Purple Bacteria Research Articles

Environmental life cycle assessment of polyhydroxyalkanoates production by purple phototrophic bacteria mixed cultures

Bioplastics offer a promising sustainable alternative to petroleum-based plastics due to their biodegradability as well as favourable thermal and mechanical properties. Among different types of biobased polymers, the production of polyhydroxyalkanoates (PHA) using purple phototrophic bacteria (PPB) and low-value substrates has gained increasing interest. Despite the momentum, challenges regarding the scalability and environmental feasibility of this biopolymer production pathway remain. In response, this study employs an exploratory LCA approach to quantitatively assesses the potential environmental implications of PHA production in powder form and the joint management of the organic fraction of municipal solid waste (OFMSW) through a novel photobiorefinery system that uses PPB mixed cultures. Environmental impacts were tested under multiple improvement scenarios and benchmarked against the production of conventional fossil-based granulate or unprocessed plastics, including low density polyethylene (LDPE), polyethylene terephthalate (PET) and polyurethane (PU). The photobiorefinery stage was found to have the greatest contribution to the impact categories, particularly due to direct emissions, consumption of electricity and production of extractive chemical agents used. These factors accounted for over 70% of the photobiorefinery impact in all cases. Avoided impacts provided net favourable outcomes in terms of carbon footprint and fossil resources when comparing PHA production to conventional plastics, especially PET and PU, with impact reductions ranging from 30% to 60%, respectively. However, when considering other impact categories like eutrophication, this situation was less favourable. The exploration of alternative scenarios offered significant impact reductions, especially when renewable electricity or an environmentally friendly extraction agent is used. Moreover, minimizing methane losses or co-producing hydrogen in the photobiorefinery had a notably positive effect on the carbon footprint, reducing the impact by more than 2 t of CO2 eq per t of PHA powder compared to the base case. Therefore, the implementation of feasible improvement measures in the short term can position PHA produced by mixed cultures as a sustainable alternative to petroleum-based plastics.

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

Genes and Pathway Reactions Related to Carotenoid Biosynthesis in Purple Bacteria.

In purple bacteria, the genes of the carotenoid pathways are part of photosynthesis gene clusters which were distributed among different species by horizontal gene transfer. Their close organisation facilitated the first-time cloning of carotenogenic genes and promoted the molecular investigation of spheroidene and spirilloxanthin biosynthesis. This review highlights the cloning of the spheroidene and spirilloxanthin pathway genes and presents the current knowledge on the enzymes involved in the carotenoid biosynthesis of purple sulphur and non-sulphur bacteria. Mostly, spheroidene or spirilloxanthin biosynthesis exists in purple non-sulphur bacteria but both pathways operate simultaneously in Rubrivivax gelatinosus. In the following years, genes from other bacteria including purple sulphur bacteria with an okenone pathway were cloned. The individual steps were investigated by kinetic studies with heterologously expressed pathway genes which supported the establishment of the reaction mechanisms. In particular, the substrate and product specificities revealed the sequential order of the speroidene and spiriloxanthin pathways as well as their interactions. Information on the enzymes involved revealed that the phytoene desaturase determines the type of pathway by the formation of different products. By selection of mutants with amino acid exchanges in the putative substrate-binding site, the neurosporene-forming phytoene desaturase could be changed into a lycopene-producing enzyme and vice versa. Concerning the oxygen groups in neurosporene and lycopene, the tertiary alcohol group at C1 is formed from water and not by oxygenation, and the C2 or C4 keto groups are inserted differently by an oxygen-dependent or oxygen-independent ketolation reaction, respectively.

Harnessing redox power for biotechnological application in purple non-sulphur bacteria

Harnessing redox power for biotechnological application in purple non-sulphur bacteria Here we explore purple non-sulfur bacteria (PNSB) and some of its biotech applications, with a focus on how these applications have been enhanced by manipulating the flow of reducing power. Cellular metabolism refers to the sum of chemical reactions that cells use to obtain energy and material to live, grow, and make products. Many of these reactions are performed by specialized proteins called enzymes, which act on a specific substrate, or input, to produce a specific effect or product, also called a metabolite. To make metabolites, you need three major inputs: carbon, energy, and electrons. Depending on the organism, carbon can be fixed from atmospheric CO2, or can be obtained from organic compounds like carbohydrates or lipids. Energy can be generated in a variety of ways, but typically results in the formation of phosphate bonds in molecules like adenosine triphosphate (ATP) for widespread use in the cell.

