Bacterium Rhodopseudomonas Palustris Research Articles

Evaluating the Sulfur Oxidation Capability of a Rhodopseudomonas palustris Strain by Gene and Enzyme Analyses for Potential Applications in Environmental Bioremediation

Sulfur-oxidizing bacteria are capable of oxidizing various sulfur compounds including sulfide and thiosulfate in the environment. Hydrogen sulfide (H2S), a product of anaerobic decomposition, is highly corrosive. With the typical smell of rotten eggs, the toxic gas H2S causes many harmful effects on the environment and human health. Sulfide:quinone oxidoreductase enzyme (SQR) produced by sulfur-oxidizing bacteria plays an important role in the sulfide oxidation process, contributing to minimizing sulfide toxicity in the environment. In this study, the presence of sqr gene and SQR enzyme in the photosynthetic purple bacterium Rhodopseudomonas palustris (R. palustris) isolated from domestic wastewater was determined by gene amplification (PCR) using specific primer pairs following by sequencing and denaturing SDS-PAGE method combined with sulfide oxidase assay that measures BaSO4 turbidity, respectively. The results showed that the sqr gene has 1254 bp in length, encoding for the SQR protein containing 417 amino acids with a molecular weight of 45.87 kDa and locating on the cell membrane. The enzyme operated optimally under the condition of pH 6.5 and temperature at 30 °C with sulfide oxidation activity recorded up to 20-21 U/ml. The study has initially shown the ability of the R. palustris bacterial strain to oxidize sulfide for further research using sulfur-oxidizing bacteria applied in environmental treatment.

Photocatalytic scaffolds enhance anticancer performances of bacterial consortium AUN

Microbial metabolism and function are regulated by ambient microenvironment, yet no functional microporous materials are known to regulate anticancer activity of complex bacterial systems. Here we show that a simple scaffold-mediated bacterial culture is useful for enhancing the anticancer therapeutic abilities of a microbial consortium designated as AUN and composed of intratumoural bacteria Proteus mirabilis (A-gyo) and purple photosynthetic bacteria Rhodopseudomonas palustris (UN-gyo). Our discovery of the mechanism by which porous scaffolds influence the bacterial activities of AUN will facilitate further designs of artificial scaffold material for effective treatment against drug-resistant, triple-negative breast cancer models. A polydimethylsiloxane composite artificial scaffold encapsulating light-activatable TiO2 particularly enhanced the therapeutic outcomes of AUN in syngeneic and orthotopic models. These strong anticancer efficacies were synergistically expressed in tumours because of the potent oncolytic ability of AUN with the help of various immune cells. AUN administration to canines showed well tolerability in an exploratory study.

Trace metal interaction with thermophilic phototrophic anaerobic bacterium Chloroflexus aurantiacus

Towards improving the knowledge of possible paleo-microorganisms interaction with trace metals (micro-nutrients and toxicants), we studied adsorption of Mn, Zn, Sr, Cd, and Pb onto modern Chloroflexus aurantiacus, thermophilic anoxygenic phototrophic bacterium which could be highly abundant in the Precambiran aquatic environments. Acid-base surface titrations allowed quantifying the number of proton-active surface groups, whereas non-electrostatic linear programming method (LPM) was used to assess the surface site concentrations and adsorption reaction constants between divalent cations (Zn, Mb, Sr, Cd, Pb) and bacterial surface, based on results of pH-dependent adsorption edge and constant-pH ‘langmuirian’ adsorption experiments. The total proton/hydroxyl binding site number of Chl. aurantiacus surfaces was sizably lower than that of other phototrophic anaerobic bacteria studied previously using similar experimental and modeling approach. Divalent metals exhibited a decreasing order of adsorption affinity (Pb > Cd ≥ Zn ≥ Mn > Sr), which reflected the order of cation hydrolysis and was similar to adsorption order on other phototrophic bacteria. At the same time, adsorption of Zn increased with increasing of temperature, from 4 °C to 60 °C and was stronger under light compared to the darkness. This suggested some active metabolic control involved in this metal interaction with bacterial surfaces. Overall, Chl. aurantiacus exhibited trace metal adsorption parameters (site number and binding constants) which were lower compared to other anoxygenic phototrophic bacteria (Rhodopseudomonas palustris; Rhodobacter blasticus) and cyanobacteria. This may reflect different bioavailability of trace metals in the paleo-ocean, given that thermophilic Chl. aurantiacus are among the oldest phototrophs on the planet.

