ABSTRACT
The primary photochemical reaction of photosynthesis in green sulfur bacteria occurs in the homodimer PscA core proteins by a special chlorophyll pair. The light induced excited state of the special pair producing P840+ is rapidly reduced by electron transfer from one of the two PscC subunits. Molecular dynamics (MD) simulations are combined with bioinformatic tools herein to provide structural and dynamic insight into the complex between the two PscA core proteins and the two PscC subunits. The microscopic dynamic model involves extensive sampling at atomic resolution and at a cumulative time-scale of 22µs and reveals well defined protein-protein interactions. The membrane complex is composed of the two PscA and the two PscC subunits and macroscopic connections are revealed within a putative electron transfer pathway from the PscC subunit to the special pair P840 located within the PscA subunits. Our results provide a structural basis for understanding the electron transport to the homodimer RC of the green sulfur bacteria. The MD based approach can provide the basis to further probe the PscA-PscC complex dynamics and observe electron transfer therein at the quantum level. Furthermore, the transmembrane helices of the different PscC subunits exert distinct dynamics in the complex.
Subject(s)
Bacterial Proteins , Chlorobi , Molecular Dynamics Simulation , Electron Transport , Chlorobi/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/chemistry , Protein Subunits/metabolism , Protein Subunits/chemistry , Photosynthesis , Chlorophyll/metabolism , Light-Harvesting Protein Complexes/metabolism , Light-Harvesting Protein Complexes/chemistryABSTRACT
The largest light-harvesting antenna in nature, the chlorosome, is a heterogeneous helical BChl self-assembly that has evolved in green bacteria to harvest light for performing photosynthesis in low-light environments. Guided by NMR chemical shifts and distance constraints for Chlorobaculum tepidum wild-type chlorosomes, the two contrasting packing modes for syn-anti parallel stacks of BChl c to form polar 2D arrays, with dipole moments adding up, are explored. Layered assemblies were optimized using local orbital density functional and plane wave pseudopotential methods. The packing mode with the lowest energy contains syn-anti and anti-syn H-bonding between stacks. It can accommodate R and S epimers, and side chain variability. For this packing, a match with the available EM data on the subunit axial repeat and optical data is obtained with multiple concentric cylinders for a rolling vector with the stacks running at an angle of 21° to the cylinder axis and with the BChl dipole moments running at an angle ß â¼ 55° to the tube axis, in accordance with optical data. A packing mode involving alternating syn and anti parallel stacks that is at variance with EM appears higher in energy. A weak cross-peak at -6 ppm in the MAS NMR with 50 kHz spinning, assigned to C-181, matches the shift of antiparallel dimers, which possibly reflects a minor impurity-type fraction in the self-assembled BChl c.
Subject(s)
Chlorobi , Chlorobi/metabolism , Light-Harvesting Protein Complexes/chemistry , Light-Harvesting Protein Complexes/metabolism , Magnetic Resonance Spectroscopy , Models, MolecularABSTRACT
Chlorosomes, the photosynthetic antenna complexes of green sulfur bacteria, are paradigms for light-harvesting elements in artificial designs, owing to their efficient energy transfer without protein participation. We combined magic angle spinning (MAS) NMR, optical spectroscopy and cryogenic electron microscopy (cryo-EM) to characterize the structure of chlorosomes from a bchQ mutant of Chlorobaculum tepidum. The chlorosomes of this mutant have a more uniform composition of bacteriochlorophyll (BChl) with a predominant homolog, [8Ethyl, 12Ethyl] BChl c, compared to the wild type (WT). Nearly complete 13C chemical shift assignments were obtained from well-resolved homonuclear 13C-13C RFDR data. For proton assignments heteronuclear 13C-1H (hCH) data sets were collected at 1.2 GHz spinning at 60 kHz. The CHHC experiments revealed intermolecular correlations between 132/31, 132/32, and 121/31, with distance constraints of less than 5 Å. These constraints indicate the syn-anti parallel stacking motif for the aggregates. Fourier transform cryo-EM data reveal an axial repeat of 1.49 nm for the helical tubular aggregates, perpendicular to the inter-tube separation of 2.1 nm. This axial repeat is different from WT and is in line with BChl syn-anti stacks running essentially parallel to the tube axis. Such a packing mode is in agreement with the signature of the Qy band in circular dichroism (CD). Combining the experimental data with computational insight suggests that the packing for the light-harvesting function is similar between WT and bchQ, while the chirality within the chlorosomes is modestly but detectably affected by the reduced compositional heterogeneity in bchQ.
