Factors affecting the photoproduction of ammonia from dinitrogen and water by the cyanobacterium Anabaena sp. strain ATCC 33047

Synthesis of ammonia from dinitrogen and water by suspensions of Anabaena sp. Strain ATCC 33047 treated with the glutamine synthetase inhibitor L-methionine-D,L-sulfoximine is strictly dependent on light. Under otherwise optimal conditions, the yield of ammonia production is influenced by irradiance, as well as by the density, depth, and turbulence of the cell suspension. The interaction among these factors seems to determine the actual amount of light available to each single cell or filament in the suspension for the photoproduction process. Under convenient illumination, the limiting factor in the synthesis of ammonia seems to be the cellular nitrogenase activity level, but under limiting light conditions the limiting factor could, however, be the assimilatory power required for nitrogen fixation. Photosynthetic ammonia production from atmospheric nitrogen and water can operate with an efficiency of ca. 10% of its theoretical maximum, representing a remarkable process for the conversion of light energy into chemical energy.     

Effect of sewage wastewater treated byNostoc muscorum andAnabaena subcylindrica on the growth ofZea mays andPhaseolus vulgaris

Zea mays (Sakha 321 ) andPhaseolus vulagris (Giza 6) were irrigated with sewage wastewater treated withNostoc muscorum andAnabaena subcylindrica. The results showed that irrigation of plants by untreated or treated wastewater caused stimulation in the measured growth parameters of both plants. In addition pigmentation as well as protein and carbohydrate contents were stimulated. This stimulation was attributed to the presence of high levels of essential nutrients such as nitrogen, phosphorus, and organic matters in wastewater. On the other side, there was a slight inhibitory effect of wastewater on some measured growth parameters of the plants. This effect may be due to the presence of some heavy metals in wastewater.     

Construction of multiple herbicide resistant ammonia excreting strains of cyanobacteriumNostoc muscorum

Summary Machete resistant (Mat ^r), basalin resistant (Bas ^r), 3(3,4 dichlorophenyl)-1,1-dimethyl urea resistant (DCMU ^r), atrazine resistant (Atr ^r) and propanil resistant (Prp ^r) phenotypes ofGloeocapsa sp. were cotransformed toNostoc muscorum at high frequency. Spontaneously occurring mutants of the multiple herbicide resistant transformant containing L-methionine-DL-sulfoximine resistant (Msx ^r), ethylene diamine resistant (Eda ^r), or phosphinothricin resistant (Ppt ^r) glutamine synthetase (GS) showed extracellular liberation of ammonia resulting from fixation of N₂ under photosynthetic conditions. Results suggest a definite role of GS activity in regulation of extracellular ammonia.     

Inactivation by ammonia of the photosynthetic reduction of nitrate in Nostoc muscorum particles.

Ammonia, as well as other uncouplers of photophosphorylation, strongly inhibit the photosynthetic reduction of nitrate by particles of the blue-green alga Nostoc muscorum. The enzyme responsible for nitrate reduction, nitrate reductase, can be reversibly inactivated by reduction in a ferredoxin-dependent reaction. Nitrate protects against this inactivation, and molecular oxygen restores the original activity.     

Relationship of ammonia excretion,dinitrogen fixation and ammonia assimilatory enzymes to encystation in a continuous culture ofAzospirillum brasilense

Continuous culture studies have been carried out on a strain ofA. brasilense to study ammonia excretion, dinitrogen fixation and ammonia assimilatory enzymes (glutamate-ammonia ligase and glutamate synthase (NADPH)) in encysted conditions. High glutamate synthase (NADPH) and low ammonia excretion was observed at the time of induction of cyst formation. Low and oscillating nitrogenase activity was observed throughout the experiment.     

