Photosynthetic Green Sulfur Bacteria Research Articles

Reconstructing the palaeoecology of a middle Permian alkaline lake using molecular fossils, case study of the Lucaogou Formation in the Junggar Basin, NW China

The lacustrine Lucaogou Formation was deposited during the middle Permian in the Junggar Basin, northwestern China. The depositional environment was an alkaline, brackish to saline lake. The biological diversity of the Lucaogou Formation was reconstructed using biologically informative molecules such as n-alkanes, steranes, and hopanes, and the δ13C composition of organic matter. C30 4α-methyl-24-ethylsterane, C31 lanostane, and abundant C30 − C31 3β-methylhopanes were detected in the organic matter, suggesting the presence of type Ⅰ aerobic methanotrophs including the Methylococcaceae. C30 − C31 2α-methylhopanes and 7- and 8-methylheptadecanes are suggestive of the presence of cyanobacteria. Cyanobacteria and green algae are proposed to have made the dominant contribution to the organic matter. The presence of more 13C-enriched hopanoids in the lower unit compared to the upper unit of the Lucaogou Formation suggest a greater contribution of cyanobacteria to the organic matter in the lower unit. The high abundances of C28 and C29 steranes relative to C27 steranes are likely due to green algae from the Chlorophyta, including the class Chlorophyceae. Dinosteranes that are diagnostic for dinoflagellates were not detected. The low concentrations of C40 isorenieratane, renieratane, β-isorenieratane, renierapurpurane, and β-renierapurpurane that are indicative of photosynthetic green and purple sulfur bacteria indicate a minor contribution from Chlorobiaceae and Chromatiaceae to the organic matter. It is inferred that low to moderate amounts of ciliates grew at or below the relatively unstable chemocline in the lake. The presence of C19-norisopimarane, 8β(H)-labdane, and 4β(H)-19-norisopimarane suggests the presence of coniferous plants growing around the lake, probably including Pinaceae, Araucariaceae, Cupressaceae.

Living on the edge: light-harvesting efficiency and photoprotection in the core of green sulfur bacteria.

Photosynthetic green sulfur bacteria are able to survive under extreme low light conditions. Nevertheless, the light-harvesting efficiencies reported so far, in particular for Fenna-Matthews-Olson (FMO) protein-reaction center complex (RCC) supercomplexes, are much lower than for photosystems of other species. Here, we approach this problem with a structure-based theory. Compelling evidence for a light-harvesting efficiency around 95% is presented for native (anaerobic) conditions that can drop down to 47% when the FMO protein is switched into a photoprotective mode in the presence of molecular oxygen. Light-harvesting bottlenecks are found between the FMO protein and the RCC, and the antenna of the RCC and its reaction center (RC) with forward energy transfer time constants of 39 ps and 23 ps, respectively. The latter time constant removes an ambiguity in the interpretation of time-resolved spectra of RCC probing primary charge transfer and provides strong evidence for a transfer-to-the trap limited kinetics of excited states. Different factors influencing the light-harvesting efficiency are investigated. A fast primary electron transfer in the RC is found to be more important for a high efficiency than the site energy funnel in the FMO protein, quantum effects of nuclear motion, or variations in the mutual orientation between the FMO protein and the RCC.

Molecular and carbon isotopic evidence of pigments indicating a dynamic oceanic chemocline 1.4 billion years ago in northern China

