Chapter 20

The Calvin–Benson
Cycle and the
Pentose Phosphate
Pathway

© 2023 W. H. Freeman and Company

 CHAPTER 20
The Calvin–Benson Cycle and the
Pentose Phosphate Pathway
 Ch.20 Learning Goals
By the end of this chapter, you should be able to:
1. Explain the function of the Calvin–Benson cycle.
2. Describe how the light reactions and the Calvin–
Benson cycle are coordinated.
3. Identify the two stages of the pentose phosphate
pathway and explain how the pathway is coordinated
with glycolysis and gluconeogenesis.
4. Identify the regulation of the oxidative phase of the
pentose phosphate pathway, including the key
enzyme and its importance in health and disease.
 Ch.20 Outline
• 20.1 The Calvin–Benson Cycle Synthesizes Hexoses from
Carbon Dioxide and Water

• 20.2 The Activity of the Calvin–Benson Cycle Depends on

Environmental Conditions

• 20.3 The Pentose Phosphate Pathway Generates NADPH

and Synthesizes Pentoses

• 20.4 The Metabolism of Glucose 6-Phosphate by the

Pentose Phosphate Pathway Is Coordinated with
Glycolysis

• 20.5 Glucose 6-Phosphate Dehydrogenase Plays a Key

Role in Protection Against Reactive Oxygen Species
The Dark Reactions and the Pentose
Phosphate Pathway
• light reactions = transform light energy into ATP and
biosynthetic reducing power, NADPH
• dark reactions = use the ATP and NADPH produced by
the light reactions to reduce carbon atoms from CO2 to a
more reduced state as a hexose
– called the Calvin–Benson cycle

• the pentose phosphate pathway = pathway that oxidizes

glucose to generate NADPH
– mirror image of the Calvin–Benson cycle in many ways
 Section 20.1 The Calvin–Benson
Cycle Synthesizes Hexoses from
Carbon Dioxide and Water
• Photosynthetic organisms use the Calvin–Benson cycle
to synthesize glucose from CO2 and H2O using sunlight
as an energy source.
• Studies using unicellular algae exposed to 14CO2
revealed the Calvin–Benson cycle has three stages.
– Stage 1: fixation of CO2 to ribulose 1,5-bisphosphate to
form two molecules of 3-phosphoglycerate
– Stage 2: reduction of 3-phosphoglycerate to form hexose
sugars
– Stage 3: The regeneration of ribulose 1,5-bisphosphate
The Calvin–Benson Cycle Consists
of Three Stages
 Stage 1: Carbon Dioxide Reacts with
Ribulose 1,5-Bisphosphate to Form
Two Molecules of 3-Phosphoglycerate
• ribulose 1,5-bisphosphate carboxylase/oxygenase =
catalyzes the highly exergonic reaction that yields two
molecules of 3-phosphoglycerate from ribulose 1,5-
bisphosphate and CO2
– also called Rubisco or RuBisCo
– located on the stromal surface of the thylakoid membrane
– most abundant enzyme in the biosphere
– maximal catalytic rate is slow at only 3 s–1
Rubisco Catalyzes the Formation of
3-Phosphoglycerate
• This reaction is the rate-limiting step in hexose
synthesis.
 The Enzyme Rubisco Comprises
16 Subunits
• Rubisco consists of:
– eight large subunits,
each with a catalytic
site and a regulatory
site.
– eight small subunits
that enhance the
catalytic activity of the
large subunits.
 Rubisco Activity Depends on
Magnesium and Carbamate
• Rubisco requires a bound
divalent metal ion for activity.
– typically Mg2+
– activates a bound substrate
molecule by stabilizing a
negative charge
– binds to a carbamate formed
upon reaction of CO2 with Lys
201 on Rubisco
 Magnesium-Coordinated Ribulose
1,5-Bisphosphate Facilitates CO2
Fixation
• Mg2+ assists in
the binding and
activating
ribulose 1,5-
bisphosphate so
it reacts with
CO2.
 In Stage 1 of the Calvin–Benson
Cycle, 3-Phosphoglycerate Is Formed
 Rubisco Also Catalyzes a Wasteful
Oxygenase Reaction
• The reactive enediolate intermediate generated on the
Mg2+ ion sometimes reacts with O2 instead of CO2 to
form a hydroperoxide intermediate.
– yields phosphoglycolate and 3-phosphoglycerate
– requires Lys 201 be in the carbamate form such that
Rubisco cannot catalyze the oxygenase reaction in the
absence of CO2
– rate is four times less than the carboxylase reaction
 The Enediolate on Rubisco Can
React with Oxygen in a Wasteful Side
Reaction
 The Salvage Pathway
• Phosphoglycolate is not a versatile metabolite, but it can
be salvaged to recover part of its carbon skeleton.
• photorespiration = process of recycling three of the four
carbon atoms of two molecules of glycolate
– occurs in the chloroplast, peroxisome, and mitochondria
– called photorespiration because O2 is consumed and CO2
is released
– wasteful process because organic carbon is converted into
CO2 without the production of ATP, NADPH, or other
energy-rich metabolites
Photorespiration Consumes O2 and
Produces CO2
Peroxisomes Are Organelles Defined
by a Single Membrane
Which reaction species is analogous
to the zinc-hydroxide species in
carbonic anhydrase? (1 of 2)
Which reaction species is analogous
to the zinc-hydroxide species in
carbonic anhydrase? (2 of 2)
 Stage 2: Hexose Phosphates Are
Made from Phosphoglycerate
• The 3-phosphoglycerate product of
Rubisco is converted into fructose
6-phosphate.
– Pathway is parallel to that of
gluconeogenesis.
– F-6P can be readily converted to
G-1P and then G-6P.

