20| Photosynthesis and Carbohydrate Synthesis

in Plants
 Oxidative Phosphorylation versus
Photophosphorylation
Oxidative Phosphorylation Photophosphorylation
• Electrons from the reduced • Water is the source of
cofactors NADH and FADH2 electrons that are passed via
are passed to proteins in the a chain of protein
respiratory chain. transporters to the ultimate
electron acceptor, NADP+.
• In eukaryotes, oxygen is the
ultimate electron acceptor • Light energy is used to
for these electrons. separate charges on
chlorophyll to generate
• Energy of oxidation is used to NADPH and phosphorylate
phosphorylate ADP. ADP.
• Oxygen is the byproduct of
water oxidation.
 Oxidative Phosphorylation versus
Photophosphorylation
Oxidative Phosphorylation Photophosphorylation
Light Energy Is Converted to ATP
in Plant Chloroplasts
 Photopigments Absorb Different
Wavelengths of Light
The energy is transferred to the photosynthetic reaction center.
Organization of Light-Absorbing
Molecules in Chloroplasts
Harvested Light Energy Excites Electrons
in the Reaction Center
Bacterial Photosynthesis Involve One of
Two Types of Reaction Centers
In Plants, Two Reaction Centers Work
Together
Photosystem II Evolves Oxygen
from Water
Water Splitting Allows Single Electron
Transfer to the Photosystem

The water-splitting complex

donates single electrons to
photosystem II but releases a
single O2 after four electrons
are freed from two waters.

Overall reaction:
2 H2O  4 e− + 4 H+ + 1 O2
Photosystem II Transfers Electrons to
Plastoquinones
Plastoquinones are
structurally and
functionally similar to
ubiquinones found in
the mitochondria.

These lipid-soluble
small molecules
transport two electrons
to the cytochrome b6f
complex.
Cytochrome b6f Complex Links PS II and I
and Translocates Protons into the Lumen

• Structurally and functionally

similar to Complex III in the
electron-transport chain
• Protons are removed from
plastoquinones as
plastoquinones are oxidized.
• The PQ cycle is similar to the
Q cycle in the ETC.
• Electrons are funneled to
plastocyanin, a single
electron carrier, which
carries the electron to PS1.
Photosystem I Results in Reduced NADPH
 Light-Induced Redox Reactions
Cause Acidification of Lumen

• In contrast to oxidative phosphorylation, protons are

pumped into the lumen, building up a large
concentration gradient quickly in the small space.
• The proton-motive force across the thylakoid membrane
drives the synthesis of ATP.
• Synthesis of ATP has the same 3H+ per 1 ATP cost as in
oxidative phosphorylation
Cyclic Photosynthesis Can Be Utilized to
Generate Excess ATP
Organization of Photosyntheic Machinery
in the Thylakoid Membrane
 Summary of Photosynthesis
• Photosynthetic organisms capture energy from light at a
variety of wavelengths using accessory pigments and
funnel it via excitons to reaction centers.
• In plants, electrons are freed from H2O, which generates
O2 and H+. These electrons are excited and moved
through the photosynthetic electron chain.
• Per water, 6 H+ are mobilized into the lumen.
• The final electron acceptor for linear photosynthesis is
NADP+, which requires two electrons for conversion to
NADPH.
• When ATP stores are low, the electrons can be
transported to cytochrome b6f and cycled through the
second half of photosynthesis, resulting in an increased
proton gradient.
 Introduction to Anabolic Pathways

• The previous chapters were mainly concerned

with catabolism: how to extract energy from
biomolecules.