Subject(s)
Bacteriochlorophylls , Chlorobi , Chlorobi/genetics , Chlorobi/metabolism , Bacteriochlorophylls/chemistry , Mutation , Light-Harvesting Protein Complexes/chemistry , Light-Harvesting Protein Complexes/metabolism , Light-Harvesting Protein Complexes/genetics , Cryoelectron Microscopy , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/metabolismABSTRACT
A Type I reaction center (RC) (Fe-S type, ferredoxin reducing) is found in several phyla containing anoxygenic phototrophic bacteria. These include the heliobacteria (HB), the green sulfur bacteria (GSB), and the chloracidobacteria (CB), for which high-resolution homodimeric RC-photosystem (PS) structures have recently appeared. The 2.2-Å X-ray structure of the RC-PS of Heliomicrobium modesticaldum revealed that the core PshA apoprotein (PshA-1 and PshA-2 homodimeric pair) exhibits a structurally conserved PSI arrangement comprising five C-terminal transmembrane α-helices (TMHs) forming the RC domain and six N-terminal TMHs coordinating the light-harvesting (LH) pigments. The Hmi. modesticaldum structure lacked quinone molecules, indicating that electrons were transferred directly from the A0 (81-OH-chlorophyll (Chl) a) acceptor to the FX [4Fe-4S] component, serving as the terminal RC acceptor. A pair of additional TMHs designated as Psh X were also found that function as a low-energy antenna. The 2.5-Å resolution cryo-electron microscopy (cryo-EM) structure for the RC-PS of the green sulfur bacterium Chlorobaculum tepidum included a pair of Fenna-Matthews-Olson protein (FMO) antennae, which transfer excitations from the chlorosomes to the RC-PS (PscA-1 and PscA-2) core. A pair of cytochromes cZ (PscC) molecules was also revealed, acting as electron donors to the RC bacteriochlorophyll (BChl) a' special pair, as well as PscB, housing the [4Fe-4S] cluster FA and FB, and the associated PscD protein. While the FMO components were missing from the 2.6-Å cryo-EM structure of the Zn- (BChl) a' special pair containing RC-PS of Chloracidobacterium thermophilum, a unique architecture was revealed that besides the (PscA)2 core, consisted of seven additional subunits including PscZ in place of PscD, the PscX and PscY cytochrome c serial electron donors and four low mol. wt. subunits of unknown function. Overall, these diverse structures have revealed that (i) the HB RC-PS is the simplest light-energy transducing complex yet isolated and represents the closest known homolog to a common homodimeric RC-PS ancestor; (ii) the symmetrically localized Ca2+-binding sites found in each of the Type I homodimeric RC-PS structures likely gave rise to the analogously positioned Mn4CaO5 cluster of the PSII RC and the TyrZ RC donor site; (iii) a close relationship between the GSB RC-PS and the PSII Chl proteins (CP)43 and CP47 was demonstrated by their strongly conserved LH-(B)Chl localizations; (iv) LH-BChls of the GSB-RC-PS are also localized in the conserved RC-associated positions of the PSII ChlZ-D1 and ChlZ-D2 sites; (v) glycosylated carotenoids of the GSB RC-PS are located in the homologous carotenoid-containing positions of PSII, reflecting an O2-tolerance mechanism capable of sustaining early stages in the evolution of oxygenic photosynthesis. In addition to the close relationships found between the homodimeric RC-PS and PSII, duplication of the gene encoding the ancestral Type I RC apoprotein, followed by genetic divergence, may well account for the appearance of the heterodimeric Type I and Type II RCs of the extant oxygenic phototrophs. Accordingly, the long-held view that PSII arose from the anoxygenic Type II RC is now found to be contrary to the new evidence provided by Type I RC-PS homodimer structures, indicating that the evolutionary origins of anoxygenic Type II RCs, along with their distinct antenna rings are likely to have been preceded by the events that gave rise to their oxygenic counterparts.