The redox state of the marine photic zone is crucial to the evolution of life, which is further linked to the degree of oxygenation of the atmosphere and ocean systems. Hydrocarbon molecular fossils (biomarkers) and isotopic fingerprints of anoxygenic photoautotrophs can provide direct evidence for seawater redox conditions within the photic zone. In this study, we identified a series of 2,3,6-trimethyl aryl isoprenoids (2,3,6-TMAI) from low-maturity sedimentary organic matter in the Xiamaling Formation (XML), deposited 1.4 billion years ago in the Xiahuayuan area of northern China. The δ13C values of C18 and C19 TMAI ranged from −19.8‰ to −21.3‰. These 13C-enriched values indicate their origin in anaerobic photosynthetic green sulfur bacteria (GSB). The episodic enrichments of 2,3,6-TMAI in the XML Formation suggest dynamic marine geochemistry. Specifically, low concentrations of 2,3,6-TMAI at the bottom of the studied profile reflect an oxygenated deposition environment (unit 4), where the organic matter was almost all decomposed in the oxic water column during deposition, while the gradually increasing 2,3,6-TMAI concentrations from the middle to the upper XML reflects alteration of oceanic redox condition from an oxygen minimum zone (OMZ) in unit 3 through periods of euxinia during the deposition of units 2 and 1. The absence of co-variation between total organic carbon (TOC) and enriched 13C values indicates that anoxygenic photoautotroph activities might have contributed little to sedimentary organic matter burial in the Mesoproterozoic XML sediments.

Annual phytoplankton cycle in a meromictic anoxic basin of a Rhode Island (USA) estuarine river

Estuarine Pettaquamscutt River is a unique habitat 10 km in length with physical and biogeochemical characteristics analogous to a miniature fjord. Its meromictic upper basins are characterized by a permanent oxic–anoxic vertical gradient in which a well-oxygenated upper layer overlies a deeper, anoxic reservoir, with persistent blooms of phototrophic anoxygenic bacteria (Chromatium, Chlorobium) at the oxic–anoxic transition layer. A diverse assemblage of nanoplanktonic, centric diatoms (Cyclotella caspia, Thalassiosira pseudonana, cf. Cyclotella cryptica) dominated the seasonal phytoplankton cycle in the aerobic layer, similar to comparable meromictic habitats elsewhere. This assemblage of nano-centric diatoms appears to be trait-separated from other species clusters and is potentially useful as a functional group flora with ecophysiology diagnostic of marine estuarine rivers and meromictic habitat niche structure. The most conspicuous phytoplankton feature, however, was the year-round occurrence of the photoautotrophic euglenid Euglenaformis (Euglena) proxima, restricted in its upper water column distribution to the O2–H2S boundary layer where it formed a consortium with the photosynthetic green and purple sulfur bacteria community. The association of E. proxima with meromixis, H2S, and phototrophic anoxygenic bacteria is similar to that reported previously in a brackish Norwegian oyster poll and a brackish loch in Scotland.

Draft Whole-Genome Sequence of the Green Sulfur Photosynthetic Bacterium Chlorobaculum sp. Strain 24CR, Isolated from the Carmel River.

Green sulfur bacteria are in the family Chlorobiaceae, which is composed of four distinct genera, namely, Chlorobaculum, Chlorobium, Prosthecochloris, and Chloroherpeton, with Chlorobium species being the most commonly represented in genome studies. We have now sequenced only the fourth species of Chlorobaculum, which established Chlorobaculum sp. 24CR as a separate species and should help characterize the genus.

Geochemical features and genetic mechanism of deep-water source rocks in the Senegal basin, West Africa

This paper discusses the maturity of source rocks of the Senegal basin through basin simulation, so as to get a better understanding of oil-source correlation. Based on the analysis of pyrolysis chromatography and total organic carbon (TOC) data of core samples taken from 11 wells, the model of Cenomanian-Turonian marine sediment-organic facies was established, and the genetic mechanism of high-quality source rocks was clarified. The results show that source rocks in the Senegal Basin may occur in the Aptian-Albian of Lower Cretaceous and Cenomanian-Turonian of Upper Cretaceous. One is hybrid organic facies in the shallow carbonate platforms in the shelf area and is characterized by moderate to high TOC (<3%) and hidrogen index ? HI, (100-400 mg HC/g). The other is well-preserved marine facies in continental slope to abyssal sea, with high TOC (>3%) and high HI (max 900 mg HC/g TOC). Molecular fossils originating from aryl carotene are the indicator of the existence of euphotic zone in the ancient ocean. The compounds of aryl isoprenes and sulfurous aryl isoprenes are detected in the black shale samples of well DSDP 367. They rooted in photosynthetic green sulfur bacteria and the carbon isotope value of these compounds is higher 10?~15? than those of fossil molecules of algae and cyanobacteria. Two packages of oil-prone source rocks separately occurr in the Aptian-Albian of Lower Cretaceous and Cenomanian-Turonian of Upper Cretaceous. High-graded marine source rocks of the Senegal Basin may occur in a sulfurous, anoxic deep-water environment with sufficient carbon sequestration.

