A study of chlorophyll-like and phycobilin pigments in the C endosymbiont of the apple- snail <i>Pomacea canaliculata</i>

Pigments present in the brown-greenish C morph of an intracellular endosymbiont of Pomacea canaliculata were investigated. Acetone extracts of the endosymbiotic corpuscles showed an absorption spectrum similar to that of chlorophylls. Three fractions obtained from silica gel column chromatography of the acetone extracts (C_I , C_II and C_III ), were studied by positive ion fast atom bombardment-mass spectrometry (FAB–MS) and hydrogen-nuclear magnetic resonance (H-NMR). Results indicated the presence of (1) a sterol in the yellow colored C_I fraction; (2) a mixture of pheophorbides a and b in the major green fraction, C_II; and (3) a modified pheophorbide a in the smaller green fraction, C_III . Aqueous extracts of the C endosymbiont did not show evidence of the occurrence of C-phycocyanin, allophycocyanin or phycoerithrin (light absorption, fluorescence emission, and electrophoresis of the protein moieties) while cyanobacterial cells (Nostoc sp.) showed evidence of C-phycocyanin and allophycocyanin. The possible phylogenetic and functional significance of the pigments present in the C endosymbiont is discussed.     

Kinetic and optimisation studies on ultrasonic intensified photo-autotrophic ethanol production from Synechocystis sp.

A novel approach to the intensification of renewable and sustainable production of photoautotrophic ethanol from microalgae was investigated using periodic ultrasonication. The effect of utilizing ultrasonic pulsing during ethanol production using a metabolically engineered ethanol producing cyanobacterium Synechocystis PCC6803 strain NAV001 was analysed with an ultrasonic frequency of 20 kHz. Ultrasonic treatment resulted in enhancement of ethanol yields. The ethanol yield was found to be higher when ultrasonic pulsing was initiated during the exponential phase of growth. The optimum ultrasonic conditions for enhanced yield of ethanol within this model system was found to be 30 °C, 15% of power input (97.5 W) and 10 min pulse time. The cytotoxic effect of ultrasonic pulsation was investigated by analysing the survival percentage and auto-fluorescence of the cyanobacterium via imaging by confocal laser scanning microscopy. The overall effect of ultrasonic dose on ethanol production and growth rate of microalgae was analysed by using Haldane inhibition kinetics.     

Effect of cyanobacteria Synechococcus PCC 7942 on carbonation kinetics of olivine at 20 °C

By accelerating the naturally-occurring carbonation of magnesian silicates, it would be possible to sequester some of the anthropogenic excess of CO₂ in more geologically-stable solid magnesium carbonates. Reaction rates can be accelerated by decreasing the particle size, raising the reaction temperature, increasing the pressure, using a catalyst, and hypothetically, by bacterial addition. We aimed here at assessing quantitatively the added value of photosynthetic microbial activity on the efficiency of Mg-silicates carbonation processes. Synechococcus PCC 7942 (freshwater cyanobacteria) was selected for this study. Two magnesian silicate minerals (substrates) were chosen: a synthetic forsterite with nanometer-sized grains and an industrial ultramafic slag (scoria). All tests were performed at 20 ± 1 °C in closed and sterile 1L Schott® glass bottle reactors. With the aim to elucidate the interaction between mineral phases and bacteria, we used pH and concentration measurements, scanning and transmission electron microscopy along with Raman spectroscopy. The results show that, at ambient temperature, cyanobacteria Synechococcus can accelerate silicate dissolution (i.e. Mg²⁺ release) and then magnesium carbonate nucleation and precipitation by adsorption on the produced exopolymeric substances and local pH increase during photosynthesis, respectively.     