Subject(s)
Chlorobi , Photosynthetic Reaction Center Complex Proteins , Chlorobi/chemistry , Chlorobi/metabolism , Photosynthetic Reaction Center Complex Proteins/metabolism , Cryoelectron Microscopy , Bacteria/metabolism , Apoproteins/metabolism , Bacterial Proteins/metabolismABSTRACT
Anaerobic pig wastewater treatment commonly generates high sulfide concentrations in the treated wastewater. This study aims to apply phototrophic green sulfur bacteria (PGB) to promote sulfide removal in lighting-anaerobic digestion (lighting-AD) treating pig wastewater. Initially, batch AD tests of pig wastewater with/without PGB addition were carried out under dark (D) and light (L) conditions. The results showed that the lighting-AD with PGB gave a higher growth rate of PGB (0.056 h-1) and the highest COD/sulfide removals as compared to the dark-AD with PGB and lighting-AD solely. More experiments under various light intensities were performed in order to find an optimal intensity for PGB growth concurrently with metagenomic changes concerning treatment performance. It appeared that sulfide removal rates had increased as increasing light intensity up to 473 lx by giving the highest rate of 12.5 mg L-1 d-1 with the highest sulfur element content in the biomass. Contrastingly, many PGB species disappeared at 1350 lx exposure subsequently sharply decreasing the rate of sulfide removal. In sum, the application of low light intensities of 400-500 lx with bioaugmented PGB could promote PGB growth and activity in sulfide removal in pig wastewater in the lighting of the AD process.
Subject(s)
Chlorobi , Wastewater , Animals , Swine , Anaerobiosis , Lighting , Sulfides , Sulfur , Bioreactors/microbiologyABSTRACT
The exciton states on the smallest type-I photosynthetic reaction center complex of a green sulfur bacterium Chlorobaculum tepidum (GsbRC) consisting of 26 bacteriochlorophylls a (BChl a) and four chlorophylls a (Chl a) located on the homodimer of two PscA reaction center polypeptides were investigated. This analysis involved the study of exciton states through a combination of theoretical modeling and the genetic removal of BChl a pigments at eight sites. (1) A theoretical model of the pigment assembly exciton state on GsbRC was constructed using Poisson TrESP (P-TrESP) and charge density coupling (CDC) methods based on structural information. The model reproduced the experimentally obtained absorption spectrum, circular dichroism spectrum, and excitation transfer dynamics, as well as explained the effects of mutation. (2) Eight BChl a molecules at different locations on the GsbRC were selectively removed by genetic exchange of the His residue, which ligates the central Mg atom of BChl a, with the Leu residue on either one or two PscAs in the RC. His locations are conserved among all type-I RC plant polypeptide, cyanobacteria, and bacteria amino acid sequences. (3) Purified mutant-GsbRCs demonstrated distinct absorption and fluorescence spectra at 77 K, which were different from each other, suggesting successful pigment removal. (4) The same mutations were applied to the constructed theoretical model to analyze the outcomes of these mutations. (5) The combination of theoretical predictions and experimental mutations based on structural information is a new tool for studying the function and evolution of photosynthetic reaction centers.