Light-driven extracellular electron transfer accelerates microbiologically influenced corrosion by Rhodopseudomonas palustris TIE-1

This study investigates the microbiologically influenced corrosion (MIC) of X80 steel accelerated by the phototrophic bacterium Rhodopseudomonas palustris TIE-1. The photorespiration plays a key role in promoting extracellular electron transfer (EET)-induced MIC. In the early corrosion stage, unstable localized corrosion dominated in the dark, while intense diffusion-controlled corrosion occurs in light. Compared to the sterile anaerobic medium, R. palustris TIE-1 accelerated corrosion of X80 steel, with a significantly higher corrosion rate under light conditions, approximately three times that of dark conditions. Inhibition of photosynthetic electron transfer or cessation of photostimulation resulted in pronounced reduction in the corrosion rate.

Growth of electroautotrophic microorganisms using hydrovoltaic energy through natural water evaporation

It has been previously shown that devices based on microbial biofilms can generate hydrovoltaic energy from water evaporation. However, the potential of hydrovoltaic energy as an energy source for microbial growth has remained unexplored. Here, we show that the electroautotrophic bacterium Rhodopseudomonas palustris can directly utilize evaporation-induced hydrovoltaic electrons for growth within biofilms through extracellular electron uptake, with a strong reliance on carbon fixation coupled with nitrate reduction. We obtained similar results with two other electroautotrophic bacterial species. Although the energy conversion efficiency for microbial growth based on hydrovoltaic energy is low compared to other processes such as photosynthesis, we hypothesize that hydrovoltaic energy may potentially contribute to microbial survival and growth in energy-limited environments, given the ubiquity of microbial biofilms and water evaporation conditions.

Improving nitrogen content in soil and lemon balm (Melissa officinalis L.) yield by purple nonsulfur bacteria Rhodopseudomonas palustris in two consecutive seasons.

The study was conducted to assess the effects of nitrogen (N)-fixing purple nonsulfur bacteria (PNSB) Rhodopseudomonas palustris TLS06, VNW02, VNW64, and VNS89 on soil fertility, N uptake, essential oil (EO) content, growth, and yield of lemon balm. The experiment followed a completely randomized block design with 9 treatments and 3 replications. The treatments consisted of (i) applying 100% N as the recommended fertilizer rate (RFR), (ii) applying 85% N as RFR, (iii) applying 70% N as RFR, (iv) applying 55% N as RFR, (v) the treatment ii combined with N-PNSB, (vi) the treatment iii combined with N-PNSB, (vii) the treatment iv combined with N-PNSB, (viii) 0% as RFR combined with N-PNSB, and (ix) 0% N as RFR. The results showed that applying N-PNSB increased the plant height, and the number of primary branches in both seasons. In addition, the treatment without N fertilizer combined with N-PNSB increased stem leaf biomass by 41.2 and 50.3% in both seasons as compared with the treatment without neither N fertilizer nor N-PNSB. For soil properties, among treatments without N fertilizer, the treatment with N-PNSB increased concentrations of NH4+, soluble P, and exchangeable K+ by 41.3, 41.4, and 26.8%, respectively, as compared with the treatment without N-PNSB at the end of the second season. Applying 85% N as RFR combined with N-PNSB had a greater yield by 5.78-11.8% as compared with the treatment with 100% N as RFR, and a greater EO content by 23% as compared with the treatment with 85% N as RFR.