Microbial composition, potential functional roles and genetic novelty in gypsum-rich and hypersaline soils of Monegros and Gallocanta (Spain)

Soil microbial communities (both Bacteria and Archaea) were studied after 16S rRNA genes massive sequencing in two hypersaline and gypsum-rich contrasted sites located in NE Spain. Soil microbial communities were also locally analysed according to environmental variables, including geological, physico-chemical, biogeochemically, and climatic data. Typical soil characteristics, climate data, and plant composition clearly split the two sites and major differences among the microbial communities for the areas were initially expected. Overall, high values of microbial species richness (up to 2300 taxa) and ecological diversity was detected in both sites. High genetic novelty levels were found mostly to environmental sequences, highlighting the high potential for microbiological studies. In contrast to the initial expectations, a substantial overlapping between Monegros and Gallocanta microbes was observed, indicating a high similarity despite of the geographical, botanical and environmental distances between sites, in agreement with both high dispersal and local selection inherent to the microbial world. The potential biogeochemical cycling showed small differences between sites, with presence of photosynthetic green and purple sulfur bacteria, cyanobacteria and aerobic and anaerobic chemolitotrophs. Potential for aerobic methane oxidation and anaerobic methanogenesis was observed in both sites, with predominance of potential nitrification mostly by ammonia-oxidizing archaea, nitrite oxidation and denitrification, and minor contribution for nitrate reduction and nitrate ammonification. The predicted functions based on the taxonomic composition showed high overlapping between the two studied regions, despite their difference in gypsum richness.

The Arnon-Buchanan cycle: a retrospective, 1966-2016.

For the first decade following its description in 1954, the Calvin-Benson cycle was considered the sole pathway of autotrophic CO2 assimilation. In the early 1960s, experiments with fermentative bacteria uncovered reactions that challenged this concept. Ferredoxin was found to donate electrons directly for the reductive fixation of CO2 into alpha-keto acids via reactions considered irreversible. Thus, pyruvate and alpha-ketoglutarate could be synthesized from CO2, reduced ferredoxin and acetyl-CoA or succinyl-CoA, respectively. This work opened the door to the discovery that reduced ferredoxin could drive the Krebs citric acid cycle in reverse, converting the pathway from its historical role in carbohydrate breakdown to one fixing CO2. Originally uncovered in photosynthetic green sulfur bacteria, the Arnon-Buchanan cycle has since been divorced from light and shown to function in a variety of anaerobic chemoautotrophs. In this retrospective, colleagues who worked on the cycle at its inception in 1966 and those presently working in the field trace its development from a controversial reception to its present-day inclusion in textbooks. This pathway is now well established in major groups of chemoautotrophic bacteria, instead of the Calvin-Benson cycle, and is increasingly referred to as the Arnon-Buchanan cycle. In this retrospective, separate sections have been written by the authors indicated. Bob Buchanan wrote the abstract and the concluding comments.

