Photosynthetic organisms harvest light for energy. Some eukaryotic algae have specialized in harvesting far-red light by tuning chlorophyll a absorption through a mechanism still to be elucidated. Here, we combined optically detected magnetic resonance and pulsed electron paramagnetic resonance measurements on red-adapted light-harvesting complexes, rVCP, isolated from the freshwater eustigmatophyte alga Trachydiscus minutus to identify the location of the pigments responsible for this remarkable adaptation. The pigments have been found to belong to an excitonic cluster of chlorophylls a at the core of the complex, close to the central carotenoids in L1/L2 sites. A pair of structural features of the Chl a403/a603 binding site, namely the histidine-to-asparagine substitution in the magnesium-ligation residue and the small size of the amino acid at the i-4 position, resulting in a [A/G]xxxN motif, are proposed to be the origin of this trait. Phylogenetic analysis of various eukaryotic red antennae identified several potential LHCs that could share this tuning mechanism. This knowledge of the red light acclimation mechanism in algae is a step towards rational design of algal strains in order to enhance light capture and efficiency in large-scale biotechnology applications.

• MeSH
• Chlorophyll A * metabolism   chemistry   MeSH
• Chlorophyll metabolism   MeSH
• Electron Spin Resonance Spectroscopy MeSH
• Phylogeny MeSH
• Light MeSH
• Light-Harvesting Protein Complexes * metabolism   genetics   chemistry   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll A * MeSH
• Chlorophyll MeSH
• Light-Harvesting Protein Complexes * MeSH

Carotenoids are conjugated linear molecules built from the repetition of terpene units, which display a large structural diversity in nature. They may, in particular, contain several types of side or end groups, which tune their functional properties, such as absorption position and photochemistry. We report here a detailed experimental study of the absorption and vibrational properties of allene-containing carotenoids, together with an extensive modeling of these experimental data. Our calculations can satisfactorily explain the electronic properties of vaucheriaxanthin, where the allene group introduces the equivalent of one C═C double bond into the conjugated C═C chain. The position of the electronic absorption of fucoxanthin and butanoyloxyfucoxanthin requires long-range corrections to be found correctly on the red side of that of vaucheriaxanthin; however, these corrections tend to overestimate the effect of the conjugated and nonconjugated C═O groups in these molecules. We show that the resonance Raman spectra of these carotenoids are largely perturbed by the presence of the allene group, with the two major Raman contributions split into two components. These perturbations are satisfactorily explained by modeling, through a gain in the Raman intensity of the C═C antisymmetric stretching mode, induced by the presence of the allene group in the carotenoid C═C chain.

• MeSH
• Alkadienes * MeSH
• Electronics MeSH
• Carotenoids * chemistry   MeSH
• Spectrum Analysis, Raman MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Alkadienes * MeSH
• Carotenoids * MeSH
• propadiene MeSH Browser

Survival of phototrophic organisms depends on their ability to collect and convert enough light energy to support their metabolism. Phototrophs can extend their absorption cross section by using diverse pigments and by tuning the properties of these pigments via pigment-pigment and pigment-protein interaction. It is well known that some cyanobacteria can grow in heavily shaded habitats by utilizing far-red light harvested with far-red-absorbing chlorophylls d and f. We describe a red-shifted light-harvesting system based on chlorophyll a from a freshwater eustigmatophyte alga Trachydiscus minutus (Eustigmatophyceae, Goniochloridales). A comprehensive characterization of the photosynthetic apparatus of T. minutus is presented. We show that thylakoid membranes of T. minutus contain light-harvesting complexes of several sizes differing in the relative amount of far-red chlorophyll a forms absorbing around 700 nm. The pigment arrangement of the major red-shifted light-harvesting complex is similar to that of the red-shifted antenna of a marine alveolate alga Chromera velia. Evolutionary aspects of the algal far-red light-harvesting complexes are discussed. The presence of these antennas in eustigmatophyte algae opens up new ways to modify organisms of this promising group for effective use of far-red light in mass cultures.