• hexose monophosphate pool = the

mixture of three phosphorylated
hexoses
Stage 3: Ribulose 1,5-Bisphosphate
Is Regenerated
• Ribulose 1,5-bisphosphate, the acceptor of carbon
dioxide in Stage 1, is regenerated.
• requires a transketolase and an aldolase to rearrange
carbon atoms to yield a five-carbon sugar from six-
carbon and three-carbon sugars
 The Transketolase Reaction
• transketolase = transfers a two-carbon unit (CO–CH2OH)
from a ketose to an aldose
– requires the coenzyme triamine pyrophosphate (TPP)
– converts fructose 6-phosphate and glyceraldehyde 3-
phosphate into erythrose 4-phosphate and xylulose 5-
phosphate
 The Aldolase Reaction
• aldolase = catalyzes an aldol condensation between
dihydroxyacetone phosphate (DHAP) and an aldehyde
– highly specific for DHAP, but accepts a wide variety of
aldehydes
– converts erythrose 4-phosphate and DHAP into
sedoheptulose 1,7-bisphosphate
The Formation of Ribose 5-Phosphate
and Xylulose 5-Phosphate
• sedoheptulose 1,7-bisphosphate phosphatase =
removes a phosphate from sedoheptulose 1,7-
bisphosphosphate to form sedoheptulose 7-phosphate
• In the final step, transketolase converts sedoheptulose
7-phosphate and glyceraldehyde 3-phosphate into
erythrose 4-phosphate and xylulose 5-phosphate.
The Formation of Five-Carbon Sugars
In Stage 3 of the Calvin–Benson Cycle,
Ribulose 1,5-Bisphosphate Is
Regenerated
 The Sum of the Stage 3 Reactions
• The sum of the reactions for the regeneration of ribulose
1,5-bisphosphate from fructose 6-phosphate and
glyceraldehyde 3-phosphate is

Fructose 6-phosphate + 2 glyceraldehyde 3-phosphate

+ dihydroxyacetone phosphate + 3 ATP →
3 ribulose 1,5-bisphosphate + 3 ADP
 The Full Calvin–Benson Cycle
Process
• The cycle must
take place twice
because two
molecules of
DHAP are
required to yield
a hexose
monophosphate.
How many molecules of ATP are
needed to yield a hexose
monophosphate? (1 of 2)

© Macmillan Learning, 2023

How many molecules of ATP are
needed to yield a hexose
monophosphate? (2 of 2)

18

© Macmillan Learning, 2023

18 ATP and 12 NADPH Molecules Are
Used to Bring Six Carbon Dioxide to
the Level of a Hexose
• The balanced equation for the net reaction of the Calvin–
Benson cycle is