Inoculants associated with fertilizers: agronomic increases in bean, soybean and lettuce crops

The objective of the study was to prove the efficiency of inoculation associated with synthetic fertilizers in the agronomic potential of bean, soybean and lettuce. Experiments were carried out in a randomized block design to evaluate the efficiency of inoculation associated with synthetic fertilizers in bean, soybean and lettuce crops. The experimental design used was randomized blocks organized in a factorial scheme, with six replications. In beans, ten treatments were evaluated in four environments. In soybean cultivation, 13 treatments were evaluated in four environments. While in lettuce, six treatments were evaluated in five environments. In each crop, the agronomic and nutritional characteristics were evaluated. Inoculation associated with fertilizers enhances the agronomic and nutritional performance of beans, soybeans and lettuce. Rhodopseudomonas palustris combined with phosphorus and nitrogen fertilization improves bean grain yield. Bradyrhizobium japonicum, Azospirillum brasiliense and Pseudomonas fluorescensare crucial for soybean. Addition of the bacterium Rhodopseudomonas palustris maximized the lettuce biomass.

Overexpression of ATP Synthase Subunit Beta (Atp2) Confers Enhanced Blast Disease Resistance in Transgenic Rice.

Previous research has shown that the pathogenicity and appressorium development of Magnaporthe oryzae can be inhibited by the ATP synthase subunit beta (Atp2) present in the photosynthetic bacterium Rhodopseudomonas palustris. In the present study, transgenic plants overexpressing the ATP2 gene were generated via genetic transformation in the Zhonghua11 (ZH11) genetic background. We compared the blast resistance and immune response of ATP2-overexpressing lines and wild-type plants. The expression of the Atp2 protein and the physiology, biochemistry, and growth traits of the mutant plants were also examined. The results showed that, compared with the wild-type plant ZH11, transgenic rice plants heterologously expressing ATP2 had no significant defects in agronomic traits, but the disease lesions caused by the rice blast fungus were significantly reduced. When infected by the rice blast fungus, the transgenic rice plants exhibited stronger antioxidant enzyme activity and a greater ratio of chlorophyll a to chlorophyll b. Furthermore, the immune response was triggered stronger in transgenic rice, especially the increase in reactive oxygen species (ROS), was more strongly triggered in plants. In summary, the expression of ATP2 as an antifungal protein in rice could improve the ability of rice to resist rice blast.

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.

Effect of Rhodopseudomonas palustris as a Bioremediation on Reducing Ammonia and Nitrite Levels of Catfish Nursery Pond Water in District Kebakkramat Regency Karanganyar

Catfish nursery pond water generally contains ammonia and nitrites. Ammonia and nitrite in this fish farming system start with uneaten catfish feed residue, feces, and metabolic products that enter the waters. Utilization of catfish feed which contains high protein can absorbed by fish as much as 20-30% of nutrients. The rest will be excreted in the form of feces. Feces and food scraps that settle in this water contain ammonia and nitrites, which affect the quality of the water. Efforts to maintain water quality are by reducing and disposing of pond water in the river. This is categorized as water pollution. Efforts to control this water pollution are the technique of bioremediation. The aim of bioremediation techniques is to restore the ecological balance in waters by utilizing microorganisms so that a process of degradation of harmful and complex compounds occurs into harmless compounds. The microorganisms used in this study were bacteria Rhodopseudomonas palustris which can to oxidize and decompose organic matter effectively reducing water quality. This research was conducted to examine the effect of bacterial efficiency Rhodopseudomonas palustris as a bioremediator for catfish pond water waste based on reducing levels of ammonia and nitrite. Testing the ammonia and nitrite levels using a spectrophotometric method with different wavelengths. Observations in this test were carried out for three days under anaerobic conditions and light at a temperature of 33°C. Observations showed that bacteria Rhodopseudomonas palustris had an effect on reducing nitrite levels 12.65% greater than the control. While the addition of bacteria to the observation of ammonia levels did not have a decreasing effect as indicated by the value F <F crit.