Globally distributed Myxococcota with photosynthesis gene clusters illuminate the origin and evolution of a potentially chimeric lifestyle

Photosynthesis is a fundamental biogeochemical process, thought to be restricted to a few bacterial and eukaryotic phyla. However, understanding the origin and evolution of phototrophic organisms can be impeded and biased by the difficulties of cultivation. Here, we analyzed metagenomic datasets and found potential photosynthetic abilities encoded in the genomes of uncultivated bacteria within the phylum Myxococcota. A putative photosynthesis gene cluster encoding a type-II reaction center appears in at least six Myxococcota families from three classes, suggesting vertical inheritance of these genes from an early common ancestor, with multiple independent losses in other lineages. Analysis of metatranscriptomic datasets indicate that the putative myxococcotal photosynthesis genes are actively expressed in various natural environments. Furthermore, heterologous expression of myxococcotal pigment biosynthesis genes in a purple bacterium supports that the genes can drive photosynthetic processes. Given that predatory abilities are thought to be widespread across Myxococcota, our results suggest the intriguing possibility of a chimeric lifestyle (combining predatory and photosynthetic abilities) in members of this phylum.

A novel mechanistic modelling approach for microbial selection dynamics: Towards improved design and control of raceway reactors for purple bacteria

Purple phototrophic bacteria (PPB) show an underexplored potential for resource recovery from wastewater. Raceway reactors offer a more affordable full-scale solution on wastewater and enable useful additional aerobic processes. Current mathematical models of PPB systems provide useful mechanistic insights, but do not represent the full metabolic versatility of PPB and thus require further advancement to simulate the process for technology development and control. In this study, a new modelling approach for PPB that integrates the photoheterotrophic, and both anaerobic and aerobic chemoheterotrophic metabolic pathways through an empirical parallel metabolic growth constant was proposed. It aimed the modelling of microbial selection dynamics in competition with aerobic and anaerobic microbial community under different operational scenarios. A sensitivity analysis was carried out to identify the most influential parameters within the model and calibrate them based on experimental data. Process perturbation scenarios were simulated, which showed a good performance of the model.

Generation of Electric Potential Difference by Chromatophores from Photosynthetic Bacteria in the Presence of Trehalose under Continuous Illumination.

Measurement of electrical potential difference (Δψ) in membrane vesicles (chromatophores) from the purple bacterium Rhodobacter sphaeroides associated with the surface of a nitrocellulose membrane filter (MF) impregnated with a phospholipid solution in decane or immersed into it in the presence of exogenous mediators and disaccharide trehalose demonstrated an increase in the amplitude and stabilization of the signal under continuous illumination. The mediators were the ascorbate/N,N,N'N'-tetramethyl-p-phenylenediamine pair and ubiquinone-0 (electron donor and acceptor, respectively). Although stabilization of photoelectric responses upon long-term continuous illumination was observed for both variants of chromatophore immobilization, only the samples immersed into the MF retained the functional activity of reaction centers (RCs) for a month when stored in the dark at room temperature, which might be due to the preservation of integrity of chromatophore proteins inside the MF pores. The stabilizing effect of the bioprotector trehalose could be related to its effect on both the RC proteins and the phospholipid bilayer membrane. The results obtained will expand current ideas on the use of semi-synthetic structures based on various intact photosynthetic systems capable of converting solar energy into its electrochemical form.

Thiorhodovibrio frisius and Trv. litoralis spp. nov., Two Novel Members from a Clade of Fastidious Purple Sulfur Bacteria That Exhibit Unique Red-Shifted Light-Harvesting Capabilities.

