6 CO2 + 6 H2O + Light energy

TOPIC 6: PHOTOSYNTHESIS C6H12O6 + 6 O2

(the equation is simplified by indicating only the net
Subtopics:
consumption of water)
6.1 Overview of photosynthesis
• For photosynthesis, green plants use
6.2 Absorption spectrum of photosynthetic pigments
– sunlight as an energy source,
6.3 Light dependent reaction
– CO2 & H2O as raw materials.
6.4 Light independent reaction
• The light energy trapped by green plant
6.5 Alternative mechanisms of carbon fixation: Hatch-Slack /C4 and
– is converted to chemical energy &
Crassulacean Acid Metabolism/CAM pathways
– stored in the bonds of organic molecules.
• O2 is released as a byproduct.
6.1 Overview of photosynthesis
• Photosynthesis occurs in the chloroplasts.
Objective:
• Chloroplasts are found mainly in mesophyll cells.
a) Outline the complete process of photosynthesis: Light dependent
and light independent reactions • O2 exits & CO2 enters the leaf through microscopic pores,
stomata, in the leaf.
• A typical mesophyll cell has 30 - 40 chloroplasts.
Definition:
• The conversion of light energy to chemical energy that is stored
in sugar/glucose or other organic compounds. (Campbell 11th
edition, pg.259)
– Occurs in plants, algae & certain prokaryotes.

• Using molecular formulas, the complex series of chemical

reactions can be summarized with this chemical equation
(Campbell 11th edition, pg. 262):

6 CO2 + 12 H2O+ Light energy C6H12O6 + 6 O2 + 6 H2O

(water appears on both sides of equation because 12 molecules
are consumed and 6 molecules are newly formed during
photosynthesis)
Photosynthesis occurs in the chloroplasts.
• Structure of Chloroplast
– The thylakoid membranes & grana increase the surface area
for attachment of chlorophyll molecules, accessory pigments
& electron carriers involved in the light dependent reactions.
– The thylakoid space contains H+ involved in chemiosmotic
synthesis of ATP.
– The thylakoid membranes are surrounded by semi-fluid;
stroma.
– Stroma contains circular DNA, ribosomes & enzymes which
catalyse light-independent reactions, starch granules & lipid
granules.

• Photosynthetic pigments are embedded in the thylakoid

membranes.
– function of the pigments: to trap / absorb the light energy.
  Two types of photosystem:
• Photosystem I (PSI)
• Photosystem II (PSII)

Photosystem
• A large complex into which the light-absorbing pigments are
organized with proteins and other molecules”
• Located in the thylakoid membrane of the chloroplast
• Involved in the light-dependent reaction P700

Reaction centre:
• Composed of a reaction center-complex chlorophyll a,
 The reaction-center complex is an organized association of P700
proteins holding a special pair of chlorophyll a molecules and a
Photosystem I
primary electron acceptor.
Photosystem
 Each light-harvesting complex consists of various pigment
molecules (which may include chlorophyll a, chlorophyll b,and
multiple carotenoids) bound to proteins
 light-harvesting complexes act as an antenna for the reaction- P680
center complex Reaction centre:
chlorophyll a,
P680

Photosystem II
 6.2 Absorption Spectrum of Photosynthetic Pigments

PHOTOSYNTHESIS Objective:
a) State the photosynthetic pigments involved in photosynthesis.

Stage 1 Introduction
Light dependent reaction • Chlorophyll is the green pigment that absorbs light energy during
photosynthesis.
– gives the green colour to the leaves.
 Solar energy  chemical energy • When light meets matter, it may be reflected, transmitted, or
absorbed.
 Chemical energy: NADPH & ATP
 Involve photoactivation of PSI and
PSII
Stage 2
Light independent reaction

 ATP & NADPH (from light dependent

reaction) are used to convert CO2 
• Different pigments absorb photons of different wavelengths.
sugar – A leaf looks green because chlorophyll (dominant
pigment)
• absorbs red & blue light, while transmitting &
reflecting green light.
– Photon: small particles, packets of energy in the light.
Photosynthetic pigments Chromatography
• Chlorophyll a • Photosynthetic pigments can be separated using a technique
– blue-green pigment callled chromatography.
– absorbs best in the red & blue-violet light and reflecting green • Pigments are extracted from the leaf using acetone & separated
light on a chromatography paper using specific solvent
– The most abundant photosynthetic pigment Among pigments identified:
i. chlorophyll a (blue-green)
– Exists in several forms; depend on its red absorption peak; may ii. chlorophyll b (yellow-green)
be at 680nm or 700nm iii. xanthophyll (yellow) carotenoids
– act as reaction center iv. carotene (orange)
v. phaeophytin (grey)
• Chlorophyll b
– yellow-green pigment
Electromagnetic Spectrum
– absorbs best in the blue wavelength
– acts as accessory / antenna molecule
• absorbs light energy but have to transfer the energy to
chlorophyll a
• Carotenoids
– yellow, orange, red or brown pigments
– absorb violet and blue green light
– acts as accessory pigments / antenna molecules
• because they pass the light energy to chlorophyll a in the
reaction centre.
• The entire range of electromagnetic radiation is the
electromagnetic spectrum.
• The segment most important for life is the narrow band between
380 to 750 nm.
– visible light: because it is detected as various colours by human
eye.
6.3 Light Dependent Reaction
Objective
a) Explain the cyclic and noncyclic photophosphorylation.