Stability and biomineralization of cadmium sulfide nanoparticles biosynthesized by the bacterium Rhodopseudomonas palustris under light

Cadmium (Cd) pollution is regarded as a potent problem due to its hazard risks to the environment, making it crucial to be removed. Compared to the physicochemical techniques (e.g., adsorption, ion exchange, etc.), bioremediation is a promising alternative technology for Cd removal, due to its cost-effectiveness, and eco-friendliness. Among them, microbial-induced cadmium sulfide mineralization (Bio-CdS NPs) is a process of great significance for environmental protection. In this study, microbial cysteine desulfhydrase coupled with cysteine acted as a strategy for Bio-CdS NPs by Rhodopseudomonas palustris. The synthesis, activity, and stability of Bio-CdS NPs-R. palustris hybrid was explored under different light conditions. Results show that low light (LL) intensity could promote cysteine desulfhydrase activities to accelerate hybrid synthesis, and facilitated bacterial growth by the photo-induced electrons of Bio-CdS NPs. Additionally, the enhanced cysteine desulfhydrase activity effectively alleviated high Cd-stress. However, the hybrid rapidly dissolved under changed environmental factors, including light intensity and oxygen. The factors affecting the dissolution were ranked as follows: darkness/microaerobic ≈ darkness/aerobic < LL/microaerobic < high light (HL)/microaerobic < LL/aerobic < HL/aerobic. The research provides a deeper understanding of Bio-CdS NPs-bacteria hybird synthesis and its stability in Cd-polluted water, allowing advanced bioremediation treatment of heavy metal pollution in water.

Quorum Sensing Is Required for the Colony Establishment of a Plant Phyllosphere Bacterium Rhodopseudomonas palustris Strain GJ-22.

The phyllosphere presents a hostile environment for many biocontrol agents; however, it is as significant as is the rhizosphere for plant health. Deploying biocontrol bacteria into the phyllosphere can efficiently suppress diseases; however, the lack of knowledge on the phyllosphere adaptive traits of biocontrol bacteria poses challenges. In this study, we demonstrated that Rhodopseudomonas palustris GJ-22 colonizes the phyllosphere by forming cell aggregates. The formation of cell aggregates required the production of exopolysaccharides (EPS), which depended on the function of the rpaI-rpaR quorum sensing (QS) mechanism, mediated by the signaling molecule p-coumaroyl-HSL (pC-HSL). The mutation of the EPS biosynthesis gene Exop1 or the signaling molecule biosynthesis gene rpaI compromised the ability of GJ-22 to tolerate reactive oxygen intermediates (ROIs), such as H2O2, in vitro and to form cell aggregates in vivo. Collectively, the results revealed that QS mediates EPS production and consequently leads to bacterial cell aggregation. IMPORTANCE Quorum sensing is used by various bacteria for coordinating the multiplication of bacterial cells in a group and for modulating the behaviors of surrounding microbial species. Host plants can benefit from this interspecies modulation, as it can disrupt the QS circuits of pathogenic bacteria. Some N-acyl homoserine lactone- (AHL-) producing bacteria that were introduced into the phyllosphere as biocontrol agents may establish AHL-based crosstalk with indigenous microbes to steer the nutritional and microecological conditions toward their own and the host plant's benefit. Here, we showed that biocontrol bacteria introduced into the phyllosphere require a functioning QS circuit to establish colonies and suppress pathogens. Furthermore, our findings provoked a broader investigation into the role of the QS circuit in beneficial microorganism-plant interactions.

