5

On the wild side

A* Extension 5.1: Finding out more about photosynthesis
The discovery that photosynthesis is a two-step reaction
It was the investigation of the effect of temperature on the rate of photosynthesis that led to the discovery of the two steps (reactions) to photosynthesis. This arose from the observation that the impact on photosynthesising cells of increasing temperature (for example, from 10 C to 30 C) depends on the light intensity, as the graph below shows.
5

I A* Extensions 5.1 to 5.3

A rate of photosynthesis/arbitrary units 4

at high light intensity temperature rise increases rate of photosynthesis

B 1

at low light intensity temperature rise does not affect rate of photosynthesis

0 0 10 20 30 temperature/C

You can see from the graph that the effect of temperature is quite different at high and low light intensities. At high light intensity, a rise in temperature increases the rate markedly, but at low light intensity, there is hardly any increase in rate at all. The reasoning that lead to the discovery of the two-step nature of photosynthesis was as follows. If photosynthesis includes a photochemical reaction (that is, a temperature-insensitive change, brought about by light energy) then when this reaction is rate limiting (at low light intensity), a rise in temperature would not have a significant effect. This is what we observe (curve B in the graph). If photosynthesis also includes enzymic reactions (that is, temperature-sensitive changes involving enzymes) then when these reactions are rate limiting (at high light intensity), a rise in temperature should have a significant effect. This, too, is what happens (curve A). So the outcome of the investigation was explained by photosynthesis consisting of two sequential reactions:
I

a light-dependent reaction a photochemical step that, like all photochemical reactions, is unaffected by temperature (this might involve the splitting of water by light energy, releasing oxygen as a waste product) a light-independent reaction biochemical reactions that are catalysed by enzymes, and are highly temperature sensitive (this might involve the fixing of carbon dioxide to form sugar).

This hypothesis has since been confirmed by biochemical investigations.

Edexcel Biology for A2 Dynamic Learning

Hodder Education 2009

ON THE WILD SIDE: A* EXTENSIONS 5.1 TO 5.3

Hydrogen acceptor molecules and how they work

In addition to being the building blocks of the nucleic acids (DNA and RNA), certain nucleotides and molecules made from nucleotides are important in the metabolism of cells. These include the hydrogen acceptor molecules NAD and NADP. NAD (nicotinamide adenine dinucleotide) consists of two nucleotides combined with two phosphate groups. In one of the combined nucleotides, the organic base is replaced by a substance called nicotinamide. Nicotinamide is derived from a vitamin of the B complex, called nicotinic acid. Nicotinamide is a ring molecule that accepts hydrogen ions and electrons. When NAD accepts electrons it is said to be reduced, but it can pass them to other acceptor molecules with relative ease. When reduced NAD passes electrons on, it becomes oxidised again. The transport of electrons like this is an important aspect of respiration. NAD is a substance known as a coenzyme, which works with different enzymes of respiration. It also takes part in other processes in cells.
NAD is a carrier of hydrogen ions and electrons in respiration

H CONH2 N action end (hydrogen ion transport)

+

adenine
O

nicotinamide O ribose

ribose

Pi

Pi

rest of molecule, recognised by enzymes with which NAD is a coenzyme

mode of action summary: NAD + 2H + 2e changes at action end:

+ +

NADH + H

H CONH2 + 2H + 2e N
+ +

H CONH2 + H N
+

A second substance, NADP (nicotinamide adenine dinucleotide phosphate), which contains an additional phosphate group, has a similar role in photosynthesis.

Edexcel Biology for A2 Dynamic Learning

Hodder Education 2009

ON THE WILD SIDE: A* EXTENSIONS 5.1 TO 5.3

A* Extension 5.2: The source of oxygen in photosynthesis

The explanation of the light-dependent reaction tells us that the oxygen given off in photosynthesis is derived exclusively from the photolysis of water (rather than from carbon dioxide, for example). An experiment using water containing the isotope oxygen-18 confirmed that this was correct. Illuminated suspensions of photosynthetic cells were provided with either carbon dioxide enriched with C18O2 or water enriched with H218O, as shown in the diagrams below. From both, the oxygen evolved was analysed to determine the oxygen-18 concentration.
two identical cultures, one supplied with C18O2 and the other with H218O C18O2 O2 CO2
18

O2

LIGHT

LIGHT

suspension of single-celled algae (Chlorella sp.) H2O H218O

magnetic stirrer

pathway of photolysis of water 4 H2O 4(H+ + e) + 4OH 2 H2O + O2 the oxygen evolved comes from water hydrogen acceptor reduced hydrogen acceptor

Incidentally, this experiment established that the traditional balanced equation for photosynthesis is incorrect because it implies that some or all of the oxygen evolved comes from carbon dioxide: 6CO2 6H2O light energy C6H12O6 6O2

Can you see why? This is because it shows 12 oxygen atoms are produced, but the six water molecules on the lefthand side of the equation contain only six. The summary equation for photosynthesis is less misleading when written as: 6CO2 12H2O light energy C6H12O6 6O2 6H2O

Edexcel Biology for A2 Dynamic Learning

Hodder Education 2009

ON THE WILD SIDE: A* EXTENSIONS 5.1 TO 5.3

A* Extension 5.3: CO2-limiting conditions and cyclic photophosphorylation

This extension material addresses HSW Criterion 2c. At any time that the carbon dioxide concentration around a green plant becomes very low (so low we describe it as a limiting factor), then the light-independent reactions will be slowed down, relative to the light-dependent reactions. An outcome is that NADPH will accumulate in the stroma and the concentration of NADP will fall. Then, without an adequate supply of oxidised hydrogen acceptor (NADP ), photosystems II and I are unable to operate together, as they do in non-cyclic photophosphorylation (Figure 5.6, on page 5 in Edexcel Biology for A2). Now it has been found that another type of ATP formation occurs under these conditions, called cyclic photophosphorylation. In cyclic photophosphorylation, excited electrons from the reaction centre in photosystem I fall back to their point of origin. They do this via electron carriers that transfer energy from those electrons and pump H into the thylakoid space. ATP synthesis by chemiosmosis follows. The Z-diagram below illustrates this.
normal flow of electrons (from water) Photosystem II + electron carriers Photosystem I + electron carriers NADP+ is inhibited Photosystem I Photosystem II ATPase excited electrons captured by electron acceptor but further passage is blocked H+ H+ ADP + Pi ATP

H+

H+

H+ H+ H+ H+ H+ H+ H+ H+

thylakoid compartment thylakoid membrane stroma

Cyclic photophosphorylation occurs when the CO2 concentration is the limiting factor (and with relatively high light intensity), so that the light-independent reactions are slowed relative to the light-dependent reactions. NADPH accumulates in the stroma.

electron has taken a circular path but causes photophosphorylation to continue as H+ flow out via ATPase hence the process is called cyclic photophosphorylation

energy causes H+ to be pumped into thylakoid space, then electron falls back to ground-state conditions in Photosystem I

H+ accumulate in thylakoid space

This type of photophosphorylation is called cyclic because the electrons have returned to the photosystem from which they originated (a cyclic path, rather than a linear one). The outcome is that ATP synthesis continues, using light energy, despite the fact that sugar production due to photosynthesis is (temporarily) terminated. Question for discussion In what circumstances or conditions would the facility for cyclic photophosphorylation be advantageous to green plants, and why (HSW Criterion 2c)?

Edexcel Biology for A2 Dynamic Learning

Hodder Education 2009