• This chapter and those that follow are concerned

with anabolism: how to build biomolecules.
 Assimilation of CO2 by Plants
• Animal cells: use 3-C intermediates (pyruvate, lactate,
etc.) for synthesis of carbohydrates, amino acids, and
lipids, but the 3-C intermediates must be made from
degradation of a larger molecule
• Plant cells: can also make 3-C intermediates for further
synthesis
– Glyceraldehyde 3-phosphate (G3P) is an important
one.
– made from CO2, H2O, plus ATP and NADPH from
photosynthesis
• Making CO2 into intermediates is CO2 assimilation.
Assimilation of CO2 into Biomass
 CO2 Assimilation Occurs in Plastids
• Organelles in plants and
algae
– includes chloroplasts (for
photosynthesis), amyloplasts
(for starch storage), and so on
• Enclosed by a double
membrane
– As in mitochondria, the inner
membrane is impermeable to
ions and polar molecules.
• Have their own small
genome
– again, like mitochondria
Amyloplasts―Filled with Starch
(and Stained with Iodine)
 CO2 Assimilation

• It occurs in the stroma of chloroplasts via a cyclic

process known as the Calvin cycle.
• Key intermediate ribulose 1,5-bisphosphate is
constantly regenerated using energy of ATP.
• It produces 3-phosphoglycerate, then
glyceraldehyde 3-phosphate (G3P) in eq with
dihydroxyacetone phosphate (DHAP)
• The net result is the reduction of CO2 with NADPH
that was generated in the light reactions of
photosynthesis.
The Three Stages of the Calvin Cycle
of CO2 Assimilation
 Three Stages of the Calvin Cycle
1. CO2 fixation: 3 ribulose 1,5-bisphosphate + 3 CO2 
6 3-phosphoglycerate
– catalyzed by rubisco

2. 3-phosphoglycerate reduced to glyceraldehyde 3-

phosphate using NADPH and ATP from photosynthesis
– Catalyzed by chloroplast isoforms of 3-phosphoglycerate
kinase and glyceraldehyde 3-phosphate dehydrogenase from
glycolysis

3. Regeneration of ribulose 1,5-bisphosphate

Overall: 3 CO2 + 6 NADPH + 5 H2O + 9 ATP → glyceraldehyde 3-
phosphate (G3P) + 2 H+ + 6 NADP+ + 9 ADP + 8 Pi
 The First Stage (CO2 Fixation) Is
Catalyzed by Rubisco

• Most plentiful enzyme on Earth

• Rubisco = ribulose 1,5-bisphosphate
carboxylase/oxygenase (also RuBisCo)
• Large Mg2+ dependent enzyme
• Catalyzes the reaction:
ribulose 1,5-bisphosphate + CO2  2 3-phosphoglycerate
 Rubisco Exists in Two Major Forms
and Is Abundant
• Form 1: plants, algae, cyanobacteria
– 8 large catalytic subunits (encoded by plastid genome) +
8 small subunits (encoded by nucleus)
• Form II: photosynthetic bacteria only
– only 2 catalytic subunits; resemble plant subunits

Low turnover # of 3 sec–1 @ 25° C means

a LOT of the enzyme needed!
50% of plant enzymes are rubisco.
Structure of Form II Rubisco
Mg2+ in Rubisco
 Rubisco
Carboxylase
Mechanism―
Overview
 1. Creation of Enediolate
Intermediate

Mg2+ facilitates interaction of carbamoylated Lys to ribulose

1,5-bisphosphate  forms enediolate intermediate
 2. Nucleophilic
Attack to Create
-Keto Acid
Intermediate

Mg2+ polarizes CO2

to accept
nucleophilic attack
from enediolate,
forming 6-C -keto
acid intermediate
3. Hydroxylation at C-3

Hydroxylation at C-3
of ribulose-1,5-
bisphosphate by
water
4. Cleavage to Yield First
3-Phosphoglycerate
 5. Protonation and
Release of Second
3-Phosphoglycerate

Deprotonation of Lys175 is
easily reversed at physiological
pH, allowing complete reset of
the active site.
 Catalytic Role of Mg2+ in Rubisco’s
Carboxylase Activity

