Photosynthesis
Photosynthesis is a fundamental process in which photosynthetic organisms, primarily plants and algae, convert light energy into chemical energy
Thylakoid Reactions (Light-Dependent Reactions)
The thylakoid reactions, also known as the light reactions, are the initial stage of photosynthesis. They take place specifically within the thylakoids of the chloroplast. The primary function of these reactions is to convert light energy into chemical energy, which is stored in two high-energy compounds: ATP and NADPH.
ATP (Adenosine triphosphate): This molecule serves as the primary energy currency for the cell. In the thylakoid reactions, ATP is produced through a process called photophosphorylation, which is driven by the movement of protons across the thylakoid membrane.
NADPH (Nicotinamide adenine dinucleotide phosphate): This is a crucial electron carrier that provides the reducing power necessary for the synthesis of sugars.
The ATP and NADPH generated during the thylakoid reactions are then used in the next stage of photosynthesis, the carbon fixation reactions (also known as the Calvin cycle), which occur in the stroma of the chloroplast. These reactions use the stored chemical energy to synthesize sugars from carbon dioxide.
The Nature of Light
Light exhibits characteristics of both a wave and a particle
Wave Properties
A wave is defined by its wavelength (λ), which is the distance between consecutive wave crests
. The frequency (ν) is the number of wave crests that pass a given point in a specific time
. The relationship between these is given by the equation
c = λν
- Light is a transverse electromagnetic wave.
Particle Properties
Light can also be considered a stream of particles called photons
. Each photon contains a discrete amount of energy, known as a quantum
. The energy (E) of a photon is directly proportional to the light's frequency, as described by Planck's law:
The human eye can only perceive a small range of frequencies within the electromagnetic spectrum, known as the visible-light region
.
Light Absorption and Pigments
Absorption and Action Spectra
An absorption spectrum measures how much light energy a substance absorbs at different wavelengths
. For example, the absorption spectrum of chlorophyll a show the portion of solar output that plants utilize . Chlorophyll appears green because it primarily absorbs light in the red and blue parts of the spectrum, while reflecting green wavelengths (around 550 nm)
. An action spectrum measures the effectiveness of different wavelengths of light in driving a biological response, such as oxygen evolution during photosynthesis
. If the pigment responsible for a response is the same as the one whose absorption is being measured, the absorption and action spectra will be similar
. However, a slight mismatch occurs in the carotenoid absorption region (450–550 nm), suggesting that energy transfer from carotenoids to chlorophylls is less efficient than between chlorophylls themselves
.
Absorption spectrum and action spectrum for photosynthesis does not overlap in the 450-550 nm region
Chlorophyll has high absorption peaks in the blue (430 nm) and red (662 nm) regions of the spectrum, while carotenoids absorb light in the blue-green region (450-550 nm).
The overall photosynthetic activity, measured by the action spectrum, is lower in the region of carotenoids absorption compared to the peaks of chlorophyll absorption.
This is because the energy absorbed by the carotenoids must be transferred to chlorophylls to be used in photosynthesis. This energy transfer is not 100% efficient, meaning some of the absorbed light energy is lost during the transfer process as heat.
As a result, the action spectrum shows a less pronounced effect in the 450-550 nm range, even though significant absorption is occurring in that same range.
The Red Drop and Emerson's Enhancement Effect
The Red Drop: When measuring the quantum yield of photosynthesis (the rate of a photochemical event relative to the rate of photon absorption), Emerson observed a sharp decrease in efficiency in the far-red region (wavelengths greater than 680 nm)
. This indicates that light with a wavelength longer than 680 nm is far less effective at driving photosynthesis compared to shorter wavelengths . The Enhancement Effect: When Emerson provided red and far-red light simultaneously, the rate of photosynthesis was greater than the sum of the rates from each light source individually
. This discovery led to the understanding that two distinct photochemical complexes, Photosystem I (PSI) and Photosystem II (PSII), work in tandem . PSII absorbs red light (680 nm), while PSI preferentially absorbs far-red light (greater than 680 nm) .
