2 ATP
2 NADH +2 H + 2 ATP 2 ATP

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© 2017 Pearson Education, Ltd.
How Is Glycolysis Regulated?
§ Glycolysis is regulated by feedback inhibition
§ Reaction 3:
Ø Phosphofructokinase (PFK), an allosteric enzyme, is
inhibited by high levels of ATP and activated by
high levels of ADP and AMP.
Ø The most important control point! All 10 reactions
of glycolysis occur
in the cytosol investment phase
Dihydroxyacetone phosphate
What goes in: ATP ATP

Enzyme
Glucose Glucose- Fructose- Fructose-
6-phosphate 6-phosphate 1,6-bisphosphate

Glycolysis begins with an

energy-investment phase: 2 ATP→ 2 ADP
What comes out: ADP ADP Glyceraldehyde-3-phosphate
 Phosphofructokinase Has Two Binding Sites for ATP

When ATP binds here, the

ATP at reaction rate slows dramatically
regulatory
site

Fructose-6-
phosphate ATP at
at active site active site

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Regulation of Glycolysis
The other control points are:
§ Reaction 1:
§ Hexokinase is inhibited by the reaction product; high
levels of glucose-6-phosphate, which prevents the
phosphorylation of glucose.

§ Reaction 10:
§ Pyruvate kinase, another allosteric enzyme is inhibited by
high levels of ATP or acetyl CoA.
 Hormonal Control
§ In well-fed state;
insulin initiates an increase in the of
glucokinase,
phosphofructokinase, and
pyruvate kinase

§ Conversely in fasting or diabetes;

Synthesis of these enzymes are
decreased since glucagon is high
and insulin is low

20
 2)Processing Pyruvate to Acetyl CoA

§ Pyruvate processing takes place inside an enormous

enzyme complex called pyruvate dehydrogenase
Cristae are sacs of inner
membrane joined to the
rest of the inner membrane
by short tubes

Mitochondrial
matrix
Cristae

Inner
membrane
Intermembrane
space
Outer membrane

100 nm

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 Pyruvate Is Oxidized to Acetyl CoA

Coenzyme A
NAD+ (CoA — SH)

CO2

Pyruvate NADH Acetyl CoA

pyruvate dehydrogenase

Regulatory point in glucose oxidation!

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 3) The Citric Acid Cycle

§ The citric acid cycle is located in the mitochondrial matrix

 The two blue carbons enter
the cycle via acetyl CoA

All 8 reactions
of the citric acid
NADH cycle occur in
the mitochondrial
matrix
Acetyl CoA Citrate Isocitrate
In each turn of
the cycle, the
two red carbons
are converted to
CO2

α-Ketoglutarate

Oxaloacetate
NADH
The CITRIC ACID CYCLE
runs twice for each
glucose molecule oxidized
NADH

In the next cycle, this

blue carbon becomes
a red carbon Succinyl CoA
Malate

ATP

GTP
Each reaction is catalyzed
by a different enzyme Succinate
Fumarate
FADH2

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The Citric Acid Cycle: Oxidizing Acetyl CoA
to CO2
§ In the citric acid cycle, each acetyl CoA from
pyruvate processing is oxidized into two CO2
§ The citric acid cycle is located in the
§ Mitochondrial matrix in eukaryotes
§ Cytosol in prokaryotes
§ The reactions are organized in a cycle
§ Starts by moving the acetyl group from acetyl CoA
to oxaloacetate to form citrate
§ At the end, oxaloacetate is regenerated
The Citric Acid Cycle: Oxidizing Acetyl
CoA to CO2
§ Some of the potential energy released is used to
1. Reduce three NAD+ to NADH
2. Reduce one FAD to FADH2
3. Phosphorylate ADP (or GDP) to form ATP (or
GTP)
§ The cycle turns twice for each glucose molecule,
since two pyruvate are produced by glycolysis
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 How Is the Citric Acid Cycle
Regulated?
The Citric Acid Cycle Is Regulated by Feedback Inhibition

These steps are

also regulated via
feedback inhibition,
This step is by NADH and ATP
regulated by ATP Citrate

Acetyl CoA
Oxaloacetate

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 6 NADH
2 NADH 2 NADH 2 FADH2

GLYCOLYSIS PYRUVATE PROCESSING CITRIC

Glucose 2 Pyruvate 2 Acetyl CoA ACID 4 CO2
CYCLE

2 CO2
2 ATP 2 ATP

Cytosol Mitochondrial matrix

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 What Happens to the NADH and FADH2?
GLYCOLYSIS PYRUVATE CITRIC ACID CYCLE
Free-energy change relative to glucose (kcal/mol)