LIGHT DEPENDENT REACTION

– Occurs in the thylakoid membrane

– Involve photoactivation of chlorophyll resulting in the
conversion of light energy into ATP through a complex and
produce NADPH + H+
– ATP is produced through photophosphorylation
What exactly happen when chlorophyll and other pigments absorb
light?
Light absorption
• Photons are absorbed by accessory photosynthetic pigments
molecules (antenna).
• The energy of the photon is converted to the potential energy of
an electron raised from its ground state to an excited state.
– photoexcited electrons
• When any antenna molecule absorbs a photon,
– it is transmitted from molecule to molecule until it reaches
reaction centre chlorophyll a molecule.
• Each photosystem consists of:
i. reaction center complex (chlorophyll a and primary electron
acceptor) surrounded by
ii. antenna complex, functions in the chloroplast as a light
harvesting unit.
• Photoactivation of Photosystem I & II
– Thylakoid membrane consists of 2 types of photosystems:
• Photosystem I
• Photosystem II
– The reaction center chlorophyll a of Photosystem I is P700,
– Because P700 is best at absorbing light with wavelength of
700nm (the far red part of the spectrum)
– Whereas for Photosystem II is P680, because its absorption
spectrum is 680nm (in the red part of the spectrum)

2 routes for electron flow
Noncyclic Photophosphorylation
(noncyclic/linear electron flow)
Cyclic Photophosphorylation
(cyclic electron flow)
I. Noncyclic Photophosphorylation (Linear electron flow)
1. Light energy/photon is absorbed by photosynthetic pigments of PS
II. The energy is transferred from one pigment to another until it is
passed to the pair of chlorophyll a molecules in the reaction-center
complex of PSII.
It excites an electron in this pair of chlorophylls to a higher energy
state (photoactivation occurs)
2. Photoactivated electron is elevated and captured by primary
electron acceptor. P680 becomes deficient/unstable (P680+)
3. Photolysis of water occurs - H2O  ½O2 + 2e- + 2H+
An enzyme catalyzes the splitting of a water molecule into two
electrons, two hydrogen ions (H+), and an oxygen atom.
The electrons are supplied one by one to the P680+ pair, each
electron replacing one transferred to the primary electron
acceptor.
4. Each photoexcited electron passes from the primary electron
acceptor of PS II to PS I via an electron transport chain
The electron transport chain between PS II and PS I is made up of
the electron carrier plastoquinone (Pq), a cytochrome complex,
and a plastocyanin (Pc).
Each component carries out redox reactions as electrons flow
down the electron transport chain, releasing free energy that is
used to pump protons (H+ ) into the thylakoid space, contributing
to a proton gradient across the thylakoid membrane.
5. The potential energy stored in the proton gradient is used to
make ATP in a process called chemiosmosis
6. Meanwhile, light energy has been transferred via light harvesting
complex pigments to the PS I reaction-center complex, exciting an
electron of the P700 pair of chlorophyll a molecules located there.
The photoexcited electron is then transferred to PS I’s primary
electron acceptor, creating an electron “hole” in the P700—which
now can call P700+.
 The missing electron in P700+ can now act as an electron acceptor, Why Cyclic Photophosphorylation is needed??
accepting an electron that reaches the bottom of the electron • Noncyclic electron flow produces ATP and NADPH in roughly
transport chain from PS II. equal quantity

7. Photoexcited electrons are passed in a series of redox reactions
from the primary electron acceptor of PS I down a second electron
transport chain through the protein ferredoxin (Fd). (This chain
does not create a proton gradient and thus does not produce ATP.)
8. The enzyme NADP+ reductase catalyzes the transfer of electrons
from Fd to NADP+. NADP+ is reduced to NADPH
• But the Calvin cycle consumes more ATP than NADPH.
• The concentration of NADPH will help regulate which pathway;
noncyclic or cyclic.
• If the chloroplast runs low on ATP for the Calvin cycle NADPH
will begin to accumulate.
• The rise of NADPH will stimulate a temporary shift from
noncyclic to cyclic electron flow
• Until ATP supply is enough to generate Calvin cycle.

