Purple Photosynthetic Bacteria Research Articles

Multidimensional engineering of Rhodobacter sphaeroides for enhanced photo-fermentative hydrogen production

The photo-fermentative hydrogen production by purple non-sulfur photosynthetic bacteria (PNSB) is regarded as a promising approach for sustainable hydrogen production. In PNSB, biohydrogen synthesis is catalyzed by nitrogenase which requires two essential cofactors of adenosine triphosphate (ATP) and electron. To avoid the insufficient ATP supply that limits the nitrogenase activity for hydrogen production, we systematically engineered Rhodobacter sphaeroides to improve the hydrogen production by manipulating the light harvesting (LH) complexes, the electron transfer chain (ETC), the adenosine diphosphate (ADP) synthetic pathway, the F0F1-ATPase expression, and the nitrogenase expression. To improve the photosynthetic efficiency, we engineered the LH complexes by eliminating pucBA and overexpressing pufQ and spbA. Then, the ETC was boosted by deleting qxt-type oxidase and integrating the electron carrier cytochrome cy at the site of cyt cbb3 oxidase. In addition, we increased the supply of precursor ADP by overexpressing adk, and the abundance of F0F1-ATPase was efficiently enhanced through overexpressing atpXF. Further transposon engineering with the integration of nifHDK and atpXF resulted in a mutant YH27 with a substantial improvement of performance over the base strain, and its nitrogenase activity and hydrogen production were enhanced by 404 % and 93 %, respectively. Taken together, the multidimensional strategies enabled the engineered R. sphaeroides with a hydrogen productivity of 61.2 mL/(Lh) and a yield of 3.0 mol/mol acetate under the fed-batch condition, which represents one of the most efficient biohydrogen-producing strains.

Genome of Lamprobacter modestohalophilus ShNLb02, a moderate halophilic photosynthetic purple bacterium of the Chromatiaceae family.

The genome sequence of the moderately halophilic Lamprobacter modestohalophilus ShNLb02 was compared to those of other Lamprobacter and Halochromatium species. It revealed an average nucleotide identity of 94% to Lpb. modestohalophilus DSM 25653 and of 89.7% to Halochromatium roseum DSM 18859, underscoring their close phylogenetic relationship.

Bioinspired polymeric supramolecular columns as efficient yet controllable artificial light-harvesting platform

Light-harvesting is an indispensable process in photosynthesis, and researchers have been exploring various structural scaffolds to create artificial light-harvesting systems. However, achieving high donor/acceptor ratios for efficient energy transfer remains a challenge as excitons need to travel longer diffusion lengths within the donor matrix to reach the acceptor. Here, we report a polymeric supramolecular column-based light-harvesting platform inspired by the natural light-harvesting of purple photosynthetic bacteria to address this issue. The supramolecular column is designed as a discotic columnar liquid crystalline polymer and acts as the donor, with the acceptor intercalated within it. The modular columnar design enables an ultrahigh donor/acceptor ratio of 20000:1 and an antenna effect exceeding 100. Moreover, the spatial confinement within the supramolecular columns facilitates control over the energy transfer process, enabling dynamic full-color tunable emission for information encryption applications with spatiotemporal regulation security.

Enhancing solar spectrum utilization in photosynthesis: exploring exciton and site energy shifts as key mechanisms

Photosynthesis is a critical process that harnesses solar energy to sustain life across Earth's intricate ecosystems. Central to this phenomenon is nuanced adaptation to a spectrum spanning approximately from 300 nm to nearly 1100 nm of solar irradiation, a trait enabling plants, algae, and phototrophic bacteria to flourish in their respective ecological niches. While the Sun’s thermal radiance and the Earth’s atmospheric translucence naturally constrain the ultraviolet extent of this range, a comprehension of how to optimize the utilization of near-infrared light has remained an enduring pursuit. This study unveils the remarkable capacity of the bacteriochlorophyll b-containing purple photosynthetic bacterium Blastochloris viridis to harness solar energy at extreme long wavelengths, a property attributed to a synergistic interplay of exciton and site energy shift mechanisms. Understanding the unique native adaptation mechanisms offers promising prospects for advancing sustainable energy technologies of solar energy conversion.