Subject(s)
Chlorobi , Cyanobacteria , Photosynthetic Reaction Center Complex Proteins , Photosynthetic Reaction Center Complex Proteins/chemistry , Chlorobi/chemistry , Mutation , Cyanobacteria/metabolism , Sulfur/metabolism , Bacteriochlorophylls/chemistry , Bacterial Proteins/chemistryABSTRACT
Two sulfate-reducing wetland bioreactors (SRB-1 filled with lignocellulosic wastes and SRB-2 with river sand) were applied for synthetic acid mine drainage treatment with bio-waste fermentation liquid as electron donor, and the influence of filling substrates on sulfate reduction, sulfur transformation and microbial community was studied. The presence of lignocellulosic wastes (mixture of cow manure, bark, sawdust, peanut shell and straw) in SRB-1 promoted sulfate reduction efficiency (68.9%), sulfate reduction rate (42.1 ± 11 mg S/(L·d)), dissolved sulfide production rate (27.4 ± 7 mg S/(L·d)), and particularly caused high conversion ratio of sulfate reduction into dissolved sulfide (66.4%). In comparison, the relatively low sulfate reduction efficiency (42.9%), sulfate reduction rate (27.0 ± 10 mg S/(L·d)), dissolved sulfide production rate (5.6 ± 3 mg S/(L·d)) and low dissolved sulfide conversion efficiency (21.2%) occurred in SRB-2. Mixed organic substrates including easily assimilated electron donors (in manure) and lignocellulosic matter were effective to promote quick start and long-term microbial sulfate reduction. More than 98% of produced dissolved sulfide was oxidized dominantly by photoautotrophic green sulfur bacteria (genera Chlorobium and Chlorobaculum), of which 64.6% and 54.5% was converted into elemental sulfur for SRB-1 and SRB-2. The oxidation of sulfide into elemental sulfur for potential recovery rather than sulfate is preferred. Diverse sulfate reducing bacteria and sulfide oxidizing bacteria co-existed in the treatment system, which led to a sustainable sulfur transformation. High metal removal efficiency for Fe (99.6%, 92.5%), Cd (99.9%, 99.9%), Zn (99.4%, 98.5%), Cu (94.5%, 94.6%) except for Mn (9.3%, 3.6%) was achieved, and effluent pH increased to 6.5-7.7 and 6.7-7.7 for SRB-1 and SRB-2, respectively. Microbial community was regulated by filling substrates. Synergism between lignocellulosic decomposing bacteria and sulfate reducing bacteria played a vital role in lignocellulosic bioreactor treating AMD, in addition to fermentation liquid serving as effective electron donor.
Subject(s)
Chlorobi , Microbiota , Biodegradation, Environmental , Sulfates , Manure , Wetlands , Bioreactors/microbiology , Bacteria , Acids , Sulfides , Sulfur , Oxidation-ReductionABSTRACT
Bacteria and archaea play a fundamental role in the biogeochemical cycles of organic matter, pollutants, and nutrients to maintain the trophic state of aquatic ecosystems. However, very little is known about the composition patterns of microbial communities in vertical distribution (water column) in freshwater lakes and their relationship with the physicochemical properties of water. "La Encantada" lake in the Lagunas de Montebello National Park (LMNP) is a site of interest due to the anthropogenic impact received and the little information about it. In this study, 3 sites were evaluated; samples were collected using 0-15 m deep water columns and analyzed using Illumina MiSeq sequencing technology based on the 16S rRNA gene. The physical parameters of pH, temperature, dissolved oxygen, electrolytic conductivity, and PO-4 were determined. The results revealed clear differences in the microbial composition of the water throughout the column; the most abundant phyla in bacterial communities were Proteobacteria (23.2%), Cyanobacteria (17.3%), and Bacteroidetes (17.2%), and for archaea were Crenarchaeota (35.9%) and Euryarchaeota (33.2%). PICRUSt metabolic inference analysis revealed that the main functional genes were related to cellular processes and biodegradation of xenobiotics, indicating an increasing trend of contaminants and residual discharges that may act as a precursor to alter microbial communities and stability of the lakes. At depths of 10 and 15 m, the microbial diversity was greater; likewise, the correlation between the physicochemical parameters and the microbial communities at the genus level showed that Chlorobaculum, Desulfomonile, and Candidatus Xiphinematobacter were favored by an increase in dissolved phosphates and by the decrease in pH and temperature. These results highlight that the microbial communities exhibit variation in their composition due to the effect of depth and physicochemical parameters, which could play a role as biological factors in the trophic states of a lake.
