Chlorophyll

Chlorophyll at different scales

Chlorophyll is responsible for the green color of many plants and algae.

Seen through a microscope, chlorophyll is concentrated within organisms in structures called chloroplasts.

There are several types of chlorophyll, but all share the chlorin magnesium ligand which forms the right side of this diagram.

Chlorophyll (also chlorophyl) is any of several closely related green pigments found in cyanobacteria and the chloroplasts of algae and plants.^[1] Its name is derived from the Greek words χλωρός, chloros ("green") and φύλλον, phyllon ("leaf").^[2] Chlorophyll is essential in photosynthesis, allowing plants to absorb energy from light.

Chlorophyll absorbs light most strongly in the blue portion of the electromagnetic spectrum, followed by the red portion.^[3] Conversely, it is a poor absorber of green and near-green portions of the spectrum, which it reflects, producing the green color of chlorophyll-containing tissues. Chlorophyll molecules are specifically arranged in and around photosystems that are embedded in the thylakoid membranes of chloroplasts. Two types of chlorophyll exist in the photosystems of green plants: chlorophyll a and b.^[4]

Chlorophyll was first isolated and named by Joseph Bienaimé Caventou and Pierre Joseph Pelletier in 1817.^[5]

Photosynthesis

Absorption maxima of chlorophylls against the spectrum of white light. Chlorophyll absorbs energy from light strongly at visible frequencies.

Chlorophyll is vital for photosynthesis, which allows plants to absorb energy from light.^[6]

Chlorophyll molecules are specifically arranged in and around photosystems that are embedded in the thylakoid membranes of chloroplasts.^[7] In these complexes, chlorophyll serves two primary functions. The function of the vast majority of chlorophyll (up to several hundred molecules per photosystem) is to absorb light and transfer that light energy by resonance energy transfer to a specific chlorophyll pair in the reaction center of the photosystems. The two currently accepted photosystem units are photosystem II and photosystem I, which have their own distinct reaction centres, named P680 and P700, respectively. These centres are named after the wavelength (in nanometers) of their red-peak absorption maximum. The identity, function and spectral properties of the types of chlorophyll in each photosystem are distinct and determined by each other and the protein structure surrounding them. Once extracted from the protein into a solvent (such as acetone or methanol),^[8][9][10] these chlorophyll pigments can be separated into chlorophyll a and chlorophyll b.

The function of the reaction center of chlorophyll is to absorb light energy and transfer it to other parts of the photosystem. The absorbed energy of the photon is transferred to an electron in a process called charge separation. The removal of the electron from the chlorophyll is an oxidation reaction. The chlorophyll donates the high energy electron to a series of molecular intermediates called an electron transport chain. The charged reaction center of chlorophyll (P680⁺) is then reduced back to its ground state by accepting an electron stripped from water. The electron that reduces P680⁺ ultimately comes from the oxidation of water into O₂ and H⁺ through several intermediates. This reaction is how photosynthetic organisms such as plants produce O₂ gas, and is the source for practically all the O₂ in Earth's atmosphere. Photosystem I typically works in series with Photosystem II; thus the P700⁺ of Photosystem I is usually reduced as it accepts the electron, via many intermediates in the thylakoid membrane, by electrons coming, ultimately, from Photosystem II. Electron transfer reactions in the thylakoid membranes are complex, however, and the source of electrons used to reduce P700⁺ can vary.

The electron flow produced by the reaction center chlorophyll pigments is used to pump H⁺ ions across the thylakoid membrane, setting up a chemiosmotic potential used mainly in the production of ATP (stored chemical energy) or to reduce NADP⁺ to NADPH. NADPH is a universal agent used to reduce CO₂ into sugars as well as other biosynthetic reactions.

Reaction center chlorophyll–protein complexes are capable of directly absorbing light and performing charge separation events without the assistance of other chlorophyll pigments, but the probability of that happening under a given light intensity is small. Thus, the other chlorophylls in the photosystem and antenna pigment proteins all cooperatively absorb and funnel light energy to the reaction center. Besides chlorophyll a, there are other pigments, called accessory pigments, which occur in these pigment–protein antenna complexes.