6CO2 + 18 ATP + 12 NADPH + 12 H2O→

C6H12O6 + 18 ADP + 18 Pi + 12 NADP+ + 6H+

• Six rounds of the Calvin–Benson cycle are required

because one carbon atom is reduced in each round.
 Starch and Sucrose Are the Major
Carbohydrate Stores in Plants
• starch = a polymer of glucose residues that is
synthesized are stored in chloroplasts
– less branched that its animal counterpart glycogen
because it contains a smaller proportion of α-1,6-glycosidic
linkages
– ADP-glucose, not UDP-glucose, is the activated precursor

• sucrose (common table sugar) = a disaccharide that is

synthesized in the cytoplasm
– readily transportable and mobilizable sugar
– stored in many plant cells, such as those in sugar beets
and sugar cane
 The Formation of Sucrose
• Triose phosphate intermediates cross from the
chloroplast into the cytoplasm in exchange for phosphate
through an antiporter.
• Fructose 6-phosphate forms from triose phosphates.
• Fructose 6-phosphate joins the glucose unit of UDP-
glucose to form sucrose 6(F)-phosphate.
• Sucrose phosphatase hydrolyzes the phosphate ester to
yield sucrose.
Carbohydrates Are Stored as
Sucrose
Inspired by the Calvin–Benson Cycle,
Scientists Are Developing New
Methods for Fixing Carbon Dioxide
• the CETCH cycle = catalyzes the conversion of 2
molecules of CO2 to 1 molecule of glyoxylate
– involves 12 steps and 15 enzymes

2 CO2 + 4 NADPH + 2 FADH2 + 4 ATP + 2 O2 + 3 H+ →

–O + 4 NADP+ + 2 FAD + 4 ADP + 2 Pi + 5 H2O
2C-CHO
Enoyl-CoA Carboxylases/ Reductases
Catalyze Carbon Fixation
• more efficient at fixing CO2 than is Rubisco and do not
react with oxygen
 Scientists Have Engineered Artificial
Photosynthesis Using Alternative CO2
Fixation Pathways
• synthetic biology
= using
engineering
principles to
develop
potentially useful
devices
 Section 20.2 The Activity of the
Calvin–Benson Cycle Depends on
Environmental Conditions
• The principle means of regulation the fixation of CO2 into
biomolecules is alteration of the stromal environment by
the light reactions
• Light reactions lead to increases in stromal pH and
stromal concentrations of Mg2+, NADPH, and reduced
ferredoxin.
– all activate the enzymes of the Calvin–Benson cycle in the
stroma
Light Regulates the Calvin–Benson
Cycle
Rubisco Is Activated by Light-Driven
Changes in Proton and Magnesium
Ion Concentrations
• In the stroma, pH increases (7 to 8) and Mg2+ levels rise.
– results from the light-driven pumping of protons into
the thylakoid membrane
• Rubisco activity increases markedly on illumination
because:
– carbamate formation is favored at alkaline pH.
– Mg2+ binds to the carbamate to generate the active
form of the enzyme.
 Thioredoxin Plays a Key Role in
Regulating the Calvin–Benson Cycle
• Reduced ferredoxin and NADPH signal that conditions
are right for biosynthesis.
• thioredoxin = activates biosynthetic enzymes including
chloroplast ATP synthase when in its reduced form
– neighboring Cys residues cycle between a reduced
sulfhydryl and an oxidized disulfide form
– acts by reducing disulfide bridges that control enzyme
activity
– also inhibits several degradative enzymes by the same
means
Thioredoxin Has a Redox-Active
Disulfide Bond
Enzymes Regulated by Thioredoxin

TABLE 20.1 Enzymes regulated by thioredoxin

Enzyme Pathway
Rubisco Carbon fixation in the Calvin-Benson cycle
Fructose 1,6-bisphosphatase Gluconeogenesis
Glyceraldehyde 3-phosphate dehydrogenase Calvin-Benson cycle, gluconeogenesis, glycolysis
Sedoheptulose 1,7-bisphosphatase Calvin-Benson cycle
Glucose 6-phosphate dehydrogenase Pentose phosphate pathway
Phenylalanine ammonia lyase Lignin synthesis
Phosphoribulokinase Calvin-Benson cycle
NADP+-malate dehydrogenase C4 pathway
CF1-CF0 ATP synthase The light reactions
 Thioredoxin Can Activate Enzymes
by Reducing Them
• In chloroplasts, ferredoxin–
thioredoxin reductase
reduces oxidized thioredoxin
that is reduced by ferredoxin.
• The activities of the light and
dark reactions are
coordinated through electron
transfer from reduced
ferredoxin to thioredoxin and
then to component enzymes
containing regulatory
disulfide bonds.
 Regulation by NADPH