Subject(s)
Archaea , Chlorobi , Archaea/genetics , Lakes , Ecosystem , Mexico , RNA, Ribosomal, 16S , Bacteria/genetics , WaterABSTRACT
The reverse tricarboxylic acid (rTCA) cycle is touted as a primordial mode of carbon fixation due to its autocatalytic propensity and oxygen intolerance. Despite this inferred antiquity, however, the earliest rock record affords scant supporting evidence. In fact, based on the chimeric inheritance of rTCA cycle steps within the Chlorobiaceae, even the use of the chemical fossil record of this group is now subject to question. While the 1.64-billion-year-old Barney Creek Formation contains chemical fossils of the earliest known putative Chlorobiaceae-derived carotenoids, interferences from the accompanying hydrocarbon matrix have hitherto precluded the carbon isotope measurements necessary to establish the physiology of the organisms that produced them. Overcoming this obstacle, here we report a suite of compound-specific carbon isotope measurements identifying a cyanobacterially dominated ecosystem featuring heterotrophic bacteria. We demonstrate chlorobactane is 13C-depleted when compared to contemporary equivalents, showing only slight 13C-enrichment over co-existing cyanobacterial carotenoids. The absence of this diagnostic isotopic fingerprint, in turn, confirms phylogenomic hypotheses that call for the late assembly of the rTCA cycle and, thus, the delayed acquisition of autotrophy within the Chlorobiaceae. We suggest that progressive oxygenation of the Earth System caused an increase in the marine sulfate inventory thereby providing the selective pressure to fuel the Neoproterozoic shift towards energy-efficient photoautotrophy within the Chlorobiaceae.
Subject(s)
Chlorobi , Cyanobacteria , Chlorobi/chemistry , Chlorobi/metabolism , Tricarboxylic Acids/metabolism , Ecosystem , Carbon Isotopes , Carbon Cycle , Carotenoids/metabolismABSTRACT
Microbes are an important part of the vineyard ecosystem, which significantly influence the quality of grapes. Previously, we identified a bud mutant variety (named 'Fengzao') from 'Kyoho' grapes. The variation of microbial communities in grape and its bud mutant variety has not been studied yet. So, in this study, with the samples of both 'Fengzao' and 'Kyoho', we conducted high-throughput microbiome sequencing and investigated their microbial communities in different tissues. Obvious differences were observed in the microbial communities between 'Fengzao' and 'Kyoho'. The fruit and the stem are the tissues with relatively higher abundance of microbes, while the leaves contained less microbes. The fruit and the stem of 'Kyoho' and the stem of 'Fengzao' had relatively higher species diversity based on the alpha diversity analysis. Proteobacteria, Enterobacteriaceae and Rhodobacteraceae had significantly high abundance in 'Fengzao'. Firmicutes and Pseudomonas were highly abundant in the stems of 'Kyoho', and family of Spirochaetaceae, Anaplasmataceae, Chlorobiaceae, and Sphingomonadaceae, and genera of Spirochaeta, Sphingomonas, Chlorobaculum and Wolbachia were abundant in the fruits of 'Kyoho'. These identified microbes are main components of the microbial communities, and could be important regulators of grapevine growth and development. This study revealed the differences in the microbial compositions between 'Kyoho' and its bud mutant, and these identified microbes will be significant resources for the future researches on the quality regulation and disease control of grapevines.
Subject(s)
Anaplasmataceae , Chlorobi , Microbiota , Vitis , Microbiota/genetics , EnterobacteriaceaeABSTRACT
Fenna-Mathews-Olson complexes participate in the photosynthetic process of Sulfur Green Bacteria. These biological subsystems exhibit quantum features which possibly are responsible for their high efficiency; the latter may comprise multipartite entanglement and the apparent tunnelling of the initial quantum state. At first, to study these aspects, a multidisciplinary approach including experimental biology, spectroscopy, physics, and math modelling is required. Then, a global computer modelling analysis is achieved in the computational biology domain. The current work implements the Hierarchical Equations of Motion to numerically solve the open quantum system problem regarding this complex. The time-evolved states obtained with this method are then analysed under several measures of entanglement, some of them already proposed in the literature. However, for the first time, the maximum overlap with respect to the closest separable state is employed. This authentic multipartite entanglement measure provides information on the correlations, not only based on the system bipartitions as in the usual analysis. Our study has led us to note a different view of FMO multipartite entanglement as tiny contributions to the global entanglement suggested by other more basic measurements. Additionally, in another related trend, the initial state, considered as a Förster Resonance Energy Transfer, is tracked using a novel approach, considering how it could be followed under the fidelity measure on all possible permutations of the FMO subsystems through its dynamical evolution by observing the tunnelling in the most probable locations. Both analyses demanded significant computational work, making for a clear example of the complexity required in computational biology.
Subject(s)
Bacterial Proteins , Chlorobi , Bacterial Proteins/chemistry , Light-Harvesting Protein Complexes/metabolism , Fluorescence Resonance Energy Transfer , Computer Simulation , Quantum TheoryABSTRACT
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.
