Chapter.

one Photosynthesis

Contents to read in this chapter

i. History of photosynthesis.
ii. Nature and units of light.
iii. Determination of oxygenic and non-oxygenic photosynthesis.
iv. Ultrastructure of thylakoid vesicle.
v. Various pigments and photosynthetic activity.
vi. Ultrastructure and composition of Photosystem-I and II.
vii. Absorption and action spectra of different pigments.
viii. Mechanism of photosynthesis - light absorption, charge separation or oxidation of water (water
oxidizing clock), electron and proton transport through thylakoid protein-pigment complexes.
ix. Photophosphorylation and its mechanism.
x. CO2 reduction (dark reactions) - C 3 pathway and Photorespiration, Regulation of C 3 pathway,
C 4 pathway and its different forms, C 3 -C 4 intermediates, CAM pathway.
xi. Methods of measurement of photosynthesis.

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 (ii) Nature and units of light

The term ‘‘light’’ describes that portion of the electromagnetic spectrum that
causes the physiological sensation of vision in humans. In other words, light is defined by
the range of wavelength between 400 and approximately 700 nanometers—capable of
stimulating the receptors located in the retina of the human eye. Strictly speaking, those
regions of the spectrum we perceive as red, green, or blue are called light, whereas the
ultraviolet and infrared regions of the spectrum, which our eyes cannot detect (although they
may have significant biological effects), are referred to as ultraviolet or infrared radiation,
respectively.
Physicists of the late nineteenth and early twentieth centuries resolved that light has
attributes of both continuous waves and discrete particles. Both of these attributes are
important in understanding the biological nature of light.

 Light as a Wave Phenomenon

The propagation of light through space is characterized by regular and repetitive changes,
or waves, in its electrical and magnetic properties. Electromagnetic radiation actually consists
of two waves—one electrical and one magnetic—that oscillate at 90◦ to each other and to the
direction of propagation (Figure6.2).
The wave properties of light may be characterized by either wavelength or frequency.
Wavelength: The distance in space between wave crests is known as the wavelength and is
represented by the Greek letter lambda (λ). Biologists commonly express wavelengths in units
of nanometers (nm), where 1nm= 10−9 m.

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Frequency: It is the number of wave crests, or cycles, passing a point in space in one second.
Frequency is represented by the Greek letter nu (ν),it is thus related to wavelength in the
following way: ν =c/λ where c is the speed of light (3×108 ms−1)
Biologists most commonly use wavelength to describe light and other forms of radiation,
although frequency is useful in certain situations. Wavelengths of primary interest to
photobiologists fall into three distinct ranges: ultraviolet, visible, and infrared (Table 6.1).

 Light as A Stream of Discrete Particles

When light is emitted from a source or interacts with matter, it behaves as though its energy
is divided into discrete units or particles called photons.The energy carried by a photon is
called a quantum (pl. quanta), to reflect the fact that the energy can be quantized, that is, it can
be divided into multiple units. The energy carried by a photon (Eq) is related to wavelength
and frequency in accordance with the following relationship: Eq =hc/λ =hν
where h is a proportionality constant, called Planck’s constant. Its value is 6.62 × 10−34 J s
photon −1. Accordingly, the quantum energy
of radiation is inversely proportional to its
wavelength or directly proportional to its
frequency. The symbol hν (pronounced ‘‘h
nu’’) is commonly used to represent a photon
in figures and diagrams.The energy carried by
a mole of photons of red light is 181 kJ mol−1,
and for blue light is correspondingly 274
kJmol-1.

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 III. Determination of oxygenic and non-oxygenic

Oxygenic Photosynthesis: -
The photosynthesis that occurs under aerobic conditions in which oxygen is produced
as by product and the final electron accepter is water.

Anoxygenic Photosynthesis: -
The photosynthesis that occurs under anaerobic conditions in which Sulphur is
produced as by product and the final electron accepter is other than water as H2S.