Chemical structure

Space-filling model of the chlorophyll a molecule

Chlorophyll is a chlorin pigment, which is structurally similar to and produced through the same metabolic pathway as other porphyrin pigments such as heme. At the center of the chlorin ring is a magnesium ion. This was discovered in 1906,^[11] and was the first time that magnesium had been detected in living tissue.^[12] For the structures depicted in this article, some of the ligands attached to the Mg²⁺ center are omitted for clarity. The chlorin ring can have several different side chains, usually including a long phytol chain. There are a few different forms that occur naturally, but the most widely distributed form in terrestrial plants is chlorophyll a. After initial work done by German chemist Richard Willstätter spanning from 1905 to 1915, the general structure of chlorophyll a was elucidated by Hans Fischer in 1940. By 1960, when most of the stereochemistry of chlorophyll a was known, Robert Burns Woodward published a total synthesis of the molecule.^[12][13] In 1967, the last remaining stereochemical elucidation was completed by Ian Fleming,^[14] and in 1990 Woodward and co-authors published an updated synthesis.^[15] Chlorophyll f was announced to be present in cyanobacteria and other oxygenic microorganisms that form stromatolites in 2010;^[16][17] a molecular formula of C₅₅H₇₀O₆N₄Mg and a structure of (2-formyl)-chlorophyll a were deduced based on NMR, optical and mass spectra.^[18] The different structures of chlorophyll are summarized below:

Structure of chlorophyll a	Structure of chlorophyll b	Structure of chlorophyll d
Structure of chlorophyll c1	Structure of chlorophyll c2	Structure of chlorophyll f

When leaves degreen in the process of plant senescence, chlorophyll is converted to a group of colourless tetrapyrroles known as nonfluorescent chlorophyll catabolites (NCC's) with the general structure:

These compounds have also been identified in several ripening fruits.^[19]

Measurement of chlorophyll content

Absorbance spectra of free chlorophyll a (blue) and b (red) in a solvent. The spectra of chlorophyll molecules are slightly modified in vivo depending on specific pigment-protein interactions.

Because it absorbs red and blue light strongly but is transparent to green light, pure chlorophyll has a strong green colour.

Measurement of the absorption of light is complicated by the solvent used to extract the chlorophyll from plant material, which affects the values obtained,

• In diethyl ether, chlorophyll a has approximate absorbance maxima of 430 nm and 662 nm, while chlorophyll b has approximate maxima of 453 nm and 642 nm.^[20]
• The absorption peaks of chlorophyll a are at 665 nm and 465 nm. Chlorophyll a fluoresces at 673 nm (maximum) and 726 nm. The peak molar absorption coefficient of chlorophyll a exceeds 10⁵ M⁻¹ cm⁻¹, which is among the highest for small-molecule organic compounds.^[21]
• In 90% acetone-water, the peak absorption wavelengths of chlorophyll a are 430 nm and 664 nm; peaks for chlorophyll b are 460 nm and 647 nm; peaks for chlorophyll c1 are 442 nm and 630 nm; peaks for chlorophyll c2 are 444 nm and 630 nm; peaks for chlorophyll d are 401 nm, 455 nm and 696 nm.^[22]

By measuring the absorption of light in the red and far red regions, it is possible to estimate the concentration of chlorophyll within a leaf.^[23]

Ratio fluorescence emission can be used to measure chlorophyll content. By exciting chlorophyll “a” fluorescence at a lower wavelength, the ratio of chlorophyll fluorescence emission at 705 nm +/- 10 nm and 735 nm +/-10 nm can provide a linear relationship of chlorophyll content when compared to chemical testing. The ratio F735/F700 provided a correlation value of r² 0.96 compared to chemical testing in the range from 41 mg m⁻² up to 675 mg m⁻². Gitelson also developed a formula for direct readout of chlorophyll content in mg m⁻². The formula provided a reliable method of measuring chlorophyll content from 41 mg m⁻² up to 675 mg m⁻² with a correlation r² value of 0.95.^[24]

Biosynthesis

In plants, chlorophyll may be synthesized from succinyl-CoA and glycine, although the immediate precursor to chlorophyll a and b is protochlorophyllide. In Angiosperm plants, the last step, the conversion of protochlorophyllide to chlorophyll, is light-dependent and such plants are pale (etiolated) if grown in darkness. Non-vascular plants and green algae have an additional light-independent enzyme and grow green even in darkness.

