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HMM/SCM1414_Biology 1

CHAPTER 7
RESPIRATION
• Living cells require energy from outside
sources to perform tasks.
• Energy enters most ecosystems as sunlight
and leaves as heat.
(Figure 9.2, Campbell, page 160)
• Photosynthesis generates O2 and organic
molecules that mitochondria of eukaryotes
use as fuel for cellular respiration.
• Cells harvest chemical energy stored in
organic molecules and use it to regenerate
ATP, the molecule that drives most cellular
work.
• 3 pathways of respiration: glycolysis, citric
acid cycle, and oxidative phosphorylation.

7.1 ATP (Adenosine Triphosphate)

• Immediate source of energy that drives


most cellular work.
• Cells manage their energy resources to do
this work by energy coupling - use of an
exergonic process to drive an endergonic
one.

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• Endergonic
Endergonic Reaction - reaction that ends
with a net gain in energy
○ Products have more energy than

reactants, e.g., photosynthesis


• Exergonic Reaction - reaction that ends
with a net loss in energy
○ Reactants have more energy than

products, e.g., cellular respiration

Structure and hydrolysis of ATP

• ATP = Nucleotide with unstable phosphate


bonds that cell hydrolyzes for energy to
drive endergonic reactions.
• Consists of adenine, ribose, & chain of
three phosphate groups.

• Unstable bonds between phosphate groups


can be hydrolyzed in an exergonic reaction.

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○ When terminal phosphate bond is


hydrolyzed, a phosphate group is removed
producing ADP (adenosine diphosphate).
ATP + H2O → ADP + Pi
○ Under standard lab conditions, reaction

releases -31 kJ/mol (-7.3 kcal/mol).


○ In living cell, reaction releases -55 kJ/mol

(-13 kcal/mol) - 77% more than under


standard conditions.
• Terminal phosphate bonds of ATP are
unstable, so:
○ Products of hydrolysis reaction are more

stable than reactants.


○ Hydrolysis of phosphate bonds is thus

exergonic as system shifts to a more


stable state.

How ATP performs work

• Exergonic hydrolysis of ATP is coupled with


endergonic processes by transferring a
phosphate group to another molecule.
° Phosphate transfer is enzymatically
controlled.
° Molecule receiving phosphate
(phosphorylated or activated
intermediate) becomes more reactive.

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• Example, conversion of glutamic acid to


glutamine:

Glu + NH3 → Gln


(Glutamic acid) (Ammonia) (Glutamine)
∆G = +14.2 kJ/mol (+3.4 kcal/mol)
(endergonic)

• Two step process of energy coupling with


ATP hydrolysis:
1. Hydrolysis of ATP and phosphorylation of
glutamic acid.
Glu + ATP → Glu-(Pi) + ADP
Unstable
phosphorylated
intermediate
2. Replacement of the phosphate with the
reactant ammonia.
Glu-(Pi) + NH3 → Gln + (Pi)
Overall ∆G:
Glu + NH3 → Gln ∆G = +14.2 kJ/mol
ATP → ADP + Pi ∆G = -31.0 kJ/mol
Net ∆G = -16.8 kJ/mol
(Overall process is exergonic)

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The regeneration of ATP

• ATP is continually regenerated by cell.


° Process is rapid (107 molecules used and
regenerated/sec/cell).
° Reaction is endergonic.

ADP + Pi → ATP
∆G = + 31 kJ/mol (+7.3 kcal/mol)

° Energy to drive endergonic regeneration


of ATP comes from exergonic process of
cellular respiration.

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7.2 Aerobic
Aerobic Respiration

Preview of cellular respiration


(See Figure 9.6, Campbell, page 164)

• Stages of respiration: glycolysis, citric acid


cycle, and electron transport chain and
oxidative phosphorylation.

• Glycolysis - in cytoplasm.
° Glucose broken down into two molecules
of pyruvate.
• Citric acid cycle - in mitochondrial matrix.
° Completes breakdown of glucose by
oxidizing a derivative of pyruvate to CO2.

