Sodium dependent acetate formation from CO2 in Peptostreptococcus productus (strain Marburg)

Abstract Washed cells of Peptostreptococcus productus (strain Marburg), which were incubated in the presence of CO/CO₂/N₂ (50%/ 17%/ 33%; 200 kPa) catalyzed the synthesis of acetate from carbon monoxide. The rate of acetate formation from CO was stimulated more than threefold by the addition of sodium (10 mM); potassium did not effect acetate synthesis. The degree of stimulation was dependent on the sodium concentration; the dependence followed simple Michaelis-Menten kinetics. The apparent K _m for sodium was determined to be about 2 mmol/1. Sodium also stimulated acetate synthesis from H₂ plus CO₂. In the absence of added sodium the formation of formate as an intermediate in methyl group synthesis was stimulated. It is suggested that the sodium dependent reaction(s) is one (or more) of the reactions involved in methyl group synthesis from CO₂.     

Sodium bioenergetics in methanogens and acetogens

Abstract Recent investigations with Methanosarcina barkeri elucidated the role of sodium ions in the energy metabolism of methanogenic bacteria and provided evidence for a novel mechanism of energy transduction with Na⁺ as the coupling ion. During methanogenesis from methanol, an eletrochemical sodium gradient generated by a Na⁺/H⁺ antiporter is used as the driving force for the thermodynamically unfavourable oxidation of methanol to the formal redox level of formaldehyde. During methanogenesis from H₂+ CO₂, the reverse reaction, the reduction of formaldehyde to the level of methanol, is accompanied by a primary, electron transport-driven sodium extrusion. Acetogenesis from H₂+ CO₂ as carried out by Acetobacterium woodii is a sodium-dependent process and is accompanied by the generation of a transmembrane sodium gradient with the reduction of formaldehyde to the level of methanol as the sodium-dependent step.     

Electron transfer reactions in methanogens

Abstract Methanogenic bacteria comprise a specialized group of obligately anaerobic microorganisms able to reduce a limited number of substrates to CH₄. The intermediates involved in this reduction process remain bound to a series of typical C₁-carriers. Reducing equivalents are either obtained from the oxidation of H₂ or from oxidation of carbon substrates to CO₂. Electron transfer reactions thus constitute the very essence of the process of methanogenesis.
In recent years much progress has been made in the elucidation of the special metabolic pathways and the nature of the C₁-carriers involved in methanogenic bacteria. The energy generated at the oxidoreduction reactions, notably at the methylreductase step, is conserved by ATP synthesis. The energy is used for cell carbon synthesis and, in catalytic amounts, for the reductive activation of some methanogenic enzymes. Before the condensing reaction resulting in the formation of acetyl-CoA takes place, 2 C₁-units are reduced or oxidized depending on the substrate to a carbonyl and a -CH₃ group. Formation of the latter proceeds via the methanogenic route. Intermediary cell carbon synthesis starting from acetyl-CoA involves reductive carboxylations and oxidoreductions by the participation of the enzymes of the tricarboxylic acid cycle.     

Leaf isoprene emission rate as a function of atmospheric CO2 concentration

There is considerable interest in modeling isoprene emissions from terrestrial vegetation, because these emissions exert a principal control over the oxidative capacity of the troposphere. We used a unique field experiment that employs a continuous gradient in CO₂ concentration from 240 to 520 ppmv to demonstrate that isoprene emissions in Eucalyptus globulus were enhanced at the lowest CO₂ concentration, which was similar to the estimated CO₂ concentrations during the last Glacial Maximum, compared with 380 ppmv, the current CO₂ concentration. Leaves of Liquidambar styraciflua did not show an increase in isoprene emission at the lowest CO₂ concentration. However, isoprene emission rates from both species were lower for trees grown at 520 ppmv CO₂ compared with trees grown at 380 ppmv CO₂. When grown in environmentally controlled chambers, trees of Populus deltoides and Populus tremuloides exhibited a 30–40% reduction in isoprene emission rate when grown at 800 ppmv CO₂, compared with 400 ppmv CO₂. P. tremuloides exhibited a 33% reduction when grown at 1200 ppmv CO₂, compared with 600 ppmv CO₂. We used current models of leaf isoprene emission to demonstrate that significant errors occur if the CO₂ inhibition of isoprene is not taken into account. In order to alleviate these errors, we present a new model of isoprene emission that describes its response to changes in atmospheric CO₂ concentration. The model logic is based on assumed competition between cytosolic and chloroplastic processes for pyruvate, one of the principal substrates of isoprene biosynthesis.     

