Reductive tricarboxylic acid cycle enzymes and reductive amino acid synthesis pathways contribute to electron balance in a Rhodospirillum rubrum Calvin-cycle mutant.

Purple non-sulfur bacteria (PNSB) use light for energy and organic substrates for carbon and electrons when growing photoheterotrophically. This lifestyle generates more reduced electron carriers than are required for biosynthesis, even during consumption of some of the most oxidized organic substrates like malate and fumarate. Reduced electron carriers not used in biosynthesis must still be oxidized for photoheterotrophic growth to occur. Diverse PNSB commonly rely on the CO2-fixing Calvin cycle to oxidize reduced electron carriers. Some PNSB also produce H2 or reduce terminal electron acceptors as alternatives to the Calvin cycle. Rhodospirillum rubrum Calvin-cycle mutants defy this trend by growing phototrophically on malate or fumarate without H2 production or access to terminal electron acceptors. We used 13C-tracer experiments to examine how a Rs. rubrum Calvin-cycle mutant maintains electron balance under such conditions. We detected the reversal of some tricarboxylic acid cycle enzymes, carrying reductive flux from malate or fumarate to αKG. This pathway and the reductive synthesis of αKG-derived amino acids are likely important for electron balance, as supplementing the growth medium with αKG-derived amino acids prevented Rs. rubrum Calvin-cycle-mutant growth unless a terminal electron acceptor was provided. Flux estimates also suggested that the Calvin-cycle mutant preferentially synthesized isoleucine using the reductive threonine-dependent pathway instead of the less-reductive citramalate-dependent pathway. Collectively, our results suggest that alternative biosynthetic pathways can contribute to electron balance within the constraints of a relatively constant biomass composition.

Effects of combined magnetic fields on bacteria Rhodospirillum rubrum VKM B-1621.

Bacteria are the simplest model of living organisms and thus are a convenient object for magnetobiological research. This paper describes some effects of combined magnetic fields (CMFs) on the bacterium Rhodospirillum rubrum strain VKM B-1621, which is not a pathogen but was selected due to its wide spectrum of growth abilities. The authors chose magnetic field-resonant phosphorus and iron (Fe3+ ) because P-containing biochemical compounds (standard abbreviations PP1 , AMP, ADP, ATP) provide energy flows in bacteria while iron could take part in formation of magnetosensitive intracellular inclusions. CMFs were produced by interaction of a geomagnetic field (ВDС ) and an alternating electromagnetic field (ВАС ), which were similar in their intensities. Their magnetic characteristics were as follows: (CMF-1) ВDC = 46.80 µÐ¢, ВАС = 86.11 µT, f = 807.0 Hz; (CMF-2) ВDC = 46.80 µÐ¢, ВАС = 86.11 µT, f = 38.3 Hz; that is, the frequencies of applied alternating electromagnetic fields coincided with cyclotron frequencies of phosphorus or ferric ions, respectively. The blank variants were exposed to the geomagnetic field. The CMFs increased bacterial consumption of dissolved iron as measured by residual concentrations of iron in the medium (P > 99%). An increase of bacterial nitrate reduction in the CMFs was statistically insignificant (P > 90%) when measured by residual concentrations of nitrate. Application of CMFs can influence bacterial activity and metabolism. Bioelectromagnetics. 2018;39:485-490, 2018. © 2018 Wiley Periodicals, Inc.

Shedding light on selenium biomineralization: proteins associated with bionanominerals.

Selenium-reducing microorganisms produce elemental selenium nanoparticles with particular physicochemical properties due to an associated organic fraction. This study identified high-affinity proteins associated with such bionanominerals and with nonbiogenic elemental selenium. Proteins with an anticipated functional role in selenium reduction, such as a metalloid reductase, were found to be associated with nanoparticles formed by one selenium respirer, Sulfurospirillum barnesii.

Tobacco biomass hydrolysate enhances coenzyme Q10 production using photosynthetic Rhodospirillum rubrum.