Biohydrogen production by novel cyanobacterial strains isolated from rice paddies in Kazakhstan

A limited supply of oil prompts the search for non-traditional energy sources to replace traditional ones. This makes hydrogen gas an appealing alternative source. Photosynthetic organisms capture sunlight very efficiently and convert it into organic molecules. A promising wild strain was isolated for the first time, from the rice paddies of Kazakhstan (Kyzylorda and Almaty regions), which can be considered as one of the most active hydrogen producers compared to the literature. The result showed that among the 13 isolated and collection cyanobacterial strains, Synechocystis sp. S-1 is the most active H₂ producer (2.35 μmol H₂ mg^?1 Chl a h^?1) in the light. In contrast, the wild-type cyanobacterium Anabaena variabilis A-1 had higher productivity, nitrogenase activity, and a stronger capacity to produce hydrogen in the dark (8.67 μmol H₂ mg^?1 Chl a h^?1), which matched the maximum yield obtained in the research. The metabolic modulation performed significantly increased hydrogen production. The highest photohydrogen production rate was observed in cells incubated with 25 μmol HEPES and 50 μmol sodium bicarbonate (NaHCO₃).     

Dark hydrogen production in nitrogen atmosphere – An approach for sustainability by marine cyanobacterium Leptolyngbya valderiana BDU 20041

Biological hydrogen production is an ideal system for three main reasons i) forms a renewable energy source, ii) gives clean fuel and iii) serves as a good supplement to oil reserves. The major challenges faced in biological hydrogen production are the presence of uptake hydrogenase and lack of sustainability in the cyanobacterial hydrogen production system. Three different marine cyanobacterial species viz. Leptolyngbya valderiana BDU 20041, Dichothrix baueriana BDU 40481 and Nostoc calcicola BDU 40302 were studied for their potential use in hydrogen production. Among these, L. valderiana BDU 20041, was found to produce hydrogen even in 100% nitrogen atmosphere which was 85% of the hydrogen produced in argon atmosphere. This is the first report of such a high rate of production of hydrogen in a nitrogen atmosphere by a cyanobacterium, which makes it possible to develop sustained hydrogen production systems. L. valderiana BDU 20041, a dark hydrogen producer uses the reductant essentially supplied by the respiratory pathway for hydrogen production. Using inhibitors, this organism was found to produce hydrogen due to the activities of both nitrogenase and bidirectional hydrogenase, while it had no ‘uptake’ hydrogenase activity. The other two organisms though had low levels of bidirectional hydrogenase, possessed considerable ‘uptake’ hydrogenase activity and hence could not release much hydrogen either in argon or nitrogen atmosphere.     

Photoproduction of H2 by wildtype Anabaena PCC 7120 and a hydrogen uptake deficient mutant: from laboratory experiments to outdoor culture

We have analyzed the filamentous cyanobacterium Anabaena PCC 7120 (wildtype) containing one nitrogenase, one uptake hydrogenase and one bidirectional hydrogenase and its hydrogen uptake deficient mutant AMC 414 for their H₂ production capacities. Anabaena PCC 7120 and AMC 414 had similar growth rates in turbidostat mode with increased growth rates at higher light intensity. Rates of C₂H₂ reduction were similar for both strains. In contrast to the wildtype, AMC 414 produced H₂ in a PhotoBioReactor (PhBR) using air as the lifting gas. The rate of H₂ production increased with light intensity and was not even saturated at 456 μEm⁻² s⁻¹. H₂ production increased significantly when replacing the air with argon. The maximal H₂ production during outdoor conditions was recorded using AMC 414 with a peak at 14.9 ml H₂ h⁻¹ l⁻¹. Despite the relatively high production, maximal efficiency of solar energy to H₂ conversion was only 0.042%. A molecular method was developed to analyze the relative abundancies of weight and mutant in competition experiments in the PhBR.     

Determination of the potential of cyanobacterial strains for hydrogen production

Hydrogen (H₂) is a renewable, abundant, and nonpolluting source of energy. Photosynthetic organisms capture sunlight very efficiently and convert it into organic molecules. Cyanobacteria produce H₂ by breaking down organic compounds and water. In this study, biological H₂ was produced from various strains of cyanobacteria. Moreover, H₂ accumulation by Synechocystis sp. PCC 6803 was as high as 0.037 μmol/mg Chl/h within 120 h in the dark. The wild-type, filamentous, non-heterocystous cyanobacterium Desertifilum sp. IPPAS B-1220 was found to produce a maximum of 0.229 μmol/mg Chl/h in the gas phase within 166 h in the light, which was on par with the maximum yield reported in the literature. DCMU at 10 μM increased H₂ production by Desertifilum sp. IPPAS B-1220 by 1.5-fold to 0.348 μmol H₂/mg Chl/h. This is the first report on the capability of Desertifilum cyanobacterium to produce H₂.     