Properties Oxygenic photosynthesis Anoxygenic photosynthesis

Occurance Plants,Algae,Cyanobacteria Green sulphur and non-sulphur

bacteria, purple bacteria,
heliobacteria and acidobacteria
Photosystems PS-I(P-700) and PS-II(P-680) Only PS-I (P-870) is present
are present
Electron Source Water(H2O) is electron source H2S or Ferrous ions

By-product Oxygen Sulphur or other than oxygen

Photosynthetic pigments chlorophylls bacteriochlorophylls

Generation of NADPH NADP act as terminal electron NADPH not produced ,electrons
accepter, producing NADPH cycled back to the system
ATP production ATP produced by non-cyclic ATP produced by cyclic
photophosphorylation photophosphorylation

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 (iv) Ultrastructure of Thylakoid Vesicle

The thylakoids (thylakoid = sac-like) consists of flattened and closed vesicles arranged as a
membranous network. The outer surface of the thylakoid is in contact with the stroma, and its
inner surface encloses an intrathylakoid space (the third compartment). Thylakoids may be
stacked like a neat pile of coins, forming grana or they may be unstacked, inter-granal, or
stromal thylakoids, forming a system of anastomosing tubules that are joined to the grana
thylakoids. There may be 40 to 80
grana in the stroma of a chloroplast.
The number of thylakoids per granum
may vary from 1 to 50 or more. For
example, there may be
 Single thylakoid (red alga)
 Paired thylakoids (Chrysophyta)
 triple thylakoids and multiple
thylakoids (green algae and
higher plants).

 Molecular Organization of Thylakoids

Molecular organization of the membrane of thylakoids is based on the fluid-mosaic model
of the membrane which represents following main characteristics: fluidity, asymmetry and
economy (i.e., lack of movement in the third dimension).
 Lipid content in the thylakoid membrane
Lipids represent about 50 per cent of the thylakoid membrane. There are two types of lipids,
 Functional lipids: The lipids that directly involved in photosynthesis called functional
lipids such as chlorophylls, carotenoids and plastoquinone.
 Structural lipids: The lipids that involved in the formation of membrane are structural
lipids. For example, Glycolipids, Sulpholipids and a few Phospholipids. Most of these
structural lipids are highly unsaturated which confer to the membrane of thylakoids a high
degree of fluidity.

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 The protein content in thylakoid membraneThe protein components of thylakoid
membrane are represented by 30 to 50 polypeptides which are disposed in the following
five major supramolecular complexes which can be isolated with mild detergent.
1. Photosystem I (PS-I)
This complex contains a reactive center composed of P700 (Type of pigment which is
bleached at the wavelength of 700 nm), several polypeptides, a lower chlorophyll a/b ratio and
β-carotene. It acts as a light trap and is present in unstacked thylakoid membranes (stromal
thylakoids). In it light induced reduction of NADP+ takes place.

2. Photosystem II (PS-II)
This complex comprises two intrinsic proteins that bind to the reaction center of
chlorophyll P680 (The pigment that bleaches when absorbing light at 680 nm). It contains a
high ratio of chlorophyll a, b and β-carotene. Frequently, the PS-II is associated with the light
harvesting complex (LHC-II) and are involved in light induced release of O2 from H2O (i.e.,
photolysis of water). PS II works as a light trap in photosynthesis and is mainly present in the
stacked thylakoid membranes of grana.
3. Cytochrome b/f
This complex contains one cytochrome F, two cytochromes of b-563, one FeS center and
a polypeptide. It is uniformly distributed in the grana and acts as the electron carrier. These
three complexes are related to the electron transport and are linked by mobile electron carriers
(i.e. plastoquinone, plastocyanin and ferredoxin). Electron transport through PS-II and PS-I
finally results in the reduction of the coenzyme NADP+. Simultaneously, the transfer of
protons from the outside to the inside of the thylakoid membrane occurs.

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4. ATP synthetase
This complex consists of a CF-0 (CF=coupling factor) hydrophobic portion, a
proteolipid that makes a proton channel, and a CF-1 that synthesizes ATP from ADP and Pi,
using the proton gradient provided by the electron transport. ATP synthetase complexes are
located in stacked membrane (grana).
5. Light harvesting complex (LHC)
The main function of LHC is to capture solar energy. It contains two main polypeptides
and both chlorophylls a and b. LH complex is mainly associated with PS-II, but may also be
associated with PS-I (Anderson, 1975). LHC is localized in stacked membranes and lacks
photochemical activity.