Roadmap of electrons from donor side to the reaction center of photosynthetic purple bacteria with mutated cytochromes

In photosynthetic bacteria, the absorbed light drives the canonical cyclic electron transfer between the reaction center and the cytochrome bc1 complexes via the pools of mobile electron carriers. If kinetic or structural barriers hinder the participation of the bc1 complex in the cyclic flow of electrons, then the pools of mobile redox agents must supply the electrons for the multiple turnovers of the reaction center. These conditions were achieved by continuous high light excitation of intact cells of bacterial strains Rba. sphaeroides and Rvx. gelatinosus with depleted donor side cytochromes c2 (cycA) and tetraheme cytochrome subunit (pufC), respectively. The gradual oxidation by ferricyanide further reduced the availability of electron donors to pufC. Electron transfer through the reaction center was tracked by absorption change and by induction and relaxation of the fluorescence of the bacteriochlorophyll dimer. The rate constants of the electron transfer (~ 3 × 103 s‒1) from the mobile donors of Rvx. gelatinosus bound either to the RC (pufC) or to the tetraheme subunit (wild type) were similar. The electrons transferred through the reaction center dimer were supplied entirely by the donor pool; their number amounted to about 5 in wild type Rvx. gelatinosus and decreased to 1 in pufC oxidized by ferricyanide. Fluorescence yield was measured as a function of the oxidized fraction of the dimer and its complex shape reveals the contribution of two competing processes: the migration of the excitation energy among the photosynthetic units and the availability of electron donors to the oxidized dimer. The experimental results were simulated and rationalized by a simple kinetic model of the two-electron cycling of the acceptor side combined with aperiodic one-electron redox function of the donor side.

Multidimensional optimization for accelerating light-powered biocatalysis in Rhodopseudomonas palustris

BackgroundWhole-cell biocatalysis has been exploited to convert a variety of substrates into high-value bulk or chiral fine chemicals. However, the traditional whole-cell biocatalysis typically utilizes the heterotrophic microbes as the biocatalyst, which requires carbohydrates to power the cofactor (ATP, NAD (P)H) regeneration.ResultsIn this study, we sought to harness purple non-sulfur photosynthetic bacterium (PNSB) as the biocatalyst to achieve light-driven cofactor regeneration for cascade biocatalysis. We substantially improved the performance of Rhodopseudomonas palustris-based biocatalysis using a highly active and conditional expression system, blocking the side-reactions, controlling the feeding strategy, and attenuating the light shading effect. Under light-anaerobic conditions, we found that 50 mM ferulic acid could be completely converted to vanillyl alcohol using the recombinant strain with 100% efficiency, and > 99.9% conversion of 50 mM p-coumaric acid to p-hydroxybenzyl alcohol was similarly achieved. Moreover, we examined the isoprenol utilization pathway for pinene synthesis and 92% conversion of 30 mM isoprenol to pinene was obtained.ConclusionsTaken together, these results suggested that R. palustris could be a promising host for light-powered biotransformation, which offers an efficient approach for synthesizing value-added chemicals in a green and sustainable manner.Graphical

Cancer immunotheranostics using bioactive nanocoated photosynthetic bacterial complexes

Developing new modalities that allow multidimensional therapy with optical and immunological regulations is a challenge ahead of pharmacology and drug discovery for effective cancer treatment. To this end, superior purple photosynthetic bacteria, which show unique near-infrared (NIR) fluorescence, strong photothermal conversion ability, and high biocompatibility, were screened from various bacterial strains. To improve the therapeutic performances of screened bacteria, we further designed and developed simple chemical bacterial membrane functionalization with bioactive molecules and polyethylene glycol derivatives. Exploring the physicochemical and physiological properties of the chemically functionalized bacteria complexes achieved excellent tumor targeting and strong anticancer efficacies. The highly biocompatible functional bacteria showed cytotoxic immunological effects, strong NIR fluorescence, and powerful photothermal conversion in the targeted tumor milieu under biologically penetrating NIR light irradiation.