• CP12 = an intrinsically disordered protein that inhibits

enzymes through association
• NADPH activates two biosynthetic enzymes by
disrupting their association with CP12:
– phosphoribulokinase
– glyceraldehyde 3-phosphate dehydrogenase
The C4 Pathway of Tropical Plants and
Grasses Accelerates Photosynthesis
by Concentrating Carbon Dioxide
• Because oxygenase activity of Rubisco increases more
rapidly with temperature than does the carboxylase
activity, hot climate plants concentrate CO2 at the site of
the Calvin–Benson cycle.
• C4 pathway (Hatch–Slack pathway) = metabolic pathway
that concentrates CO2 in bundle-sheath cells, the major
sites of photosynthesis
• C4 plants include tropical plants, grasses, and sedges.
 The C4 Pathway
• Four-carbon (C4) compounds carry CO2 from mesophyll
cells to bundle-sheath cells.
• Decarboxylation of the C4 compound maintains a high
concentration of CO2 at the site of the Calvin–Benson
cycle.
• The three-carbon product returns to the mesophyll cell
for another round of carboxylation.
The C4 Pathway Can Concentrate CO2
 The Net Reaction of the C4 Pathway
• The net reaction of the C4 pathway is

CO2 (in mesophyll cell) + ATP + 2 H2O →

CO2 (in bundle-sheath cell) + AMP + 2 Pi + 2 H+

• The C4 pathway requires an extra 12 ATP when

compared to the C3 pathway.
 The Net Reaction of the C4 Pathway
and the Calvin–Benson Cycle
• The net reaction of the C4 pathway and Calvin–Benson
cycle is

6 CO2 + 30 ATP + 12 NADPH + 24 H2O →

C6H12O6 + 30 ADP + 30 Pi + 12 NADP+ + 18 H+

• 30 ATP are consumed per hexose in the C4 pathway,

whereas 18 ATP are consumed per hexose in the C3
pathway.
 Stomata Can Open to Facilitate Gas
Exchange
• stomata = openings
in the leaves that
allow gas exchange
• When Rubisco is
adequately supplied
with CO2, stomata do
not have to open
completely.
– prevents water loss
Both C3 and C4 Plants Are Important
 Crassulacean Acid Metabolism
Permits Growth in Arid Ecosystems
• Many plants, including succulents, keep their stomata
closed in the heat of the day to prevent water loss.
– CO2 cannot be absorbed during the day.

• Instead, CO2 enters the leaf when the stomata open at

night.
• crassulacean acid metabolism (CAM) = adaption that
plants use to store CO2 until it can be used during the day
– At night, CO2 is fixed by the C4 pathway into malate and
stored in vacuoles.
– During the day, malate is decarboxylated, and the CO2
becomes available to the Calvin–Benson cycle.
 Crassulacean Acid Metabolism
Facilitates Growth in Arid Climates
• Because
malate is the
sole source of
CO2 and
malate storage
is limited, the
growth rate of
CAM plants is
slower than
that of C3 and
C4 plants.
 Which statement is true of plants
with a C4 pathway? (1 of 2)
a. CO2 is concentrated in mesophyll cells of these plants by
the expenditure of ATP in bundle-sheath cells.
b. Tropical plants with a C4 pathway do high levels of
photorespiration.
c. Plants with a C4 pathway have a disadvantage in a hot
environment and under high illumination.
d. Decarboxylation of four-carbon compounds in bundle-
sheath cells maintains a high concentration of CO2.
e. C4 plants consume 18 molecules of ATP per hexose
molecule formed in the absence of photorespiration.

© Macmillan Learning, 2023

 Which statement is true of plants
with a C4 pathway? (2 of 2)
a. CO2 is concentrated in mesophyll cells of these plants by
the expenditure of ATP in bundle-sheath cells.
b. Tropical plants with a C4 pathway do high levels of
photorespiration.
c. Plants with a C4 pathway have a disadvantage in a hot
environment and under high illumination.
*d. Decarboxylation of four-carbon compounds in bundle-
sheath cells maintains a high concentration of CO2.
e. C4 plants consume 18 molecules of ATP per hexose
molecule formed in the absence of photorespiration.

