CARBON REACTIONS OF PHOTOSYNTHESIS

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 Photosynthetic Processes

1. Light absorbtion
light
2H2O O2 + 4H+ +4 e-

2. Electron transport
light
2H2O + 2NADP+ 2H+ + 2 NADPH + O2

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3. ATP formation

H+ + ADP 3- + Pi 2- ATP 4- + H2O

4. CO2 fixation to form a carbohydrate

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 The light reactions of photosynthesis show how
water (H2O) is oxidised into O2 couples with ATP
and NADPH formations via the reactions occur in
the thylakoid membrane

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• The reactions which catalyse CO2 reduction to
form carbohydrate, couple with ATP and
NADPH uses, occur in the Stroma

• These reactions used to be refferred as the dark

reactions of photosynthesis

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 •lumen

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• Now, it is known that those reactions need light.

• Therefore, the conversion of CO2 into

carbohydrates, has to be called as the carbon
reactions of photosynthesis

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Every photosynthetic organism which reduces
CO2 into carbohydrate, has the same basic
mechanism known as Calvin cycle
or C3 cycle
or reductive pentose phosphate cycle

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 CALVIN CYCLE

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 • In Calvin cycle, CO2 and H2O from the
environment are enzymatically combined with a
5-carbon acceptor molecule to produce 2
molecule of 3-carbon intermediate (3-
phosphoglycerate)
• This intermediate is subsequently reduced into
carbohydrate using the ATP and NADPH

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 The Calvin cycle occurs in three stages :

1. Carboxylation
2. Reduction
3. Regeneration

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 1. Carboxylation

• is the addition of CO2 molecule into Ribulose-1,5-

bisphosphate (RuBP) to form 2 molecule of 3-
phosphoglycerate, catalysed by a Ribulose-1,5-
bisphosphate carboxylase/oxygenase (Rubisco)

• RuBP + CO2 + H2O Rubisco 2(3-

phosphoglycerate/ PGA) + 2 H+ …………(*)

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 The carboxylation reaction consists of two
reactions:
a. Carboxylation: CO2 + RuBP resulting in the
formation of unstable intermediate: 2-
carboxy-3-ketoarabinitol-1,5-bisphosphate
b. Hidrolysis: of 2-carboxy-3-ketoarabinitol-1,5-
bisphosphate into 2(phosphoglycerate/ PGA)

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 Notes:

• Rubisco can catalyse both carboxylation (+ CO2) and

oxygenation (+ O2) of the same substrate (RuBP) ,
because both reactions occur in the same catalytic site
of the rubisco
• Rubisco is the most abundant dissolve protein in the
plant (40%)
• Rubisco affinity towards CO2 is very high
• Right bound reaction (*) is easily occur

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 2. Reduction

During the reduction,

• PGA is firstly phosphorylated (using ATP) to
produce 1,3-bisphosphoglecerate + ADP
• The 1,3-bisphosphoglecerate is subsequently
reduced (using NADPH) to produce
Glyceraldehyde-3-phosphate

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 Notes:

The reduction processes can be traced by looking

at the atomic charge of the carbon atom :
• CO2: +4
• PGA: +3
• Glyceraldehyde-3-phosphate: +1

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 3. Regeneration

• As a consequence of the continuous absorbtion

of CO2, RuBP should also be regenerated
continuously
• The carboxylation and subsequent reduction of
3 RuBP molecules result in the formation of 6
glyceraldehyde–3-phosphate

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• From those 6 molecules of glyceraldehyde-3-
phosphate, 1 molecule will be used to form
sucrose/starch.
• The remaining 5 molecules of glyceraldehyde-
3-phosphate (3C) will be transformed to 3
molecules of RuBP (5C)

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 Reaction starts
Ribulose-1,5 bsphosphate
ADP CO2 + H2O

1. Carboxylation

3. Regeneration
3-Phosphoglycerate

ATP + NADPH

2. Reduction

ATP
Glyceraldehyde- ADP + Pi, NADP+
3-phosphate

Sucrose, starch

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 Summary

• 3 RuBP + 3 CO2 + 3 H2O 6(3-PGA)

(Carboxylation)

• 6PGA + 6 ATP + 6 NADPH

• 6 Glyceraldehyde-3-phosphate+ 6 ADP + 6 NADP+
(Reduction)

• 6 Glyceraldehyde-3-phosphate + 2 H2O 3 RuBP +

Glyceraldehyde-3-phosphate (for sucrose/starch synthesis)
(Regeneration)

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 Note:

• The Calvin cycle has an autocatalytic properties: its rate of

operation can be speed up by adding its intermediate

• The Calvin is able to produce more substrate, as far as the

Glyceraldehyde-3-phosphate was not used in other processes:

5 RuBP 4- + 5 CO2 + 9 H2O + 16 ATP 4- + 10 NADPH

6 RuBP 4- + 14HOPO3 2- + 6 H+ + 16 ADP 3- +
10 NADP+

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The Calvin cycle reactions:

6 CO2 + 11 H2O + 12 NADPH + 18 ATP

Fructose-6-phosphate + 12NADP+ + 6 H+ + 18
ADP + 17 Pi

• For each CO2 molecule fixed to form

carbohydrate, 2 NADPH and 3 ATP molecules are
needed

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Light-Dependent Enzyme Activation Regulates
the Calvin Cycle

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 Light regulates the following enzymes:

1) Rubisco

2) NADP Glyceraldehyde-3-phosphate
dehydrogenase
3) Fructose-1,6-bisphosphate phosphatase
4) Sedoheptulose-1,7-phosphatase
5) Ribulose-5-phosphate kinase
(via a feredoxin-thioredoxin system)

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 Light controls the activity four
enzymes via the ferredoxin–thioredoxin system

• In the dark these residues exist in the oxidized state

(—S—S—), which renders the enzyme inactive or
subactive

• In the light the —S—S— group is reduced to the

sulfhydryl state (—SH HS—), resulted in the
activation of the enzyme

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 h
PSI

Ferredoxin Ferredoxin
Oks Red

Red Oks

Thioredoxin Thioredoxin

SH-HS S-S

Oks Red

Enzim target Enzim target

S-S
SH-HS

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• The rubisco activity increases by the light
• Inhibitor which bound to rubisco in the dark,
will be release after rubisco receive light
• The increase of [H+] in the lumen caused by
light resulted in the increase of stromal pH
(from 7 to 8), in which rubisco will be more
active

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 THE C2 OXIDATIVE PHOTOSYNTHETIC
CARBON CYCLE

• Rubisco catalyses both carboxylation and

oxygenation.
• Oxygenation is the primary reaction of
photorespiration
• The oxygenation of RuBP produces
2-phosphoglycolate + 3-PGA + H+

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• Photorespiration resulted in the nett loss of CO2
from the cell.
• The lost CO2 is scavenged by a Photorespiratory
Carbon Oxidation (PCO) Cycle
• The Photorespiratory Carbon Oxidation (PCO)
Cycle invoves 3 organels: chloroplas, peroxisome,
and mitochondrion

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• 2 (2-phosphoglycolate ) 3-PGA + CO2
•

• 3 C are successfully scavenged by PCO cycle

(75 %)
• the real photosynthetic rate should be: 120 –
125 %

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• The Calvin cycle is independent
• PCO cycle is a Calvin cycle dependent, as
source of RuBP

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The balance betwee Calvin and PCO cycles is
controlled by 3 factors

• Kinetic properties of Rubisco (Carboxylation is

easier than oxygenation)
• [CO2] and [O2]
• Temperature

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• By the increase of temperature, [CO2] decrease
is far faster than that of [O2], leading to the
decrease of [CO2]:[O2]. As a consequence
oxygenation reaction will be easier to occur.

• Moreover, the increase of temperature

stimulates the oxygenation reaction

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 Note:

Photorespiration occurs in the low [CO2]

and high temperature conditions

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 Photorespiration functions

• The biological functions of photorespiration has

not been fully understood.

• One possibility is that the PCO cycle is an effort to

scavenge the lost CO2 during photorespiration

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• Another possibility is that photorespiration is used
to release the excess ATP and NADPH, therefore
avoiding both photooxydation and photoinhibition

• The most recent finding, using genetic

engineering, showed that photorespiration protects
C3 plants from photooxydation and photoinhibition

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 CO2 Concentrating Mechanism

• In several plants their photorespiration is severely

inhibited or even no photorespiration occurred.