Subject(s)
Carcinoma, Renal Cell , Chlorobi , Kidney Neoplasms , Humans , Light-Harvesting Protein Complexes/metabolism , Bacterial Proteins/metabolismABSTRACT
Due to the high concentration of pollutants, swine wastewater needs to be treated prior to disposal. The combination of anaerobic and aerobic technologies in one hybrid system allows to obtain higher removal efficiencies compared to those achieved via conventional biological treatment, and the performance of a hybrid system depends on the microbial community in the bioreactor. Here, we evaluated the community assembly of an anaerobic-aerobic hybrid reactor for swine wastewater treatment. Sequencing of partial 16S rRNA coding genes was performed using Illumina from DNA and retrotranscribed RNA templates (cDNA) extracted from samples from both sections of the hybrid system and from a UASB bioreactor fed with the same swine wastewater influent. Proteobacteria and Firmicutes were the dominant phyla and play a key role in anaerobic fermentation, followed by Methanosaeta and Methanobacterium. Several differences were found in the relative abundances of some genera between the DNA and cDNA samples, indicating an increase in the diversity of the metabolically active community, highlighting Chlorobaculum, Cladimonas, Turicibacter and Clostridium senso stricto. Nitrifying bacteria were more abundant in the hybrid bioreactor. Beta diversity analysis revealed that the microbial community structure significantly differed among the samples (p < 0.05) and between both anaerobic treatments. The main predicted metabolic pathways were the biosynthesis of amino acids and the formation of antibiotics. Also, the metabolism of C5-branched dibasic acid, Vit B5 and CoA, exhibited an important relationship with the main nitrogen-removing microorganisms. The anaerobic-aerobic hybrid bioreactor showed a higher ammonia removal rate compared to the conventional UASB system. However, further research and adjustments are needed to completely remove nitrogen from wastewater.
Subject(s)
Chlorobi , Microbiota , Water Purification , Animals , Swine , Wastewater , Sewage/chemistry , Waste Disposal, Fluid , Anaerobiosis , Chlorobi/genetics , RNA, Ribosomal, 16S/genetics , DNA, Complementary , Bioreactors/microbiologyABSTRACT
In the biosynthetic pathway of bacteriochlorophyll(BChl)-a/b/c/d/e molecules, BchF and BchV enzymes catalyze the hydration of a C3-vinyl to C3-1-hydroxyethyl group. In this study, the in vitro reactions catalyzed by BchF and BchV partially afforded a C31-epimeric mixture of the hydrated products (secondary alcohols), with the primary recovery of the C3-vinylated substrate. The stereoselectivity and substrate specificity for the in vitro reverse enzymatic dehydration were examined using zinc chlorophyll analogs as model substrates by BchF and BchV, which were obtained from extracts of Escherichia coli overexpressing the respective genes from Chlorobaculum tepidum and used without further purification. Both BchF and BchV preferred dehydration of the (31R)-epimers over the (31S)-epimers. The (31R)-epimer was directly dehydrated by BchF and BchV to give the C3-vinylated product. By contrast, two reaction pathways for BchF and BchV dehydrations of the (31S)-epimer were proposed: (1) the (31S)-epimer would be directly dehydrated to C3-vinyl group. (2) the (31S)-epimer would be epimerized to the (31R)-epimer, and the resulting epimer was dehydrated. The results indicated that both BchF and BchV did function as a hydratase/dehydratase and could play a role in the C31-epimerization. An increase in the alkyl size at the C8-position gradually suppressed the BchF and BchV-catalyzed dehydration in vitro, while the C121- and C20-methylation only slightly affected the reaction. Using the BchF dehydration, a large amount of 3-vinyl-bacteriochlorophyllide-a was successfully prepared, with the retention of the chemically labile, central magnesium atom.