Amphiphilic zinc bacteriochlorophyll a derivatives that function as artificial energy acceptors in photosynthetic antenna complexes chlorosomes of the green sulfur photosynthetic bacterium Chlorobaculum limnaeum

Abstract Green photosynthetic bacteria have extramembranous light-harvesting complexes called chlorosomes. Light-energy captured by bacteriochlorophyll (BChl) self-aggregates inside chlorosomes is transferred to BChl a -peptide complexes called baseplates on the chlorosome surface. We substitute the baseplate complex in chlorosomes from a green sulfur photosynthetic bacterium Chlorobaculum limnaeum with amphiphilic Zn BChl a derivatives. Zn BChl a derivatives 1 – 3 , which were esterified with polyethylene glycol (PEG) 600, PEG400, and diethylene glycol, respectively, were successfully hybridized with chlorosomes without baseplates. These derivatives in the hybridized chlorosomes emitted by predominant excitation of BChl e self-aggregates at 460 nm, indicating that they were able to accept the excitation energy from BChl e aggregates inside chlorosomes. Zn BChl a derivatives 1 and 2 emitted at 787 nm by the excitation energy transfer. In contrast, fluorescence from 3 esterified with a short hydrophilic chain, diethylene glycol, was broadened around 785 and 820 nm. The broad emission bands would consist of at least two bands, which were ascribable to the monomeric and aggregated forms. These results indicate that PEG600 and PEG400 provide appropriate amphiphilicity to Zn BChl a derivatives for dispersion in the membranous region of chlorosomes. The present study will be helpful for not only elucidation of the energy transfer mechanism in chlorosomes but also development of artificial photofunctional supramolecular complexes by hybridization of natural photosynthetic systems with synthetic molecules.

Effects of exogenous isoprenoid diphosphates on in vivo attachment to bacteriochlorophyllide c in the green sulfur photosynthetic bacterium Chlorobaculum tepidum

Metabolic substitution of the esterifying chain in bacteriochlorophyll (BChl) c in green photosynthetic bacteria grown by supplementation of exogenous alcohols has attracted attentions to study supramolecular structures and biogenesis of major antenna complexes chlorosomes in these bacteria as well as BChl pigment biosynthesis. Actual substrates in the enzymatic attachment of the esterifying moieties to the precursor of BChl c, namely bacteriochlorophyllide (BChlide) c, in these bacteria are believed to be diphosphate esters of alcoholic substrates, although only intact alcohols have so far been supplemented in the bacterial cultures. We report herein BChl c compositions in the green sulfur photosynthetic bacterium Chlorobaculum tepidum by supplementation with geranyl and geranylgeranyl diphosphates. The supplementation of these diphosphates hardly produced BChl c derivatives esterified with geraniol and geranylgeraniol in Cba. tepidum, whereas these BChl c derivatives were accumulated by supplementation of intact geraniol and geranylgeraniol. The sharp contrast of the incorporation efficiency of the supplemental isoprenoid moieties in BChl c using the isoprenoid diphosphates to that by the isoprenoid alcohols was mainly ascribable to less penetration abilities of the diphosphate substrates into Cba. tepidum cells because of their anionic and polar diphosphate moiety.

Probing the excitonic landscape of the Chlorobaculum tepidum Fenna-Matthews-Olson (FMO) complex: a mutagenesis approach

In this paper we report the steady-state optical properties of a series of site-directed mutants in the Fenna-Matthews-Olson (FMO) complex of Chlorobaculum tepidum, a photosynthetic green sulfur bacterium. The FMO antenna complex has historically been used as a model system for energy transfer due to the water-soluble nature of the protein, its stability at room temperature, as well as the availability of high-resolution structural data. Eight FMO mutants were constructed with changes in the environment of each of the bacteriochlorophyll a pigments found within each monomer of the homotrimeric FMO complex. Our results reveal multiple changes in low temperature absorption, as well as room temperature CD in each mutant compared to the wild-type FMO complex. These datasets were subsequently used to model the site energies of each pigment in the FMO complex by employing three different Hamiltonians from the literature. This enabled a basic approximation of the site energy shifts imparted on each pigment by the changed amino acid residue. These simulations suggest that, while the three Hamiltonians used in this work provide good fits to the wild-type FMO absorption spectrum, further efforts are required to obtain good fits to the mutant minus wild-type absorption difference spectra. This demonstrates that the use of FMO mutants can be a valuable tool to refine and iterate the current models of energy transfer in this system.