Mapping charge-transfer excitations in Bacteriochlorophyll dimers from first principles

Photoinduced charge-transfer excitations are key to understand the primary processes of natural photosynthesis and for designing photovoltaic and photocatalytic devices. In this paper, we use Bacteriochlorophyll dimers extracted from the light harvesting apparatus and reaction center of a photosynthetic purple bacterium as model systems to study such excitations using first-principles numerical simulation methods. We distinguish four different regimes of intermolecular coupling, ranging from very weakly coupled to strongly coupled, and identify the factors that determine the energy and character of charge-transfer excitations in each case. We also construct an artificial dimer to systematically study the effects of intermolecular distance and orientation on charge-transfer excitations, as well as the impact of molecular vibrations on these excitations. Our results provide design rules for tailoring charge-transfer excitations in Bacteriochloropylls and related photoactive molecules, and highlight the importance of including charge-transfer excitations in accurate models of the excited-state structure and dynamics of Bacteriochlorophyll aggregates.

Discovery of Intratumoral Oncolytic Bacteria Toward Targeted Anticancer Theranostics.

Unveiling biomedical functions of tumor-resident microbiota is challenging for developing advanced anticancer medicines. This study demonstrates that isolated intratumoral bacteria, associated with natural purple photosynthetic bacteria, have inherent biocompatibility and strong immunogenic anticancer efficacies. They preferentially grow and proliferate within a targeted tumor milieu, which effectively causes immune cells to infiltrate the tumor and provoke strong anticancer responses in various syngeneic mouse models, including colorectal cancer, sarcoma, metastatic lung cancer, and extensive drug-resistant breast cancer. Furthermore, these functional bacteria-treated mice exhibit excellent anticancerous responses and have significantly prolonged survival rates with effective immunological memory. Light-harvesting nanocomplexes of microbial consortia of intratumoral bacteria and purple photosynthetic bacteria can diagnose tumors using bio-optical-window near-infrared light, making them useful theranostic agents for highly targeted immunological elimination of the tumor and for precisely marking tumor location.

A novel route for anoxygenic polymerization of dopamine via purple photosynthetic bacteria metabolism

Dopamine is an efficient building block to produce a versatile coating polymer able to adhere on a vast repertoire of material surfaces. Polydopamine, a dark-bioinspired polymer, is produced by the self-assembly of the dopamine under aerobic conditions in an alkaline environment. The presence of oxygen is crucial for self-polymerization of dopamine in aqueous solution. In this manuscript we show that is possible to drive the polymerization in absence of oxygen exploiting the metabolism of anaerobic photosynthetic purple bacteria.Graphical

Conversion of light into electricity in a semi-synthetic system based on photosynthetic bacterial chromatophores

Chromatophores (Chr) from photosynthetic nonsulfur purple bacterium Rhodobacter sphaeroides immobilized onto a Millipore membrane filter (MF) and sandwiched between two semiconductor indium tin oxide (ITO) electrodes (termed ITO|Chr – MF|ITO) have been used to measure voltage (ΔV) induced by continuous illumination. The maximum ΔV was detected in the presence of ascorbate / N,N,N’N'-tetramethyl-p-phenylenediamine couple, coenzyme UQ0, disaccaride trehalose and antimycin A, an inhibitor of cytochrome bc1 complex. In doing so, the light-induced electron transfer in the reaction centers was the major source of photovoltages. The stability of the voltage signal upon prolonged irradiation (>1 h) may be due to the maintenance of a conformation that is optimal for the functioning of integral protein complexes and stabilization of lipid bilayer membranes in the presence of trehalose. Retaining ∼70 % of the original photovoltage performance on the 30th day of storage at 23 °C in the dark under air was achieved after re-injection of fresh buffer (∼40 μL) containing redox mediators into the ITO|Chr – MF|ITO system. The approach we use is easy and can be extended to other biological intact systems (cells, thylakoid membranes) capable of converting energy of light.

Atomic force microscopic analysis of the light-harvesting complex 2 from purple photosynthetic bacterium Thermochromatium tepidum.