Subject(s)
Chlorobi , Chlorophyll , Humans , Chlorophyll/metabolism , Chlorobi/genetics , Substrate Specificity , Zinc , DehydrationABSTRACT
Chlorobaculum tepidum is an anaerobic green sulfur bacterium which oxidizes sulfide, elemental sulfur, and thiosulfate for photosynthetic growth. It can also oxidize sulfide to produce extracellular S0 globules, which can be further oxidized to sulfate and used as an electron donor. Here, we performed label-free quantitative proteomics on total cell lysates prepared from different metabolic states, including a sulfur production state (10 h post-incubation [PI]), the beginning of sulfur consumption (20 h PI), and the end of sulfur consumption (40 h PI), respectively. We observed an increased abundance of the sulfide:quinone oxidoreductase (Sqr) proteins in 10 h PI indicating a sulfur production state. The periplasmic thiosulfate-oxidizing Sox enzymes and the dissimilatory sulfite reductase (Dsr) subunits showed an increased abundance in 20 h PI, corresponding to the sulfur-consuming state. In addition, we found that the abundance of the heterodisulfide-reductase and the sulfhydrogenase operons was influenced by electron donor availability and may be associated with sulfur metabolism. Further, we isolated and analyzed the extracellular sulfur globules in the different metabolic states to study their morphology and the sulfur cluster composition, yielding 58 previously uncharacterized proteins in purified globules. Our results show that C. tepidum regulates the cellular levels of enzymes involved in sulfur metabolism in response to the availability of reduced sulfur compounds.
Subject(s)
Chlorobi , Proteomics , Sulfur , Chlorobi/metabolism , Oxidation-Reduction , Proteomics/methods , Sulfides/metabolism , Sulfur/metabolism , Thiosulfates/metabolism , PhotosynthesisABSTRACT
The fully reduced flavin cofactor (FADred) in ferredoxin-NADP+ oxidoreductase (FNR) is a functional intermediate that displays different catalytic and steady-state spectral properties for enzymes from Bacillus subtilis (BsFNR), Chlorobaculum tepidum (CtFNR), and Rhodopseudomonas palustris (RpFNR). Using ultrafast spectroscopy, we reveal that at physiological pH, photoexcited FADred in BsFNR and RpFNR exhibits unprecedentedly fast decays (dominantly in 6 and 8 ps, respectively), whereas in CtFNR the decay is much slower (â¼400 ps), as in other flavoproteins. Correlating these observations with the protonation states of FADred and the dynamic properties of the protein environment, we conclude that the excited state of neutral FADred can be intrinsically short-lived even in proteins, contrasting with the well-documented behavior of the anionic form that systematically displays markedly increased excited-state lifetime upon binding to proteins. This work provides new insight into the photochemistry of fully reduced flavins, which are emerging as functional initial states in bioengineered photocatalysts.
Subject(s)
Chlorobi , Ferredoxins , Ferredoxins/metabolism , Chlorobi/metabolism , NADP/metabolism , Flavins/metabolism , Ferredoxin-NADP Reductase/chemistry , Ferredoxin-NADP Reductase/metabolism , Oxidation-Reduction , KineticsABSTRACT
Light energy absorption and transfer are very important processes in photosynthesis. In green sulfur bacteria light is absorbed primarily by the chlorosomes and its energy is transferred via the Fenna-Matthews-Olson (FMO) proteins to a homodimeric reaction center (RC). Here, we report the cryogenic electron microscopic structure of the intact FMO-RC apparatus from Chlorobaculum tepidum at 2.5 Å resolution. The FMO-RC apparatus presents an asymmetric architecture and contains two FMO trimers that show different interaction patterns with the RC core. Furthermore, the two permanently bound transmembrane subunits PscC, which donate electrons to the special pair, interact only with the two large PscA subunits. This structure fills an important gap in our understanding of the transfer of energy from antenna to the electron transport chain of this RC and the transfer of electrons from reduced sulfur compounds to the special pair.