Coenzyme Q10 (CoQ10), a potent antioxidative dietary supplement, was produced using a photosynthetic bacteria Rhodospirillum rubrum ATCC 25852 by submerged fermentation supplemented with tobacco biomass hydrolysate (TBH) in comparison with media supplemented with hydrolysates prepared with alfalfa (ABH) or spinach (SBH). Growth medium supplemented with 20% (v/v) TBH was found favorable with regard to cell density and CoQ10 concentration. The stimulation effects on cell growth (shortened lag phase, accelerated exponential growth, and elevated final cell concentration) and CoQ10 production (enhanced specific CoQ10 content per unit cell weight) could be attributed to the presence of solanesol, the precursor of CoQ10, in the tobacco biomass. The final yield of CoQ10 reached 20.16 mg/l in the fermentation medium supplemented with 20% TBH.

Rhodospirillum rubrum: utilization of condensed corn solubles for poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) production.

AIMS: This study sought to develop a less expensive medium for growth of the polyhydroxyalkanoate-producing bacterium Rhodospirillum rubrum from the ethanol production coproduct, condensed corn solubles (CCS). METHODS AND RESULTS: Small-scale trials using R. rubrum were performed in aerated or anaerobic stoppered serum bottles filled with media. The CCS (240 g l(-1)) achieved a maximum cell density and growth rate comparable with the defined supplemented malate-ammonium medium (mSMN) or tryptic soy broth. Microaerophilic solubles medium cultures exhibited significantly higher maximum cell densities and growth rates than did strictly anaerobic cultures; while illumination, nickel or biotin addition had no effect. Growth of R. rubrum in a pH controlled bioreactor was significantly better in CCS (240 g l(-1)) than in mSMN medium and supported production of 0.36% (cell dry weight) poly-(3-hydroxybutyrate-Co-3-hydroxyvalerate) after 24 h. CONCLUSIONS: A CCS medium was devised that supported R. rubrum growth for biopolymer production as effective as the defined medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that a more economical medium can be developed for biopolymer production using a low value coproduct from ethanol production. The impact is that this inexpensive solubles medium may make it more economical to produce the biopolymer on a commercial scale.

Factors affecting hydrogen production from cassava wastewater by a co-culture of anaerobic sludge and Rhodospirillum rubrum.

Series of batch experiments were used to investigate the effects of environmental factors, i.e., total nitrogen and total phosphorus concentrations, initial pH, illumination pattern and stirring conditions on hydrogen production from cassava wastewater by a co-culture of anaerobic sludge and Rhodospirillum rubrum. The maximum of the hydrogen yield of 150.46 and 340.19 mL g-COD(-1) was obtained at the total nitrogen and total phosphorus concentrations of 0.2 and 0.04 M, respectively. An effect of initial pH was investigated at COD:N:P ratio of 100:10:1. Results indicated that an optimum initial pH for hydrogen production was pH 7 with a high hydrogen yield of 158.78 mL g-COD(-1) was obtained. No significantly different (p < 0.05) in the effect of illumination pattern (24 h of light and 12 h dark/light cycle) on hydrogen production were observed under continuous-illumination and periodic-illumination with hydrogen yield of 131.84 and 126.92 mL g-COD(-1), respectively. Therefore, a periodic-illumination was applicable in hydrogen fermentation due to its cost-effective. Hydrogen fermentation with a stirring at 100 rpm provided more effective hydrogen production (164.83 mL g-COD(-1)) than static-fermentation (93.93 mL g-COD(-1)). The major soluble products from hydrogen fermentation were acetic and butyric acids, in the ranges of 28.33-48.30 and 35.23-66.07%, respectively, confirming an ability of a co-culture to produce hydrogen from cassava wastewater.

Similarities between the abiotic reduction of selenite with glutathione and the dissimilatory reaction mediated by Rhodospirillum rubrum and Escherichia coli.