• Notice that Mg2+ is held by negatively charged side

chains of:
• Glu, Asp, and carbamoylated Lys
• Mg2+ brings together the reactants in a correct
orientation and stabilizes the negative charge that forms
upon the nucleophilic attack of enediolate to CO2.
 Rubisco Is Activated via Covalent
Modification of the Active Site Lysine
• Rubisco is highly regulated (as first step of CO2
assimilation).
• Inactive until Lys201 is carbamoylated by CO2 (Lys201
inaccessible).
• Rubisco activase (an enzyme sometimes triggered by
light) changes rubisco conformation to expose Lys201.
• Rubisco activase requires ATP.
How Rubisco Activase Exposes Lys201
 Rubisco Is also Inhibited by a
“Noctural” Inhibitor
• 2-carboxyarabinitol 1-phosphate inhibits
carbamoylated rubisco.
– transition state analog of -keto acid intermediate
– synthesized in the dark in some plants
 Stage 2: 3-phosphoglycerate Reduced
to Glyceraldehyde 3-Phosphate
Six 3-phosphoglycerate + 6 ATP + 6 NADPH + 6 H+  6
glyceraldehyde 3-phosphate + 6 ADP + 6 NADP+ + 6 Pi
• Requires NADPH and ATP from photosynthesis
• Part of gluconeogenesis (except NADPH used rather
than NADH)
• uses enzymes 3-phosphoglycerate kinase and
glyceraldehyde 3-phosphate dehydrogenase
• Driven forward by high concentration of NADPH and
ATP in the chloroplast stroma
Stage 3: Regeneration of Ribulose 1,5-
Bisphosphate
• Six glyceraldehyde-3-phosphate are produced
during the Calvin cycle.
• Ribulose-1,5-bisphosphate must be regenerated in
order to restart the cycle.
• 5/6 glyceraldehyde-3-phosphate products are
regenerated into ribulose-1,5-bisphosphate.
• 1/6 glyceraldehyde-3-phosphate products are pulled
out of the cycle and used for building carbohydrates.
Stage 3: Fate of One
Glyceraldehyde-3-
Phosphate in
Carbohydrate
Biosynthesis
Glyceraldehyde-3-
phosphate is an
intermediate in glycolysis
and gluconeogenesis and
can be used in anabolic
processes to create
starch and sucrose as
well as catabolic
processes to create
pyruvate and acetyl-CoA.
 Stage 3: Regeneration Steps for 5/6 of
Glyceraldehyde-3-Phosphate Products
of the Calvin Cycle
1. G3P converted to DHAP by triose phosphate
isomerase
2. G3P and DHAP converted to fructose 6-
phosphate (F6, 6C) via aldolase and fructose 1,6-
bisphosphatase
3. 2C removed from F6P to yield erythrose 4-
phosphate (E4P, 4C); catalyzed by transketolase
4. Transketolase transfers 2C from step 3 to G3P,
yielding xylulose 5-phosphate (Xu5P, 5C)
5. E4P and DHAP combine to sedoheptulose 1,7-
bisphosphate (S1,7BP; 7C) by aldolase.
6. Sedoheptulose 1,7-bisphosphate dephosphorylated
to sedoheptulose 7-phosphate (S7P) via
sedoheptulose 1,7-bisphosphatase
– This enzyme is unique to plants.
7. S7P has 2C removed by transketolase to yield ribose
5-phosphate (R5P); 2C on transketolase transferred
to G3P to yield another Xu5P
8. R5P converted into ribulose 5-phosphate (Ru5P)
by phosphopentose isomerase
9. Also, from step 7, Xu5P converted to RuP by
phosphopentose epimerase
10. Finally, RuP phosphorylated to ribulose 1,5-
bisphosphate by phosphoribulokinase
– This enzyme is also unique to plants.
 Stage 3:
Regeneration
Steps for 5/6 of
Glyceraldehyde-
3-phosphate
Products of the
Calvin Cycle
(Overview)
Stage 3: Addition of Phosphate from ATP to Ribulose-5-
Phosphate Commits Product to Calvin Cycle
 Transketolase Uses Thiamine
Pyrophosphate as the Cofactor