Identification of a chondroitin synthase from an unexpected source, the green sulfur bacterium Chlorobium phaeobacteroides.

Glycosaminoglycans (GAGs) are known to be present in all animals as well as some pathogenic microbes. Chondroitin sulfate is the most abundant GAG in mammals where it has various structural and adhesion roles. The Gram-negative bacteria Pasteurella multocida Type F and Escherichia coli K4 produce extracellular capsules composed of unsulfated chondroitin or a fructosylated chondroitin, respectively. Such polysaccharides that are structurally related to host molecules do not generally provoke a strong antibody response thus are thought to be employed as molecular camouflage during infection. We observed a sequence from the photosynthetic green sulfur bacteria, Chlorobium phaeobacteroides DSM 266, which was very similar (~62% identical) to the open reading frames of the known bifunctional chondroitin synthases (PmCS and KfoC); some segments are strikingly conserved amongst the three proteins. Recombinant E. coli-derived Chlorobium enzyme preparations were found to possess bona fide chondroitin synthase activity in vitro. This new catalyst, CpCS, however, has a more promiscuous acceptor usage than the prototypical PmCS, which may be of utility in novel chimeric GAG syntheses. The finding of such a similar chondroitin synthase enzyme in C. phaeobacteroides is unexpected for several reasons including (a) a free-living nonpathogenic organism should not "need" an animal self molecule for protection, (b) the Proteobacteria and the green sulfur bacterial lineages diverged ~2.5-3 billion years ago and (c) the ecological niches of these bacteria are not thought to overlap substantially to facilitate horizontal gene transfer. CpCS provides insight into the structure/function relationship of this class of enzymes.

Biosynthesis of unnatural glycolipids possessing diyne moiety in the acyl chain in the green sulfur photosynthetic bacterium Chlorobaculum tepidum grown by supplementation of 10,12-heptadecadiynic acid

Unnatural glycolipids possessing the diyne moiety in their acyl groups were successfully biosynthesized in the green sulfur photosynthetic bacterium Chlorobaculum (Cba.) tepidum by cultivation with supplementation of 10,12-heptadecadiynic acid. Monogalactosyldiacylglycerol (MGDG) and rhamnosylgalactosyldiacylglycerol (RGDG) esterified with one 10,12-heptadecadiynic acid were primarily formed in the cells, and small amounts of glycolipids esterified with the two unnatural fatty acids can also be detected. The relative ratio of these unnatural glycolipids occupied in the total glycolipids was estimated to be 49% based on HPLC analysis using a evaporative light scattering detector. These results indicate that the acyl groups in glycolipids, which play important roles in the formation of extramembranous antenna complexes called chlorosomes, can be modified in vivo by cultivation of green sulfur photosynthetic bacteria with exogenous synthetic fatty acids. Visible absorption and circular dichroism spectra of Cba. tepidum containing the unnatural glycolipids demonstrated the formation of chlorosomes, indicating that the unnatural glycolipids in this study did not interfere with the biogenesis of chlorosomes.

Nonlinear network model analysis of vibrational energy transfer and localisation in the Fenna-Matthews-Olson complex.

Collective protein modes are expected to be important for facilitating energy transfer in the Fenna-Matthews-Olson (FMO) complex of photosynthetic green sulphur bacteria, however to date little work has focussed on the microscopic details of these vibrations. The nonlinear network model (NNM) provides a computationally inexpensive approach to studying vibrational modes at the microscopic level in large protein structures, whilst incorporating anharmonicity in the inter-residue interactions which can influence protein dynamics. We apply the NNM to the entire trimeric FMO complex and find evidence for the existence of nonlinear discrete breather modes. These modes tend to transfer energy to the highly connected core pigments, potentially opening up alternative excitation energy transfer routes through their influence on pigment properties. Incorporating localised modes based on these discrete breathers in the optical spectra calculations for FMO using ab initio site energies and excitonic couplings can substantially improve their agreement with experimental results.