In the pursuit of cultivating anaerobic anoxygenic phototrophs with unusual absorbance spectra, a purple sulfur bacterium was isolated from the shoreline of Baltrum, a North Sea island of Germany. It was designated strain 970, due to a predominant light harvesting complex (LH) absorption maximum at 963-966 nm, which represents the furthest infrared-shift documented for such complexes containing bacteriochlorophyll a. A polyphasic approach to bacterial systematics was performed, comparing genomic, biochemical, and physiological properties. Strain 970 is related to Thiorhodovibrio winogradskyi DSM 6702T by 26.5, 81.9, and 98.0% similarity via dDDH, ANI, and 16S rRNA gene comparisons, respectively. The photosynthetic properties of strain 970 were unlike other Thiorhodovibrio spp., which contained typical LH absorbing characteristics of 800-870 nm, as well as a newly discovered absorption band at 908 nm. Strain 970 also had a different photosynthetic operon composition. Upon genomic comparisons with the original Thiorhodovibrio strains DSM 6702T and strain 06511, the latter was found to be divergent, with 25.3, 79.1, and 97.5% similarity via dDDH, ANI, and 16S rRNA gene homology to Trv. winogradskyi, respectively. Strain 06511 (=DSM 116345T) is thereby described as Thiorhodovibrio litoralis sp. nov., and the unique strain 970 (=DSM 111777T) as Thiorhodovibrio frisius sp. nov.

Achieving Discharge Limits in Single-Stage Domestic Wastewater Treatment by Combining Urban Waste Sources and Phototrophic Mixed Cultures.

This work shows the potential of a new way of co-treatment of domestic wastewater (DWW) and a liquid stream coming from the thermal hydrolysis of the organic fraction of municipal solid waste (OFMSW) mediated by a mixed culture of purple phototrophic bacteria (PPB) capable of assimilating carbon and nutrients from the medium. The biological system is an open single-step process operated under microaerophilic conditions at an oxidative reduction potential (ORP) < 0 mV with a photoperiod of 12/24 h and fed during the light stage only so the results can be extrapolated to outdoor open pond operations by monitoring the ORP. The effluent mostly complies with the discharge values of the Spanish legislation in COD and p-values (<125 mg/L; <2 mg/L), respectively, and punctually on values in N (<15 mg/L). Applying an HRT of 3 d and a ratio of 100:7 (COD:N), the presence of PPB in the mixed culture surpassed 50% of 16S rRNA gene copies, removing 78% of COD, 53% of N, and 66% of P. Furthermore, by increasing the HRT to 5 d, removal efficiencies of 83% of COD, 65% of N, and 91% of P were achieved. In addition, the reactors were further operated in a membrane bioreactor, thus separating the HRT from the SRT to increase the specific loading rate. Very satisfactory removal efficiencies were achieved by applying an HRT and SRT of 2.3 and 3 d, respectively: 84% of COD, 49% of N, and 93% of P despite the low presence of PPB due to more oxidative conditions, which step-by-step re-colonized the mixed culture until reaching >20% of 16S rRNA gene copies after 49 d of operation. These results open the door to scaling up the process in open photobioreactors capable of treating urban wastewater and municipal solid waste in a single stage and under microaerophilic conditions by controlling the ORP of the system.

Characterization of the Pyrroloquinoline Quinone Producing Rhodopseudomonas palustris as a Plant Growth-Promoting Bacterium under Photoautotrophic and Photoheterotrophic Culture Conditions.

Rhodopseudomonas palustris is a purple non-sulfide bacterium (PNSB), and some strains have been proven to promote plant growth. However, the mechanism underlying the effect of these PNSBs remains limited. Based on genetic information, R. palustris possesses the ability to produce pyrroloquinoline quinone (PQQ). PQQ is known to play a crucial role in stimulating plant growth, facilitating phosphorous solubilization, and acting as a reactive oxygen species scavenger. However, it is still uncertain whether growth conditions influence R. palustris's production of PQQ and other characteristics. In the present study, it was found that R. palustris exhibited a higher expression of genes related to PQQ synthesis under autotrophic culture conditions as compared to acetate culture conditions. Moreover, similar patterns were observed for phosphorous solubilization and siderophore activity, both of which are recognized to contribute to plant-growth benefits. However, these PNSB culture conditions did not show differences in Arabidopsis growth experiments, indicating that there may be other factors influencing plant growth in addition to PQQ content. Furthermore, the endophytic bacterial strains isolated from Arabidopsis exhibited differences according to the PNSB culture conditions. These findings imply that, depending on the PNSB's growing conditions, it may interact with various soil bacteria and facilitate their infiltration into plants.

