Calvin Cycle (Dark Reactions /

Light Independent Reaction)

 Calvin Cycle

Biochemical pathway that allows for

carbon fixation and occurs in the stroma.

Carbon enters in the form of CO2 and

leaves in the form of sugar.

Builds sugar from smaller molecules by

using ATP and the energy of electrons
carried by NADPH.
The actual sugar product of the Calvin cycle is
not glucose, but a three-carbon sugar,
glyceraldehyde-3-phosphate (G3P).

Each turn of the Calvin cycle fixes one carbon.

For the net synthesis of one G3P molecule, the

cycle must take place three times, fixing three
molecules of CO2.

To make one glucose molecules would require

six cycles and the fixation of six CO2 molecules.
 grana disks 2 outer
(thylakoids) membranes

stroma
compartment

Chloroplast

Calvin Cycle earlier was designated as the

photosynthetic “dark reactions”.
The ATP and NADPH are used to fix and reduce CO2 to
form carbohydrate (starch).
Enzymes of the Calvin Cycle are located in the stroma.
• Using CO2 + ATP + NADPH
• ATP = phosphate donor
• NADPH = hydrogen and electron donor
• CO2 = carbon donor forming sugar
• Starting material, Ribulose-1,5-bisphosphate is
regenerated at the end of Calvin Cycle
• Sugar produced = glyceraldehyde-3-phosphate (G3P)
• Calvin cycle must use 3 CO2 to produce 1 G3P
• Involved 3 stages:
– Carbon fixation
– Reduction
– Regeneration of Ribulose-1,5-bisphosphate
 Summary of the process:

12 NADPH C6H12O6 + 12NADP+

+ 18 ATP + 18 ADP++ 18 Pi + 6 H2O
+ 6CO2
Calvin Cycle
 Stage 1:
Carbon fixation
Carbon fixation
The Calvin cycle has three phases.

Phase 1: Carbon Fixation

- each CO2 molecule attach to a 5-carbon sugar,

Ribulose biphosphate (RuBP).

- catalyzed by enzyme RuBP carboxylase or

Rubisco.

- produce 6-carbon intermediate 2 molecules

of 3-phosphoglycerate (3-PG) for each CO2
fixed.
 2-
H2C OPO3 -O O
O C C
+ 3CO2
H C OH H C OH
H C

H2C
OH

OPO32-
 H2C OPO3
2-

6 3-Phosphoglycerate
3 Ribulose-1,5-bisphosphate
(3PG)
(RuBP)

Ribulose Bisphosphate Carboxylase (enzyme Rubisco),

catalyzes CO2 fixation:
Ribulose-1,5-bisphosphate + CO2  2 X 3-phosphoglycerate
Rubisco is the most abundant enzyme on earth.
 Stage 2:
Reduction
Reduction
Phase 2: Reduction

- 3-PG + P (from ATP) 1,3 biphosphoglycerate.

- a pair of electron from NADPH reduces 1,3

biphosphoglycerate G3P.

- for every three molecules of CO2 builds six

molecules of G3P.

- only one G3P can be counted as a net gain of

carbohydrate.
 Reduction Phase
•G3P may be converted to organic compounds:
• metabolites (e.g., fructose-6-P, glucose-1-P)
• energy stores (e.g., sucrose, starch)
• cell wall constituents (e.g., cellulose).
•G3P can also be utilized by plant cells as carbon
source for synthesis of other compounds such
as fatty acids & amino acids
 Stage 3:
Regeneration
Phase 3: Regeneration of the CO2 acceptor
(RuBP)

- the other 5 molecules of G3P are rearranged into

3 molecules of RuBP.

- the cycle spends three more molecules of ATP.

- RuBP is now prepared to receive CO2 again and

the cycle continues.
 Regeneration of RuBP
The three
molecules
of RuBP
will start a
new Calvin
Cycle.

Melvin Ellis Calvin
(April 8, 1911 - January 8, 1997)
Award: 1961 Nobel Prize in
Chemistry.
Andrew Benson
Award: Sugar Research Foundation
Award in 1950 and the Stephen
Hales Prize of the American
Society of Plant Biologists in 1972
for his discovery of ribulose.
James Alan Bassham: Associate Director
for Calvin Cycle Research.
Photorespiration
 Photorespiration
• Enzyme rubisco responsible for catalyzing the
combination of CO2 with RuBP.

• Active site of rubisco also accept O2

– O2 is the competitive inhibitor for CO2.

