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Utilization of glucose and the effect of organic compounds on the chemolithotroph Thiobacillus ferrooxidans.

The utilization of glucose by the chemolithotroph Thiobacillus ferrooxidans results in a repression of the ability to oxidize iron, the substrate for autotrophic growth. An assay with resting cells was used to measure iron oxidation rates. Concomitant with the decreased iron oxidation rates, the enzyme responsible for carbon dioxide fixation, ribulose diphosphate (RuDP) carboxylase, was also repressed. Maximum iron oxidation rates precede peak RuDP carboxylase levels, consistent with the role of these processes in autotrophic metabolism in nonrepressed cells. The degree of iron oxidation repression depends on the organic substrate supplied, as does the level of RuDP carboxylase. The uptake of glucose parallels an increase in synthesis of glucose-6-phosphate dehydrogenase and the accumulation in cells of poly-beta-hydroxybutyrate. The organism is also capable of growing on glucose and other organic supplements in the absence of its inorganic energy source; growth rates depend on the organic substrate supplied.

Carbon Dioxide↗

Start up of deammonification process in one single SBR system.

A process for autotrophic nitrogen removal named aerobic/anoxic deammonification wherein NH4+ is oxidized by nearly 50% to NO2- and subsequently the ammonia is converted together with the nitrite to molecular nitrogen (N2 gas), has come to full-scale application within the last few years. In this research, sludge from a biological rotation disk located at a landfill leachate plant at Mechernich, Germany, which is capable of performing the deammonification process, was used as seed sludge for acclimating deammonification activities in laboratory scale batch-reactors. In parallel, the same tests were performed with normal activated sludge. Research results indicated that deammonification activities could be obtained from the seeded reactor and also, with limited performance, from normal activated sludge in a single SBR system after several months acclimation. It was also seen that oxygen is an important factor that influences the deammonification from both the acclimatization process and process running. Further results were approved that report an impact of nitrite as a process intermediate on the closely related process of anaerobic ammonia oxidation ("Anammox"). However, limiting concentrations on a bacteria population performing deammonification were found to be different to those reported for a pure Anammox-culture. Also the influence of another intermediate, hydrazine, was tested for speeding up the acclimating process by inducing the deammonification activities and recovering the activities of deammonification from nitrite inhibition.

Bacteria, Anaerobic↗

Sulfide oxidation under chemolithoautotrophic denitrifying conditions.

Chemolithoautotrophic denitrifying microorganisms oxidize reduced inorganic sulfur compounds coupled to the reduction of nitrate as an electron acceptor. These denitrifiers can be applied to the removal of nitrogen and/or sulfur contamination from wastewater, groundwater, and gaseous streams. This study investigated the physiology and kinetics of chemolithotrophic denitrification by an enrichment culture utilizing hydrogen sulfide, elemental sulfur, or thiosulfate as electron donor. Complete oxidation of sulfide to sulfate was observed when nitrate was supplemented at concentrations equal or exceeding the stoichiometric requirement. In contrast, sulfide was only partially oxidized to elemental sulfur when nitrate concentrations were limiting. Sulfide was found to inhibit chemolithotrophic sulfoxidation, decreasing rates by approximately 21-fold when the sulfide concentration increased from 2.5 to 10.0 mM, respectively. Addition of low levels of acetate (0.5 mM) enhanced denitrification and sulfate formation, suggesting that acetate was utilized as a carbon source by chemolithotrophic denitrifiers. The results of this study indicate the potential of chemolithotrophic denitrification for the removal of hydrogen sulfide. The sulfide/nitrate ratio can be used to control the fate of sulfide oxidation to either elemental sulfur or sulfate.

Acetates↗

Complementary limiting factors of astaxanthin synthesis during photoautotrophic induction of Haematococcus pluvialis: C/N ratio and light intensity.

We investigated the effect of carbon/nitrogen (C/N) ratio on astaxanthin synthesis in Haematococcus pluvialis during photoautotrophic induction by continuous input of both CO(2)-air mixture and intense light. When H. pluvialis was induced by constant irradiance induction at 200 micromol photon m(-2) s(-1), there was a positive correlation with astaxanthin content and C/N ratio, which was similar to the case for heterotrophic induction. Lower C/N ratios did not retard Haematococcus encystment, but did increase culture biomass, resulting in a decrease in astaxanthin production because of light limitation. However, induction using variable irradiance showed that reduction of astaxanthin production at low C/N ratios was successfully overcome by simply increasing the light intensity from 200 to 300 micromol photon m(-2) s(-1) to overcome the light limitation. This resulted in a greatly enhanced astaxanthin synthesis in proportion to cell density in cultures with low C/N ratios. Our results indicate that light intensity is more critical than C/N ratio in astaxanthin production by H. pluvialis during photoautotrophic induction.

Autotrophic Processes↗

Microbial diversity in natural environments: focusing on fundamental questions.