In situ high-resolution structure of the baseplate antenna complex in Chlorobaculum tepidum.

Photosynthetic antenna systems enable organisms harvesting light and transfer the energy to the photosynthetic reaction centre, where the conversion to chemical energy takes place. One of the most complex antenna systems, the chlorosome, found in the photosynthetic green sulfur bacterium Chlorobaculum (Cba.) tepidum contains a baseplate, which is a scaffolding super-structure, formed by the protein CsmA and bacteriochlorophyll a. Here we present the first high-resolution structure of the CsmA baseplate using intact fully functional, light-harvesting organelles from Cba. tepidum, following a hybrid approach combining five complementary methods: solid-state NMR spectroscopy, cryo-electron microscopy, isotropic and anisotropic circular dichroism and linear dichroism. The structure calculation was facilitated through development of new software, GASyCS for efficient geometry optimization of highly symmetric oligomeric structures. We show that the baseplate is composed of rods of repeated dimers of the strongly amphipathic CsmA with pigments sandwiched within the dimer at the hydrophobic side of the helix.

Speciation and ecological success in dimly lit waters: horizontal gene transfer in a green sulfur bacteria bloom unveiled by metagenomic assembly.

A natural planktonic bloom of a brown-pigmented photosynthetic green sulfur bacteria (GSB) from the disphotic zone of karstic Lake Banyoles (NE Spain) was studied as a natural enrichment culture from which a nearly complete genome was obtained after metagenomic assembly. We showed in situ a case where horizontal gene transfer (HGT) explained the ecological success of a natural population unveiling ecosystem-specific adaptations. The uncultured brown-pigmented GSB was 99.7% identical in the 16S rRNA gene sequence to its green-pigmented cultured counterpart Chlorobium luteolum DSM 273T. Several differences were detected for ferrous iron acquisition potential, ATP synthesis and gas vesicle formation, although the most striking trait was related to pigment biosynthesis strategy. Chl. luteolum DSM 273T synthesizes bacteriochlorophyll (BChl) c, whereas Chl. luteolum CIII incorporated by HGT a 18-kbp cluster with the genes needed for BChl e and specific carotenoids biosynthesis that provided ecophysiological advantages to successfully colonize the dimly lit waters. We also genomically characterized what we believe to be the first described GSB phage, which based on the metagenomic coverage was likely in an active state of lytic infection. Overall, we observed spread HGT and we unveiled clear evidence for virus-mediated HGT in a natural population of photosynthetic GSB.

Introduction of perfluoroalkyl chain into the esterifying moiety of bacteriochlorophyll c in the green sulfur photosynthetic bacterium Chlorobaculum tepidum by pigment biosynthesis

The green sulfur photosynthetic bacterium Chlorobaculum (Cba.) tepidum was grown in liquid cultures containing perfluoro-1-decanol, 1H,1H,2H,2H-heptadecafluoro-1-decanol [CF3(CF2)7(CH2)2OH] or 1H,1H-nonadecafluoro-1-decanol [CF3(CF2)8CH2OH], to introduce rigid and fluorophilic chains into the esterifying moiety of light-harvesting bacteriochlorophyll (BChl) c. Exogenous 1H,1H,2H,2H-heptadecafluoro-1-decanol was successfully attached to the 172-carboxy group of bacteriochlorophyllide (BChlide) c in vivo: the relative ratio of the unnatural BChl c esterified with this perfluoroalcohol over the total BChl c was 10.3%. Heat treatment of the liquid medium containing 1H,1H,2H,2H-heptadecafluoro-1-decanol with β-cyclodextrin before inoculation increased the relative ratio of the BChl c derivative esterified with this alcohol in the total BChl c in Cba. tepidum. In a while, 1H,1H-nonadecafluoro-1-decanol was not attached to BChlide c in Cba. tepidum, which was grown by its supplementation. These results suggest that the rigidity close to the hydroxy group of the esterifying alcohol is not suitable for the recognition by the BChl c synthase called BchK in Cba. tepidum. The unnatural BChl c esterified with 1H,1H,2H,2H-heptadecafluoro-1-decanol participated in BChl c self-aggregates in chlorosomes.