• Several steps in glycolysis and citric acid


cycle are redox reactions - dehydrogenase
enzymes transfer electrons from substrates
to NAD+, forming NADH.
• NADH passes electrons to electron
transport chain (ETC).
• Electrons then move from molecule to
molecule until they combine with molecular
O2 and H+ to form water.

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• As they are passed along chain, energy


carried by electrons is transformed in
mitochondrion into a form that can be used
to synthesize ATP via oxidative
phosphorylation.

• Inner membrane of mitochondrion is site of


electron transport and chemiosmosis,
processes that together constitute
oxidative phosphorylation.
° Oxidative phosphorylation produces
almost 90% of ATP generated by
respiration.
• Some ATP is formed directly during
glycolysis and citric acid cycle by
substrate-
substrate-level phosphorylation.
° Enzyme transfers phosphate group from
an organic substrate to ADP, forming
ATP. (See Figure 9.7, Campbell, page 164)

• For each molecule of glucose degraded to


CO2 & H2O by respiration, cell makes up to
38 ATP, each with 7.3 kcal/mol of free
energy.
• Respiration uses small steps in respiratory
pathway to break large denomination of
energy contained in glucose into the small
change of ATP.

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° Quantity of energy in ATP is more


appropriate for level of work required in
cell.

7.2.1 Glycolysis
(See Figure 9.9, Campbell, page 166 - 167)

• Glucose is split into two 3C sugars.


• 3C sugars oxidized and rearranged to form
two molecules of pyruvate, ionized form of
pyruvic acid.
• Two phases of glycolysis:
(Figure 9.8, Campbell, page 165)

1. Energy investment phase


° Cell invests ATP to provide activation
energy by phosphorylating glucose.
° Requires 2 ATP per glucose.
2. Energy payoff phase
° ATP produced by substrate-level
phosphorylation.
° NAD+ reduced to NADH by electrons
released by oxidation of glucose.

• Net yield from glycolysis is 2 ATP and 2


NADH per glucose.
° No CO2 is produced during glycolysis.
• Glycolysis can occur in presence or
absence of O2.

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The Glycolytic Pathway


(Figure 9.9, Campbell, page 166 – 167)
http://www.db.uth.tmc.edu/faculty/alevine/1521_2000/glydetail.htm

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• Overall reaction showing all reactants and


products resulting from glycolysis.

Glucose + 2ATP + 2 Pyruvate + 2ADP +


2Pi + 4ADP + 2NAD+ 2ATP + 2NADH + 2H+ +
2H2O

• Equation showing net reaction of glycolytic


pathway.

Glucose + 2Pi + 2 Pyruvate + 2ATP +


2ADP + 2NAD+ 2NADH + 2H+ + 2H2O

7.2.2 Pyruvate oxidation


(Figure 9.10, Campbell, page 168)

• More than three-quarters of original energy


in glucose is still present in the two
molecules of pyruvate.
• If O2 is present, pyruvate enters
mitochondrion where enzymes of citric acid
cycle complete its oxidation to CO2.
• After pyruvate enters mitochondrion via
active transport, it is converted to acetyl
coenzyme A (acetyl
acetyl CoA).
CoA

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• This is accomplished by multienzyme


complex that catalyzes three reactions:

1. Carboxyl group is removed as CO2.


2. Remaining 2C fragment is oxidized to
acetate. An enzyme transfers the pair of
electrons to NAD+ to form NADH.
3. Acetate combines with coenzyme A to
form the very reactive molecule acetyl
CoA.

• Acetyl CoA is now ready to feed its acetyl


group into the citric acid cycle for further
oxidation.

7.2.3 The Krebs cycle/ Citric Acid Cycle


(Figure 9.12, Campbell, page 169)

• Cycle oxidizes organic fuel derived from


pyruvate.
° Acetyl group of acetyl CoA joins cycle by
combining with oxaloacetate, forming
citrate.
° Citrate regeneraterd back (via several
steps) to OAA.
° Three CO2 molecules are released,
including the one released during the
conversion of pyruvate to acetyl CoA.