PROCESSING
ATPATP

2 NADH
Glucose 2 ATP
2 ATP

2 NADH
Pyruvate

2 NADH
Acetyl CoA

2 NADH
In each of these drops, 2 ATP
energy is transferred to
energy-storing molecules
2 FADH2
ATP, NADH, or FADH2

2 NADH
Oxaloacetate

© 2017 Pearson Education, Ltd. Oxidation of glucose →

§ Two questions remain:
1. What happens to the energy that is released as
electrons are transferred?
2. How is this energy used to make ATP?
 Electron Transport Chain
§ The molecules that oxidize NADH
and FADH2 are called the
electron transport chain (ETC)
ETC and Oxidative Phosphorylation
Electron Transport and Chemiosmosis: Building
a Proton Gradient to Produce ATP

§ Molecules in the inner mitochondrial

membrane could cycle between oxidized and
reduced states
§ What are these molecules, and how do
they work?
The Electron Transport Chain
(ETC)
§ They have different ability to accept electrons, called
their redox potential: Complex I–IV

As electrons move from one molecule to another in

the ETC
§ Electrons move from the highest redox
potential complex to the lowest redox
potential.
 The Electron Transport Chain
§ Inner mitochondrial membrane can be disrupted into five separate
protein complexes;
Ø Complexes I, II, III, IV (and V)
Electron Carriers of ETC
§ NADH
§ Flavoproteins
§ Coenzyme Q (Ubiquinone)
§ Cytochromes
§ Iron-sulfur proteins
 NADH

Complexes I–IV FADH2

Complex I
Complex

Free-energy change relative to O2 (kcal/mol)

ETC reactions
II
take place in the
inner membrane
and cristae of the
mitochondrion

Complex III

Complex
IV

FMN: flavin-containing prosthetic

group in flavoprotein
Fe•S: protein with an iron–sulfur
Requirement for oxygen cofactor
makes the electron transport Cyt: protein with a heme
prosthetic group
process the respiratory
Q: ubiquinone, a nonprotein
chain, which accounts for the coenzyme
greatest portion of the body’s
use of oxygen.
Reduction-oxidation reactions →
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© 2017 Pearson Education, Ltd.
 Proton electrochemical gradient
The electron transport
chain occurs in the
inner membrane
of the mitochondrion

Intermembrane
space

Complex
Inner Complex IV
membrane I

Complex
II Complex
III

Mitochondrial
matrix

NADH FADH2 O2

Complex I Complex II Complex III Complex IV

What goes in:
What comes out:

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e- Transport Coupling and Oxidative
Phosphorylation
This coupling was a mystery for many years

§ Many biochemists squandered careers searching for

the elusive "high energy intermediate”.
§ Peter Mitchell proposed a novel idea -a proton
gradient (chemiosmosis) across the inner membrane
could be used to drive ATP synthesis.
 ATP Synthase and the Chemiosmosis
Hypothesis

Intermembrane
space

Fo unit

Stator Rotor Mitochondrial

shaft matrix

F1 unit

ADP + Pi
ATP
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 The discovery of ATP
synthase
Knobs project
“inside-out” of
mitochondrial
membrane

50 nm

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 Chemiosmotic model
§ H+ make inner mitochondria acidic.
§ Produces different proton gradient.
§ H+ pass through ATP synthase
ATP Yield from Cellular
Respiration
§ The energy to produce ATP comes from a proton
gradient: oxidative phosphorylation
§ Substrate-level phosphorylation occurs during
glycolysis and the citric acid cycle
Web Activity: Glucose Metabolism
Aerobic vs Anaerobic Respiration
§ Oxygen as the final electron acceptor for the ETC:
aerobic respiration

§ In oxygen-poor environments other electron

acceptors: anaerobic respiration
NAD+ Recovery

§ Two major mechanisms for oxidizing NADH are:

1) the NADH-linked conversion of pyruvate to lactate
(anaerobic)
2) oxidation of NADH via the respiratory chain
(aerobic)

48
Fermentation
§ Fermentation is a metabolic pathway that regenerates
NAD+ from NADH

ØIf the cells are lack of mitochondria such as in lens and

cornea of the eye, leukocytes and red blood cells.

49
 (a) Lactic acid fermentation occurs in humans.

2 ADP + 2 Pi 2 ATP

Glucose

2 NAD+ 2 NADH 2 Pyruvate

No intermediate;
pyruvate accepts
electrons from NADH
2 Lactate

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 (b) Alcohol fermentation occurs in yeast.

2 ADP + 2 Pi 2 ATP

Glucose

2 NAD+ 2 NADH 2 Pyruvate

2 Ethanol 2 Acetylaldehyde

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Pathways for Pyruvate: Summary