Photochemical Reactions and Energy Transfer
When a chlorophyll molecule absorbs a photon, it becomes energized, transitioning from its stable "ground state" to an unstable "excited state." This high-energy state is fleeting and must quickly release its energy. Absorption of blue light transitions chlorophyl to higher excited state than red light. In the higher excited state, it quickly emits some energy as heat and enters lower excited state. This excited state has four possible pathways to release its energy:
Fluorescence: Re-emitting a photon and returning to the ground state. The emitted photon has a longer wavelength and lower energy than the absorbed one
. Chlorophyll fluoresces in the red region of the spectrum. Heat Dissipation: Converting the excitation energy directly into heat without emitting a photon
. Energy Transfer: Transferring the energy to an adjacent molecule
. This is how antenna pigments transfer energy to the reaction center. Photochemistry: Using the absorbed energy to drive a chemical reaction
. The excited chlorophyll molecule transfers an electron to an electron-acceptor molecule, initiating a chain of redox (reduction-oxidation) reactions. This is the primary event that converts light energy into chemical energy.
Photosynthetic Pigments
Chlorophyll
Chlorophyll molecules have a porphyrin-like ring with a central magnesium () ion
. This ring is the site of electron changes during excitation, oxidation, and reduction . Chlorophylls differ in the substituents on their rings, which gives them different properties
. For example, chlorophyll a has a -CH₃ group, while chlorophyll b has a -CHO group . The molecule has a long hydrocarbon tail that anchors it in the photosynthetic membrane.
Carotenoids
Carotenoids are linear molecules with multiple conjugated double bonds found in photosynthetic organisms
. They absorb light in the 400-500 nm range, which gives them their characteristic orange color . They act as accessory pigments by absorbing light and transferring the energy to chlorophyll for photosynthesis
. Carotenoids also provide photoprotection
. They act as a safety valve, dissipating excess energy as heat before it can damage the photosynthetic machinery . If unquenched, excited chlorophyll can react with oxygen to form highly reactive singlet oxygen (¹O₂*), which can harm cellular components .
Photoprotection by Carotenoids
- Carotenoids are essential accessory pigments in photosynthesis that provide crucial protection against the damaging effects of excess light.
Their primary photoprotective function is to dissipate surplus light energy as heat, a process known as non-photochemical quenching (NPQ). - When light intensity is high, the light-harvesting complex can absorb more energy than the photosystems can use. This excess energy can lead to the formation of a highly reactive and toxic molecule called singlet oxygen (¹O₂)*.
Singlet oxygen can cause oxidative damage to cellular components, particularly lipids in the thylakoid membranes, leading to a condition called photoinhibition. - Carotenoids, such as zeaxanthin, antheraxanthin, and violaxanthin, act as the plant's first line of defense. They are involved in the xanthophyll cycle, a process that regulates the dissipation of excess light energy. In high light conditions, violaxanthin is converted to zeaxanthin. Zeaxanthin binds to proteins in the light-harvesting antenna complex, inducing a conformational change that promotes the safe release of excess energy as heat.
- Furthermore, carotenoids can also directly quench the excited state of chlorophyll molecules, preventing them from transferring energy to oxygen and forming damaging singlet oxygen.
The excited state of carotenoids has a lower energy level than that of singlet oxygen, so it cannot transfer energy to form it. Instead, the carotenoid returns to its ground state by releasing the energy as harmless heat, thus protecting the photosynthetic apparatus from damage.
The Photosynthetic Apparatus and Units
Location
Photosynthesis takes place in chloroplasts
. The essential proteins for photosynthesis are embedded in the thylakoid membranes
. PSII is primarily located in the grana lamellae, while PSI and ATP synthase are mostly in the stroma lamellae and at the edges of the grana
. The cytochrome b₆f complex is distributed between both the stroma and grana lamellae
.