• Contains thiazolium anion for nucleophilic attack

on carbonyls
• Also used by:
– pyruvate dehydrogenase in acetyl CoA formation
– pyruvate decarboxylase in ethanol metabolism
– -ketoglutarate dehydrogenase in CAC
– transketolase in pentose phosphate pathway
Stoichiometry of CO2 Assimilation
in the Calvin Cycle

• Fixation of three CO2

molecules yields one
glyceraldehyde 3-
phosphate for use in
anabolic processes.
• Nine ATP molecules
and six NADPH
molecules are
consumed.
 Fate of the Pi from ATP Hydrolysis
in Stage 2
• Eight of the nine Pi combine with ADP to
regenerate ATP.
• The ninth is incorporated into G3P.
– requires that Pi be transferred from cytosol
– uses a special Pi-triose antiport
 The Pi-Triose Phosphate Antiporter
Is Needed for Sucrose Synthesis
• Unlike starch (made in the chloroplast stroma),
sucrose is made in the cytosol!
– used for transport to distant plant tissues
• But the inner chloroplast membrane is
impermeable to phosphorylated compounds.
• So, the antiporter exchanges G3P or DHAP for
one phosphate (Pi), thus sending the triose
phosphates to the cytosol for sucrose synthesis.
Photosynthesis: From Light and CO2 to
Glyceraldehyde 3-Phosphate

• The assembly of one molecule of glyceraldehyde 3-

phosphate requires the capture of roughly 24 photons.
– H+ move from the stroma to the thylakoid
– creates alkaline conditions in the stroma
• Accompanied by Mg2+ transport from thylakoid to
stroma
• Enzyme for photosynthesis and CO2 assimilation more
active in the alkaline, high [Mg2+] conditions of the
stroma
How H+ and Mg2+
Movement
Assist in
Activation of
Photosynthesis
and CO2
Assimilation
 Light Activates Four Enzymes
via Electron-Driven Reduction
of Cys-Cys Crosslinks
• Target enzymes are:
– ribulose 5-phosphate kinase
– fructose 1,6-bisphosphatase
– seduloheptose 1,7-bisphosphatase
– glyceraldehyde 3-phosphate dehydrogenase

• If oxidized (Cys residues in Cys-Cys disulfide form) 

enzymes are inactive.
• In light, photosystem I sends e– to ferredoxin, which
sends them to thioredoxin (Fig. 20-19), which donates
them to disulfide bonds to reduce them to free Cys.
Light Activation of Calvin Cycle
Enzymes
 Mitochondrial Respiration and
Photorespiration
• So far, we have seen that plants oxidize water to O2 and
reduce CO2 to carbohydrates.
CO2 + H2O  O2 + (CH2O)
• Plants also carry out the reverse in mitochondria: O2 is
oxidized to water, whereas substrates are reduced to CO2.
• In addition, a wasteful side reaction catalyzed by rubisco
occurs in mitochondria that consume O2 and yield CO2.
– This photorespiration reaction is driven by light and is coupled
with a costly salvage pathway.
– Unlike mitochondrial respiration, it does not yield energy.
 Oxygenase Activity of
Rubisco
• O2 competes with CO2 for the
active site.
~1 in every 3 or 4 turnovers, O2 binds
• The reactive nucleophile in the
rubisco reaction is the electron-
rich enediol form of ribulose 1,5-
bisphosphate.
• The nucleophile adds to O2 to
form 3-phosphoglycerate (same
as in Calvin cycle) and
2-phosphoglycerate (Fig. 20-12).
2-PG is metabolically useless.
Salvaging its C requires energy.
The Glycolate Pathway

• Complex ATP-consuming
process for the recovery of
C2 fragments from
photorespiration.
• Uses three organelles
• Loss of C as CO2 by
mitochondrial
decarboxylation of glycine
• Ultimately, two 2PGs are
converted to Ser + CO2.
Mechanism of Glycine Decarboxylase
 Glycine Decarboxylase Is Abundant
in Photosynthetic Parts of Plants

• Flux through rubisco reaction and glycolate

pathway can be high in bright sun, so plants need
a lot of glycine decarboxylase.
– makes up 50% of protein in some plant leaves
– but nonphotosynthetic parts of plants (tubers,
etc.) have little glycine decarboxylase
 C4 versus C3 Plants

• Most crop plants are C3.