• Keywords
• Chromatic acclimation, Eustigmatophyta, Light-harvesting protein, Oligomeric LHC, Red-shifted LHC, Violaxanthin,
• MeSH
• Pigments, Biological metabolism   MeSH
• Diuron MeSH
• Spectrometry, Fluorescence MeSH
• Stramenopiles metabolism   radiation effects   MeSH
• Membrane Proteins metabolism   MeSH
• Fresh Water * MeSH
• Light * MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Temperature MeSH
• Thylakoids metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Pigments, Biological MeSH
• Diuron MeSH
• Membrane Proteins MeSH
• Light-Harvesting Protein Complexes MeSH

The soil chromophyte alga Xanthonema (X.) debile contains only non-carbonyl carotenoids and Chl-a. X. debile has an antenna system denoted Xanthophyte light-harvesting complex (XLH) that contains the carotenoids diadinoxanthin, heteroxanthin, and vaucheriaxanthin. The XLH pigment stoichiometry was calculated by chromatographic techniques and the pigment-binding structure studied by resonance Raman spectroscopy. The pigment ratio obtained by HPLC was found to be close to 8:1:2:1 Chl-a:heteroxanthin:diadinoxanthin:vaucheriaxanthin. The resonance Raman spectra suggest the presence of 8-10 Chl-a, all of which are 5-coordinated to the central Mg, with 1-3 Chl-a possessing a macrocycle distorted from the relaxed conformation. The three populations of carotenoids are in the all-trans configuration. Vaucheriaxanthin absorbs around 500-530 nm, diadinoxanthin at 494 nm and heteroxanthin at 487 nm at 4.5 K. The effective conjugation length of heteroxanthin and diadinoxanthin has been determined as 9.4 in both cases; the environment polarizability of the heteroxanthin and diadinoxanthin binding pockets is 0.270 and 0.305, respectively.

• Keywords
• Algae, Carotenoids, Chl-a, Diadinoxanthin, Heteroxanthin, Light-harvesting complex, Resonance Raman,
• MeSH
• Stramenopiles chemistry   MeSH
• Carotenoids chemistry   MeSH
• Protein Conformation MeSH
• Spectrum Analysis, Raman MeSH
• Light-Harvesting Protein Complexes chemistry   MeSH
• Chromatography, High Pressure Liquid MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Carotenoids MeSH
• Light-Harvesting Protein Complexes MeSH

We have used time-resolved absorption and fluorescence spectroscopy with nanosecond resolution to study triplet energy transfer from chlorophylls to carotenoids in a protective process that prevents the formation of reactive singlet oxygen. The light-harvesting complexes studied were isolated from Chromera velia, belonging to a group Alveolata, and Xanthonema debile and Nannochloropsis oceanica, both from Stramenopiles. All three light-harvesting complexes are related to fucoxanthin-chlorophyll protein, but contain only chlorophyll a and no chlorophyll c. In addition, they differ in the carotenoid content. This composition of the complexes allowed us to study the quenching of chlorophyll a triplet states by different carotenoids in a comparable environment. The triplet states of chlorophylls bound to the light-harvesting complexes were quenched by carotenoids with an efficiency close to 100%. Carotenoid triplet states were observed to rise with a ~5 ns lifetime and were spectrally and kinetically homogeneous. The triplet states were formed predominantly on the red-most chlorophylls and were quenched by carotenoids which were further identified or at least spectrally characterized.

• Keywords
• Algae, Energy transfer, Light harvesting, Photoprotection, Photosynthesis, Transient spectroscopy,
• MeSH
• Anaerobiosis MeSH
• Time Factors MeSH
• Chlorophyll metabolism   MeSH
• Spectrometry, Fluorescence MeSH
• Photochemical Processes * MeSH
• Stramenopiles metabolism   MeSH
• Carotenoids metabolism   MeSH
• Kinetics MeSH
• Chlorophyll Binding Proteins metabolism   MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll MeSH
• Carotenoids MeSH
• Chlorophyll Binding Proteins MeSH
• Light-Harvesting Protein Complexes MeSH