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v. Various pigments and photosynthetic activity

Photoreceptors are defined as pigment molecules that process the energy and informational
content of light into a form that can be used by the plant. A pigment that contains protein as
an integral part of the molecule is known as a chromoprotein. Thus, photoreceptors typically
are chromoproteins. The chromophore (Gr. phoros, bearing) is that portion of the
chromoprotein molecule responsible for absorbing light and, hence, color. The protein portion
of a chromoprotein molecule is called the apoprotein. The complete molecule, or holochrome,
consists of the chromophore plus the protein. The principal photoreceptors found in plants are
described here.
 Chlorophylls
Chlorophylls are the pigments primarily responsible for harvesting light energy used in
photosynthesis. The chlorophyll molecule consists of two parts; -
 Porphyrin Head
A porphyrin is a cyclic tetra-pyrrole, made up of four
nitrogen-containing pyrrole rings arranged in a cyclic
fashion. Porphyrins are universal in living organisms and
include the heme group found in mammalian hemoglobin
and the photosynthetic and respiratory pigments,
cytochromes.
The porphyrin ring of chlorophyll actually absorbs the
light and not the phytol tail. Note also that chlorophyll
does not absorb strongly in the green region of the visible
light spectrum (490–550nm). The strong absorbance in the
blue and red and transmission in the green is what gives
chlorophyll its characteristic green color.
 Phytol Tail
Esterified to ring-IV of the porphyrin in chlorophyll is
a 20-carbon alcohol, phytol. This long, lipid-soluble
hydrocarbon tail is a derivative of the 5-carbon isoprene.
Isoprene is the precursor to a variety of important
molecules, including other pigments (carotenes),
hormones (the gibberellins), and steroids. The presence of

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the long hydrocarbon phytol tail exerts a dominant effect on the solubility of chlorophyll,
interpreting it virtually insoluble in water.
 Completely the chlorophyll molecule has a magnesium ion chelated to the four nitrogen
atoms in the center of ring.
 Loss of magnesium ion from chlorophyll resulted in the formation of non-green product,
pheophytin.
 Pheophytin is readily formed during extraction under acidic conditions, but small amounts
are also found naturally in the chloroplast where it serves as an early electron acceptor.

Chlorophyll a: C55 H72 O5 N4 Mg

Chlorophyll b: C55 H70 O6 N4 Mg
 Species of Chlorophyll
Four species of chlorophyll, designated chlorophyll a, b, c, and d, are known. Chlorophyll
e is rare in nature and found in some algae. Chlorophyll a is dominating among others so they
can be described in comparison with Chl a.
1) Chlorophyll a
The chemical structure of chlorophyll a has both Head and Tail with (-CH=CH2) group
attached at Ring-I and a (CH3) methyl group at Ring-II. It is a primary photosynthetic
pigment in all higher plants, algae and the cyanobacteria.
Chlorophyll Structural difference with Chl a Occurance
2) Chl b Chlorophyll b replaces the methyl Also found in all higher plants,
group(-CH3) with formyl group(- algae and the cyanobacteria like
CHO) at ring-II Chl a
3) Chl c Phytol tail is not present It is found in diatoms,
dinoflagellates and brown algae
4) Chl d A (—O—CHO) group replaces the It is found only in the red algae
(—CH=CH2) group on ring I

 Carotenoids
 Carotenoids also called tetraterpenoids, are yellow, orange, and red organic pigments that
are produced by plants and algae, as well as several bacteria and fungi.
 Carotenoids give the characteristic color to pumpkins, carrots, corn, tomatoes, canaries,
flamingos, and daffodils.
 In general, carotenoids absorb wavelengths ranging from 400–550nanometers (violet to
green light).