Cytochrome P450-catalyzed oxidation of halogen-containing substrates

Cytochrome P450 (CYP) enzymes are heme-thiolate monooxygenases which catalyze the oxidation of aliphatic and aromatic C-H bonds and other reactions. The oxidation of halogens by cytochrome P450 enzymes has also been reported. Here we use CYP199A4, from the bacterium Rhodopseudomonas palustris strain HaA2, with a range of para-substituted benzoic acid ligands, which contain halogens, to assess if this enzyme can oxidize these species or if the presence of these electronegative atoms can alter the outcome of P450-catalyzed reactions. Despite binding to the enzyme, there was no detectable oxidation of any of the 4-halobenzoic acids. CYP199A4 was, however, able to efficiently catalyze the oxidation of both 4-chloromethyl- and 4-bromomethyl-benzoic acid to 4-formylbenzoic acid via hydroxylation of the α‑carbon. The 4-chloromethyl substrate bound in the enzyme active site in a similar manner to 4-ethylbenzoic acid. This places the benzylic α‑carbon hydrogens in an unfavorable position for abstraction indicating a degree of substrate mobility must be possible within the active site. CYP199A4 catalyzed oxidations of 4-(2′-haloethyl)benzoic acids yielding α-hydroxylation and desaturation metabolites. The α-hydroxylation product was the major metabolite. The desaturation pathway is significantly disfavored compared to 4-ethylbenzoic acid. This may be due to the electron-withdrawing halogen atom or a different positioning of the substrate within the active site. The latter was demonstrated by the X-ray crystal structures of CYP199A4 with these substrates. Overall, the presence of a halogen atom positioned close to the heme iron can alter the binding orientation and outcomes of enzyme-catalyzed oxidation.

ATP Is a Major Determinant of Phototrophic Bacterial Longevity in Growth Arrest.

How bacteria transition into growth arrest as part of stationary phase has been well-studied, but our knowledge of features that help cells to stay alive in the following days and weeks is incomplete. Most studies have used heterotrophic bacteria that are growth-arrested by depletion of substrates used for both biosynthesis and energy generation, making is difficult to disentangle the effects of the two. In contrast, when grown anaerobically in light, the phototrophic bacterium Rhodopseudomonas palustris generates ATP from light via cyclic photophosphorylation, and builds biomolecules from organic substrates, such as acetate. As such, energy generation and carbon utilization are independent from one another. Here, we compared the physiological and molecular responses of R. palustris to growth arrest caused by carbon source depletion in light (energy-replete) and dark (energy-depleted) conditions. Both sets of cells remained viable for 6 to 10 days, at which point dark-incubated cells lost viability, whereas light-incubated cells remained fully viable for 60 days. Dark-incubated cells were depleted in intracellular ATP prior to losing viability, suggesting that ATP depletion is a cause of cell death. Dark-incubated cells also shut down measurable protein synthesis, whereas light-incubated cells continued to synthesize proteins at low levels. Cells incubated in both conditions continued to transcribe genes. We suggest that R. palustris may completely shut down protein synthesis in dark, energy-depleted, conditions as a strategy to survive the nighttime hours of day/night cycles it experiences in nature, where there is a predictable source of energy in the form of sunlight only during the day. IMPORTANCE The molecular and physiological basis of bacterial longevity in growth arrest is important to investigate for several reasons. Such investigations could improve treatment of chronic infections, advance use of non-growing bacteria as biocatalysts to make high yields of value-added products, and improve estimates of microbial activities in natural habitats, where cells are often growing slowly or not at all. Here, we compared survival of the phototrophic bacterium Rhodopseudomonas palustris under conditions where it generates ATP (incubation in light), and where it does not generate ATP (incubation in dark) to directly assess effects of energy depletion on long-term viability. We found that ATP is important for long-term survival over weeks. However, R. palustris survives 12 h periods of ATP depletion without loss of viability, apparently in anticipation of sunrise and restoration of its ability to generate ATP. Our work suggests that cells respond to ATP depletion by shutting down protein synthesis.

Mode of autophosphorylation in bacteriophytochromes RpBphP2 and RpBphP3.