© Macmillan Learning, 2023

Section 20.3 The Pentose Phosphate
Pathway Generates NADPH and
Synthesizes Pentoses
• pentose phosphate pathway = pathway generating a
crucial source of NADPH for use in reductive
biosynthesis and for protection against oxidative stress
– occurs in the cytoplasm of all organisms
 The Pentose Phosphate Pathway
Consists of Two Phases
• The first phase is the oxidative generation of NADPH.
– occurs when glucose 6-phosphate is oxidized to ribulose
5-phosphate

Glucose 6-phosphate + 2 NADP+ + H2O →

ribulose 5-phosphate + 2 NADPH + 2 H++ CO2

• The second phase is the nonoxidative interconversion of

three-, four-, five-, six-, and seven-carbon sugars.
– Interconversions rely on the same reactions leading to
regeneration of ribulose 1,5-bisphosphate in the Calvin–
Benson cycle.
Pathways Requiring NADPH
TABLE 20.2 Pathways requiring NADPH

Reductive biosynthesis
Fatty acid biosynthesis
Cholesterol biosynthesis
Neurotransmitter biosynthesis
Nucleotide biosynthesis
Protection from oxidative stress
Reduction of oxidized glutathione
Cytochrome P450 monooxygenases
The Pentose Phosphate Pathway
Assuming three molecules of ribulose
5-phosphate enter the nonoxidative
phase of the pentose phosphate
pathway and are converted to
fructose 6-phosphate and
glyceraldehyde 3-phosphate, how
many carbons are present at the end?
(1 of 2)

© Macmillan Learning, 2023

Assuming three molecules of ribulose
5-phosphate enter the nonoxidative
phase of the pentose phosphate
pathway and are converted to
fructose 6-phosphate and
glyceraldehyde 3-phosphate, how
many carbons are present at the end?
(2 of 2)
15

© Macmillan Learning, 2023

 Two Molecules of NADPH Are
Generated in the Conversion of Glucose
6-Phosphate into Ribulose 5-Phosphate
• glucose 6-phosphate dehydrogenase = initiates the
oxidative phase of the pentose phosphate pathway with the
conversion of glucose 6-phosphate into 6-phosphoglucono-
δ-lactone, yielding NADPH and H+
– highly specific for NADP+

• lactonase = hydrolyzes 6-phosphoglucono-δ-lactone to 6-

phosphogluconate
• 6-phosphogluconate dehydrogenase = oxidatively
decarboxylates 6-phosphogluconate to ribulose phosphate,
yielding CO2 and NADPH
– NADP+ is the electron acceptor
Oxidative Phase of the Pentose
Phosphate Pathway Yields Two
Molecules of NADPH
The Pentose Phosphate Pathway and
Glycolysis Are Linked by
Transketolase and Transaldolase
• Phosphopentose isomerase isomerizes ribulose 5-
phosphate into ribose 5-phosphate.
 The Pentose Phosphate Pathway
and Glycolysis Are Reversibly
Linked
• Transketolase and transaldolase catalyze three
successive reactions:

• The net result of these reactions is the conversion of

three pentoses into two hexoses and one triose:
The Formation of Glyceraldehyde 3-
Phosphate and Sedoheptulose
7-Phosphate
• Transketolase catalyzes the formation of glyceraldehyde
3-phosphate and sedoheptulose 7-phosphate from two
pentoses.
– first of three linking reactions
 The Phosphopentose Epimerase
Reaction
• The transketolase substrate is a ketose with its hydroxyl
group at C-3 having the configuration of xylulose rather
than ribulose.
• phosphopentose epimerase = catalyzes the conversion of
xylulose 5-phosphate into its epimer ribulose 5-phosphate
 The Formation of Fructose 6-
Phosphate and Erythrose 4-Phosphate
• Transaldolase catalyzes the formation of a six-carbon
and a four-carbon sugar from a three-carbon and a
seven-carbon sugar.
– second of three linking reactions
The Formation of Fructose 6-Phosphate
and Glyceraldehyde 3-Phosphate
• Transketolase catalyzes the formation of a six-carbon
and a three-carbon sugar from a four-carbon and a five-
carbon sugar.
– third of three linking reactions
 The Sum of the Transketolase,
Epimerase, and Transaldolase Reactions
• The sum of the reactions catalyzed by the transketolase,
phosphopentose epimerase, and transaldolase is