Interactions with Gijs Kuenen and other Dutch scientists have led my lab to fundamental insights into the composition, structure and function of a hot spring cyanobacterial mat community that should influence our thinking about all microbial communities. By focusing on the distribution of molecular sequence variants of predominant mat phototrophs, we have discovered that small-scale sequence variation can be ecologically meaningful. By applying novel cultivation approaches, we have been able to obtain genetically relevant community members and thus to test the hypothesis that closely related sequence variants arose via adaptive evolutionary radiation. By applying the analytical tools of organic geochemistry we have gained insight into the metabolisms of major phototrophic members of the mat community as well as interactions between phototrophic guilds. These observations challenge traditional paradigms about prokaryotic species and cause us to consider evolutionary ecology theory as we develop genome-based methods for high-resolution analysis of the species-like fundamental units comprising microbial communities, and for investigating how such units coordinate the physiological activities within guilds of the community.

Autotrophic Processes↗

Photosynthesis in the Archean era.

The earliest reductant for photosynthesis may have been H2. The carbon isotope composition measured in graphite from the 3.8-Ga Isua Supercrustal Belt in Greenland is attributed to H2-driven photosynthesis, rather than to oxygenic photosynthesis as there would have been no evolutionary pressure for oxygenic photosynthesis in the presence of H2. Anoxygenic photosynthesis may also be responsible for the filamentous mats found in the 3.4-Ga Buck Reef Chert in South Africa. Another early reductant was probably H2S. Eventually the supply of H2 in the atmosphere was likely to have been attenuated by the production of CH4 by methanogens, and the supply of H2S was likely to have been restricted to special environments near volcanos. Evaporites, possible stromatolites, and possible microfossils found in the 3.5-Ga Warrawoona Megasequence in Australia are attributed to sulfur-driven photosynthesis. Proteobacteria and protocyanobacteria are assumed to have evolved to use ferrous iron as reductant sometime around 3.0 Ga or earlier. This type of photosynthesis could have produced banded iron formations similar to those produced by oxygenic photosynthesis. Microfossils, stromatolites, and chemical biomarkers in Australia and South Africa show that cyanobacteria containing chlorophyll a and carrying out oxygenic photosynthesis appeared by 2.8 Ga, but the oxygen level in the atmosphere did not begin to increase until about 2.3 Ga.

Autotrophic Processes↗

Reversible, cell-heritable changes during the development of tobacco pith tissues.

Cytokinin requiring cells of Nicotiana tabacum L. cv "Havana 425" can be induced in culture to become cytokinin autotrophic. This process is known as cytokinin habituation. Earlier we showed that pith parenchyma tissue consists of inducible cells, which habituate at high rates when treated with cytokinin, and noninducible cells, which remain cytokinin requiring under these conditions. The inducible and noninducible phenotypes are determined states that arise during the development of the tobacco plant and are inherited by individual cells. Here we show that pith tissue of plants regenerated from cloned lines of noninducible cells exhibits the inducible phenotype indicating that noninducible cells, or their descendants, can become inducible. This change in competence for habituation appears to have an epigenetic basis; it is reversible, occurs at high rates, and depends on the developmental state of the cells. The habituated state occurs in two forms that can be distinguished by their difference in developmental potential. Habituated cells derived from inducible pith cells give rise to normal plants whose leaf and pith tissues require cytokinin for growth in culture. In contrast, habituated cells obtained by transferring noninducible cells on media with progressively lower cytokinin concentrations give rise to plants whose leaf and pith tissues exhibit a cytokinin-habituated phenotype in culture.

Cell Line↗

Hydrogen-dependent denitrification in a two-reactor bio-electrochemical system.

An autotrophic biological process was developed for the treatment of nitrate-contaminated drinking water. The system comprised of two steps: the water to be treated was first enriched with hydrogen (energy source) in the cathodic chamber of an electrochemical cell, and then denitrified in the bioreactor. The bioreactor was a packed bed of granulated activated carbon, and the water flow was directed in an upward continuous mode. The system was operated for one year, at various water velocities and current intensities. Denitrification rates up to 0.25 kg N m-3 d-1 were obtained at the hydraulic residence time of 1 h. The system was stable. When detected in the effluent, the concentration of nitrite was low, even under conditions that resulted in the elution of very high concentrations of nitrate.

Bioreactors↗

Application of calorimetric measurements for biokinetic characterisation of nitrifying population in activated sludge.

A preliminary investigation is described on the application of calorimetry as a sensitive technique to evaluate nitrifying activity in activated sludge. Calorimetric profiles (thermograms) related to heat dissipation due to biological nitrification reactions (ammonia or nitrite consumption) have been interpreted. Correlations between calorimetric data and the main process variables, i.e. ammonia and nitrite concentration and oxygen uptake, have been verified, and confirm the potential of calorimetry to investigate, monitor and control even weakly exothermic biological processes like autotrophic nitrification. Heat yields (Y(Q/i)) for ammonia, nitrite, and oxygen, defined as the heat released per unit amount of converted reactant, have been separately evaluated. Moreover, calorimetric experiments on activated sludge from a full-scale nitrogen removal wastewater treatment plant have been carried out and kinetic parameters for both ammonia and nitrite oxidising bacteria have been estimated.