Structural information on the circular arrangements of repeating pigment-polypeptide subunits in antenna proteins of purple photosynthetic bacteria is a clue to a better understanding of molecular mechanisms for the ring-structure formation and efficient light harvesting of such antennas. Here, we have analyzed the ring structure of light-harvesting complex 2 (LH2) from the thermophilic purple bacterium Thermochromatium tepidum (tepidum-LH2) by atomic force microscopy. The circular arrangement of the tepidum-LH2 subunits was successfully visualized in a lipid bilayer. The average top-to-top distance of the ring structure, which is correlated with the ring size, was 4.8 ± 0.3nm. This value was close to the top-to-top distance of the octameric LH2 from Phaeospirillum molischianum (molischianum-LH2) by the previous analysis. Gaussian distribution of the angles of the segments consisting of neighboring subunits in the ring structures of tepidum-LH2 yielded a median of 44°, which corresponds to the angle for the octameric circular arrangement (45°). These results indicate that tepidum-LH2 has a ring structure consisting of eight repeating subunits. The coincidence of an octameric ring structure of tepidum-LH2 with that of molischianum-LH2 is consistent with the homology of amino acid sequences of the polypeptides between tepidum-LH2 and molischianum-LH2.

Cryo-EM structure of a monomeric RC-LH1-PufX supercomplex with high-carotenoid content from Rhodobacter capsulatus

In purple photosynthetic bacteria, the photochemical reaction center (RC) and light-harvesting complex 1 (LH1) assemble to form monomeric or dimeric RC-LH1 membrane complexes, essential for bacterial photosynthesis. Here, we report a 2.59-Å resolution cryoelectron microscopy (cryo-EM) structure of the RC-LH1 supercomplex from Rhodobacter capsulatus. We show that Rba. capsulatus RC-LH1 complexes are exclusively monomers in which the RC is surrounded by a 15-subunit LH1 ring. Incorporation of a transmembrane polypeptide PufX leads to a large opening within the LH1 ring. Each LH1 subunit associates two carotenoids and two bacteriochlorophylls, which is similar to Rba. sphaeroides RC-LH1 but more than one carotenoid per LH1 in Rba. veldkampii RC-LH1 monomer. Collectively, the unique Rba. capsulatus RC-LH1-PufX represents an intermediate structure between Rba. sphaeroides and Rba. veldkampii RC-LH1-PufX. Comparison of PufX from the three Rhodobacter species indicates the important residues involved in dimerization of RC-LH1.

Nanodiscs as a novel approach to resolve inter-protein energy transfer within the photosynthetic membrane of purple bacteria.

Photosynthetic purple bacteria capture sunlight and convert it to chemical energy with almost 100% quantum efficiency: almost every photon absorbed leads to charge separation. Such high efficiency is achieved through a series of ultrafast inter-protein energy transfer events within an antenna network of light-harvesting proteins located in the photosynthetic membrane. Understanding how the organization of proteins within the membrane leads to efficient energy transfer is imperative to designing efficient artificial solar harvesting techniques. Determining inter-protein energy transfer between light-harvesting complex 2 (LH2) proteins, the most common light-harvesting protein in vivo, has proven challenging due to the heterogeneous organization of proteins within the membrane environment. In this work, we introduce model membrane nanodiscs as a novel technique to reconstruct the complex membrane environment in a controlled manner. By forming nanodiscs large enough to incorporate two variants of LH2, we can directly resolve inter-protein energy transfer. By controlling nanodisc size, we can change the inter-protein distance and resolve the effect of membrane organization on energy transfer rate. Using a combination of ultrafast transient absorption spectroscopy, cryogenic electron microscopy, and quantum chemical calculations, we find that LH2 complexes prefer to associate closely in the membrane (25 Å), with an energy transfer rate of 5.7 ps. The results suggest that these tightly-packed LH2s are important for long-distance energy transfer, as the 25 Å distance is similar to the most common inter-protein distance in vivo. Overall, our work introduces nanodiscs as a platform to study complex energy transfer events and the effect of membrane organization on critical biological processes.