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 Carotenoids are the dominant pigment in autumn leaf coloration of about 15-30% of tree
species.
 Carotenoids serve two key roles in plants and algae: (1) they absorb light energy for use in
photosynthesis, (2) they protect chlorophyll from photo-damage.
 There are over 1100 known carotenoids which can be further categorized into two classes,
xanthophylls (which contain oxygen) and carotenes (which are purely hydrocarbons, and
contain no oxygen).
 Xanthophylls
Xanthophylls (originally phylloxanthins) are yellow pigments that occur widely in nature
and form one of two major divisions of the carotenoid group. The name is derived from
Greek xanthos "yellow" and phyllon "leaf" due to their formation of the yellow band seen in
early chromatography of leaf pigments.
 Molecular structure
Xanthophylls typically contain oxygen as a hydroxyl group. Plant xanthophylls form the
bright yellow band next to the green. As both are carotenoids, xanthophylls and carotenes are
similar in structure, but xanthophylls contain oxygen atoms while carotenes are purely
hydrocarbons, which do not contain oxygen. Their content of oxygen causes xanthopylls to
be more polar (in molecular structure) than carotenes.
 Occurrence
Like other carotenoids, xanthophylls are found in highest quantity in the leaves of greenest
plants, where they act to modulate light energy and perhaps serve as a non-photochemical
reducing agent to deal with triplet chlorophyll (an excited form of chlorophyll), which is
overproduced at high light levels in photosynthesis. The group of xanthophylls includes
(among many other compounds) lutein, zeaxanthin, neoxanthin, violaxanthin, flavoxanthin,
and α- and β-cryptoxanthin.
 Carotene
The term carotene also carotin, derived from the Latin carota, "carrot". Carotenes are
photosynthetic pigments important for photosynthesis. Carotenes contain no oxygen atoms.
They absorb ultraviolet, violet, and blue light and scatter orange or red light, and (in low
concentrations) yellow light. Carotenes are also responsible for the orange (but not all of the
yellow) colours in dry foliage. Carotenes contribute to photosynthesis by transmitting the light
energy they absorb to chlorophyll. They also protect plant tissues by helping to absorb the

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energy from singlet oxygen, an excited form of the oxygen molecule (O2) which is formed
during photosynthesis.
 Molecular structure
Chemically, carotenes are polyunsaturated hydrocarbons containing 40 carbon atoms
per molecule, variable numbers of hydrogen atoms, and no other elements. Some carotenes are
terminated by hydrocarbon rings, on one or both ends of the molecule. Since they are
hydrocarbons, and therefore contain no oxygen, carotenes are fat-soluble and insoluble in water
(in contrast with other carotenoids, the xanthophylls, which contain oxygen and thus are less
chemically hydrophobic).
 Occurrence
Carotenes are found in plants in two primary forms designated by characters from the
Greek alphabet: alpha-carotene (α-carotene) and beta-carotene (β-carotene).

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 VI. Ultrastructure and Composition of Photosystem-I and II

Photosystems as Major Components of the Photosynthetic ETC

The key to the photosynthetic electron transport chain is the presence of two large,
multi-molecular, pigment-protein complexes known as photosystem I (PSI) and photosystem
II (PSII) (Figure 7.3) PS-I consists of 18 distinct subunits whereas PS-II consists of 31
individual subunits! These two
photosystems operate in series linked
by a third multiprotein aggregate
called the cytochrome complex.
Overall, the effect of the chain is to
extract low-energy electrons from
water and, using light energy trapped
by chlorophyll, raise the energy level
of those electrons to produce a strong reductant NADPH.
Composition of Photosystems
Studies have revealed that PSI and PSII each contain several different proteins together
with a collection of chlorophyll and
carotenoid molecules that absorb photons. The
bulk of the chlorophyll in the photosystem
functions as antenna chlorophyll(Figure7.4).
The association of chlorophyll with specific
proteins forms a number of different
chlorophyll-protein (CP) complexes, which
can be separated by gel electrophoresis and
identified on the basis of their molecular mass
and absorption spectrum.
The core antenna for photosystem II, for example, consists of two chlorophyll-proteins
(CP) known as CP43 and CP47 (Figure 7.6). These two CP complexes each contain 20 to 25
molecules of chlorophyll a. The core antenna chlorophyll a absorb light but do not participate
directly in photochemical reactions. The reaction center chlorophyll a of PSI and PSII are

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designated as P700 and P680, respectively. These designations identify the reaction center
chlorophyll a, or pigment (P), with an absorbance maximum at either 700nm (PSI) or 680nm
(PSII). Tightly associated with the reaction centers, P680 and P700, are core antenna
complexes. CP-47 and CP-43 are the core antenna of PSII whereas CP1 is the core complex
of PSI(Figure7.6). There are also two additional chlorophyll-protein complexes, depicted in
close association with PSII and PSI—light-harvesting complex II (LHCII) and light-
harvesting complex I (LHCI), respectively. LHCII is associated with PSII and LHCI is
associated with PSI. As their name simply, the light-harvesting complexes function as extended
antenna systems for harvesting additional light energy. LHCI and LHCII together contain as
much as 70 percent of the total chloroplast pigment, including virtually all of the chlorophyll
b. LHCI is relatively small, has a chlorophyll a/b ratio of about 4/1, and appears rather tightly
bound to the core photosystem.
A schematic of the photosynthetic electron transport chain representing the
arrangement of PSI, PSII, and the cytochrome b6/f complex in the thylakoid membrane is
presented in (Figure7.6). A fourth complex—the CF0-CF1 coupling factor or ATP synthase—
is also shown. All four complexes are membrane-spanning, integral membrane proteins with a
significant portion of their structure buried in the hydrophobic lipid bilayer.