The Efficacy of 5-Aminolevulinic Acid-Producing Luteovulum sphaeroides Strains on Saline Soil Fertility, Nutrient Uptakes, and Yield of Rice

Saline soils negatively affect and cause serious problems for rice cultivation. This study aimed to evaluate the efficacy of the purple nonsulfur bacteria (PNSB) capable of secreting 5-aminolevulinic acid (ALA) to reduce soil salinity, improve soil fertility, and enhance rice growth and yield. A two-factorial experiment was conducted in a randomized complete block design with four replications. Factor one was the salinity of the irrigated water, and factor two was the supplementation of the ALA-producing PNSB. The results indicated that watering with saline water above 3‰ led to decreases in plant growth and rice yield compared to the treatments watered with tap water. Application of either an individual strain or the mixture of W01, W14, and W22 ameliorated soil properties and increased total NPK uptake, whereas treatments supplied with the mixed strains reduced total Na uptake (9.50 mg Na pot−1). Supplying the W01, W14, and W22 strains individually or in a mixture enhanced the plant height by 3.51–5.45% and rice grain yield by 14.7–26.2%, compared with those of the control treatment. From the study, the combination of the L. sphaeroides W01, W14, and W22 strains is promising for application in saline or salt-contaminated regions to aid the damages caused by salinity on cultivars there, especially rice. Furthermore, this is a biological approach to ease an environmental problem and improve crop performance, which is supposed to be a trend in the sustainable agriculture.

Cultivation of Non-Sulphur bacteria at a Research Laboratory Level

Purple non-sulphur bacteria (PNSB) are flagellated, gram-negative proteobacteria. The preparation of purple non sulphur bacteria is a crucial step in their utilization for various applications. This review aimed to develop an optimized protocol for the isolation, culturing, and growth of purple non sulphur bacteria. Key factors such as nutrient sources, environmental conditions, and light intensity were systematically evaluated to enhance biomass production and pigment synthesis. The interpretation from previous work highlighted the importance of specific nutrient compositions and provided insights into the optimal growth parameters, including temperature and pH. The findings contribute to advancing the understanding of purple non sulphur bacteria cultivation and offer valuable information for future research and potential industrial applications. Keywords: purple non sulphur bacteria, cultivation, isolation, culturing.

Removal of pollutants and accumulation of high-value cell inclusions in a batch reactor containing Rhodopseudomonas for treating real heavy oil refinery wastewater

Treating wastewater using purple non-sulfur bacteria (PNSB) is an environmentally friendly technique that can simultaneously remove pollutants and lead to the accumulation of high-value cell inclusions. However, no PNSB system for treating heavy oil refinery wastewater (HORW) and recovering high-value cell inclusions has yet been developed. In this study, five batch PNSB systems dominated by Rhodopseudomonas were used to treat real HORW for 186 d. The effects of using different hydraulic retention times (HRT), sludge retention times (SRT), trace element solutions, phosphate loads, and influent loads were investigated, and the bacteriochlorophyll, carotenoid, and coenzyme Q10 concentrations were determined. The community structure and quantity of Rhodopseudomonas in the systems were determined using a high-sequencing technique and quantitative polymerase chain reaction technique. The long-term results indicated that phosphate was the limiting factor for treating HORW in the PNSB reactor. The soluble chemical oxygen demand (SCOD) removal rates were 67.03% and 85.26% without and with phosphate added, respectively, and the NH4+-N removal rates were 32.18% and 89.22%, respectively. The NO3−-N concentration in the effluent was stable at 0–3 mg/L with or without phosphate added. Adding phosphate increased the Rhodopseudomonas relative abundance and number by 13.21% and 41.61%, respectively, to 57.35% and 8.52 × 106 gene copies/μL, respectively. The SRT was the limiting factor for SCOD removal, and the bacteria concentration was the limiting factor for nitrogen removal. Once the inflow load had been increased, the total nitrogen (TN) removal rate increased as the HRT increased. Maximum TN removal rates of 64.46%, 68.06%, 73.89%, 82.15%, and 89.73% were found at HRT of 7, 10, 13, 16, and 19 d, respectively. The highest bacteriochlorophyll, carotenoid, and coenzyme Q10 concentrations were 2.92, 4.99, and 4.53 mg/L, respectively. This study provided a simple and efficient method for treating HORW and reutilizing resources, providing theoretical support and parameter guidance for the application of Rhodopseudomonas in treating HORW.