Subject(s)
Chlorobi , Photosynthetic Reaction Center Complex Proteins , Photosynthetic Reaction Center Complex Proteins/chemistry , Chlorobi/metabolism , Cryoelectron Microscopy , Bacterial Proteins/metabolism , Sulfur/metabolism , Light-Harvesting Protein Complexes/metabolismABSTRACT
The photosynthetic reaction center complex (RCC) of green sulfur bacteria (GSB) consists of the membrane-imbedded RC core and the peripheric energy transmitting proteins called Fenna-Matthews-Olson (FMO). Functionally, FMO transfers the absorbed energy from a huge peripheral light-harvesting antenna named chlorosome to the RC core where charge separation occurs. In vivo, one RC was found to bind two FMOs, however, the intact structure of RCC as well as the energy transfer mechanism within RCC remain to be clarified. Here we report a structure of intact RCC which contains a RC core and two FMO trimers from a thermophilic green sulfur bacterium Chlorobaculum tepidum at 2.9 Å resolution by cryo-electron microscopy. The second FMO trimer is attached at the cytoplasmic side asymmetrically relative to the first FMO trimer reported previously. We also observed two new subunits (PscE and PscF) and the N-terminal transmembrane domain of a cytochrome-containing subunit (PscC) in the structure. These two novel subunits possibly function to facilitate the binding of FMOs to RC core and to stabilize the whole complex. A new bacteriochlorophyll (numbered as 816) was identified at the interspace between PscF and PscA-1, causing an asymmetrical energy transfer from the two FMO trimers to RC core. Based on the structure, we propose an energy transfer network within this photosynthetic apparatus.
Subject(s)
Carcinoma, Renal Cell , Chlorobi , Kidney Neoplasms , Photosynthetic Reaction Center Complex Proteins , Photosynthetic Reaction Center Complex Proteins/chemistry , Photosynthetic Reaction Center Complex Proteins/metabolism , Chlorobi/chemistry , Chlorobi/metabolism , Cryoelectron Microscopy , Bacterial Proteins/metabolismABSTRACT
Biogas desulfurization based on anoxygenic photosynthetic processes represents an alternative to physicochemical technologies, decreasing the risk of O2 and N2 contamination. This work aimed at assessing the potential of Allochromatium vinosum and Chlorobium limicola for biogas desulfurization under different light intensities (10 and 25 klx) and H2S concentrations (1 %, 1.5 % and 2 %) in batch photobioreactors. In addition, the influence of rising biogas flow rates (2.9, 5.8 and 11.5 L d-1 in stage I, II and III, respectively) on the desulfurization performance in a 2.3 L photobioreactor utilizing C. limicola under continuous mode was assessed. The light intensity of 25 klx negatively influenced the growth of A. vinosum and C. limicola, resulting in decreased H2S removal capacity. An increase in H2S concentrations resulted in higher volumetric H2S removal rates in C. limicola (2.9-5.3 mg L-1 d-1) tests compared to A. vinosum (2.4-4.6 mg L-1 d-1) tests. The continuous photobioreactor completely removed H2S from biogas in stage I and II. The highest flow rate in stage III induced a deterioration in the desulfurization activity of C. limicola. Overall, the high H2S tolerance of A. vinosum and C. limicola supports their use in H2S desulfurization from biogas.
Subject(s)
Chlorobi , Hydrogen Sulfide , Biofuels , PhotobioreactorsABSTRACT
The Fenna-Matthews-Olson (FMO) complex of green sulfur bacteria has been serving as a prototypical light-harvesting protein for studying excitation energy transfer (EET) dynamics in photosynthesis. The most widely used Frenkel exciton model for FMO complex assumes that each excited bacteriochlorophyll site couples to an identical and isolated harmonic bath, which does not account for the heterogeneous local protein environment. To better describe the realistic environment, we propose to use the recently developed multistate harmonic (MSH) model, which contains a globally shared bath that couples to the different pigment sites according to the atomistic quantum mechanics/molecular mechanics simulations with explicit protein scaffold and solvent. In this work, the effects of heterogeneous protein environment on EET in FMO complexes from Prosthecochloris aestuarii and Chlorobium tepidum, specifically including realistic spectral density, site-dependent reorganization energies, and system-bath couplings are investigated. Semiclassical and mixed quantum-classical mapping dynamics were applied to obtain the nonadiabatic EET dynamics in several models ranging from the Frenkel exciton model to the MSH model and their variants. The MSH model with realistic spectral density and site-dependent system-bath couplings displays slower EET dynamics than the Frenkel exciton model. Our comparative study shows that larger average reorganization energy, heterogeneity in spectral densities, and low-frequency modes could facilitate energy dissipation, which is insensitive to the static disorder in reorganization energies. The effects of the spectral densities and system-bath couplings along with the MSH model can be used to optimize EET dynamics for artificial light-harvesting systems.