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 VII. Mechanism of Photosynthesis

(light absorption, charge separation or oxidation of water (water oxidizing clock), electron and
proton transport through thylakoid protein-pigment complexes.)
Role of photosystem II
Electron transport actually begins with the arrival of excitation energy(sunlight) at the
photosystem II reaction center chlorophyll, P680, which is located near the luminal side of
the reaction center. As illustrated in Figure 7.7, this excitation energy is required to change the
redox potential of P680 from +0.8eV to about −0.4eV for P680*, the excited form of P680. As
a consequence of this initial endergonic excitation process, P680* can rapidly (within
picoseconds, 10−12 s) transfer electrons exergonically to Pheophytin (Pheo).
Role of Pheophytin
Pheophytin considered the primary electron acceptor in PSII, is a form of chlorophyll
a in which the magnesium ion has been replaced by two hydrogens. Since this initial oxidation
of P680 is light dependent, this is called a photo-oxidation event, which results in the
formation of P680+ and Pheo−, a charge separation. This charge separation effectively stores
light energy as redox potential energy and represents the actual conversion of light energy to
chemical energy. It is essential that
this charge separation be stabilized by
the rapid movement of the electron
from P680 at the lumen side of the
PSII reaction center to an electron
acceptor molecule localized at the
stromal side of the PSII reaction
center. If the electron were permitted
to recombine with P680+, there would Figer-7.7
be no forward movement of electrons, the energy would be wasted, and ultimately, carbon
could not be reduced.
Role of Plastoquinone
Within picoseconds, pheophytin passes one electron on to a quinone electron acceptor
called QA, resulting in the formation of [P680+ Pheo Q− A]. Now the PSII reaction center is
considered to be ‘‘closed,’’ that is, it is unable to undergo another photo-oxidation event
(Figure 7.8). On a slower time scale of microseconds, the electron is passed from QA to

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plastoquinone(PQ), resulting in the formation of [P680+ Pheo QA]. PQ is a quinone that binds
transiently to a binding site (QB) that is on the stromal side of the D1 reaction center protein
(Figure 7.6). The reduction of PQ to plastoquinol (PQH2) decreases its affinity for the binding
site on the D1 polypeptide. The plastoquinol is thus released from the reaction center, to be
replaced by another molecule of PQ.
PQ requires two electrons to become
fully reduced to PQH2, reduction at
the QB site is considered a two-
electron gate (Figure 7.8). Second, the
initial charge separation is further
stabilized because P680+ is a very
strong oxidant (perhaps the strongest
known in biological systems) and is
able to ‘‘extract’’ electrons from water. Thus P680+ is rapidly reduced, again within
picoseconds, to P680, resulting in the formation of [P680 Pheo QA]. Now the PSII reaction
center is said to be ‘‘open,’’ that is, it is ready to receive another excitation (Figure 7.8).
Role of Oxygen Evolving Complex
The electrons that reduce P680+ are most immediately supplied by a cluster of four
manganese ions associated with a small complex of proteins called the oxygen-evolving
complex (OEC). As the name implies, the OEC is responsible for the splitting (oxidation) of
water and the consequent evolution of molecular oxygen. The OEC is located on the lumen
side of the thylakoid membrane. The OEC is bound to the D1 and D2 proteins of the PSII
reaction center and functions to stabilize the manganese cluster. It also binds Cl− which is
necessary for the water-splitting function.
2H2O → O2 +4H+ + 4e
According to equation 7.8, the oxidation of two moles of water generates one mole of oxygen,
four moles of protons, and four moles of electrons. It has been determined that only one PSII
reaction center and OEC is involved in the release of a single oxygen molecule. Thus, in order
to complete the oxidation of two water molecules, expel four protons, and produce a single
molecule of O2, the PSII reaction center must be ‘‘closed’’ and then ‘‘opened’’ four times
(Figure 7.8). OEC has the capacity to store charges and the organisms containing PSII and the
OEC exhibit oxygenic photosynthesis, that is, a photosynthetic process that generates
molecular oxygen (O2). Since water is generally very abundant in our biosphere, the evolution