Cyclic Photophosphorylation
• Under certain condition, photoactivated electron takes an Light Dependent Reaction
alternative path called cyclic electron flow.
• Involves only PS I.
• The electron from ferredoxin (Fd) is transferred to cytochrome Convert LIGHT ENERGY
to CHEMICAL ENERGY
complex and plastocyanin which returns to PS I to stabilize it.
• Only ATP is generated.
Through NONCYCLIC
ATP PHOTOPHOSPHORYLATION
NADPH (PS II & PS I)

If ATP not enough,

Cannot proceed Calvin cycle
So, need short cut to have more ATP

ThroughCYCLIC
PHOTOPHOSPHORYLATION
(PS I)
 6.4 Light independent reaction/Calvin Cycle
Objective
a) Explain Calvin cycle involving carbon fixation, reduction and
regeneration of CO2 acceptor (RuBP)

Calvin cycle
• Uses ATP and NADPH to convert CO2 to glucose/sugar
 ATP: source of energy
 NADPH: reducing agent
• Occurs in the stroma of the chloroplast and each stage is
mediated by an enzyme
• Consist of three stages:
1. Carbon fixation

The light reactions: organization of the thylakoid membrane. 2. Reduction

3. Regeneration of CO2 acceptor (RuBP)

Differentiate between Cyclic and Noncyclic Photophosphorylation

Criteria Cyclic Non-cyclic
photophosphorylation photophosphorylation
Photosystem involved

Products

Electron flow

Photolysis of water

Oxygen

Electron donor

Final electron acceptor

Stage 1: Carbon Fixation
• Each CO2 molecule, one at a time, by attaching it to a five-
carbon sugar ribulose bisphosphate (RuBP) catalyzes by RuBP
carboxylase-oxygenase, or rubisco.
• The product of the reaction is unstable six-carbon
intermediate (short-lived)
• The intermediate splits into two molecules of 3-
phosphoglycerate (PGA) per one CO2 molecule.
Stage 2: Reduction
• Each molecule of 3-phosphoglycerate receives an additional
phosphate group from ATP, becoming 1,3-bisphosphoglycerate.
• Next, a pair of electrons donated from NADPH reduces 1,3-
bisphosphoglycerate which also loses a phosphate group in the
process, becoming glyceraldehyde 3- phosphate (G3P)
• The NADP+ and ADP formed in this process return to the
thylakoids to regenerate NADPH and ATP in the light dependent
reactions
• For every 3 molecules of CO2 that enter the cycle, 6 molecules of
G3P formed. But only 1 molecule of G3P can be counted as a net
gain of glucose. Another 5 molecules of G3P will be used in the
next phase
Stage 3: Regeneration of CO2 acceptor (RuBP)
• The remaining 5 molecules of G3P are rearranged into 3
molecules of RuBP
– 3 molecules of ATP are required
• Now, RuBP molecules are prepared to receive CO2 again and the
cycle continues.
Summary
For every 3 CO2 fixed into Calvin Cycle
• 1 molecule of G3P is formed (3 CO2 : 1 G3P).
• The remaining 5 molecules of G3P re-enter Calvin cycle to
produce 3 molecules of RuBP.
• G3P molecules formed in the Calvin cycle can remain in the
chloroplast; converted to starch.
• Alternatively, G3P molecules can be exported from the chloroplast
(where it is converted to sucrose) to cytosol.
• Out of 6 molecules of G3P (produced in phase 2), only 1 G3P exits
the Calvin cycle
• Consumes a total of 9 ATP & 6 NADPH.
• Therefore, to produce one molecule of glucose,
• 2 molecules of G3P are needed from the Calvin cycle. That
consumes:
 6 CO2
 18 ATP & 12 NADPH
C3 Plants and Photorespiration • We naturally view photorespiration as wasteful.
• Most plants use Calvin cycle to fix CO2 from environment. • The environmental conditions that allow photorespiration are:
hot, dry, bright days.
– And the 1st molecule to form from CO2 fixation is 3C molecule,
PGA. • In certain plant species, alternate modes of carbon fixation
(minimise photorespiration), even in hot, arid climate have
– This type of plants is called C3 plants.
evolved.
• On a hot and dry day, most plants close their stomata to
• The 2 most important of these photosynthetic adaptations are
conserve water.
i. C4 pathway
• When stomata closed,
ii. Crassulacean acid metabolism (CAM) pathway
– CO2 concentration in the air spaces within the leaf decreases.
– O2 concentration (from light dependent reaction) begins to
increase. 6.5 Alternative mechanism of carbon fixation: Hatch-Slack
(C4) and Crassulacean Acid Metabolism (CAM) pathways
• This condition within the leaf will lead to wasteful process called
photorespiration.
• When O2 concentration overtake CO2 concentration (within the Objective
air space in the leaf), a) Explain photorespiration and state the alternative mechanism of
– RuBP carboxylase (Rubisco) now add O2 to the Calvin cycle carbon fixation (C4 and Crassulacean Acid Metabolism (CAM)
instead of CO2. pathways.
b) Compare carbon fixation in C4 and CAM plants.
RuBP + O2 phosphoglycolate + PGA