Antenna and Reaction Centers
Pigments like chlorophylls and carotenoids form an antenna complex that collects light and transfers the energy to the reaction center
. - The antenna system is a physical phenomenon where energy is transferred via fluorescence resonance energy transfer (FRET)
. This transfer is highly efficient (95-99%) . The reaction center, which contains special chlorophylls (P700 and P680), is where the photochemical redox reactions occur, converting light energy into chemical energy
. The reaction center includes 4-6 chlorophyll a molecules (P700 and P680), and associated proteins and cofactors.
The reaction center chlorophyll is an energy sink that absorbs light of longest wavelength and lowest energy in the complex.
- Energy transfer that occurs in the antenna complex is a physical phenomenon while electron transfer (redox reactions) at the reaction center is a chemical phenomenon.
The energy trapping mechanism is directional and irreversible
The directionality of energy transfer in the antenna complex is maintained by a carefully arranged sequence of pigments
. - Energy Gradient: The pigments within the antenna complex are organized in a way that creates an "energy gradient." The pigments that are further away from the reaction center absorb light at shorter wavelengths, which corresponds to higher energy
.
- Progressive Energy Loss: As the excitation energy is transferred from one pigment molecule to the next, closer to the reaction center, it moves to pigments that absorb at progressively longer, redder wavelengths and therefore have lower energy excited states
.
- Irreversibility: The small difference in energy between each consecutive excited chlorophyll is lost to the surroundings as heat
. This minor energy loss makes the transfer process energetically favorable in one direction (towards the reaction center) and largely irreversible, ensuring that the collected light energy is efficiently funneled to the reaction center .
Photosynthetic Unit (Quantasome)
A photosynthetic unit is the minimum number of collaborating pigment molecules required for a photochemical event
. The evolution of one oxygen molecule requires 8 quanta of light and approximately 2500 chlorophyll molecules
. This means about 300 chlorophyll molecules are needed to process each light quantum . Within this unit, only one specialized chlorophyll molecule participates in the photochemical reaction, while the others serve to absorb and transfer light to it
.
Photosystem I (PSI) and Photosystem II (PSII) Reaction Centers
The reaction centers of PSI and PSII are crucial pigment-protein complexes embedded within the thylakoid membranes of chloroplasts that convert light energy into chemical energy during photosynthesis
Photosystem II (PSII) Reaction Center
It is designated as P680 as its core chlorophyll pigment absorbs light most efficiently at a wavelength of 680 nm.
Location: Primarily found in the grana lamellae, which are the stacked regions of the thylakoid membranes
. Structure: The core of the reaction center consists of two membrane proteins, D1 and D2
. It also contains a special pair of chlorophyll-a molecules (P680), bound one to each of these two proteins, along with additional chlorophylls, pheophytins (colorless chlorophyll lacking Mg2+), carotenoids, and electron acceptors like plastoquinones . Additionally, the oxygen evolving complex is also a part of PSII. Function: PSII acts as a powerful oxidant
. Its primary role is to split water molecules (photolysis) to replace the electron it loses . This process releases oxygen as a byproduct . The electrons from the water are used to re-reduce the oxidized P680 . The splitting of water and subsequent electron transfer also pumps protons into the thylakoid lumen, contributing to the proton motive force needed for ATP synthesis .
Photosystem I (PSI) Reaction Center
It is designated as P700 as its core chlorophyll pigment absorbs light most effectively at a wavelength of 700 nm
. Location: Predominantly located in the stroma lamellae and at the edges of the grana lamellae
. Structure: It is a large, multi-subunit complex
. Its core consists of a heterodimer of two membrane proteins, PsaA and PsaB, each binding a reaction center chlorophyll molecule (P700) . Another pair of chlorophyll a molecules (A0) also remains associated with the heterodimer and accepts electron from P700. There are additional proteins PsaC to PsaN and a pair of vitamin K1 molecules called phylloquinone bound to the heterodimer, one per subunit. It also contains a core antenna of about 100 chlorophyll molecules and a series of 3 membrane associated iron sulfur clusters - FeSX, FeSA and FeSB. A soluble iron sulfur protein, ferredoxin (Fd) is also a part of this large complex. Function: PSI produces a strong reductant
. The electron it receives from the electron transport chain (via plastocyanin) is excited by light and transferred to acceptors, eventually being used to reduce NADP+ to NADPH . NADPH is a high-energy compound used in the Calvin cycle .