Performance of fish sludge solubilization and phototrophic bioconversion by purple phototrophic bacteria for nutrient recovery in aquaponic system

Nutrient recovery from fish sludge in aquaponics is crucial to improve the economic output of a system sustainably and hygienically. Currently, fish sludge is treated using conventional anaerobic and aerobic mineralization, which does not allow the recovery of valuable nutrients in fish wastes. In this study, a two-stage approach (named as solubilization process and phototrophic bioconversion) is proposed to convert fish sludge into mineral nutrients and biomass nutrients using purple phototrophic bacteria (PPB), thereby promoting the growth of plants and fish simultaneously in aquaponics. Anaerobic and aerobic solubilization methods are tested to pretreat the fish sludge, generating substrates for PPB. Anaerobic solubilization yields 2.1 times more soluble chemical oxygen demand (SCOD) and 3.7 times more total volatile fatty acid (t-VFA) from fish sludge compared with aerobic solubilization. The anaerobic solubilization effluent indicates a CODt-VFA/SCOD of 60% and a VFA comprising 13.3% acetate and 49.0% propionate for PPB. The phototrophic bioconversion using anaerobic solubilization effluent under the light-anaerobic condition results in the highest biomass yield (0.94 g CODbiomass/g CODremoved) and the highest PPB dominance (Ectothiorhodospira, 58.7%). The anaerobic solubilization and light-anaerobic phototrophic bioconversion achieves 54.1% of carbon recovery efficiency (CRE) (in terms of COD), as well as 44.8% and 91.3% of nutrient recovery efficiency (NRE) for N and P. A novel multiloop aquaponic system combined with PPB-based nutrient recovery is proposed for the reuse of mineral nutrients and PPB biomass generated from fish sludge.

Role of hydrogen-bond networks on the donor side of photosynthetic reaction centers from purple bacteria.

For the last decades, significant progress has been made in studying the biological functions of H-bond networks in membrane proteins, proton transporters, receptors, and photosynthetic reaction centers. Increasing availability of the X-ray crystal and cryo-electron microscopy structures of photosynthetic complexes resolved with high atomic resolution provides a platform for their comparative analysis. It allows identifying structural factors that are ensuring the high quantum yield of the photochemical reactions and are responsible for the stability of the membrane complexes. The H-bond networks are known to be responsible for proton transport associated with electron transfer from the primary to the secondary quinone as well as in the processes of water oxidation in photosystem II. Participation of such networks in reactions proceeding on the periplasmic side of bacterial photosynthetic reaction centers is less studied. This review summarizes the current understanding of the role of H-bond networks on the donor side of photosynthetic reaction centers from purple bacteria. It is discussed that the networks may be involved in providing close association with mobile electron carriers, in light-induced proton transport, in regulation of the redox properties of bacteriochlorophyll cofactors, and in stabilization of the membrane protein structure at the interface of membrane and soluble phases.

The genome-scale metabolic model for the purple non-sulfur bacterium Rhodopseudomonas palustris Bis A53 accurately predicts phenotypes under chemoheterotrophic, chemoautotrophic, photoheterotrophic, and photoautotrophic growth conditions.

The purple non-sulfur bacterium Rhodopseudomonas palustris is recognized as a critical microorganism in the nitrogen and carbon cycle and one of the most common members in wastewater treatment communities. This bacterium is metabolically extremely versatile. It is capable of heterotrophic growth under aerobic and anaerobic conditions, but also able to grow photoautotrophically as well as mixotrophically. Therefore R. palustris can adapt to multiple environments and establish commensal relationships with other organisms, expressing various enzymes supporting degradation of amino acids, carbohydrates, nucleotides, and complex polymers. Moreover, R. palustris can degrade a wide range of pollutants under anaerobic conditions, e.g., aromatic compounds such as benzoate and caffeate, enabling it to thrive in chemically contaminated environments. However, many metabolic mechanisms employed by R. palustris to breakdown and assimilate different carbon and nitrogen sources under chemoheterotrophic or photoheterotrophic conditions remain unknown. Systems biology approaches, such as metabolic modeling, have been employed extensively to unravel complex mechanisms of metabolism. Previously, metabolic models have been reconstructed to study selected capabilities of R. palustris under limited experimental conditions. Here, we developed a comprehensive metabolic model (M-model) for R. palustris Bis A53 (iDT1294) consisting of 2,721 reactions, 2,123 metabolites, and comprising 1,294 genes. We validated the model using high-throughput phenotypic, physiological, and kinetic data, testing over 350 growth conditions. iDT1294 achieved a prediction accuracy of 90% for growth with various carbon and nitrogen sources and close to 80% for assimilation of aromatic compounds. Moreover, the M-model accurately predicts dynamic changes of growth and substrate consumption rates over time under nine chemoheterotrophic conditions and demonstrated high precision in predicting metabolic changes between photoheterotrophic and photoautotrophic conditions. This comprehensive M-model will help to elucidate metabolic processes associated with the assimilation of multiple carbon and nitrogen sources, anoxygenic photosynthesis, aromatic compound degradation, as well as production of molecular hydrogen and polyhydroxybutyrate.