Hatch-Slack (C4) Pathway

phosphoglycolate CO2 + H2O
• Pathway for transporting CO2 from mesophyll cells to bundle
sheath cells.
• It is called photorespiration because it occurs in the light (photo) – Once in the bundle sheath cells, the CO2 is released again and
and consumes O2 (respiration). normal C3 photosynthesis occurs.
• During photorespiration, • A group of plants has evolved by using an alternative method to
fix carbon dioxide from the environment.
– No ATP generated and no food produced
– does not rely on RuBP carboxylase
– Decreases photosynthetic output by reducing organic
material from Calvin cycle. • Carbon fixation (from CO2) forms a four-carbon (4C) compound as
its first product.
 – This type of plants is called C4 plants.
• C4 plants, have 2 different types of photosynthetic cells:
i. bundle-sheath cells
ii. mesophyll cells

Hatch-Slack (C4) Pathway

CO2

Phosphoenol
Oxaloacetate (4C)
pyruvate (3C)

ADP NADPH + H+

ATP NADP+
Pyruvate (3C) Malate (4C)

-oxygenase

Pyruvate (3C) Malate (4C)

NADP+

CO2
Calvin NADPH + H+
Glucose

cycle
Hatch-Slack (C4) Pathway Differences in the Physiology of C3 and C4 plants

First Carbon Fixation (in mesophyll cells)

1. First CO2 Fixation: CO2 first combines with 3C phosphoenol-
pyruvate (PEP) to form oxaloacetate(OAA).
2. Catalysed by PEP carboxylase.
3. OAA(4C) is reduced by NADPH into malate(4C)
4. Malate is transported to bundle sheath cells.
-oxygenase -oxygenase

Second Carbon Fixation (in bundle sheath cells)

1. In the bundle sheath cells, malate(4C) undergoes decarboxylation
to form CO2 and pyruvate(3C).
2. Pyruvate is transported back to mesophyll cells and
phosphorylated by ATP to form phosphoenolpyruvate
3. Second CO2 fixation occurs when CO2 released in decarboxylation
undergoes second CO2 fixation in Calvin cycle.
4. CO2 fix with RuBP catalyse by rubisco forming PGA and eventually
into sugar
5. The mesophyll cells of a C4 plants pump CO2 into the bundle-
sheath cells.
6. To keep CO2 concentration in the bundle-sheath cells high enough
for rubisco to accept CO2 rather than oxygen.
7. In this way, C4 plants minimizes photorespiration and enhances
sugar production.
8. Examples of C4 plant: sugarcane and corn
 1st product formed

Efficiency of
photosynthesis

Photorespiration
rate

Crassulacean Acid Metabolism/CAM Pathway

C3 plants C4 plants 1. A second strategy to minimise photorespiration.
Cells involved in Cells involved in 2. Found in succulent plants such as cactus and pineapples.
photosynthesis: photosynthesis: 3. Known as CAM plants for crassulacean acid metabolism (CAM).
mesophyll cells only mesophyll cells
4. CAM plants open stomata during the night and close them during
bundle-sheath cells usually bundle sheath cells
the day.
lack of chloroplast bundle-sheath cells have
5. At night, stomata open:
chloroplasts
– CO2 from environment enters the leaf tissue
C3 plants C4 plants – CO2 + PEP(3C)  Oxaloacetate(4C) Malate(4C)
Leaf anatomy – Malate is transported into the vacuole
6. During the day, the light dependent reactions supply ATP and
Chloroplast NADPH to the Calvin cycle and CO2 is released from the organic
acids.
Bundle sheath cells
7. During the day, stomata are closed
Carbon dioxide – Malate (from vacuole) moves into the chloroplast.
fixation – Malate (4C) converted to pyruvate(3C) and releases CO2.
Carbon dioxide – CO2 enters Calvin cycle
acceptor

Enzyme involved in
CO2 fixation