Microcystin ecotypes in a perennial Planktothrix agardhii bloom

The dynamics and microcystins (MC) concentrations of a perennial Planktothrix agardhii bloom were investigated in a eutrophic lake (Viry-Chatillon, France). A weak relationship was observed between P. agardhii population biomass and the MC concentrations in a 1-year survey. To further investigate the causes of MC concentration changes, we concurrently conducted experiments on 41 strains isolated from this lake. We first checked the clonal diversity of P. agardhii population (i) by molecular techniques, to assess the presence of MC synthetase gene (mcyB), (ii) by biochemical assay (PP2A inhibition assay), for MC production, and (iii) by mass spectrometry (MS), to identify the MC chemotypes. Our results illustrated the diversity of genotype and MC chemotypes within a P. agardhii natural population. Eleven chemotypes among the 16 possible ones were found by MS. Furthermore, we noticed major differences in the MC content of isolated strains (from 0.02 to 1.86 microg equiv. MC-LR mg DW(-1), n=25). Growth and MC production of one MC-producing strain and one non-MC-producing strain were also assessed at two temperatures (10 and 20 degrees C). We showed that growth capacities of these strains were similar at the two tested temperatures, and that the MC production rate was correlated to the growth rate for the MC-producing strain. On the basis of these results, several hypotheses are discussed to explain the weakness of relationships between natural P. agardhii biomass and MC concentration. One of the main reasons could lie in the proportion of MC-producing clones and non-MC-producing clones that may change during the sampling period. Also, the MC-producing clones may present different intracellular MC content due to (i) MC chemotypes diversity, (ii) changes in MC variants proportions within a strain, and (iii) changes in MC rate production depending on the physiological state of cells. Finally, we concluded that various biological organization levels have to be considered (population, cellular and molecular), through an integrative approach, in order to provide a better understanding of P. agardhii in situ MC production.     

Kinetics of Bis-Allylic Hydroperoxide Synthesis in the Iron-Containing Lipoxygenase 2 from Cyanothece and the Effects of Manganese Substitution

Lipoxygenases (LOX) catalyze the regio‐ and stereospecific insertion of dioxygen into polyunsaturated fatty acids. While the catalytic metal of LOX is typically a non‐heme iron, some fungal LOX contain manganese as catalytic metal (MnLOX). In general, LOX insert dioxygen at C9 or C13 of linoleic acid leading to the formation of conjugated hydroperoxides. MnLOX (EC 1.13.11.45), however, catalyze the oxygen insertion also at C11, resulting in bis‐allylic hydroperoxides. Interestingly, the iron‐containing CspLOX2 (EC 1.13.11.B6) from Cyanothece PCC8801 also produces bis‐allylic hydroperoxides. What role the catalytic metal plays and how this unusual reaction is catalyzed by either MnLOX or CspLOX2 is not understood. Our findings suggest that only iron is the catalytically active metal in CspLOX2. The enzyme loses its catalytic activity almost completely when iron is substituted with manganese, suggesting that the catalytic metal is not interchangeable. Using kinetic and spectroscopic approaches, we further found that first a mixture of bis‐allylic and conjugated hydroperoxy products is formed. This is followed by the isomerization of the bis‐allylic product to conjugated products at a slower rate. These results suggest that MnLOX and CspLOX2 share a very similar reaction mechanism and that LOX with a Fe or Mn cofactor have the potential to form bis‐allylic products. Therefore, steric factors are probably responsible for this unusual specificity. As CspLOX2 is the LOX with the highest proportion of the bis‐allylic product known so far, it will be an ideal candidate for further structural analysis to understand the molecular basis of the formation of bis‐allylic hydroperoxides.