• It happens due to the existence of CO2 concentrating

mechanism in the rubisco area, leading to the inhibition
of RuBP oxygenation reaction

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 CO2 Concentrating Mechanisms

• CO2 pumping in algae and Cyanobacteria

• C4 Carbon cycle
• Crassulacean Acid Metabolism

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 Mechanism I. CO2 pumping in algae and
Cyanobacteria

• is a pumping mechanism of CO2-HCO3 in the

plasma membrane using ATP, during the low CO2
conditions

• Outcome: the inhibition of RuBP oxygenation

(Photorespiration inhibition)

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 Mechanism II. C4 carbon cycle
(Siklus Hatch Slack)

• in a C3 plants, Chloroplast is located in the mesophyll

cells
• on the other hand, in a C4 plants, there are 2 (two)
chloroplast containing cell types i.e. mesophylls cells
and bundle sheath cells/Kranz anatomy

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 C4 Leaf C3 Leaf

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 C4 Leaf

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C4 Cycle concentrating the CO2 in bundle sheath
cells, which occur in 4 steps:

1. CO2 Fixation i.e. the carboxylation of a Phosphoenol

pyruvate (PEP), in the mesophyll cells, resulting in 4 C
acid formation (malate or aspartate)

2. Transport of a 4C molecule into bundle sheath cells

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3. Decarboxylation of the 4C acid, within the bundle
sheath cells, and resulted in CO2 formation, which will
be reduced in Calvin Cycle

4. Transport of a 3C molecule, a product of the

decarboxylation into the mesophyll cells and subsequent
regeneration of PEP

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C4 cycle

Effectively transport CO2 from the atmosphere into the

bundle sheath cells, which therefore increases [CO2] in
the carboxylation area , leading to the inhibition of
RuBP oxygenation (photorespiration) to occur

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 C4 Cycle Variations

Those variation are based on the kind of C4 transported

into the bundle sheath cells (malate/aspartate) and
the kind of C3 acid transported back into the
mesophyll cells (pyruvate/alanine) ; and the enzyme
which catalyses the carboxylation reaction

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 NADP Malic Enzyme Type

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 NAD Malic Enzyme Type

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Phosphoenolpyruvate Carboxykinase Type

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 In the C4 Cycle
• 2 ATP molecules are needed for every CO2
molecule being transferred into the bundle sheath

• Therfore, if it is combined with the Calvin Cycle

which also occurs in C4 plants, the total energy
needed to fixed 1 molecule of CO2 into
carbohydrate is 2 NADPH and 5 ATP

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Several enzymes involve in C4 cycle (PEP carboxyilase,
NADP:malate dehydrogenase, piruvate-ortoposphate
dikinase) all are affected by light

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 The advantage of C4 Cycle

1. Preventing a water lost at high temperature . The PEP

carboxylase has a very high affinity to HCO3- , which is
also not a O2 competitor, therefore plants can reduce the
stomatal aperture when they fix CO2, which in turn
reduce the water lost.
2. Preventing photorespiration: the high [CO2] in the
carboxylation area (Calvin cycle) in bundle sheath
inhibit RuBP oxygenation (photorespiration) to occur

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 Mechanism III. Crassulacean Acid Metabolism
(CAM)

• CAM occurrs not only in Crassulacea Family, but also in

Cactus and Euphorbia

• CAM increases water use efficiency (50 – 100 g of water

is lost for every 1 gram CO2 gained) (C4: 250 –300; C3: 400
– 500)

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The CAM mechanism is similar in many respects to the
C4 cycle.

• In C4 plants, the formation of the C4 acids in the

mesophyll is spatially separated from decarboxylation
of the C4 acids and from refixation of the resulting
CO2 by the Calvin cycle in the bundle sheath.

• In CAM plants, formation of the C4 acids is both

temporally and spatially separated from
decarboxylation of the C4 acids and from refixation of
the resulting CO2 by the Calvin cycle .
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At night in the cytosol

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At day time at the chloroplast

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 Sucrose and Starch Synthesis

• Sucrose (glucose-fructose chain) is the principal

form of carbohydrate translocated throughout the
plant by the phloem.

• Starch (glucose polymer) is an insoluble stable

carbohydrate reserve that is present in almost all
plants.

• Both starch and sucrose are synthesized from the

triose phosphate generated by the Calvin cycle

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• Sucrose and starch syntheses are two competing
processes, occur in different part of the cell

• Sucrose synthesis occurs in cytosol, whereas starch

synthesis and accumulation occur in chloroplast

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• The transformation of triose phosphate to either sucrose
or starch, is determined by the concentration of
orthophosphate (Pi) in the cell.

• When the [Pi] in the cytosol is high, then the triose

phosphate in the chloroplast will be transported into the
cytosol (replaced by the Pi), and sucrose is then
synthesised.

• On the other hand , when the [Pi] in the cytosol is low,

then the triose phosphate in the chloroplast is retained in
the chloroplast, and starch is synthesised

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 Summary:

The carbon reaction of photosynthesis is started by

carbon reduction in the Calvin cycle coupled with the
use of ATP and NADPH , and resulted in the
formation of triose phosphate (carbohydrate), which
then transformed to energy and carbon reserves in the
forms of sucrose and starch

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