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Osmotic adaptation and accumulation of compatible solutes is a key process for life at high osmotic pressure and elevated salt concentrations. Most important solutes that can protect cell structures and metabolic processes at high salt concentrations are glycine betaine and ectoine. The genome analysis of more than 130 phototrophic bacteria shows that biosynthesis of glycine betaine is common among marine and halophilic phototrophic Proteobacteria and their chemotrophic relatives, as well as in representatives of Pirellulaceae and Actinobacteria, but are also found in halophilic Cyanobacteria and Chloroherpeton thalassium. This ability correlates well with the successful toleration of extreme salt concentrations. Freshwater bacteria in general lack the possibilities to synthesize and often also to take up these compounds. The biosynthesis of ectoine is found in the phylogenetic lines of phototrophic Alpha- and Gammaproteobacteria, most prominent in the Halorhodospira species and a number of Rhodobacteraceae. It is also common among Streptomycetes and Bacilli. The phylogeny of glycine-sarcosine methyltransferase (GMT) and diaminobutyrate-pyruvate aminotransferase (EctB) sequences correlate well with otherwise established phylogenetic groups. Most significantly, GMT sequences of cyanobacteria form two major phylogenetic branches and the branch of Halorhodospira species is distinct from all other Ectothiorhodospiraceae. A variety of transport systems for osmolytes are present in the studied bacteria.

ABSTRACT The genus Chloroflexus is a deeply branching group of thermophilic filamentous anoxygenic phototrophic bacteria. The bacteria in this genus have been shown to grow well heterotrophically under anaerobic photosynthetic and aerobic respiratory conditions. We examined autotrophic growth in new isolates of Chloroflexus strains from hot springs in Nakabusa, Japan. The isolates belonging to Chloroflexus aggregans (98.7% identity of 16S rRNA gene sequence to the respective type strain) and Chloroflexus aurantiacus (99.9% identity to the respective type strain) grew photoautotrophically under a 24% H2 atmosphere. We also observed chemolithotrophic growth of these isolates under 80% H2 and 5% O2 conditions in the dark. This is the first report showing that Chloroflexus grew under both photoautotrophic and chemolithotrophic conditions in addition to photoheterotrophic and aerobic chemoheterotrophic conditions. This is the first report showing that Chloroflexus can grow under both photoautotrophic and chemolithotrophic conditions in addition to photoheterotrophic and aerobic chemoheterotrophic conditions.

Background Pinene is a monoterpene, that is used in the manufacture of fragrances, insecticide, fine chemicals, and renewable fuels. Production of pinene by metabolic-engineered microorganisms is a sustainable method. Purple non-sulfur photosynthetic bacteria belong to photosynthetic chassis that are widely used to synthesize natural chemicals. To date, researches on the synthesis of pinene by purple non-sulfur photosynthetic bacteria has not been reported, leaving the potential of purple non-sulfur photosynthetic bacteria synthesizing pinene unexplored. Results Rhodobacter sphaeroides strain was applied as a model and engineered to express the fusion protein of heterologous geranyl diphosphate synthase (GPPS) and pinene synthase (PS), hence achieving pinene production. The reaction condition of pinene production was optimized and 97.51 μg/L of pinene was yielded. Then, genes of 1-deoxy- d -xylulose 5-phosphate synthase, 1-deoxy- d -xylulose 5-phosphate reductoisomerase and isopentenyl diphosphate isomerase were overexpressed, and the ribosome binding site of GPPS-PS mRNA was optimized, improving pinene titer to 539.84 μg/L. Conclusions In this paper, through heterologous expression of GPPS-PS, pinene was successfully produced in R. sphaeroides , and pinene production was greatly improved by optimizing the expression of key enzymes. This is the first report on pinene produce by purple non-sulfur photosynthetic bacteria, which expands the availability of photosynthetic chassis for pinene production.

Background As an antioxidant, hydrogen (H 2 ) can selectively react with the highly toxic hydroxyl radical (·OH) in tumor cells to break the balance of reactive oxygen species (ROS) and cause oxidative stress. However, due to the high diffusibility and storage difficulty of hydrogen, it is impossible to achieve long-term release at the tumor site, which highly limited their therapeutic effect. Results Photosynthetic bacteria (PSB) release a large amount of hydrogen to break the balance of oxidative stress. In addition, as a nontoxic bacterium, PSB could stimulate the immune response and increase the infiltration of CD4+ and CD8+ T cells. More interestingly, we found that hydrogen therapy induced by our live PSB did not lead to the up-regulation of PD-L1 after stimulating the immune response, which could avoid the tumor immune escape. Conclusion Hydrogen-immunotherapy significantly kills tumor cells. We believe that our live microbial hydrogen production system provides a new strategy for cancer hydrogen treatment combining with enhanced immunotherapy without up-regulating PD-L1. Graphical Abstract