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of PSII allowed oxygenic photosynthetic organisms to survive and reproduce almost anywhere
in our biosphere. Thus, the evolution of PSII and its associated OEC was a major factor
determining the global distribution of oxygenic photosynthetic organisms that fundamentally
changed the development of all life on Earth.
Role of The Cytochrome Complex and Photosystem I
Following its release from PSII, plastoquinol diffuses laterally through the membrane
until it encounters a cytochrome b6f complex (Figure 7.6). This is another multiprotein,
membrane-spanning redox complex whose principal constituents are cytochrome b6
(Cytb6) and cytochrome f (Cytf). The cytochrome complex also contains an additional redox
component called the Rieske iron-sulfur (FeS) protein—iron-binding proteins in which the
iron complexes with sulfur residues rather than a heme group as in the case of the cytochromes.
Plastoquinol diffuses within the plane of the thylakoid membrane and passes its electrons first
to the FeS protein and then to Cyt f. The oxidation of plastoquinol is thought to be diffusion
limited, this is the slowest step in photosynthetic electron transport. The Rieske FeS protein
and the heme of Cyt f are located on the lumenal side of the thylakoid membrane. From Cyt
f, the electrons are picked up by a copper-binding protein, plastocyanin (PC).
Plastocyanin (PC) is a small peripheral protein that is able to diffuse freely along the
lumenal surface of the thylakoid membrane. In the meantime, a light-driven charge separation
similar to that involving P680 has also occurred in the reaction center of PSI. Excitation energy
transferred to the reaction center chlorophyll of PSI (P700) is used to change the redox potential
of P700 from about +0.4eV to about −0.6eV for P700*, the excited form of P700 (Figure 7.7).
As a consequence of this initial endergonic excitation, P700* is rapidly photo-oxidized to
P700+ by the primary electron acceptor (A) in PSI, a molecule of chlorophyll a (Figure 7.6);
the electron is then passed through a quinone and additional FeS centers and finally, on the
stroma side of the membrane, to ferredoxin.
Ferredoxin is another FeS-protein that is soluble in the stroma. Ferredoxin in turn is
used to reduce NADP+, a reaction mediated by the enzyme ferredoxin-NADP+-
oxidoreductase. Finally, the electron deficiency in P700+ is satisfied by withdrawing an
electron from reduced PC (Figure 7.6).
The quantum requirement for oxygen evolution is defined as the number photons
required to evolve one molecule of O2. As discussed above, two excitations, one at PSII and
one at PSI, are required to move each electron through non cyclic electron transport from H2O
to NADP +. From equation 7.8, the evolution of one molecule of O2 by P680+ generates four
electrons that are eventually transferred through PSI to reduce 2NADP+ to 2NADPH.

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 VIII. Photophosphorylation and its mechanism

The chemiosmotic synthesis of ATP in chloroplasts will going on discussion here. The ATP
required for carbon reduction and other metabolic activities of the chloroplast is synthesized
by photophosphorylation in accordance with Mitchell’s chemiosmotic mechanism.
Phosphorylation is the addition of inorganic phosphate to an ADP to form ATP.
Photophosphorylation
Light-driven production of ATP by chloroplasts is known as photophosphorylation. It is
very important because, in addition to using ATP (along with NADPH) for the reduction of
CO2, a continual supply of ATP is required to support a variety of other metabolic activities in
the chloroplast. These activities include amino acid, fatty acid, and starch biosynthesis, the
synthesis of proteins in the stroma, and the transport of proteins and metabolites across the
envelope membranes.
 Non-Cyclic Photophosphorylation
Electrons are continuously supplied from water and withdrawn as NADPH. This
flow-through form of electron transport is thus known as either noncyclic or linear electron
transport. Thus the formation of ATP in association with noncyclic electron transport is
noncyclic photophosphorylation.
 Electrons are continuously supplied from water and withdrawn as NADPH.
 Noncyclic photophosphorylation results in the production of both ATP and NADPH.
 PSI units and PSII units in the membrane are not physically linked as implied by the Z-
scheme, but are even separated into different regions of the thylakoid.
 Cyclic Photophosphorylation
The heterogeneous distribution in the membranes by which the PSI units may
transport electrons independently of PSII, through a process known as cyclic electron
transport.
In terrestrial plants, the major pathway for PSI cyclic electron transport is thought to occur
via P700 to ferredoxin (fd) which transfers the electrons back to PQ via a recently discovered
protein, PGR5 rather than to NADP+(NADP reductase). The electron then returns to P700+,
passing through the cytochrome b6/f complex and plastocyanin.
 Using a genetic approach in Arabidopsis thaliana, the gene, PGR5, was shown to encode
a small thylakoid polypeptide that is
essential for PSI cyclic electron transport.
 Cyclic electron transport will also
contribute to the establishment of the pH
gradient required to support ATP
synthesis, in cyclic photophosphorylation.
 It is thought that cyclic
photophosphorylation is a source of ATP
required for chloroplast activities over and
above that required in the carbon-
reduction cycle.