Electron Transport and Photophosphorylation
The Z-Scheme (Non-cyclic Electron Transport)
The Z-scheme illustrates how PSI and PSII work in series
. The entire chemical reaction is carried out by four integral membrane complexes that are vectorially arranged in the thylakoid membrane: - PSII
- Cytochrome b₆f complex
- PSI
- ATP Synthase
PSII: P680 in PSII absorbs a photon, gets excited (P680*), and transfers an electron to pheophytin
. The electron-deficient P680+ is then re-reduced by electrons from water oxidation . This process releases protons into the thylakoid lumen, and the reduced product is plastohydroquinone (PQH₂) . Cytochrome b₆f: This complex oxidizes PQH₂ and transfers electrons to plastocyanin (PC)
. This process also pumps protons from the stroma into the lumen, creating a proton motive force . PSI: Plastocyanin then reduces the oxidized P700+ in PSI
. P700 absorbs a photon, gets excited, and transfers an electron down a series of acceptors, ultimately reducing NADP⁺ to NADPH by the action of ferredoxin and flavoprotein-ferredoxin NADP+ reductase (FNR) . ATP Synthase: It produces ATP as protons diffuse back through it from lumen to stroma.
- The process begins when light energy is absorbed by the antenna pigments of Photosystem II (PSII) and funneled to its reaction center, P680
. This energy excites an electron in P680 to a higher energy state, and the electron is then transferred to an electron acceptor molecule, pheophytin (primary photochemical act) . Now the P680 is in oxidized state (electron deficient) and pheophytin is in reduced state (electron rich).
- This primary electron acceptor will now donate its extra electron to a secondary eelectron acceptor and so on down the ETC. On the other hand, the oxidized reaction center chlorophyll will accept the missing electron from a secondary electron donor, which in turn will regain alectron from a tertiary electron donor and so on.
- The ultimate electron donor is water and ultimate electron acceptor is NADP+.
- To replace the lost electron, PSII uses a powerful oxidant to split a water molecule (photolysis)
. This reaction releases oxygen, protons (H+), and electrons . The electrons from water are donated to P680, allowing it to return to its ground state and participate in another photochemical event .
- The electron from PSII is transferred to a mobile electron carrier, plastoquinone (PQ), which becomes reduced to plastohydroquinone (PQH2)
.
- PQH2 then moves to the cytochrome complex, where it is oxidized
. The cytochrome b6f complex acts as a proton pump, using the energy from the electron transfer to move protons from the stroma into the thylakoid lumen, which contributes to the proton motive force for ATP synthesis .
The oxygen-Evolving Complex (OEC) is a key component of PSII that is responsible for splitting water molecules during photosynthesis
The OEC is a cluster of four manganese (Mn) ions, as well as a calcium ion (Ca2+) and a chloride ion (Cl−) that are essential for its function
The OEC operates through a series of five distinct oxidation states, known as the S states (S0,S1,S2,S3,S4). Each state represents a different level of oxidation of the manganese cluster, with each transition driven by the absorption of a photon by PSII
The Five Oxidation Steps (S States):
The process of water splitting occurs through the following steps:
S₀ to S₁: P680 becomes oxidized (P680+) by light
. The OEC donates one electron to a tyrosine residue (YZ) in the D1 protein of the PSII reaction center, which then reduces the oxidized P680 . This causes the OEC to advance from the S0 state to the S1 state. S₁ to S₂: A second photon excites a P680 molecule, which is reduced by YZ. YZ then takes another electron from the OEC, moving it from the S1 to the S2 state.