Sewage lagoons and wastewater ponds from industrialised swine and poultry farms are typically hypereutrophic, auxinic and dominated by purple non-sulphur bacteria and unicellular green algae both typically growing photoheterotrophically. To manage such ponds, it is essential to know the balance between oxygenic and anoxygenic photosynthesis. Typical spectrophotometric algorithms to estimate chlorophyll use 750 nm as a zero (A750 nm) but a 750 nm zero protocol is unsuitable where substantial amounts of bacteriochlorophylls are present. Algorithms were developed to estimate chlorophylls a and b (Chl a and Chl b) and bacteriochlorophyll a (BChl a) in solvent in ethanol, 7:2 acetone/ethanol and 90% acetone. The algorithms use an 850-nm absorbance zero (A850 nm) well outside the absorbance ranges of both chlorophylls and bacteriochlorophylls in solvent. There are many habitats where the presence of anoxygenic photosynthetic bacteria is unsuspected and so using a routine A750 nm zero effectively masks their presence and leads to underestimations of Chl a and Chl b. The in-solvent red peak of bacteriochlorophyll c is too close to that of Chl a for BChl c and Chl a to be resolved spectroscopically, but its presence can be easily identified from in vivo scans. The spectroscopic advantage of 90% acetone is negated by its poor quantitative extraction of pigments. Acetone/ethanol (7:2) is an excellent solvent spectroscopically and as an extractant.

To address the slow growth rate of photosynthetic bacteria (PSB), this study introduces pantothenic acid as a biological enhancing factor. The effects of pantothenic acid on PSB proliferation and its effectiveness in treating high-concentration ammonia–nitrogen wastewater were systematically evaluated. Additionally, the effects of different culture conditions, including dark aeration, darkness, light exposure, and light aeration, on PSB growth were investigated. The results show that optimal PSB growth was achieved with 20 mg/L of pantothenic acid; however, higher concentrations of pantothenic acid inhibited bacterial growth. The addition of pantothenic acid also significantly enhanced the performance of PSB in treating high-concentration organic wastewater, increasing the removal rates of COD, ammonia nitrogen, total phosphorus, and total nitrogen to 43.0%, 94.0%, 49.7%, and 51.0%, respectively. Furthermore, a synergistic effect between dark aeration and light exposure was observed. When the time of light and dark aeration was set at 1:1, the highest PSB yield was recorded, and the removal efficiencies of COD, ammonia nitrogen, total nitrogen, and total phosphorus increased to 71.4%, 95.3%, 57.1%, and 74.7%, respectively. Through the introduction of pantothenic acid and optimization of culture mode, the rapid growth of PSB and highly efficient treatment of organic wastewater were achieved, providing a new approach for advanced wastewater treatment and resource utilization.

Sulfide detoxification can be catalyzed by ancient membrane-bound flavoproteins, sulfide:quinone oxidoreductases (Sqr), which have important roles in sulfide homeostasis and sulfide-dependent energy conservation processes by transferring electrons from sulfide to respiratory or photosynthetic membrane electron flow. Sqr enzymes have been categorized into six groups. Several members of the groups I, II, III, and V are well-known, but type IV and VI Sqrs are, as yet, uncharacterized or hardly characterized at all. Here, we report detailed characterization of a type VI sulfide:quinone oxidoreductase (TrSqrF) from a purple sulfur bacterium, Thiocapsa roseopersicina. Phylogenetic analysis classified this enzyme in a special group composed of SqrFs of endosymbionts, while a weaker relationship could be observed with SqrF of Chlorobaculum tepidum which is the only type VI enzyme characterized so far. Directed mutagenesis experiments showed that TrSqrF contributed substantially to the sulfide:quinone oxidoreductase activity of the membranes. Expression of the sqrF gene could be induced by sulfide. Homologous recombinant TrSqrF protein was expressed and purified from the membranes of a SqrF-deleted T. roseopersicina strain. The purified protein contains redox-active covalently bound FAD cofactor. The recombinant TrSqrF enzyme catalyzes sulfur-dependent quinone reduction and prefers ubiquinone-type quinone compounds. Kinetic parameters of TrSqrF show that the affinity of the enzyme is similar to duroquinone and decylubiquinone, but the reaction has substantially lower activation energy with decylubiquinone, indicating that the quinone structure has an effect on the catalytic process. TrSqrF enzyme affinity for sulfide is low, therefore, in agreement with the gene expressional analyis, SqrF could play a role in energy-conserving sulfide oxidation at high sulfide concentrations. TrSqrF is a good model enzyme for the subgroup of type VI Sqrs of endosymbionts and its characterization might provide deeper insight into the molecular details of the ancient, anoxic, energy-gaining processes using sulfide as an electron donor.