• Xylulose 5-phosphate can be formed from ribose 5-

phosphate by phosphopentose isomerase and
phosphopentose epimerase, so the net reaction starting
from ribose 5-phosphate is
 Pentose Phosphate Pathway
TABLE 20.3 Pentose phosphate pathway
Reaction Enzyme
Oxidative phase
Glucose 6-phosphate + NADP+ → 6- Glucose 6-phosphate
phosphoglucono-δ-lactone + NADPH + H+ dehydrogenase
6-Phosphoglucono-δ-lactone + H2O → 6- Lactonase
phosphogluconate + H+
6-Phosphogluconate + NADP+ → ribulose 5- 6-Phosphogluconate
phosphate + CO2 + NADPH + H+ dehydrogenase
Nonoxidative phase
Phosphopentose isomerase
Phosphopentose epimerase
Transketolase
Transaldolase
Transketolase
 Transketolase and Transaldolase
Stabilize Carbanionic Intermediates by
Different Mechanisms
• Transketolase transfers a two-carbon unit and contains a
tightly bound thiamine pyrophosphate (TPP) as its
prosthetic group.
• Transaldolase transfers a three-carbon unit and does not
contain a prosthetic group.
• Each of these carbon units is transiently attached to the
enzyme in the course of the reaction.
– characteristic of double displacement reactions
The Transketolase Reaction Takes
Place on Thiamin Pyrophosphate
 Transketolase Reaction
• Step 1: The C-2 carbon of bound TTP ionizes to give a
carbanion.

• Step 2: The carbanion of TPP attacks the carbonyl group of

the ketose substrate.

• Step 3: Cleavage of a carbon–carbon bond frees the aldose

product, yielding an activated glycolaldehyde joined to TPP.

• Step 4: The carbonyl group of an aldose acceptor condenses

with the activated glycolaldehyde, forming a new ketose.

• Step 5: The ketose is released from the enzyme, freeing TPP

for the next reaction cycle.
The Transaldolase Reaction Involves
a Schiff Base Intermediate
 Transaldolase Reaction
• Step 1: A Schiff base forms between a Lys residue in
transaldolase and the ketose substrate.
• Step 2: Protonation of the Schiff base occurs, and the
bond between C-3 and C-4 is split.
• Step 3: Deprotonation leads to the release of the aldose
product, leaving a three-carbon fragment attached to
Lys.
• Step 4: A suitable aldose binds.
• Step 5: Protonation allows the formation of new carbon–
carbon bond.
• Step 6: Deprotonation occurs.
• Step 7: hydrolysis of the Schiff base releases the ketose
For Transketolase and Transaldolase,
a Carbanion Intermediate Is Stabilized
by Resonance
 Section 20.4 The Metabolism of
Glucose 6-Phosphate by the Pentose
Phosphate Pathway Is Coordinated
with Glycolysis
• Glucose 6-phosphate is metabolized by both the
glycolytic pathway and the pentose phosphate pathway.
• The fate of glucose 6-phosphate is controlled by the
cytoplasmic concentration of NADP+.
The Rate of the Oxidative Phase of the
Pentose Phosphate Pathway Is
Controlled by the Level of NADP+
• The first reaction of the pentose phosphate pathway is
the dehydrogenation of glucose 6-phosphate by glucose
6-phosphate dehydrogenase.
– This is the rate-limiting step of the pathway.
– The reaction is essentially irreversible.