Phytochromes are red-light photoreceptors that regulate a wide range of physiological processes in plants, fungi and bacteria. Canonical bacteriophytochromes are photosensory histidine kinases that undergo light-dependent autophosphorylation, thereby regulating cellular responses to red light via two-component signaling pathways. However, the molecular mechanism of kinase activation remains elusive for bacteriophytochromes. In particular, the directionality of autophosphorylation is still an open question in these dimeric photoreceptor kinases. In this work, we perform histidine kinase assays on two tandem bacteriophytochromes RpBphP2 and RpBphP3 from the photosynthetic bacterium Rhodopseudomonas palustris. By examining the kinase activities of full-length bacteriophytochromes and two loss-of-function mutants under different light conditions, we demonstrate that RpBphP2 and RpBphP3 undergo light-dependent trans-phosphorylation between protomers in both homodimeric and heterodimeric forms. We have further determined the crystal structure of the histidine kinase domains of RpBphP2 at 3.19Å resolution. Based on structural comparisons and homology modeling, we also present a model to account for the actions of trans-autophosphorylation in bacteriophytochromes.

Dissolution rate and growth performance reveal struvite as a sustainable nutrient source to produce a diverse set of microbial protein

To provide for the globally increasing demand for proteinaceous food, microbial protein (MP) has the potential to become an alternative food or feed source. Phosphorus (P), on the other hand, is a critical raw material whose global reserves are declining. Growing MP on recovered phosphorus, for instance, struvite obtained from wastewater treatment, is a promising MP production route that could supply protein-rich products while handling P scarcity. The aim of this study was to explore struvite dissolution kinetics in different MP media and characterize MP production with struvite as sole P-source. Different operational parameters, including pH, temperature, contact surface area, and ion concentrations were tested, and struvite dissolution rates were observed between 0.32 and 4.7 g P/L/d and a solubility between 0.23 and 2.22 g P-based struvite/L. Growth rates and protein production of the microalgae Chlorella vulgaris and Limnospira sp. (previously known as Arthrospira sp.), and the purple non‑sulfur bacterium Rhodopseudomonas palustris on struvite were equal to or higher than growth on conventional potassium phosphate. For aerobic heterotrophic bacteria, two slow-growing communities showed decreased growth on struvite, while the growth was increased for a third fast-growing one. Furthermore, MP protein content on struvite was always comparable to the one obtained when grown on standard media. Together with the low content in metals and micropollutants, these results demonstrate that struvite can be directly applied as an effective nutrient source to produce fast-growing MP, without any previous dissolution step. Combining a high purity recovered product with an efficient way of producing protein results in a strong environmental win-win.

Characterization of CdSe QDs biosynthesized by a recombinant Rhodopseudomonas palustris

This study proposed a novel approach for the biosynthesis of CdSe quantum dots (QDs) by a recombinant photosynthetic bacteria Rhodopseudomonas palustris with metallothionein expression. The biosynthesis mechanisms and physico-chemical properties of the CdSe QDs were analyzed by various techniques, and its potential application was also investigated. The results showed that the biosynthesized CdSe QDs presented green fluorescence with maximum excitation and emission wavelengths at 460 and 530 nm, respectively. The quantum yield of the biosynthesized CdSe QDs was found out to be 3.85%. They were well-dispersed spherical particles with an average size of 5.37 ± 0.06 nm. Their crystal structure was cubic with the d spacings of 3.1 Å and 2.5 Å, corresponded to the planes of (111) and (220), respectively. The biosynthesis process of CdSe QDs took place in the cytoplasm and may involve some proteins, peptides, lipids or other organic substances covering the surface of CdSe QDs through several specific functional groups, such as hydroxyl or sulfhydryl groups to maintain their stability. The purified CdSe QDs had good biocompatibility and great potential for cellular imaging application. It is concluded that the process may provide a more cost-effective and eco-friendly method not only to recover harmful cadmium in the environment but also to synthesize simultaneously a clean product for various industrial applications.