S₂ to S₃: This step is identical to the previous two; a third photon leads to the transfer of a third electron from the OEC, advancing it to the S3 state.
S₃ to S₄: A fourth and final photon causes the OEC to donate a fourth electron, transitioning it to the short-lived S4 state.
S₄ to S₀ (Oxygen Evolution): In the S4 state, the OEC has accumulated enough oxidizing power to strip four electrons from two water molecules
. This reaction releases one molecule of oxygen (O2) and four protons (H+) into the thylakoid lumen. The OEC then returns to its initial S0 state, ready to begin the cycle again . This entire process is driven by four photons .
The four electrons are passed one at a time from the OEC to the P680 reaction center, as it can only accept one electron at a time
The cytochrome b6f complex contains several key components that facilitate the Q-cycle:
- Two b-type cytochromes (Cyt b)
. - A c-type cytochrome, also known as cytochrome f (Cyt f)
. - A Rieske Fe-S protein (FeSR)
. - Two quinone oxidation-reduction sites
.
The Non-Cyclic Electron Path (Oxidation of first PQH2)
The Q-cycle begins with a plastoquinone () molecule, which arrives from PSII, binding to an oxidation site near the thylakoid lumen.
The PQH2 molecule is oxidized, releasing two electrons and two protons into the thylakoid lumen.
One of the two electrons is transferred to the Rieske Fe-S protein, which then passes it to cytochrome f.
From cytochrome f, the electron is carried by plastocyanin (PC) to reduce the oxidized reaction center of PSI, thereby continuing the linear or non-cyclic electron flow of photosynthesis.
The other electron from PQH2 is passed on to one of the b-type cytochromes.
The reduced b-type cytochrome then passes on the electron to the other b-type cytochrome which then reduces plastoquinone (PQ) to plastosemiquinone (PQ-).
The Cyclic Electron Path (Oxidation of second PQH2)
During oxidation of the second PQH2 molecule, first electron follows a similar path to P700. The second electron from this PQH2 molecule follows a different, cyclic path within the cytochrome b6f complex.
The second electron is transferred from the
PQH2 to a b-type cytochrome. This electron then moves to the other b-type cytochromes and finally reduces plastosemiquinone to plastohydroquinone (PQH2) taking up two protons from the stroma.
This mode of electron transport involves only PSI
. It occurs when NADP⁺ is not available . Electrons ejected from P700 are shuttled back to the cytochrome b₆ complex and then return to the electron hole of PSI
. This process does not produce NADPH or oxygen but is coupled to the phosphorylation of ADP to ATP
. This is a significant ATP source in the bundle sheath chloroplasts of C4 plants.
Photophosphorylation (ATP Synthesis)
A part of the captured light energy is utilized for light driven ATP synthesis, known as photophosphorylation.
Photosynthetic electron flow is usually coupled with ATP synthesis. Electron flow without ATP synthesis is called uncoupled.
ATP synthesis in chloroplasts is driven by a chemiosmotic mechanism
. The vectorial electron flow results in a proton gradient across the thylakoid membrane, making the lumen acidic and the stroma alkaline
. The energy of this proton gradient, known as the proton motive force (∆p), is used by the ATP synthase (CF₀-CF₁ complex) to synthesize ATP
. The CF₀ part of the enzyme is hydrophobic and membrane bound. It acts as a proton channel. It consists of 14 subunits each of which can transport one proton per rotation.
The CF₁ part, which sticks into the stroma, contains the catalytic sites for ATP synthesis
. This enzyme functions like a tiny molecular motor, rotating as protons pass through it to synthesize ATP
. It consists of several subunits including three copies of each of alpha and beta subunits arranged alternately. Catalytic sites are on beta subunits. Other subunits have regulatory roles. Three ATP molecules are generated per rotation. Hence, the stoichiometry of proton translocated to ATP synthesized is 14/3 = 4.67.