Isolated environments such as hot springs are particularly interesting for studying the microbial biogeography. These environments create an ‘island effect’ leading to genetic divergence. We studied the phylogeographic pattern of thermophilic anoxygenic phototrophic bacteria, belonging to the Roseiflexus genus. The main characteristic of the observed pattern was geographic and geochronologic fidelity to the hot springs within Circum-Pacific and Alpine-Himalayan-Indonesian orogenic belts. Mantel test revealed a correlation between genetic divergence and geographic distance among the phylotypes. Cluster analysis revealed a regional differentiation of the global phylogenetic pattern. The phylogeographic pattern is in correlation with geochronologic events during the break up of Pangaea that led to the modern configuration of continents. To our knowledge this is the first geochronological scenario of intercontinental prokaryotic taxon divergence. The existence of the modern phylogeographic pattern contradicts with the existence of the ancient evolutionary history of the Roseiflexus group proposed on the basis of its deep-branching phylogenetic position. These facts indicate that evolutionary rates in Roseiflexus varied over a wide range. The analysis of 16S rRNA gene sequences and environmental data revealed the existence of the specific phylogeographic pattern for thermophilic phototrophic bacteria belonging to Roseiflexus genus. Graphical Abstract Figure. The analysis of 16S rRNA gene sequences and environmental data revealed the existence of the specific phylogeographic pattern for thermophilic phototrophic bacteria belonging to Roseiflexus genus.

The dark relaxation of the yield of variable BChl fluorescence in the 10⁻⁵–10 s time range is measured after laser diode (808 nm) excitation of variable duration in intact cells of photosynthetic bacteria Rba. sphaeroides, Rsp. rubrum, and Rvx. gelatinosus under various treatments of redox agents, inhibitors, and temperature. The kinetics of the relaxation is complex and much wider extended than a monoexponential function. The longer is the excitation, the slower is the relaxation which is determined by the redox states, sizes, and accessibility of the pools of cytochrome [Formula: see text] and quinone for donor and acceptor side-limited bacterial strains, respectively. The kinetics of fluorescence decay reflects the opening kinetics of the closed RC. The relaxation is controlled preferentially by the rate of re-reduction of the oxidized dimer by mobile cytochrome [Formula: see text] in Rba. sphaeroides and Rsp. rubrum and by the rate constant of the [Formula: see text] interquinone electron transfer, (350 μs)⁻¹and/or the quinol/quinone exchange at the acceptor side in Rvx. gelatinosus. The commonly used acceptor side inhibitors (e.g., terbutryn) demonstrate kinetically limited block of re-oxidation of the primary quinone. The observations are interpreted in frame of a minimum kinetic and energetic model of electron transfer reactions in bacterial RC of intact cells.

Photosynthetic bacteria exhibit significant bioremediation potential and resource recycling characteristics, rendering them valuable candidates for sustainable wastewater treatment applications. Sea cucumber boiling broth (SCBB) contains high concentrations of organic compounds and nutrient salts, whose indiscriminate discharge poses serious environmental risks. This study aimed to evaluate a photosynthetic bacterium capable of effectively treating SCBB, which was isolated from the intertidal sediment samples. The bacterial strain was identified using 16S rDNA sequencing, and optimal growth conditions, including light, pH, and temperature, were determined. Finally, a small-scale trial was conducted in a fed-batch fermenter. The results showed that 16S rDNA analysis placed this strain in the Chromatiaceae family, forming a distinct lineage from the closest related species Marichromatium purpuratum and M. gracile, and was tentatively named Marichromatium sp. DYYC01. The strain exhibited optimal growth under anaerobic conditions at 30 °C, light intensity of 100 μmol photons/m2/s, and pH 7.0. Batch culture experiments demonstrated maximum biomass accumulation (OD660 = 0.831) in SCBB medium with an initial COD loading of 3913 mg L⁻1, concomitant with significant nutrient removal efficiencies: 76.45% COD, 55.82% total nitrogen (TN), and 56.67% total phosphorus (TP). Scaling up to fed-batch fermentation enhanced bioremediation performance, achieving removal rates of 83.13% COD, 72.17% TN, and 73.07% TP with enhanced growth (OD660 = 1.2). This study reveals Marichromatium sp. DYYC01’s exceptional halotolerance in high-salinity organic wastewater treatment. The strain’s capacity for simultaneous biomass production and efficient nutrient recovery from hypersaline processing effluent positions it as a promising candidate for developing circular bioeconomy strategies.