Isolation ofNostoc muscorum cyanophages from a domestic sewage

TwoNostoc muscorum cyanophages were isolated from a domestic sewage in Kuwait. N-1L cyanophage had a hexagonal head with a long tail, while N-2S cyanophage was a short-tailed virus. N-1L cyanophage was active at 50°C and at acidic pH, compared with N-2S, which was more heat stable and active at pH 7.0. Seasonal variations in the total number of plaque-forming units ofN. muscorum cyanophages were determined for sewage samples collected at each treatment step.     

Fractionation of phycobilisomes from the blue-green alga Nostoc muscorum

Phycobilisomes (PBS) isolated from Nostoc muscorum contain eight spectral forms of phycobiliproteins. PBS partially dissociated by osmotic or temperature shock, were separated into 42S and 34S particles by centrifugation in sucrose density gradient. The 42S particles are enriched with phycocyanin, the 34S ones--with allophycocyanin. The 42S particles dissociate to free pigments by repeated osmotic or temperature shock and the 34S ones dissociate to subparticles I and II having the constant pigments composition. The integrity of PBS and isolated substructures is controlled by the yield of energy transfer between pigment molecules. PBS from Nostoc muscorum was found to contain the rods of three different types. The model PBS containing different spectral forms of biliproteins is proposed.     

Nitrogenase activity in Nostoc muscorum; recovery from desiccation

Summary Nitrogenase activity in Nostoc crusts desiccated to below 10 per cent saturation for 24 hours and 7 days recovered on rewetting. Recovery over a 3 day period was greater in crusts returned to higher saturation levels. Possible reasons for, and limitations to, recovery are considered. re]19760420     

Action Spectra for Conversions of Phycochrome c from Nostoc muscorum

The reversibly photochromic pigment, phycochrome c, was extracted from the blue-green alga Nostoc muscorum strain A. Action spectra were determined for in vitro conversions of the pigment from the short wavelength to the long wavelength form and vice versa. The action peak for the absorbance decrease at 650 nm is at 630 nm. During this decrease there is only a slight increase of the absorbance in the green region. Green and yellow light (maximum efficiency at 580 nm) completely restores absorbance at 650 nm. The observations are explained by the existence of three spectrally different forms of phycochrome c: P^c₆₃₀ and P^c₆₅₀ which equilibrate in darkness and P^c₅₈₀ which is reversibly photoconvertible to P^c₆₃₀. We have also measured the absorbance changes brought about by saturating irradiations with light of various wavelengths (“photostationary state spectrum”). Extreme photostationary states were obtained with about 650 nm and 500 nm light.     

Formation of glutamine from [13n]ammonia, [13n]dinitrogen, and [14C]glutamate by heterocysts isolated from Anabaena cylindrica.

A method is described for the isolation of metabolically active heterocysts from Anabaena cylindrica. These isolated heterocysts accounted for up to 34% of the acetylene-reducing activity of whole filaments and had a specific activity of up to 1,560 nmol of C2H4 formed per mg of heterocyst chlorphyll per min. Activity of glutamine synthetase was coupled to activity of nitrogenase in isolated heterocysts as shown by acetylene-inhibitable formation of [13N]NH3 and of amidelabeled [13N]glutamine form [13N]N2. A method is also described for the production of 6-mCi amounts of [13N]NH3. Isolated heterocysts formed [13N]glutamine from [13N]NH3 and glutamate, and [14C]glutamine from NH3 and [14C]glutamate, in the presence of magnesium adenosine 5'-triphosphate. Methionine sulfoximine strongly inhibited these syntheses. Glutamate synthase is, after nitrogenase and glutamine synthetase, the third sequential enzyme involved in the assimilation of N2 by intact filaments. However, the kinetics of solubilization of the activity of glutamate synthase during cavitation of suspensions of A. cylindrica indicated that very little, if any, of the activity of that enzyme was located in heterocysts. Concordantly, isolated heterocysts failed to form substantial amounts of radioactive glutamate from either [13N]glutamine or alph-[14C]ketoglutarate in the presence of other substrates and cofactors of the glutamate synthase reaction. However, they formed [14C]glutamate rapidly from alpha-[14C]ketoglutarate by aminotransferase reactions, with various amino acids as the nitrogen donor. The implication of these findings with regard to the identities of the substances moving between heterocysts and vegetative cells are discussed.     