• Low levels of NADP+ limit dehydrogenation of glucose 6-

phosphate because it is needed as the electron acceptor.
– ensures NADPH is not generated unless needed for
reductive biosyntheses or protection against oxidative
stress
 The Flow of Glucose 6-Phosphate
Depends on the Need for NADPH,
Ribose 5-Phosphate, and ATP
• The pentose phosphate pathway can operate in four
distinct modes:
– much more ribose 5-phosphate than NADPH is required
– the needs for NADPH and ribose 5-phosphate are
balanced
– much more NADPH than ribose 5-phosphate is required
– both NADPH and ATP are required
The Pentose Phosphate Pathway
Can Operate in Four Modes
 Mode 1—Much More Ribose 5-
Phosphate Than NADPH Is Required
• occurs, for example, when rapidly dividing cells need
ribose 5-phosphate for the synthesis of nucleotide
precursors of DNA
• Glucose 6-phosphate is converted into fructose 6-
phosphate (F-6P) and glyceraldehyde 3-phosphate
(GAP) by the glycolytic pathway.
• Transaldolase and transketolase convert two molecules
of F-6P and one molecule of GAP into three molecules
of ribose 5-phosphate.
• The stoichiometry of mode 1 is
5 Glucose 6-phosphate + ATP → 6 ribose 5-phosphate + ADP + 2 H+
 Mode 2—The Needs of Ribose 5-
Phosphate and NADPH Are Balanced
• Glucose 6-phosphate is processed to one molecule of
ribulose 5-phosphate while generating two molecules of
NADPH.
• Ribulose 5-phosphate is converted into ribose 5-
phosphate.
• The stoichiometry of mode 2 is
Glucose 6-phosphate + 2 NADP+ + H2O →
ribose 5-phosphate + 2 NADPH + 2 H+ + CO2
 Mode 3—Much More NADPH Than
Ribose 5-Phosphate Is Required
• occurs, for example, when adipose tissue requires a
high level of NADPH for the synthesis of fatty acids
• Glucose 6-phosphate is completely oxidized to CO2
through three groups of reactions:
– The oxidative phase of the pentose phosphate pathway
forms two molecules of NADPH and one molecule of
ribulose 5-phosphate.
– Ribulose 5-phosphate is converted into F-6P and GAP by
transketolase and transaldolase.
– Glucose 6-phosphate is resynthesized from fructose.
 The Stoichiometry of Mode 3
• The stoichiometries of the three sets of reactions are

6 Glucose 6-phosphate + 12 NADP+ + 6 H2O →

6 Ribose 5-phosphate + 12 NADPH + 12 H+ + 6 CO2

6 Ribose 5-phosphate →
4 Fructose 6-phosphate + 2 glyceraldehyde 3-phosphate

4 Fructose 6-phosphate + 2 glyceraldehyde 3-phosphate + H2O →

5-Glucose 6-phosphate + Pi

• The stoichiometry of mode 3 is

Glucose 6-phosphate + 12 NADP++ 7 H2O → 6 CO2 + 12 NADPH +12 H+ + Pi

Tissues with Active Pentose
Phosphate Pathways
TABLE 20.4 Tissues with active pentose phosphate pathways

Tissue Function
Adrenal gland Steroid synthesis
Liver Fatty acid and cholesterol synthesis
Testes Steroid synthesis
Adipose tissue Fatty acid synthesis
Ovary Steroid synthesis
Mammary gland Fatty acid synthesis
Red blood cells Maintenance of reduced glutathione
 Mode 4—Both Ribose 5-Phosphate
and NADPH Are Required
• Ribulose 5-phosphate formed from glucose 6-phosphate
can be converted into pyruvate.
• F-6P and GAP derived from ribose 5-phosphate enter the
glycolytic pathway rather than reverting to glucose 6-
phosphate.
• Pyruvate formed can be oxidized to generate more ATP or
used as a building block in a variety of biosynthesis.
• The stoichiometry of mode 4 is
3 Glucose 6-phosphate + 6 NADP+ + 5 NAD+ + 5 Pi + 8 ADP →
5 pyruvate + 3 CO2 + 6 NADPH + 5 NADH + 8 ATP + 2 H2O + 8 H+
 The Pentose Phosphate Pathway Is
Required for Rapid Cell Growth
• Rapidly dividing cells, such as cancer cells, require:
– ribose 5-phosphate for nucleic acid synthesis.
– NADPH for fatty acid and membrane lipid synthesis.