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 Mechanism
A key to energy conservation in photosynthetic electron transport and the accompanying
production of ATP is the light-driven accumulation of protons in the lumen. There are two
principal mechanisms that account for this accumulation of protons:
 The oxidation of water, in which two protons are deposited in to the lumen for
each water molecule oxidized.
 PQ-cytochrome proton pump.
The energy of the resulting proton gradient is then used to drive ATP synthesis in accordance
with Mitchell’s chemiosmotic hypothesis. The precise mechanism by which protons are
moved across the membrane by the cytochrome complex is not yet understood, although
several models have been proposed. The most widely accepted model is known as the Q-cycle,
based on an original proposal by Mitchell. A simplified version of the Q-cycle during steady-
state operation is shown in Figure7.10.
The Q-cycle:- When PQ is reduced by PSII, it binds temporarily to the D1 protein (QB) as a
semi-Quinone after it accepts the first electron from QA. Subsequently, the QB semiquinone
is converted to the fully reduced plastoquinol (PQH2) after it has accepted another electron
from QA, plus two protons are picked up from the surrounding stroma. PQH2 dissociates from
the PSII complex and diffuses laterally through the membrane until it encounters the lumenal
PQH2 binding site of the cytochrome b6/f complex. There, two PQH2 bind sequentially and
are reoxidized to PQ through a semiquinone intermediate (PQHz, not shown) by the combined
action of the Rieske FeS-protein and the low reduction potential form of cytochrome b6 (LP).
Along with, 4 H+ are transferred to the lumen. One of the PQ molecules returns to the thylakoid
PQ pool to be reduced again by PSII, while the other PQ molecule is transferred to the stromal
binding site of the cytochrome b6/f complex where it becomes reduced by the high reduction
potential form of cytochrome b6 (HP) and is protonated using 2H + from the stroma. This
PQH2 molecule is then released from the stromal binding site and recycled into the thylakoid
PQH2 pool.

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 XI. CO2 Reduction (Dark Reactions)/ PCR
(C 3 pathway and Photorespiration, Regulation of C 3 pathway, C 4 pathway and its different forms, C 3 -C 4
intermediates, CAM pathway.)
The pathway by which all photosynthetic eukaryotic organisms ultimately incorporate CO2
into carbohydrate is known as carbon
fixation or the photosynthetic carbon
reduction (PCR) cycle. It is also referred
to as the Calvin cycle, in honor of
Melvin Calvin, who directed the
research effort that explained the
pathway. Calvin was awarded the Nobel
Prize for chemistry in 1961.
The PCR cycle can be divided into
three primary stages (1) Carboxylation
(2) Reduction (3) Regeneration

(1) The Carboxylation (Fixation of CO2)