Unravelling the Roles of Integral Polypeptides in Excitation Energy Transfer of Photosynthetic RC-LH1 Supercomplexes.

Elucidating the photosynthetic processes that occur within the reaction center-light-harvesting 1 (RC-LH1) supercomplexes from purple bacteria is crucial for uncovering the assembly and functional mechanisms of natural photosynthetic systems and underpinning the development of artificial photosynthesis. Here, we examined excitation energy transfer of various RC-LH1 supercomplexes of Rhodobacter sphaeroides using transient absorption spectroscopy, coupled with lifetime density analysis, and studied the roles of the integral transmembrane polypeptides, PufX and PufY, in energy transfer within the RC-LH1 core complex. Our results show that the absence of PufX increases both the LH1 → RC excitation energy transfer lifetime and distribution due to the role of PufX in defining the interaction and orientation of the RC within the LH1 ring. While the absence of PufY leads to the conformational shift of several LH1 subunits toward the RC, it does not result in a marked change in the excitation energy transfer lifetime.

Ammonia impact on the selection of a phototrophic - chemotrophic consortium for polyhydroxyalkanoates production under light-feast / dark-aerated-famine conditions

Phototrophic polyhydroxyalkanoate (PHA) production is an emerging technology for recovering carbon and nutrients from diverse wastewater streams. However, reliable selection methods for the enrichment of PHA accumulating purple phototrophic bacteria (PPB) in phototrophic mixed cultures (PMC) are needed. This research evaluates the impact of ammonia on the selection of a PHA accumulating phototrophic-chemotrophic consortium, towards the enrichment of PHA accumulating PPB. The culture was operated under light-feast/dark-aerated-famine and winter simulated-outdoor conditions (13.2 ± 0.9 °C, transient light, 143.5 W/m2), using real fermented domestic wastewater with molasses as feedstock. Three ammonia supply strategies were assessed: 1) ammonia available only in the light phase, 2) ammonia always present and 3) ammonia available only during the dark-aerated-famine phase. Results showed that the PMC selected under 1) ammonia only in the light and 3) dark-famine ammonia conditions, presented the lowest PHA accumulation capacity during the light period (11.1 % g PHA/g VSS and 10.4 % g PHA/g VSS, respectively). In case 1), the absence of ammonia during the dark-aerated-famine phase did not promote the selection of PHA storing PPB, whereas in case 3) the absence of ammonia during the light period favoured cyanobacteria growth as well as purple sulphur bacteria with increased non-PHA inclusions, resulting in an overall decrease of phototrophic PHA accumulation capacity. The best PHA accumulation performance was obtained with selection under permanent presence of ammonia (case 2), which attained a PHA content of 21.6 % g PHA/g VSS (10.2 Cmmol PHA/L), at a production rate of 0.57 g PHA/L·day, during the light period in the selection reactor. Results in case 2 also showed that feedstock composition impacts the PMC performance, with feedstocks richer in more reduced volatile fatty acids (butyric and valeric acids) decreasing phototrophic performance and leading to acids entering the dark-aerated phase. Nevertheless, the presence of organic carbon in the aerated phase was not detrimental to the system. In fact, it led to the establishment of a phototrophic-chemotrophic consortium that could photosynthetically accumulate a PHA content of 13.2 % g PHA/g VSS (6.7 Cmmol PHA/L) at a production rate of 0.20 g PHA/L·day in the light phase, and was able to further increase that storage up to 18.5 % g PHA/g VSS (11.0 Cmmol PHA/L) at a production rate of 1.35 g PHA/L·day in the dark-aerated period. Furthermore, the light-feast/dark-aerated-famine operation was able to maintain the performance of the selection reactor under winter conditions, unlike non-aerated PMC systems operated under summer conditions, suggesting that night-time aeration coupled with the constant presence of ammonia can contribute to overcoming the seasonal constraints of outdoor operation of PMCs for PHA production.