Chlorosome-Inspired Synthesis of Templated Metallochlorin-Lipid Nanoassemblies for Biomedical Applications.

Chlorosomes are vesicular light-harvesting organelles found in photosynthetic green sulfur bacteria. These organisms thrive in low photon flux environments due to the most efficient light-to-chemical energy conversion, promoted by a protein-less assembly of chlorin pigments. These assemblies possess collective absorption properties and can be adapted for contrast-enhanced bioimaging applications, where maximized light absorption in the near-infrared optical window is desired. Here, we report a strategy for tuning light absorption toward the near-infrared region by engineering a chlorosome-inspired assembly of synthetic metallochlorins in a biocompatible lipid scaffold. In a series of synthesized chlorin analogues, we discovered that lipid conjugation, central coordination of a zinc metal into the chlorin ring, and a 3(1)-methoxy substitution were critical for the formation of dye assemblies in lipid nanovesicles. The substitutions result in a specific optical shift, characterized by a bathochromically shifted (72 nm) Qy absorption band, along with an increase in absorbance and circular dichroism as the ratio of dye-conjugated lipid was increased. These alterations in optical spectra are indicative of the formation of delocalized excitons states across each molecular assembly. This strategy of tuning absorption by mimicking the structures found in photosynthetic organisms may spur new opportunities in the development of biophotonic contrast agents for medical applications.

In vitro stereospecific hydration activities of the 3-vinyl group of chlorophyll derivatives by BchF and BchV enzymes involved in bacteriochlorophyll c biosynthesis of green sulfur bacteria.

The photosynthetic green sulfur bacterium Chlorobaculum (Cba.) tepidum produces bacteriochlorophyll (BChl) c pigments bearing a chiral 1-hydroxyethyl group at the 3-position, which self-aggregate to construct main light-harvesting antenna complexes, chlorosomes. The secondary alcoholic hydroxy group is requisite for chlorosomal aggregation and biosynthesized by hydrating the 3-vinyl group of their precursors. Using recombinant proteins of Cba. tepidum BchF and BchV, we examined in vitro enzymatic hydration of some 3-vinyl-chlorophyll derivatives. Both the enzymes catalyzed stereoselective hydration of zinc 3-vinyl-8-ethyl-12-methyl-bacteriopheophorbide c or d to the zinc 31 R-bacteriopheophorbide c or d homolog, respectively, with a slight amount of the 31 S-epimric species. A similar R-stereoselectivity was observed in the BchF-hydration of zinc 3-vinyl-8-ethyl- and propyl-12-ethyl-bacteriopheophorbides c, while their BchV-hydration gave a relatively larger amount of the 31 S-epimers. The in vitro stereoselective hydration confirmed the in vivo production of the S-epimeric species by BchV. The enzymatic hydration for the above 8-propylated substrate proceeded more slowly than that for the 8-ethylated, and the 8-isobutylated substrate was no longer hydrated. Based on these results, biosynthetic pathways of BChl c homologs and epimers are proposed.

Chlorobaculum tepidum, a model photosynthetic green sulfur bacterium, possesses a variety of bacteriochlorophyll c pigments in light-harvesting organelles called chlorosomes (background; TEM of negatively stained chlorosomes). Bacteriochlorophyll has a 3-

Chlorobaculum tepidum, a model photosynthetic green sulfur bacterium, possesses a variety of bacteriochlorophyll c pigments in light-harvesting organelles called chlorosomes (background; TEM of negatively stained chlorosomes). Bacteriochlorophyll has a 3-