The reaction is a carboxylation in which CO2 is added to RuBP, forming a six-carbon
intermediate. The intermediate, which is temporary and unstable, remains bound to the enzyme
and is quickly hydrolyzed to two molecules of 3-PGA(3-Phosphoglyceric acid).
Enzyme: Rubisco(ribulose-1,5-bisphosphate carboxylase-oxygenase).
Rubisco: It is the most abundant protein in the world, accounting for approximately 50
percent of the soluble protein in most leaves.The enzyme also has a high affinity for CO2 that,
together with its high concentration in the chloroplast stroma, ensures rapid carboxylation at
the normally low atmospheric concentrations of CO2. Thus, the reaction catalyzed by Rubisco
maintains the CO2 concentration gradient between the internal air spaces of a leaf and the
atmospheric air to ensure a constant supply of this substrate for the PCR cycle. The
carboxylation reaction, with a ∆G of −35 kJ/mol, is energetically very favorable. Energy is
required at two points: first for the reduction of 3-PGA and second for regeneration of the
RuBP acceptor molecule.
(2) Reduction of 3-PGA
In order for the chloroplast to continue to take up CO2, two conditions must be met. Both
require energy in the form of ATP and NADPH produced in light reactions.
 The product molecules (3-PGA) must be continually removed.
 Suitable supply of the acceptor molecule (RuBP).
The 3-PGA is removed by reduction to the triose phosphate, glyceraldehyde-3-
phosphate(G3P). This is done in two-steps,
(i) 3-PGA is first phosphorylated to 1,3-bisphosphoglycerate. (ATPADP+Pi)
(ii) Reduced to glyceraldehyde-3-phosphate (G3P).
The resulting triose sugar-phosphate, G3P, is available for export to the cytoplasm,
probably after conversion to dihydroxyacetone phosphate (DHAP).
Enzyme: 3-PGA-kinase and G3P-dehydrogenase.

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 (3) Regeneration of RuBP
In order to maintain the process of CO2 reduction, it is necessary to ensure a continuing supply
of the acceptor molecule, RuBP (3CO2 + 3RuBP 3+3x5= 18C). Five G3P molecules
produce three RuBP molecules, using up three molecules of ATP. This is accomplished by a
series of reactions involving 4-, 5-, 6-, and 7-carbon sugars.
Note: Here there is the reactions of six G3P molecules out of twelve is given. Enzymes are underlined.
Correction of carbon number is given in brackets at last. (six G3P=6x3=18)
(1) Triose phosphate isomerase converts the three G3P molecules reversibly into
dihydroxyacetone phosphate (DHAP), both are 3-carbon molecule.
-----------------
(2) Aldolase convert a G3P and a DHAP into Fructose-1,6-bisphosphate, which is converted
into Fructose 6-phosphate (6C) by Fructose-1,6-bisphosphatase. A phosphate ion is lost into
solution.
(3+3=6, two used resulting four G3P remains)
-----------------
(3) Fructose-6-phosphate(6C) combines with one G3P(3C) to form a 9C compound which later
cleaves by Transketolase to form one Erythrose-4-phosphate(4C) and one ketose Xylulose-
5-phosphate(5C).
(6+3=9C, converted to 4C and 5C, one more G3P used so three G3P remains)
-----------------
(4) Erythrose-4-phosphate (4C) and a DHAP are converted into Sedoheptulose-1,7-
bisphosphate (7C) by Aldolase enzyme.
(4+3=7C, one more 3C-DHAP used so two G3P remains)
-----------------
(5) Sedoheptulose-1,7-bisphosphatase (one of only three enzymes of the Calvin cycle that are
unique to plants) cleaves Sedoheptulose-1,7-bisphosphate into Sedoheptulose-7-phosphate,
releasing an inorganic phosphate ion into solution.
-----------------
(6) The ketose Sedoheptulose-7-phosphate(7C) combine with one G3P(3C) which on
breakdown give one ribose-5-phosphate (5C), and one Xylulose-5-phosphate(5C).
(7+3=10C one G3P more used, here total 5 out of 6 G3P used and One G3P Exported earlier)
-----------------
(7) Isomerization and epimerization of 5-carbon sugars takes place as above formed: -
One ribose-5-phosphate(5C) is isomerized and two Xylulose-5-phosphate(5C)
epimerized into a single type of three molecules of ribulose-5-phosphate by two enzymes
ribulose-5-phosphate isomerase, epimerase.
-----------------
(8) The above mentioned three molecules of ribulose-5-phosphate are phosphorylated
(3ATP3ADP+3Pi) by an enzyme ribulose-5-phosphate kinase to convert them into
ribulose-1,5-bisphosphate (RuBP-CO2 accepter) completing the Calvin cycle. This requires
the input of one ATP.
(Here resulting three RuBP regenerated by five G3P in single way process so when this reaction run double
time then it produces six RuBP and one Glucose)

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