• If CO2 is higher then the plant will continue

with photosynthesis.
• If O2 is higher, then photorespiration will
occur.
 Photorespiration
• If O2 enters Calvin Cycle, Rubisco add O2 to RuBP
to produce glycolic acid.
• Peroxisomes and mitochondria break down
glycolic acid to CO2.
• Is a wasteful process, using O2 and producing
CO2.
• No food is produced.
• It decreases the efficiency of photosynthesis.
Photorespiration
Photorespiration
Differences between cellular respiration and
photosynthesis

Cellular Respiration
In cellular respiration, O2 is
the final electron acceptor. It
accepts electron at the end
of ETC and reduced to H2O.

Photosynthesis

In photosynthesis, H2O
brings electrons to the
photosystem and releases
O2
Differences between cellular respiration and
photosynthesis

During the Calvin cycle, CO2 is being

reduced into sugar

CO2
During the Krebs cycle, sugars are
oxidized and become CO2
C3, C4 & CAM
Plants
 C3 Plants
• First organic compounds of carbon fixation
(Calvin Cycle) is a 3 carbons 3PG.
• E.g. rice, soybeans, wheat.
• Produce less food when stomata closed in hot
day.
• Plants usually found in higher latitude.
• Less effective compared to C4 plants when low
CO2.
• Photorespiration can occurs.
Leaf anatomy of C3 plant
 C4 Plants
• First organic compounds of carbon fixation is a
4 carbons oxaloacetate.
• E.g. corns, sugarcanes.
• Not effected by low CO2.
• Photorespiration does not occur.
• Found in tropics and able to adapt to higher
temperature.
Leaf anatomy of C4 plant
 C4 Plants
• C4 plants have distinct
chloroplast containing
bundle sheath cells.
• Bundle sheath cells
surrounding the veins of
leaf tightly.

• C4 pathway happen in mesophyll cells but Calvin

Cycle happen in bundle-sheath cells.
• Mesophyll cells are closely associated with bundle-
sheath cells.
 C4 Plants
• Mesophyll cells are associated closely with bundle-
sheath cells.
• C-4 pathway occurs first in mesophyll cells, the Calvin
cycle occurs later in bundle-sheath cells.
• In mesophyll cell, PEP carboxylase enzyme present has
very high affinity towards CO2 (compared to Rubisco).
• PEP carboxylase catalyze fixation of CO2 to
Phosphoenol pyruvate (PEP) (3C compound) to form
oxaloacetate (4C).
• Oxaloacetate is then converted to malate (4C) using
NADPH.
• Then malate is decarboxylated to produce pyruvate
(3C) + CO2 + NADPH in bundle sheath cells.
Fig. 10.21
 C4 Plants
• CO2 is released in the bundle-sheath cells.
• Bundle-sheath cells are impermeable to CO2
 CO2 can be retained in very high concentration
• When CO2 is in high concentration, Calvin cycle
proceeds where rubisco enzyme fix CO2 to RuBP
• In addition, pyruvate produce earlier will return
to mesophyll cells and regenerated back to PEP
• New PEP will be ready to accept another CO2 in
mesophyll cells.
 4C
3C
NADPH NADP+

4C
3C

NADP+ + H NADPH
C3 vs. C4 Plants
At high CO2concentration, photosynthesis in C3 plants is more efficient than C4 plants at all
temperatures.
ADVANTAGES OF C4 OVER C3 PLANTS
Photosynthesis occurred even at very low CO2
concentration
PEP carboxylase has higher affinity towards CO2

Stored CO2 can be used later. CO2 release from

decarboxylation of malate to pyruvate can be used
when CO2 is low. (The method usually used by
plants when supply of water is low/draught =
stomata is closed)

No photorespiration will occur.

 CAM Plants

A plant that utilizes the Crassulacean acid

metabolism (CAM) as an adaptation for arid (dry)
conditions. CO2 entering the stomata during the
night is converted into organic acids, which release
CO2 for the Calvin Cycle during the day, when the
stomata are closed.
CAM Plants: alternate to Calvin Cycle
 CAM Plants
 Usually for plants in desert / hot places.
 E.g. cactus, pineapple (stomata close at day
time and open at night)
 (Other plants open stomata at day time)
 CAM pathway occurs at night followed by
Calvin cycle during day time.
 At night, enzyme PEP carboxylase fix CO2 to
PEP  oxaloacetate  malate.
 Malate stored in vacuole.
C4 vs. CAM Plants
Sugarcane Pineapple
C4 CAM CO2
CO2
1 CO2 incorporated
Mesophyll Organic acid (carbon fixation) Organic acid Night
cell

CO2 CO2
Bundle- 2 CO2 released Day
sheath Calvin Calvin
Cycle to the Calvin Cycle
cell
cycle

Sugar Sugar
(a) Spatial separation of steps (b) Temporal separation of steps