Nitrous oxide and dinitrogen emissions from urine-affected soil under controlled conditions

A ¹⁵N labelling technique was used to measure N₂O and N₂ emissions from an undisturbed grassland soil treated with cow urine and held at 30 cm water tension and 20°C in a laboratory. Large emissions of dinitrogen were detected immediately following urine application to pasture. These coincided with a rapid and large increase in soil water-soluble carbon levels, some of this increase being attributed to solubilization of soil organic matter by high pH and ammonia concentrations. Emissions of nitrous oxide generally increased with time in contrast to dinitrogen fluxes which decreased as time progressed. Estimated losses of N₂O and N₂ over a 30 day period were between 1 to 5% and 30 to 65% of the urine N applied plus N mineralized from soil organic matter, respectively. Most of the N₂ and N₂O originated from denitrification with nitrification-denitrification being of minor significance as a source of N₂O. Comparisons of the ¹⁵N enrichments in the soil mineral N pools and the evolved N₂O suggested that much of the N₂O was produced in the 5–8 cm zone of the soil. It is concluded that established grassland soils contain large amounts of readily-oxidizable organic carbon which may be used by soil denitrifying organisms when nitrate is non-limiting and soil redox potential is lowered due to high rates of biological activity and high soil moisture contents. ei]{gnR}{fnMerckx}     

The evolution of dinitrogen fixation

It is argued that nitrogenase originated monophyletically in obligate anaerobes similar to Clostridia. The enzyme system was later inherited, without much change, by photosynthetic bacteria, by prokaryotic plants (blue-greens) and by aerobic bacteria. The hydrogenase function of the enzyme complex preceded the nitrogenase function, and was useful in hydrogen fermentations. The consumption of ATP served to assure disposal of electrons in the form of hydrogen gas. The present need of the enzyme system, whether acting as a hydrogenase or as a nitrogenase, for ATP may be a relic from the period when the biosphere was still reducing.     

Purification and Properties of Glycogen Isolated from a Blue-Green Alga, Nostoc muscorum

Alpha granules were isolated from a blue-green alga, Nostoc muscorum, in large quantities and high purity by sucrose density gradient centrifugation. Sodium deoxycholate was used to eliminate membrane contamination. Isolated alpha granules from this species have average dimension of 31 nm in width and 65 nm in length. Each alpha granule consists of two equal parts. Attempts to dissociate the intact granule under mild conditions into relatively large subunits did not succeed. Analytical centrifugation of the alpha granules yields a sedimentation coefficient of 265S (S(20, w)). Chemical analysis reveals that alpha granules contain highly branched polyglucosyl units with short external chain lengths.     

Effects of cyclic nucleotides on morphogenesis inNostoc muscorum

The filamentous cyanophyteNostoc muscorum A grew aseriately in light in a mineral salts (sugar-free) culture medium supplemented with adenosine 3:5-cyclic-monophosphate or N⁶, O²-dibutyryl adenosine 3:5-cyclic-monophosphate (1 mM). The aseriate morphology thus formed in the light on the 10th day following inoculation was similar to that formed in the dark after 20–30 days growth in cAMP-free medium containing glucose or sucrose. Inoculum previously grown in sucrose- or glucose-containing medium displayed aseriate morphology with lesser proliferation of coccoid cells as compared to inoculum grown in the absence of glucose or sucrose. cGMP, ADP, AMP and inhibitors of phosphodiesterase (theophylline and caffeine) did not have any effect on the persistence of aseriate morphology. However they stimulated cell division at the aseriate stage and delayed the release of hormogonia.Abbreviations cAMP adenosine 3:5-cyclic-monophosphate - db cAMP N⁶, O²-dibutyryl adenosine 3:5-cyclic-monophosphate - cGMP guanosine 3:5-cyclic-monophosphate - ATP adenosine 5-triphosphate - ADP adenosine5-diphosphate - AMP adenosine 5-monophosphate     