– as discussed, first step in Calvin cycle is CO2 fixation into
3-C product, 3-phosphoglycerate
• for example: wheat, barley, potatoes
• C4 plants have an earlier step (before rubisco).
– bypass the C-3 fixation step by fixing CO2 into a 4-C
compound (oxaloacetate from acetyl CoA)
– tend to grow in hotter, sunnier climates
– have high rates of growth, photosynthesis, low water
loss, special leaf structure
• for example: sugar cane, corn/maize, sorghum
 Benefits of C4 Plants: Heat and
Drought Resistance

• Physical separation of reactions:

– CO2 is captured into oxaloacetate (4C) in mesophyll cells of
the leaf.
– Oxaloacetate is derived from pyruvate.
– Oxaloacetate then passes into bundle-sheath cells (where
rubisco is located) and releases CO2.
• The C4 pathway has a higher energy cost, but also has
increased efficiency in heat.
– As temperature increases, rubisco affinity for CO2 increases.
 C4 Pathway

C4 plants spatially
separate CO2 fixation
from rubisco activity,
resulting in less
reaction of rubisco
with oxygen and
avoidance of the costly
glycolate pathway.
 CAM Plants Separate CO2 Trapping
and Fixation in Time
• Another pathway to avoid the rubisco
reaction with oxygen
• First discovered in Crassulacae, so called
Crassulacean Acid Metabolism (CAM)
• Found in plants that grow at high temps,
dry conditions
• for example: pineapple, cacti
• minimize loss of water vapor during heat of the
day
 CAM Plants Open Gas-absorbing
Stomata Only at Night
• At night, air is cooler and moister.
– potential loss of water by the plant is lower
• CO2 absorbed at night is fixed to make
oxaloacetate (4C) via PEP carboxykinase.
• Oxaloacetate is reduced to malate and stored in
vacuoles.
• In daytime, malate releases CO2 via NADP-linked
malic enzyme, and CO2 fixation proceeds.
• Since stomata are closed, [O2] is low.
 Excess Triose Phosphate Is
Converted to Starch and Sucrose
• Regardless of whether a plant is C3, C4, or CAM,
glyceraldehyde-3-phosphate can be used to build
larger carbohydrates.
• Starch is made in chloroplasts for short-term
storage.
– or made in amyloplasts of nonphotosynthetic tubers,
seeds, roots, and so on, for long-term storage
– provides the bulk of stored energy for the plant
• Sucrose is made in cytosol for transport.
– Sucrose is the “blood glucose” of the plant.
 Starch Synthesis Proceeds Similarly
to Glycogen Synthesis
• Gluconeogenesis of two glyceraldehyde-3-
phosphates will result in glucose-6-phosphate.
• Phosphoglucomutase then converts glucose-6-
phosphate to glucose-1-phosphate.
• But unlike glycogen, glucose monomers in starch
come from ADP-glucose (not UDP-glucose), which
is made by ADP-glucose pyrophosphorylase.
• Starch synthase is similar to glycogen synthase. It
creates α(14) bonds (amylose).
– amylopectin: (16) branches added by branching
enzyme
 Starch Synthesis Is Regulated at
ADP-Glucose Pyrophosphorylase

• Activated by 3-phosphoglycerate
• Inhibited by Pi
 Sucrose Synthesis Uses UDP-Glucose
and Creates an α1β2 Bond

• Synthesis of sucrose occurs in the cytosol.

