Photosynthetic Purple Nonsulfur Research Articles

Engineered Mutants of a Marine Photosynthetic Purple Nonsulfur Bacterium with Increased Volumetric Productivity of Polyhydroxyalkanoate Bioplastics.

Polyhydroxyalkanoates (PHAs) are green and sustainable bioplastics that could replace petrochemical synthetic plastics without posing environmental threats to living organisms. In addition, sustainable PHA production could be achieved using marine photosynthetic purple nonsulfur bacteria (PNSBs) that utilize natural seawater, sunlight, carbon dioxide gas, and nitrogen gas for growth. However, PHA production using marine photosynthetic PNSBs has not been economically feasible yet due to its high cost and low productivity. In this work, strain improvement, using genome-wide mutagenesis coupled with high-throughput screening via fluorescence-activated cell sorting, we were able to create Rhodovulum sulfidophilum mutants with enhanced volumetric PHA productivity, with an up to 1.7-fold increase. The best selected mutants (E6 and E6M4) reached the stationary growth phase 1 day faster and accumulated the maximum PHA content 2 days faster than the wild type. Maximizing volumetric PHA productivity before the stationary growth phase is indeed an additional advantage for R. sulfidophilum as a growth-associated PHA producer.

TCA Cycle Replenishing Pathways in Photosynthetic Purple Non-Sulfur Bacteria Growing with Acetate.

Purple non-sulfur bacteria (PNSB) are anoxygenic photosynthetic bacteria harnessing simple organic acids as electron donors. PNSB produce a-aminolevulinic acid, polyhydroxyalcanoates, bacteriochlorophylls a and b, ubiquinones, and other valuable compounds. They are highly promising producers of molecular hydrogen. PNSB can be cultivated in organic waste waters, such as wastes after fermentation. In most cases, wastes mainly contain acetic acid. Therefore, understanding the anaplerotic pathways in PNSB is crucial for their potential application as producers of biofuels. The present review addresses the recent data on presence and diversity of anaplerotic pathways in PNSB and describes different classifications of these pathways.

Optimal iron concentrations for growth-associated polyhydroxyalkanoate biosynthesis in the marine photosynthetic purple bacterium Rhodovulum sulfidophilum under photoheterotrophic condition.

Polyhydroxyalkanoates (PHAs) are a group of natural biopolyesters that resemble petroleum-derived plastics in terms of physical properties but are less harmful biologically to the environment and humans. Most of the current PHA producers are heterotrophs, which require expensive feeding materials and thus contribute to the high price of PHAs. Marine photosynthetic bacteria are promising alternative microbial cell factories for cost-effective, carbon neutral and sustainable production of PHAs. In this study, Rhodovulum sulfidophilum, a marine photosynthetic purple nonsulfur bacterium with a high metabolic versatility, was evaluated for cell growth and PHA production under the influence of various media components found in previous studies. We evaluated iron, using ferric citrate, as another essential factor for cell growth and efficient PHA production and confirmed that PHA production in R. sulfidophilum was growth-associated under microaerobic and photoheterotrophic conditions. In fact, a subtle amount of iron (1 to 2 μM) was sufficient to promote rapid cell growth and biomass accumulation, as well as a high PHA volumetric productivity during the logarithmic phase. However, an excess amount of iron did not enhance the growth rate or PHA productivity. Thus, we successfully confirmed that an optimum concentration of iron, an essential nutrient, promotes cell growth in R. sulfidophilum and also enhances PHA utilization.

Effect of pH on optimization of photofermentative hydrogen production by co-culture of Rhodobacter sphaeroides-NMBL-02 and Bacillus firmus-NMBL-03.

Rhodobacter sphaeroides NMBL-02, photosynthetic purple non sulfur (PNS) bacteria and associated Bacillus firmus NMBL-03 were isolated from water sample collected from 15-20 inches beneath the surface of ponds from Northern region of India in modified Sistrom's media (120 ml) containing 3 g/L malate and 1.2 g/L ammonium sulfate. The isolation was done in air tight serum bottles (120 ml) under tungsten bulb (1.8 kLux light intensity) at 30 oC ± 2 oC. The PNS and heterotrophic bacteria associated with the culture was purified by clonal selection method and characterized by 16S rDNA sequencing. The PNS isolate was identified as Rhodobacter sphaeroides NMBL-02 (ID: 1467407, Accession BANKIT: JN256030) and associated heterotroph as Bacillus firmus NMBL-03 (Gene Bank Accession no.: JN 256029). The effect of initial medium pH on optimization of hydrogen production was investigated in batch process. The maximum hydrogen potential and hydrogen production rate was 2310 ± 55 ml/L and 4.75 ml/L culture/h respectively using glutamate (1.7 mmol/L) as nitrogen source and malate (22.38 mmol/L) as carbon source with 76.39% malate conversion efficiency at initial medium pH 5.0. This co-culture has the ability to produce significant amount of hydrogen in the pH range of 5.0 to 10.0 with 76.39% to 35.71% malate conversion respectively.