Ammonia↗

The PsbQ protein is required in Arabidopsis for photosystem II assembly/stability and photoautotrophy under low light conditions.

RNA interference was used to simultaneously suppress the expression of the two genes that encode the PsbQ proteins of Photosystem II (PS II) in Arabidopsis thaliana, psbQ-1 (At4g21280) and psbQ-2 (At4g05180). Two independent PsbQ-deficient plant lines were examined. These plant lines produced little detectable PsbQ protein. Under normal growth light conditions, the wild type and mutant plants were visually indistinguishable. Additionally, analysis of steady state oxygen evolution rates and chlorophyll fluorescence characteristics indicated little alteration of photosynthetic capacity in the mutant plants. No loss of other PS II proteins was evident. Interestingly, flash oxygen yield analysis performed on thylakoid membranes isolated from the mutant and wild type plants indicated that the oxygen-evolving complex was quite unstable in the mutants. Furthermore, the lifetime of the S2 state of the oxygen-evolving complex appeared to be increased in these plants. Incubation of the wild type and mutant plants under low light growth conditions led to a significantly stronger observed phenotype in the mutants. The mutant plants progressively yellowed (after 2 weeks) and eventually died (after 3-4 weeks). The wild type plants exhibited only slight yellowing after 4 weeks under low light conditions. The mutant plants exhibited a large loss of a number of PS II components, including CP47 and the D2 protein, under low light conditions. Additionally, significant alterations of their fluorescence characteristics were observed, including an increased FO and decreased FV, yielding a large loss in PS II quantum efficiency (FV/FM). Analysis of QA- decay kinetics in the absence of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea indicated a defect in electron transfer from QA- to QB, whereas experiments performed in the presence of this herbicide indicated that the recombination rate between QA- and the S2 state was strongly retarded. These results indicate that the loss of the PsbQ protein induces significant changes in Photosystem II function, particularly in low light-grown plants, and that the PsbQ protein is required for photoautotrophic growth under low light conditions.

Animals↗

CO2 uptake and fixation by endosymbiotic chemoautotrophs from the bivalve Solemya velum.

Chemoautotrophic symbioses, in which endosymbiotic bacteria are the major source of organic carbon for the host, are found in marine habitats where sulfide and oxygen coexist. The purpose of this study was to determine the influence of pH, alternate sulfur sources, and electron acceptors on carbon fixation and to investigate which form(s) of inorganic carbon is taken up and fixed by the gamma-proteobacterial endosymbionts of the protobranch bivalve Solemya velum. Symbiont-enriched suspensions were generated by homogenization of S. velum gills, followed by velocity centrifugation to pellet the symbiont cells. Carbon fixation was measured by incubating the cells with (14)C-labeled dissolved inorganic carbon. When oxygen was present, both sulfide and thiosulfate stimulated carbon fixation; however, elevated levels of either sulfide (>0.5 mM) or oxygen (1 mM) were inhibitory. In the absence of oxygen, nitrate did not enhance carbon fixation rates when sulfide was present. Symbionts fixed carbon most rapidly between pH 7.5 and 8.5. Under optimal pH, sulfide, and oxygen conditions, symbiont carbon fixation rates correlated with the concentrations of extracellular CO(2) and not with HCO(3)(-) concentrations. The half-saturation constant for carbon fixation with respect to extracellular dissolved CO(2) was 28 +/- 3 microM, and the average maximal velocity was 50.8 +/- 7.1 micromol min(-1) g of protein(-1). The reliance of S. velum symbionts on extracellular CO(2) is consistent with their intracellular lifestyle, since HCO(3)(-) utilization would require protein-mediated transport across the bacteriocyte membrane, perisymbiont vacuole membrane, and symbiont outer and inner membranes. The use of CO(2) may be a general trait shared with many symbioses with an intracellular chemoautotrophic partner.

Animals↗

Simultaneous biological removal of sulphide and nitrate by autotrophic denitrification in an activated sludge system.

The feasibility of an autotrophic denitrification process in an activated sludge reactor, using sulphide as the electron donor, was tested for simultaneous denitrification and sulphide removal. The reactor was operated at nitrate (N) to sulphide (S) ratios between 0.5 and 0.9 to evaluate their effect on the N-removal efficiency, the S-removal efficiency and the product formation during anoxic oxidation of sulphide. One hundred per cent removal of both nitrate and sulphide was achieved at a NLR of 7.96 mmol N-L(-1) x d(-1) (111.44 mg NO3- -N x L(-1) x d(-1)) and at a N/S ratio of 0.89 with complete oxidation of sulphide to sulphate. The oxygen level in the reactor (10%) was found to influence the N-removal efficiency by inhibiting the denitrification process. Moreover, chemical (or biological) oxidation of sulphide with oxygen occurred, resulting in a loss of the electron donor. FISH analysis was carried out to study the microbial population in the system.

Nitrates↗