Combining the microbial agent Rhodopseudomonas palustris strain PSB-06 with fungicides for controlling rice blast

The rice blast disease caused by Magnaporthe oryzae threatens global rice production yields. Tricyclazole and isoprothiolane are widely used fungicides with high activity against rice blast, and our previous study indicated the photosynthetic bacterium Rhodopseudomonas palustris PSB-06 significantly antagonizes rice blast. However the effect of combining these two chemical fungicides with PSB-06 on rice blast control is unclear. Here we test the control effect of photosynthetic bacteria PSB-06 combined with isoprothiolane and tricyclazole on rice blast. The growth of PSB-06 was unaffected by up to 1.25 mg/L of tricyclazole and 0.3 mg/L of isoprothiolane in the photosynthetic medium, indicated the two fungicides have no inhibition on PSB-06. The control efficiency in the field test reached 76.06% when PSB-06 was combined with isoprothiolane. This value was significantly higher than the individual efficiency of PSB-06 (67.99%) and tricyclazole (65.46%) and the combined control efficiency (72.20%) of those two antifungal agents. Our current findings highlighted the potential of combining R. palustris strain PSB-06 with isoprothiolane to control rice blast, providing environmental protection and reducing the use of fungicides.

Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation.

The cytochrome P450 family of monooxygenase enzymes have essential biological roles involving the selective oxidation of carbon-hydrogen bonds. They can also catalyze other important metabolic reactions including desaturation to form alkenes. Currently the factors that control the partition between P450 hydroxylation and desaturation pathways are poorly defined. The CYP199A4 enzyme from the bacterium Rhodopseudomonas palustris HaA2 catalyzes the oxidation of 4-ethyl- and 4-isopropyl- benzoic acids with hydroxylation and desaturation occurring in significant quantities. Here we demonstrate that 4-cyclopropylbenzoic acid is regioselectively hydroxylated by CYP199A4 at the benzylic carbon. In contrast, the oxidation of 4-n-propylbenzoic acid by CYP199A4 results in three major metabolites: an alkene from desaturation and two hydroxylation products at the benzylic (Cα) and Cβ carbons in similar quantities. Extending the length of the alkyl substituent resulted in 4-n-butylbenzoic acid being oxidized at the benzylic position (45%) and desaturated (55%). In contrast, 4-isobutylbenzoic generated very little alkene (5%) but was hydroxylated at the benzylic position (54%) and at the tertiary Cβ position (41%). The oxidation of 4-n-propylbenzoic acid by the F298 V mutant of CYP199A4 occurred with no hydroxylation at Cβ and a significant increase in metabolites arising from desaturation (73%). The X-ray crystal structures of CYP199A4 with each substrate revealed that they bind in the active site with the alkyl substituent positioned over the heme. However, the longer alkylbenzoic acids were bound in a different conformation as was 4-n-propylbenzoic acid in the F298 V mutant. Overall, the changes in metabolite distribution could be ascribed to bond strength differences and the position of the alkyl group relative to the heme.

Extracellular electron uptake for CO2 fixation by Rhodopseudomonas palustris during electro-cultivation in darkness

As a vital part of the global carbon cycle, photosynthesis helps in fixing CO2 to produce diverse biomass. However, over-reliance on optical density results in inadequate photosynthesis under limited light sources. The coupling of extracellular respiration and photosynthetic chain via the quinone pool provides a possibility for electrically driven photosynthesis in darkness, which is not well understood. In this study, CO2 fixation of photosynthetic bacteria Rhodopseudomonas palustris was enhanced in the dark via extracellular electron uptake from the electrode at −0.4 V. The copy number of R. palustris increased by 35 folds during 28 days of operation, accompanied by the increase of ATP content, NADH/NAD+, and NADPH/NADP+ of cells. Especially, the activity of Rubisco, the key enzyme of the Calvin cycle, increased by 28 % during electro-cultivation. Accordingly, the electrochemical activity of R. palustris was found to increase, which might be attributed to the structural modification of protein-like substances due to the enhanced proton-coupled electron transfer (PCET) process in electro-cultivation, which was further confirmed by in situ Fourier transform infrared spectroscopy and kinetic isotope effect tests. This study indicated that extracellular respiration could be electrostimulated via PCET to maintain photosynthesis in R. palustris in the dark.