Inhibition ofCandida albicans andStaphylococcus aureus by phenolic compounds from the terrestrial cyanobacteriumNostoc muscorum

Phenolic compounds were determined in methanolic extract from the algal mass of aNostoc muscorum culture. Bioassays with two human pathogens,Candida albicans andStaphylococcus aureus indicated that algal phenolic compounds evoked significant growth inhibition for both species (89.1% and 88.2%, respectively). It is suggested that this strong inhibitory effect is of potential medicinal value.     

Comparison of Ni-sensitive and Ni-resistant strains of Nostoc muscorum

A wild-type Ni-sensitive (Ni^s) strain of Nostoc muscorum ISU spontaneously yielded mutants resistant to inhibition by 40 M Ni with a frequency of about 10^-7. A Ni-resistant (Ni^r) mutant was deficient in the activities of urease and uptake hydrogenase. Cellular Ni uptake in the Ni^s strain was dependent on concentration (40 to 120 M) and time (0 to 30 min) (V_max=0.51 nmol/g protein.min; K_m=92 M). The Ni bioconcentration factor for such cells ranged between 0.95×10³ and 1.89×10³. Ni uptake in spheroplast preparations from Ni^s cells followed almost the same trend as intact cells except that the bioconcentration factor was slightly less [(0.82 to 1.39)×10³]. In contrast, Ni uptake in the Ni^r intact cells was not concentration dependent and also the uptake was saturated, even at 40 M, within 10 min. Spheroplasts from the Ni^r strain showed a Ni bioconcentration factor of 1.19×10³ compared with 4.41×10³ for intact cells. The invariably lower Ni uptake by spheroplasts was attributed to altered membrane transport properties.R.K. Asthana, A.L. Singh and S.P. Singh are with the Algal Research Laboratory, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi-221 005, India.     

Photoevolution of hydrogen during oxygenic photosynthesis of cyanobacteriumNostoc muscorum

Summary Nitrogen fixing cultures of the cyanobacteriumNostoc muscorum lacked hydrogen evolution but cultures infected with cyanophage N-1 showed significant hydrogen evolution and inactive nitrogenase, suggesting that nitrogenase activity is not responsible for the observed oxygen-resistant photoproduction of hydrogen. Significant oxygen-resistant hydrogen production by nitrate or ammonium assimilating cultures deficient in both nitrogenase and uptake hydrogenase activity supports this conclusion. These findings suggest a role of uptake hydrogenase in blocking the production of hydrogen during aerobic photosynthetic conditions.     

Studies on poly-beta-hydroxybutyrate synthase activity of Nostoc muscorum

This study compares the PHB synthase activity of Nostoc muscorum, a N(2)-fixing cyanobacterium under control (grown in usual BG-11 medium), nitrogen (N) and phosphorus (P) deprivation and chemoheterotrophic conditions. Specific activity of PHB synthase did not depict significant variations in the latter three types of cultures, except for the control one, where a significantly lower activity was recorded. PHB synthase activity was detected only in the soluble fractions of both the control as well as cells incubated under chemoheterotrophic conditions. A K(m) of 80.2 microM DL-beta-hydroxybutyryl-CoA and V(max) of 197.5 nmol thiobenzoate (TNB) mg protein(-1)min(-1) were observed for the enzyme. PHB synthase remained insensitive to acetyl-CoA, ATP, NADP, NADPH supplementation under in vitro condition. Addition of acetyl phosphate was found to activate the enzyme and the level of activation was dependent on the concentration of acetyl phosphate supplementation. Inhibition of PHB synthase in 2,3-butanedione supplemented cultures and reactivation following acetyl phosphate addition proved the post-translational control of acetyl phosphate over PHB synthase.