Selective expression of light-harvesting complexes alters phospholipid composition in the intracytoplasmic membrane and core complex of purple phototrophic bacteria

Phospholipid–protein interactions play important roles in regulating the function and morphology of photosynthetic membranes in purple phototrophic bacteria. Here, we characterize the phospholipid composition of intracytoplasmic membrane (ICM) from Rhodobacter (Rba.) sphaeroides that has been genetically altered to selectively express light-harvesting (LH) complexes. In the mutant strain (DP2) that lacks a peripheral light-harvesting (LH2) complex, the phospholipid composition was significantly different from that of the wild-type strain; strain DP2 showed a marked decrease in phosphatidylglycerol (PG) and large increases in cardiolipin (CL) and phosphatidylcholine (PC) indicating preferential interactions between the complexes and specific phospholipids. Substitution of the core light-harvesting (LH1) complex of Rba. sphaeroides strain DP2 with that from the purple sulfur bacterium Thermochromatium tepidum further altered the phospholipid composition, with substantial increases in PG and PE and decreases in CL and PC, indicating that the phospholipids incorporated into the ICM depend on the nature of the LH1 complex expressed. Purified LH1–reaction center core complexes (LH1–RC) from the selectively expressing strains also contained different phospholipid compositions than did core complexes from their corresponding wild-type strains, suggesting different patterns of phospholipid association between the selectively expressed LH1–RC complexes and those purified from native strains. Effects of carotenoids on the phospholipid composition were also investigated using carotenoid-suppressed cells and carotenoid-deficient species. The findings are discussed in relation to ICM morphology and specific LH complex–phospholipid interactions.

Isolation and characterization of novel potassium-solubilizing purple nonsulfur bacteria from acidic paddy soils using culture-dependent and culture-independent techniques.

The current research as aimed (i) to isolate and select the purple nonsulfur bacteria (PNSB) possessing the potassium-solubilizing ability from acid paddy fields and (ii) to evaluate the ability to release the plant growth-promoting substances (PGPS) of selected PNSB. A total of 35 acid sulfate (AS) soil samples were collected in An Giang province, Vietnam. Then, 70 PNSB strains were isolated from the AS soil samples. In the current study, the isolated strains were screened and selected according to their tolerability to acidic conditions, ability to solubilize potassium, and characteristics of a plant growth promoter on basic isolation media with various incubation conditions. Therein, three strains, TT07.4, AN05.1, and AC04.1, presented the highest potassium solubilization under the microaerobic light (11.8-17.7 mg L-1) and aerobic dark (16.4-24.7 mg L-1) conditions and stresses from Al3+, Fe2+, and Mn2+ toxicity. The selected strains were identified as Rhodopseudomonas pentothenatexigens by the 16S rDNA sequence, with 99% similarity. The selected acidic-resistant strains possessed the traits of biofertilizers under both microaerobic light and aerobic dark conditions, with abilities to fix nitrogen (0.17-6.24; 7.93-11.2 mg L-1); solubilize phosphorus from insoluble compounds with 3.22-49.9 and 9.49-11.2 mg L-1 for Al-P, 21.9-25.8 and 20.2-25.1 mg L-1 for Ca-P, and 10.1-29.8 and 18.9-23.2 mg L-1 for Fe-P; produce 5-aminolevulinic acid (0.63-3.01; 1.19-6.39 mg L-1), exopolymeric substances (0.14-0.76; 0.21-0.86 mg L-1), indole-3-acetic acid (12.9-32.6; 13.6-17.8 mg L-1), and siderophores (28.4-30.3; 6.15-10.3%). The selected potassium-solubilizing strains have a great potential to apply in liquid form into rice seed and solid form in AS soils to supply nutrients and PGPS for enhancing rice growth and grain yield.