Reconstituted LH2 in multilayer membranes induced by poly-l-lysine: Structure of supramolecular and electronic states

To explore the effect of cell membrane stacking on the function of light-harvesting complex 2 (LH2) in purple nonsulfur photosynthetic bacteria, LH2 from Rhodobacter sphaeroides 2.4.1 (R. sph 2.4.1) was reconstituted into lipid bilayer vesicles (LH2@liposome) and further formed multi-layer structure by electrostatic interaction with poly-l-lysine (LH2@liposome/PLL), which was characterized by cryo-electron microscopy (cryo-EM) and TEM. When embedded in liposomes and additionally in multi-layer liposomes, the absorption band, zero-crossing point of CD signals and fluorescence emission of LH2 B850 excitons were observed to uniformly have 1–2 nm red-shifting. Combining with the corresponding fluorescence quench and the generation of shorter-living fluorescence species, a new excitonic species generated through B850 structural splitting was proposed. By FT-Raman and triplet carotenoid dynamics, the structural mechanism was deduced and discussed. Briefly, all environmental changes, including LH2 aggregating and multi-layer membrane stacking, eventually applied forces on B850 exciton molecules mainly through the hydrogen bonding between the C3-acetyl carbonyl groups of B850 BChls and Tyr44 and 45 residues at C-terminus of α-polypeptides. The effect of multi-layer structure on LH2 could be assigned as a kind of photoprotection.

Biyorafineri konseptiyle Rhodobacter sphaeroides O.U.001 ile biyolojik hidrojen ve bakteriyel karotenoid üretimi

In this study, the goal was to produce biohydrogen and bacterial carotenoids with Rhodobacter sphaeroides O.U.001, a purple non-sulfur photosynthetic bacterium, utilizing sugar beet molasses in the context of biorefinery. First, media with different sugar concentrations (10 g/L, 20 g/L, 30 g/L, 40 g/L, 50 g/L) were prepared for bacterial growth. Then, hydrogen production was carried out using these media in anaerobic conditions in 100 ml bioreactors. After hydrogen gas was collected from the bioreactors, carotenoid extraction was performed from the remaining bacteria. As a result of the analyzes, it was found that the amount of biohydrogen and the amount of bacterial carotenoids obtained were inversely proportional to the increased sugar concentrations. The maximum hydrogen formation was detected in the medium containing 10 g/L of sugar (19.18 mL). According to the results of gas chromatography analysis, the quantity of hydrogen in the total gas was found to be around 23.6%. The highest yield of carotenoids was again obtained from bacteria reproduced in a medium containing 10 g/L of sugar (3.12 mg/g, carotenoid/dry biomass). As a conclusion, this study provides an example for the successful realization of two high value-added products within a biorefinery approach by using molasses obtained at an affordable cost.

Effect and mechanism of the improvement of coastal silt soil by application of organic fertilizer and gravel combined with Sesbania cannabina cultivation.

Jiangsu Province of China has a large area of coastal silt soil (CSS) with poor permeability, high salinity, and poor nutrients, which brings great difficulties to the development and utilization of coastal zones, so that needs to be improved as a matter of urgency. In this study, river-sand, serpentine, and organic fertilizer were used as additives in CSS, and Sesbania cannabina, a salt-tolerant cash crop, was planted in these differently treated soils. Through high-throughput sequencing, analysis of soil physico-chemical properties, and detection of plant growth status, the rhizosphere bacterial diversity of S. cannabina growing in CSS under different treatments and their environmental impact factors were studied, while exploring the effect and mechanism of organic fertilizer combined with gravel as a CSS modifier. The results implied that single application of organic fertilizer could significantly increase the fertility levels of total nitrogen (TN), total organic carbon (TOC) and Avail. P in CSS; then, the application of organic fertilizer with river-sand significantly reduced salt content and alkalinity of soil; meanwhile, in the treatment of single application of organic fertilizer and application of organic fertilizer combined with river-sand, the rhizosphere of S. cannabina enriched the bacterial communities of organic matter degradation and utilization to varying degrees. The soil moisture content and indicators related to saline-alkali soil (including total salt, electrical conductivity (EC), exchangeable sodium percentage (ESP), Avail. Na and Avail. K, etc.) were further reduced significantly by the application of organic fertilizer combined with river-sand and serpentine. The method has greatly improved the growth conditions of S. cannabina and promoted the positive development of its rhizosphere bacterial community. Among them, in the treatment of organic fertilizer combined with river-sand and serpentine, a variety of plant growth-promoting rhizobacteria (PGPR, such as Sphingomonas, Ensifer, and Rhodobacter) and nitrogen-cycle-related bacteria (such as nitrate-reduction-related bacteria, nitrogen-fixing bacteria like Ensifer, and purple non-sulfur photosynthetic bacteria like Rhodobacter) were enriched in the rhizosphere of S. cannabina; moreover, the mutual association and robustness of bacterial co-occurrence networks have been significantly enhanced. The results provide a theoretical basis and reference model for the improvement of coastal saline-alkali silt soil.