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• Cycle generates one ATP per turn by


substrate-level phosphorylation.
° GTP molecule is formed by substrate-
level phosphorylation.
° GTP used to synthesize an ATP, the only
ATP generated directly by cycle.
• Most of chemical energy is transferred to
NAD+ and FAD during redox reactions.
• Reduced coenzymes NADH and FADH2 then
transfer high-energy electrons to electron
transport chain.
• Each cycle produces one ATP by substrate-
level phosphorylation, three NADH, and one
FADH2 per acetyl CoA.

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http://chemistry.gsu.edu/glactone/PDB/Proteins/Krebs/Krebs.html

Summary:

Acetyl CoA + 3 NAD+ 2 CO2 + CoA-SH +


+ FAD + ADP + Pi 3NADH + 3H+ +
+2H2O FADH2 + ATP

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7.2.4 Electron transport chain (oxidative


phosphorylation)

• Only 4 of 38 ATP produced by respiration of


glucose are produced by substrate-level
phosphorylation:
° Glycolysis – 2 ATP.
° Citric acid cycle – 2 ATP.
• NADH and FADH2 account for the majority
of energy extracted from food.
° These reduced coenzymes link glycolysis
and citric acid cycle to oxidative
phosphorylation, which uses energy
released by electron transport chain
(ETC) to power ATP synthesis.

The Pathway of Electron Transport


(Figure 9.13, Campbell, page 171)

• ETC is a collection of molecules embedded


in cristae.
° Most components of ETC are proteins
bound to prosthetic groups.
• Electrons drop in free energy as they pass
down ETC.

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• During electron transport along ETC,


electron carriers alternate between
reduced and oxidized states as they accept
and donate electrons.
° Each component of chain becomes
reduced when it accepts electrons from
its “uphill” neighbor, which is less
electronegative.
° It then returns to its oxidized form as it
passes electrons to its more
electronegative “downhill” neighbor.
• Electrons carried by NADH are transferred
to the first molecule in ETC, a flavoprotein.
• Electrons continue along chain that
includes several cytochrome proteins and
one lipid carrier.
° Prosthetic group of each cytochrome is a
heme group with an iron atom that
accepts and donates electrons.
• Last cytochrome of chain, cyt a3, passes its
electrons to oxygen, which is very
electronegative.
° Each oxygen atom also picks up a pair of
H+ from aqueous solution to form water.
° For every two electron carriers (four
electrons), one O2 molecule is reduced to
two molecules of water.

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• Electrons carried by FADH2 have lower free


energy and are added at a lower energy
level than those carried by NADH.
° ETC provides about one-third less energy
for ATP synthesis when electron donor is
FADH2 rather than NADH.
• ETC generates no ATP directly.
• Its function is to break the large free
energy drop from food to oxygen into a
series of smaller steps that release energy
in manageable amounts.

• How does the mitochondrion couple


electron transport and energy release to
ATP synthesis?
 Chemiosmosis.

Chemiosmosis: Energy-
Energy-Coupling Mechanism
(Figure 9.14, Campbell, page 171)

1. NADH delivers two electrons and two


protons to the first protein complex (I) in
cytochrome system located in cristae.
2. Complex uses energy released from
electrons to actively pump H+ from matrix
to inter-membrane space.

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3. Process is done two more time times.


Electrons are passed through two more
protein complexes (III & IV) which
transport two more H+ across membrane.
4. After passing through three protein
complexes, electrons combine with one
oxygen atom and two H+ to form water.
5. This transport across membrane
produces a concentration gradient with
more H+ on one side of membrane than
the other. The H+ gradients that results is
referred to as a proton-
proton-motive force.
force This
gradient is used to make ATP.
6. Cristae membrane is very impermeable to
H+ except through a special protein called
ATP synthase. As protons pass through
this protein, energy is obtained to make
ATP from ADP & Pi.
7. FADH2 delivers its electron via protein
complex II and so results in fewer
electrons being into the inter-membrane
space.

• Oxidative phosphorylation = Electron


transport + chemiosmosis.