• DHAP is transported out of the chloroplast
using Pi-triose phosphate transporter.
• Once in the cytosol, DHAP undergoes
gluconeogenesis to synthesize glucose-6-
phosphate and fructose-6-phosphate.
• Glucose-6-phosphate is converted to UDP-
glucose by UDP-glucose pyrophosphorylase.
 Sucrose Synthesis Involves Conversion of
DHAP + G3P to Fructose Followed by
Addition of UDP-Glucose

1. Aldolase combines DHAP + G3P to form fructose

1,6-bisphosphate (F1,6BP).
2. F1,6BP is dephosphorylated by F1,6-
bisphosphatase to form fructose 6-phosphate
(F6P).
3. Sucrose 6-phosphate synthase adds UDP-
glucose to F6P to yield sucrose 6-phosphate.
4. Sucrose 6-phosphatase cleaves a PO43− to yield
sucrose.
Sucrose Synthesis Occurs in Two
Enzymatic Steps
Regulation of Sucrose Synthesis Occurs
at Interconversion of F1,6BP and F6P
• Isoforms of PFK-1 andfFructose-6-phosphatase have specific
regulation in plants.
• Specific isoform of PFK-1 is pyrophosphate-dependent PFK-1 (PP-PFK-
1).
 FBPase-1 and PP-PFK-1 Are
Regulated by Light and Dark
• In the dark, there is no photosynthesis to fuel the
Calvin cycle, and therefore no triose phosphates,
so no sucrose synthesis… but sucrose breakdown
is needed for energy.
– FBPase-1 inhibited (like gluconeogenesis)
– PP-PFK-1 active (like glycolysis)
• In the light, triose phosphate is made, so sucrose
is synthesized.
– FBPase-1 active
– PP-PFK-1 inhibited
 FBPase-1 and PP-PFK-1 Are Also
Regulated by F26BP
• Remember that F26BP is the important
glycolysis/gluconeogenesis regulator in animal cells.
– made and broken down by bifunctional enzyme
 Regulation of Sucrose Phosphate
Synthesis by Phosphorylation
• Activated by glucose 6-
phosphate
• G6P also inhibits the
kinase that makes SPS
less active.

• SPS is inhibited by Pi.

• Pi also inhibits the
phosphatase that makes
SPS more active.
Cell Walls Are Strong Due to Cellulose
• More than 1011 metric tons made every yearr!
• Glucose polymer with (14) linkages
– Straight shape allows chains to crosslink via H-bonds
 strong microfibrils.
• Synthesized from intracellular precursors but
assembled outside plasma membrane
Cellulose Structure
 Steps in One Model of Cellulose
Synthesis
1. Glucose attaches to lipid in the cell membrane.
2. Sucrose synthase generates UDP-glucose, and
with the help of cellulose synthase, six chains are
created.
• The cellulose-protein complexes form “rosettes” (seen by electron
microsopy).
3. Cellulose synthase extends the chain.
4. Growth and laying down of the polymer appear
to be directed by micotubules.
5. Chains crystallize to form fibrils.
A Model for Cellulose Synthesis
Plants Also Use Fats
and Proteins
for Carbohydrate
Synthesis:
Glyoxylate Cycle
 Chapter 20: Summary
In this chapter, we learned that:
• ATP and NADPH from photosynthesis are needed in order to assimilate
CO2 into carbohydrates
• assimilations of three CO2 molecules via the Calvin cycle lead to the
formation of one molecule of glyceraldehyde-3-phosphate for use in
anabolic reactions
• the key enzyme of the Calvin cycle, rubisco, fixes carbon dioxide as well
as oxygen
• low selectivity of rubisco causes a wasteful incorporation of molecular
oxygen in C3 plants. C4 and CAM plants have evolved separate methods
for reducing this waste.
• plants can assemble starch for energy storage, sucrose for transport, or
cellulose for structure. Each uses unique enzymes and has specific
regulation
• plants can convert acetyl-CoA into carbohydrates via the glyoxylate cycle