Tuning photosynthetic oxygen for hydrogen evolution in synergistically integrated, sulfur deprived consortia of Coccomyxa chodatii and Rhodobium gokarnense at dim and high light

In this work, tuning oxygen tension was targeted to improve hydrogen evolution. To achieve such target, various consortia of the chlorophyte Coccomyxa chodatii with a newly isolated photosynthetic purple non-sulfur bacterium (PNSB) strain Rhodobium gokarnense were set up, sulfur replete/deprived, malate/acetate fed, bicarbonate/sulfur added at dim/high light. C. chodatii and R. gokarnense are newly introduced to biohydrogen studies for the first time. Dim light was applied to avoid the inhibitory drawbacks of photosynthetic oxygen evolution, values of hydrogen are comparable with high light or even more and thus economically feasible to eliminate the costs of artificial illumination. Particularly, the consortium of 2n− (n = 1.9 × 105 cell/ml, sulfur deprived) demonstrated its perfection for the target, i.e., the highest possible cumulative hydrogen. This consortium exhibited negative photosynthesis, i.e., oxygen uptake in the light. Most hydrogen in consortia is from bacterial origin, although algae evolved much more hydrogen than bacteria on per cell basis, but for only one day (the second 24 h), as kinetics revealed. The higher hydrogen in unibacterial culture or consortia results from higher bacterial cell density (20 times). Consortia evolved more hydrogen than their respective separate cultures, further enhanced when bicarbonate and sulfur were supplemented at higher light. The share of algae relatively increased as bicarbonate or sulfur were added at higher light intensity, i.e., PSII activity partially recovered, resulting in a transient autotrophic hydrogen evolution. The addition of acetic acid in mixture with malic acid significantly enhanced the cumulative hydrogen levels, mostly decreased cellular ascorbic acid indicating less oxidative stress and relief of PSII, relative to malic acid alone. Starch, however, decreased, indicating the specificity of acetic acid. Exudates (reducing sugars, amino acids, and soluble proteins) were detected, indicating mutual utilization. Yet, hydrogen evolution is limited; tuning PSII activity remains a target for sustainable hydrogen production.

Environmental protection via biomonitoring lead exposure by photosynthetic purple bacteria

Environmental protection via biomonitoring lead exposure by photosynthetic purple bacteria

Comparative computational study to augment UbiA prenyltransferases inherent in purple photosynthetic bacteria cultured from mangrove microbial mats in Qatar for coenzyme Q10 biosynthesis.

Coenzyme Q10 (CoQ10) is a powerful antioxidant with a myriad of applications in healthcare and cosmetic industries. The most effective route of CoQ10 production is microbial biosynthesis. In this study, four CoQ10 biosynthesizing purple photosynthetic bacteria: Rhodobacter blasticus, Rhodovulum adriaticum, Afifella pfennigii and Rhodovulum marinum, were identified using 16S rRNA sequencing of enriched microbial mat samples obtained from Purple Island mangroves (Qatar). The membrane bound enzyme 4-hydroxybenzoate octaprenyltransferase (UbiA) is pivotal for bacterial biosynthesis of CoQ10. The identified bacteria could be inducted as efficient industrial bio-synthesizers of CoQ10 by engineering their UbiA enzymes. Therefore, the mutation sites and substitution residues for potential functional enhancement were determined by comparative computational study. Two mutation sites were identified within the two conserved Asp-rich motifs, and the effect of proposed mutations in substrate binding affinity of the UbiA enzymes was assessed using multiple ligand simultaneous docking (MLSD) studies, as a groundwork for experimental studies.