Resonance Raman spectroscopy was used to evaluate pigment-binding site properties in the violaxanthin-chlorophyll-a-binding protein (VCP) from Nannochloropsis oceanica. The pigments bound to this antenna protein are chlorophyll-a, violaxanthin, and vaucheriaxanthin. The molecular structures of bound Chl-a molecules are discussed with respect to those of the plant antenna proteins LHCII and CP29, the crystal structures of which are known. We show that three populations of carotenoid molecules are bound by VCP, each of which is in an all-trans configuration. We assign the lower-energy absorption transition of each of these as follows. One violaxanthin population absorbs at 485 nm, while the second population is red-shifted and absorbs at 503 nm. The vaucheriaxanthin population absorbs at 525 nm, a position red-shifted by 2138 cm-1 as compared to isolated vaucheriaxanthin in n-hexane. The red-shifted violaxanthin is slightly less planar than the blue-absorbing one, as observed for the two central luteins in LHCII, and we suggest that these violaxanthins occupy the two equivalent binding sites in VCP at the centre of the cross-brace. The presence of a highly red-shifted vaucheriaxanthin in VCP is reminiscent of the situation of FCP, in which (even more) highly red-shifted populations of fucoxanthin are present. Tuning carotenoids to absorb in the green-yellow region of the visible spectrum appears to be a common evolutionary response to competition with other photosynthetic species in the aquatic environment.

• Keywords
• Carotenoids, Light-harvesting complex, Nannochloropsis oceanica, Resonance Raman, VCP,
• MeSH
• Chlorophyll chemistry   MeSH
• Carotenoids chemistry   MeSH
• Spectrum Analysis, Raman MeSH
• Light-Harvesting Protein Complexes chemistry   MeSH
• Carrier Proteins chemistry   MeSH
• Xanthophylls chemistry   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll MeSH
• Carotenoids MeSH
• Light-Harvesting Protein Complexes MeSH
• Carrier Proteins MeSH
• violaxanthin MeSH Browser
• Xanthophylls MeSH

Photosystem I (PSI) is a multi-subunit integral pigment-protein complex that performs light-driven electron transfer from plastocyanin to ferredoxin in the thylakoid membrane of oxygenic photoautotrophs. In order to achieve the optimal photosynthetic performance under ambient irradiance, the absorption cross section of PSI is extended by means of peripheral antenna complexes. In eukaryotes, this role is played mostly by the pigment-protein complexes of the LHC family. The structure of the PSI-antenna supercomplexes has been relatively well understood in organisms harboring the primary plastid: red algae, green algae and plants. The secondary endosymbiotic algae, despite their major ecological importance, have so far received less attention. Here we report a detailed structural analysis of the antenna-PSI association in the stramenopile alga Nannochloropsis oceanica (Eustigmatophyceae). Several types of PSI-antenna assemblies are identified allowing for identification of antenna docking sites on the PSI core. Instances of departure of the stramenopile system from the red algal model of PSI-Lhcr structure are recorded, and evolutionary implications of these observations are discussed.

• Keywords
• Electron microscopy, Light-harvesting complex, Nannochloropsis, Photosystem I, Stramenopila,
• MeSH
• Photosystem I Protein Complex metabolism   MeSH
• Plastids metabolism   MeSH
• Rhodophyta metabolism   MeSH
• Spectrophotometry, Ultraviolet MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Photosystem I Protein Complex MeSH

Eustigmatophyte algae represent an interesting model system for the study of the regulation of the excitation energy flow due to their use of violaxanthin both as a major light-harvesting pigment and as the basis of xanthophyll cycle. Fluorescence induction kinetics was studied in an oleaginous marine alga Nannochloropsis oceanica. Nonphotochemical fluorescence quenching was analyzed in detail with respect to the state of the cellular xanthophyll pool. Two components of nonphotochemical fluorescence quenching (NPQ), both dependent on the presence of zeaxanthin, were clearly resolved, denoted as slow and fast NPQ based on kinetics of their formation. The slow component was shown to be in direct proportion to the amount of zeaxanthin, while the fast NPQ component was transiently induced in the presence of membrane potential on subsecond timescales. The applicability of these observations to other eustigmatophyte species is demonstrated by measurements of other representatives of this algal group, both marine and freshwater.

• Keywords
• Chl a fluorescence, Eustigmatophyceae, Nannochloropsis, Nonphotochemical quenching, Xanthophyll cycle,
• MeSH
• Fluorescence MeSH
• Photosynthesis MeSH
• Seaweed chemistry   MeSH
• Publication type
• Journal Article MeSH