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7.2.5 Calculations of Total ATP Production


Production by
Cellular Respiration

• Products generated when cellular


respiration oxidizes a molecule of glucose
to six CO2 molecules:
(Figure 9.16, Campbell, page 173)

Conversions

• NADH in cytoplasm produces 2 or 3 ATP by


oxidative phosphorylation depending on
shuttle system used to transport electrons
from cytosol into mitochondrion:
○ If electrons are passed to FAD, e.g. brain

cells, 2 ATP are produced.


+
○ If electrons are passed to NAD , e.g. liver

cells & heart cells, 3 ATP are produced.

• NADH in mitochondria produces 3 ATP.

• FADH2 adds its electrons to the electron


transport system at a lower level than
NADH, so it produces 2 ATP.

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1. Glycolysis

• Substrate-level phosphorylation = 2 ATP


• 2 NADH = 2 x 2 ATP = 4 ATP; or
= 2 x 3 ATP = 6 ATP

2. Formation of Acetyl CoA

• 2 NADH = 2 x 3 ATP = 6ATP

3. Krebs Cycle

• 6 NADH = 6 x 3 ATP = 18 ATP


• 2 FADH2 = 2 x 2 ATP = 4 ATP
• Substrate-level phosphorylation = 2 ATP

4. Total Yield

Glycolysis = 2 ATP
Aerobic respiration = 34 or 36 ATP

5. Summary

Pathway Substrate-level Oxidative Total


phosphorylation phosphorylation ATP
Glycolysis 2 ATP 2 NADH = 4-6 ATP 6-8
Coa 2 NADH = 6 ATP 6
Krebs 2 ATP 6 NADH = 18 ATP 24
cycle 2 FADH2 = 4 ATP
Total 4 ATP 32 – 34 ATP 36-38

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• How efficient is respiration in generating


ATP?

° Complete oxidation of glucose releases


686 kcal/mol.
° Phosphorylation of ADP to form ATP
requires at least 7.3 kcal/mol.
° Efficiency of respiration
= 7.3 kcal/mol x 38 ATP/glucose x 100%
686 kcal/mol glucose
= 40%.
° ≈ 60% of energy from glucose lost as
heat.
 Some of that heat is used to maintain
our high body temperature (37°C).
• Cellular respiration is remarkably efficient
in energy conversion.

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7.3 Anaerobic respiration

• Without oxygen, oxidative phosphorylation


ceases.
• Through fermentation, some cells can
oxidize organic fuel and generate ATP
without use of O2.

• Anaerobic catabolism of sugars can occur


by fermentation.
• Fermentation generate ATP from glucose
by substrate-level phosphorylation as long
as there is a supply of NAD+ to accept
electrons.
° If NAD+ pool is exhausted, glycolysis
shuts down.
° Under aerobic conditions, NADH transfers
its electrons to ETC, recycling NAD+.
• Under anaerobic conditions, various
fermentation pathways generate ATP by
glycolysis and recycle NAD+ by transferring
electrons from NADH to pyruvate or
derivatives of pyruvate.

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7.3.1 Ethanol fermentation


(Figure 9.17 (a), Campbell, page 173)

• In alcohol fermentation,, pyruvate is


converted to ethanol in two steps.
1. Pyruvate converted to acetaldehyde
(2C), by removal of CO2.
2. Acetaldehyde reduced by NADH to
ethanol.
° Alcohol fermentation by yeast - brewing
and winemaking.

7.3.2 Lactic fermentation


(Figure 9.17 (b), Campbell, page 173)

• Pyruvate is reduced directly by NADH to


form lactate without release of CO2.
° Lactic acid fermentation by some fungi
and bacteria is used to make cheese and
yogurt.
° Human muscle cells switch from aerobic
respiration to lactic acid fermentation to
generate ATP when O2 is scarce.
 The waste product, lactate, may cause
muscle fatigue, but ultimately it is
converted back to pyruvate in the liver.

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Fermentation
Fermentation and Cellular Respiration
Compared

• Similarities – both use


1. Glycolysis to oxidize sugars to pyruvate
with a net production of 2 ATP by
substrate-level phosphorylation.
2. NAD+ as an oxidizing agent to accept
electrons from food during glycolysis.