Various mechanisms have been proposed to explain the biological dissimilatory reduction of selenite (SeO3(2-)) to elemental selenium (Se(o)), although none is without controversy. Glutathione, the most abundant thiol in the eukaryotic cells, the cyanobacteria, and the alpha, beta, and gamma groups of the proteobacteria, has long been suspected to be involved in selenium metabolism. Experiments with the phototrophic alpha proteobacterium Rhodospirillum rubrum showed that the rate of selenite reduction was decreased when bacteria synthesized lower than normal levels of glutathione, and in Rhodobacter sphaeroides and Escherichia coli the reaction was reported to induce glutathione reductase. In the latter organism superoxide dismutase was also induced in cells grown in the presence of selenite, indicating that superoxide anions (O2-) were produced. These observations led us to investigate the abiotic (chemical) reduction of selenite by glutathione and to compare the features of this reaction with those of the reaction mediated by R. rubrum and E. coli. Our findings imply that selenite was first reduced to selenodiglutathione, which reached its maximum concentration within the 1st min of the reaction. Formation of selenodiglutathione was paralleled by a rapid reduction of cytochrome c, a known oxidant for superoxide anions. Cytochrome c reduction was inhibited by superoxide dismutase, indicating that O2- was the source of electrons for the reduction. These results demonstrated that superoxide was produced in the abiotic reduction of selenite with glutathione, thus lending support to the hypothesis that glutathione may be involved in the reaction mediated by R. rubrum and E. coli. The second phase of the reaction, which led to the formation of elemental selenium (Se(o)), developed more slowly. Se(o) precipitation reached a maximum within 2 h after the beginning of the reaction. Secondary reactions leading to the degradation of the superoxide significantly decreased the yield of Se(o) in the abiotic reaction compared with that of the bacterially mediated selenite reduction. Abiotically formed selenium particles showed the same characteristic orange-red color, spherical structure, and size as particles produced by R. rubrum, again providing support for the hypothesis that glutathione is involved in the reduction of selenite to elemental selenium in this organism.

Reconstruction of photosynthetic, cyclic electron transport system from photoreaction unit, ubiquinone-10 protein, cytochrome c2 and polar lipids purified from Rhodospirillum rubrum.

It was previously reported that in chromatophores of Rhodospirillum rubrum, reaction center, which consists of three kinds of protein (Mm, about 78K), is a small fragment of a large protein complex (PRU; photoreaction unit), which contains six other kinds of protein including light-harvesting bacteriochlorophyll protein, has Mm of about 700K and is free of phospholipid [J. Biochem. 86, 1211-1224 (1979); 94, 1815-1826 (1983(]. In the present study, the photosynthetic, cyclic electron transport system sensitive to antimycin A was effectively reconstructed by incubating 60 nM PRU (which contained 1 mol of reaction center and 2 mol of ubiquinone-10 per mol) with 300 nM each of oxidized ubiquinone-10 protein, reduced cytochrome c2 and lipoamino acid (which were all purified from Rhodospirillum rubrum) in the presence of low concentrations of cholate and deoxycholate (pH 8.0). In the light, the cytochrome was oxidized while the quinone was reduced. The oxidation and reduction each progressed rapidly at first, then slowly, reaching maxima (steady states) 1-2 min after the light had been turned on. At the steady states, 30% of the cytochrome was oxidized while 11% of the total quinone was reduced. When the light was turned off, the original oxidation-reduction states of the cytochrome and quinone were restored at rapid rates initially then at slow rates. Antimycin A stimulated the slow rates in the light-on state and depressed them in the light-off state, but did not influence the fast rates. Ubiquinone-10 protein was required for the antibiotic-sensitive, slow oxidation reactions. This indicates that the slow rates were due to cyclic electron transport. Cytochrome c2 was tightly bound to PRU at a molar ratio of 1:1. This cytochrome as well as the quinone bound to PRU was responsible for the fast rates. PRU had other sites able to bind cytochrome c2 and ubiquinone-10 protein with Km of 0.4 and 0.1 microM, respectively. Of the polar lipids tested, lipoamino acid was the most effective for reconstruction, and its effect was maximal at 300 nM, which is far below its critical micelle concentration.

Regulation of nitrogenase synthesis in intact cells of Rhodospirillum rubrum: inactivation of nitrogen fixation by ammonia, L-glutamine and L-asparagine.

The synthesis of nitrogenase by intact cells of Rhodospirillum rubrum was repressed in N-free media supplemented with L-glutamine or L-asparagine, but was unaffected by the presence of L-glutamate, L-aspartate or L-histidine. Specific activities attained by cultures in supplemented media maintained under Ar-CO2 were 2 to 3 times higher than those in N-free medium under N2-CO2. A loss in total activity occurred both in cultures growing with N2 after maximum activity had been reached, and in cultures maintained under Ar when the gas phase was changed to N2. There was a rapid loss in nitrogen-fixing activity when low concentrations of NH4+, L-glutamine or L-asparagine were added to cultures with high activities, but this could be recovered in the absence of demonstrable protein synthesis. During growth, the degree of inactivation brought about by 0-5 mM-inactivator increased to 80 to 90%, and NH4+ excreted into the medium reached a maximum concentration towards the end of exponential growth.