• Rapidly dividing cells:

– switch to aerobic glycolysis to meet ATP needs.
– divert glucose 6-phosphate and glycolytic intermediates to
the nonoxidative phase of the pentose phosphate pathway
to generate NADPH and ribose 5-phosphate.
 Pyruvate Kinase Isozyme, PKM
• pyruvate kinase isozyme (PKM) = facilitates the
diversion of glycolytic intermediates into the nonoxidative
phase
– PKM has a low catalytic activity.
– Glycolytic intermediates accumulate and enter the pentose
phosphate pathway.

• The diversion of phosphorylated intermediates is further

enabled by the inhibition of triose phosphate isomerase
by phosphoenolpyruvate, the substrate of PKM.
 The Calvin–Benson Cycle and the
Pentose Phosphate Pathway Are
Essentially Mirror Images of One
Another
• The Calvin–Benson cycle begins with the fixation of CO2
and proceeds to use NADPH in the synthesis of glucose.
• The pentose phosphate pathway begins with the
oxidation of a glucose-derived carbon atom to CO2,
generating NADPH.
• In photosynthetic organisms, many enzymes are common
to the two pathways.
 Which mode of the pentose
phosphate pathway does not
generate the formation of NADPH?
(1 of 2)
 Which mode of the pentose
phosphate pathway does not
generate the formation of NADPH?
(2 of 2)

Mode 1
 Section 20.5 Glucose 6-Phosphate
Dehydrogenase Plays a Key Role in
Protection Against Reactive Oxygen
Species
• reduced glutathione (GSH) = a
tripeptide with a free sulfhydryl
group that combats oxidative stress
by reducing ROS to harmless forms

• Oxidized glutathione (GSSG) must

be reduced to regenerate GSH.
– Reducing power is supplied by
the NADPH generated by
glucose 6-phosphate
dehydrogenase in the pentose
phosphate pathway.
 Glucose 6-Phosphate
Dehydrogenase Deficiency Causes a
Drug-Induced Hemolytic Anemia
• pamaquine = the first synthetic
antimalarial drug introduced in 1926
• Some patients developed drug-
induced hemolytic anemia due to a
deficiency of glucose 6-phosphate
dehydrogenase.
• This deficiency results in a scarcity of
NADPH in all cells.
– most acute in red blood cells because
they lack mitochondria
 Primaquine
• primaquine = an antimalarial closely related to
pamaquine that is widely used in malaria-prone regions
of the world
• Drug-induced hemolytic anemia is seen in patients
deficient in glucose 6-phosphate dehydrogenase.
 Fava Beans Produce a Pyrimidine
Glycoside
• vicine = a pyrimidine glycoside of fava beans
• People deficient in glucose 6-phosphate dehydrogenase
suffer hemolysis from eating fava beans.
 Pamaquine, Primaquine, and Vicine
Generate Peroxides
• peroxides = reactive oxygen species that can damage
membranes and other biomolecules
• Pamaquine, primaquine, and vicine are oxidative agents
that generate peroxides.
 Hemolysis Caused by Pamaquine,
Primaquine, and Vicine
• glutathione peroxidase = eliminates peroxides using
glutathione as a reducing agent

2GSH  ROOH 

Glutathione peroxidase
 GSSG  H2 O  ROH

• In red blood cells, glutathione reductase uses NADPH to

regenerate the reduced form of GSH.
 Reduced Glutathione Maintains the
Structure of Hemoglobin
• Reduced form of glutathione serves as a sulfhydryl
buffer that keeps the residues of hemoglobin in the
reduced sulfhydryl form.
• Without adequate levels of reduced glutathione,
hemoglobin molecules cross-link with one another to
form aggregates called Heinz bodies on cell membranes.
• Membranes damaged by Heinz bodies and reactive
oxygen species become deformed.
– These cells are more likely to undergo lysis.
Heinz Bodies Are Made of Denatured
Hemoglobin
A Deficiency of Glucose 6-Phosphate
Dehydrogenase Confers an
Evolutionary Advantage in Some
Circumstances
• glucose 6-phosphate dehydrogenase deficiency =
characterized by a 10-fold reduction in enzymatic activity
in red blood cells
– more common among Americans of African heritage
– protects against the deadliest form of malaria, which is
prevalent in Africa's subtropical regions

• Parasites causing malaria require NADPH for growth and

infection induces oxidative stress in infected human cells.
• Because the pentose phosphate pathway is compromised,
the cells and parasite die from oxidative damage.
Remember to complete your exit poll
tonight!

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