Chlorophyll itself is bound to proteins and can transfer the absorbed energy in the required direction. Protochlorophyllide occurs mostly in the free form and, under light conditions, acts as a photosensitizer, forming highly toxic free radicals. Hence, plants need an efficient mechanism of regulating the amount of chlorophyll precursor. In angiosperms, this is done at the step of aminolevulinic acid (ALA), one of the intermediate compounds in the biosynthesis pathway. Plants that are fed by ALA accumulate high and toxic levels of protochlorophyllide; so do the mutants with the damaged regulatory system.^[25]

Chlorosis is a condition in which leaves produce insufficient chlorophyll, turning them yellow. Chlorosis can be caused by a nutrient deficiency of iron—called iron chlorosis—or by a shortage of magnesium or nitrogen. Soil pH sometimes plays a role in nutrient-caused chlorosis; many plants are adapted to grow in soils with specific pH levels and their ability to absorb nutrients from the soil can be dependent on this.^[26] Chlorosis can also be caused by pathogens including viruses, bacteria and fungal infections, or sap-sucking insects.

Complementary light absorbance of anthocyanins with chlorophylls

Superposition of spectra of chlorophyll a and b with oenin (malvidin 3O glucoside), a typical anthocyanidin, showing that, while chlorophylls absorb in the blue and yellow/red parts of the visible spectrum, oenin absorbs mainly in the green part of the spectrum, where chlorophylls don't absorb at all.

Anthocyanins are other plant pigments. The absorbance pattern responsible for the red color of anthocyanins may be complementary to that of green chlorophyll in photosynthetically active tissues such as young Quercus coccifera leaves. It may protect the leaves from attacks by plant eaters that may be attracted by green color.^[27]

Distribution

The chlorophyll maps show milligrams of chlorophyll per cubic meter of seawater each month. Places where chlorophyll amounts were very low, indicating very low numbers of phytoplankton, are blue. Places where chlorophyll concentrations were high, meaning many phytoplankton were growing, are yellow. The observations come from the Moderate Resolution Imaging Spectroradiometer (MODIS) on NASA's Aqua satellite. Land is dark gray, and places where MODIS could not collect data because of sea ice, polar darkness, or clouds are light gray.The highest chlorophyll concentrations, where tiny surface-dwelling ocean plants are thriving, are in cold polar waters or in places where ocean currents bring cold water to the surface, such as around the equator and along the shores of continents. It is not the cold water itself that stimulates the phytoplankton. Instead, the cool temperatures are often a sign that the water has welled up to the surface from deeper in the ocean, carrying nutrients that have built up over time. In polar waters, nutrients accumulate in surface waters during the dark winter months when plants cannot grow. When sunlight returns in the spring and summer, the plants flourish in high concentrations.^[28]

Culinary use

Chlorophyll is registered as a food additive (colorant), and its E number is E140. Chefs use chlorophyll to color a variety of foods and beverages green, such as pasta and absinthe.^[29] Chlorophyll is not soluble in water, and it is first mixed with a small quantity of vegetable oil to obtain the desired solution.

See also

Wikimedia Commons has media related to Chlorophyll.

• Bacteriochlorophyll, related compounds in phototrophic bacteria
• Chlorophyll a, an essential chlorophyll pigment
• Chlorophyll b, also an essential chlorophyll pigment
• Chlorophyllin, a semi-synthetic derivative of chlorophyll
• Deep chlorophyll maximum
• Grow light, a lamp that promotes photosynthesis
• Chlorophyll fluorescence, to measure plant stress

References