• Difference
1. Mechanism for oxidizing NADH to NAD+.
° Fermentation - electrons of NADH are
passed to an organic molecule to
regenerate NAD+.
° Respiration - electrons of NADH are
ultimately passed to O2, generating
ATP by oxidative phosphorylation.
2. ATP generated.
° Under aerobic respiration, a molecule
of glucose yields 36 - 38 ATP, but the
same molecule of glucose yields only 2
ATP under anaerobic respiration.

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Facultative Anaerobes

 Makes ATP by aerobic respiration if O2 is


present but can switch to fermentation I
absence of O2.
• Example: Yeast and many bacteria &
human muscle cells (at a cellular level).
° Aerobic conditions: pyruvate is converted
to acetyl CoA and oxidation continues in
the citric acid cycle.
° Anaerobic conditions: pyruvate serves as
an electron acceptor to recycle NAD+.

Evolutionary Significance of Glycolysis

• Oldest bacterial fossils are more than 3.5


billion years old, appearing long before
appreciable quantities of O2 accumulated in
atmosphere.
° Therefore, first prokaryotes may have
generated ATP exclusively from
glycolysis.
• The fact that glycolysis is the most
widespread metabolic pathway and occurs
in cytosol without membrane-enclosed
organelles suggests that glycolysis evolved
early in the history of life.

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7.4 Catabolism of Fat and Protein


(Figure 9.19, Campbell, page 177)

Catabolism

• Glycolysis accepts a wide range of


carbohydrates for catabolism.
° Polysaccharides (starch/glycogen) can be
hydrolyzed to glucose monomers that
enter glycolysis.
° Hexose sugars, such as galactose and
fructose, can also be modified to undergo
glycolysis.

• Proteins and fats, also enter respiratory


pathways used by carbohydrates.
• Proteins must first be digested to individual
amino acids.
° Amino acids that will be catabolized must
have their amino groups removed via
deamination.
° Nitrogenous waste is excreted as
ammonia, urea, or another waste product.
• The carbon skeletons are modified by
enzymes and enter as intermediaries into
glycolysis or citric acid cycle, depending on
their structure.

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• Fats must be digested to glycerol and fatty


acids.
° Glycerol can be converted to G3P, an
intermediate of glycolysis.
° Energy-rich fatty acids are split into 2C
fragments via beta oxidation.
° These molecules enter citric acid cycle
as acetyl CoA.

Biosynthesis (Anabolic Pathways)

• Intermediaries in glycolysis and citric acid


cycle can be diverted to anabolic
pathways.
° Example, human cell can synthesize
about half the 20 different amino acids by
modifying compounds from citric acid
cycle.
° Glucose can be synthesized from
pyruvate; fatty acids can be synthesized
from acetyl CoA.
• Glycolysis and citric acid cycle function as
metabolic interchanges that enable cells to
convert one kind of molecule to another.
° Example, excess carbohydrates and
proteins can be converted to fats through
intermediaries of glycolysis and citric
acid cycle.

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Feedback mechanisms
mechanisms control cellular
respiration

• Basic principles of supply and demand


regulate metabolic economy.
° If cell has excess of a certain amino acid,
it uses feedback inhibition to prevent
intermediates from citric acid cycle being
synthesized to that amino acid.

• Rate of catabolism also regulated by ATP


level in cell.
° If ATP levels drop, catabolism speeds up
to produce more ATP.

• Control of catabolism is based mainly on


regulating activity of enzymes at strategic
points in catabolic pathway.
• One strategic point occurs in 3rd step of
glycolysis, catalyzed by allosteric enzyme,
phosphofructokinase.
° Catalyzes earliest step that irreversibly
commits the substrate to glycolysis.
° Inhibited by ATP and stimulated by AMP
(derived from ADP).
 When ATP levels are high, inhibition of
this enzyme slows glycolysis.

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 As ATP levels drop and ADP and AMP


levels rise, enzyme becomes active
again and glycolysis speeds up.
• Citrate is also an inhibitor of
phosphofructokinase.
° Synchronizes rate of glycolysis and citric
acid cycle.
• If intermediaries from citric acid cycle are
diverted to other uses (e.g., amino acid
synthesis), glycolysis speeds up to replace
these molecules.

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