Metabolic engineering of Rhodobacter sphaeroides for improved hydrogen production

Rhodobacter sphaeroides 2.4.1 is a facultative photosynthetic purple nonsulfur bacterium that can generate hydrogen by using sunlight and organic carbon sources under anaerobic conditions. Hydrogen is produced by the nitrogenase. To optimize hydrogen production, we combined in a single strain several mutations known to enhance hydrogen production in anoxygenic phototrophic bacteria. The first metabolic engineering approach involved elimination of the major reductive pathways (electron sinks) that compete for electrons with the nitrogenase. A strain containing mutations in the pathways involving hydrogen uptake hydrogenase (hupSL), poly-β-hydroxybutyrate synthesis (phbC), and light harvesting complex-II (pucBA) showed significantly increased hydrogen production. The second approach involved increasing immediate electron flow to nitrogenase via the Rnf complex. This strategy resulted in modest improvements in hydrogen yield. The third approach involved upregulating nitrogenase expression and decoupling its expression from the presence of ammonium in the growth media. Mutations in the nifA gene, encoding the master transcriptional activator of nitrogen fixation genes, were found to be most effective for increasing total hydrogen yield, accelerating hydrogen production, and making this process tolerant to the presence of ammonia in the medium. The combination of beneficial mutations resulted in an additive and superior hydrogen producing strain of R. sphaeroides.

Aspects of the metabolism of hydrogen production by Rhodobacter sphaeroides

Photosynthetic bacteria are favorable candidates for biological hydrogen production due to their high conversion efficiency and versatility in the substrates they can utilize. For large-scale hydrogen production, an integrated view of the overall metabolism is necessary in order to interpret results properly and facilitate experimental design. In this study, a summary of the hydrogen production metabolism of the photosynthetic purple non-sulfur (PNS) bacteria will be presented. Practically all hydrogen production by PNS bacteria occurs under a photoheterotrophic mode of metabolism. Yet results show that under certain conditions, alternative modes of metabolism—e.g. fermentation under light deficiency—are also possible and should be considered in experimental design. Two enzymes are especially critical for hydrogen production. Nitrogenase promotes hydrogen production and uptake hydrogenase consumes hydrogen. Though a wide variety of substrates can be used for growth, only a portion of these is suitable for hydrogen production. The efficiency of a certain substrate depends on factors such as the activity of the TCA cycle, the carbon-to-nitrogen ratio, the reduction-state of that material and the conversion potential of the substrate into alternative metabolites such as PHB. All these individual components of the hydrogen production interact and are subject to strict regulatory controls. An overall scheme for the hydrogen production metabolism is presented.

Photohydrogen production using purple nonsulfur bacteria with hydrogen fermentation reactor effluent

Hydrogen production from waste using photosynthetic bacteria is an attractive methodology. A combination of purple nonsulfur photosynthetic bacteria and anaerobic bacteria is ideal for the efficient conversion of wastewater into hydrogen. In this paper, photohydrogen production using effluent from different hydrogen fermentation reactors was carried out using two strains of photosynthetic purple nonsulfur bacteria. The results indicated that the effluent from the hydrogen fermentation reactors could be used directly for photohydrogen production without aeration or dilution pretreatment. Effluent from the carbohydrate fed hydrogen fermentation reactors is more suitable for photohydrogen production than effluent from a peptone fed reactor. Among the initial dark hydrogen fermentation stage effluents from the three carbohydrate fed reactors (CSTR, ASBR, UASB), CSTR effluent was the most suitable for photohydrogen production.

Identification of the components of a putative cytochrome bc1 complex in Rhodopseudomonas viridis

Chromatophore membranes isolated from the bacteriochlorophyll b-containing, photosynthetic purple nonsulfur bacterium, Rhodopseudomonas viridis, have been shown to contain a Rieske iron-sulfur protein, a cytochrome similar to cytochrome c 1, and also at least one b-type cytochrome. These observations suggest the presence of a previously undetected cytochrome bc 1 complex in this bacterium.

Microflora related to the nitrogen cycle in the tropical paddy soils

Abstract Paddy soils collected in 103 sites of Thailand consisted of surface soil (0-1 cm) and subsurface soil (1-10 cm). Collections were made twice, in the middle of the rainy season and in the middle of the dry season. The counts of 12 groups of microbes related to the nitrogen cycle were determined. In the soils sampled in the rainy season the population level of aerobic bacteria per 1 g of dry soil ranged from 107 to 106. For other microorganisms, the values were as follows: actinomycetes—l04 to 104, anaerobic bacteria—l05, cellulose decomposer—104, denitrifier—105, ammonifier—104 to 104, nitrite oxidizer—103, ammonia oxidizer-105, Clostridium-105 to 104, nitrogen-fixing blue green algae, Azotobacter and purple nonsulfur bacteria-103, respectively. Populations of all 12 groups examined were larger in the oxidized surface soil than in the subsurface soil and, in particular, the populations of aerobic nitrite oxidizer and photosynthetic purple nonsulfur bacteria were more than 4 times higher than those...