Photoinhibition of respiratory CO2 release in the green alga Scenedesmus

¹⁴CO₂ evolution of prelabeled Scenedesmus obliquus Kütz, has been followed in the dark and in the light. In the light, no carbon dioxide is evolved. Addition of unlabeled NaHCO, leads to ¹⁴CO₂ release attaining 20 to 30% of the dark rate. Double-reciprocal plots of NaHCO₃ concentrations vs ¹⁴CO₂ release results in a straight line, indicative of competition between exogenously supplied bicarbonate and endogenously evolved carbon dioxide. With this method, it is possible to measure CO₂ evolved by respiration in the light and to show that true photoinhibition of respiration occurs in Scenedesmus . In the light. DCMU substantially increases ¹⁴CO₂ evolution; in the presence of the uncoupler carbonyl cyanide- m -chlorophenylhydrazone. ¹⁴CO₂ evolution is comparable to that in the dark. ¹⁴CO₂ release and oxygen uptake in the dark are only slightly affected by cyanide, indicative of a cyanide-resistant respiration and/or fermentation as the essential CO₂-yielding processes in the presence of cyanide. These results, compared with concurrent ATP levels, lead us to assume that energy charge is not the only factor responsible for photoinhibition of respiration.     

Microbial H2/CO2 acetogenesis in animal guts: nature and nutritional significance

Abstract The intestinal tract of invertebrate and vertebrate animals, including man, is an anoxic habitat wherein microbial formation of acetate from H₂+ CO₂ is often a major H₂-consuming reaction. This paper will discuss the magnitude and microbiology of H₂/CO₂ acetogenesis in animal guts, its impact on host animal nutrition, competition for H₂ between anaerobic microbes, and the global significance of intestinal H₂/CO₂ acetogenesis.     

Heterotrophic (Dark) CO2 Fixation by Euglena gracilis. Possible regulation by tricarboxylic acid cycle intermediates

SYNOPSIS. Heterotrophic (dark) CO₂ fixation by Euglena gracilis strain Z varies with phase of batch culture and mode of nutrition. Dark CO₂ fixation increased transiently during the growth of cells under photoautotrophic (CO₂, light) and heterotrophic (glucose, dark) conditions. Cells grown heterotrophically with acetate or ethanol had no transient increase in fixation. The addition of acetate to a heterotrophically growing culture during the period of increasing dark CO₂ fixation resulted in rapid elimination of this fixation. The results suggest that dark CO₂ fixation in Euglena functions in anaplerotic feeding of the tricarboxylic acid cycle, drained by biosyntheses during growth. Induction of the glyoxylate cycle by acetate may provide an alternate source of tricarboxylic cycle intermediates, obviating the requirement for dark CO₂ fixation as a source of the intermediates.     

Fermentation characteristics of Clostridium pasteurianum LMG 3285 grown on glucose and mannitol

Clostridium pasteurianum fermented glucose to acetate, butyrate, CO₂ and H₂. In batch cultures the fermentation pattern was only slightly affected by culture pH over the range 8·0 to 5·5. The acetate/butyrate ratio was always higher than or equal to one. Between 2·14 and 2·33 mol H₂ was produced per mol glucose fermented. At unregulated pH, more butanol and less butyrate was formed. In a carbon-limited chemostat, the steady-state acetate/butyrate ratio was always lower than one. H₂ production was approximately 1·70 mol per mol glucose consumed. Substantial amounts of extracellular protein were formed. With decreasing pH, acetate and formate production decreased, while H₂ production was highest at pH 6.0. With increasing dilution rate ( D ), the product spectrum hardly changed, but more biomass was formed. Y _glucose^max and Y _ATP^max were 55·97 and 31·48 g dry weight per mol glucose or ATP respectively. With increasing glucose input the formation of fatty acids and H₂ slightly decreased.
Continuous cultures fermented mannitol to acetate, butyrate, butanol, CO₂ and H₂. With acetate as co-substrate, butanol production and molar growth yields, Y _mannitol and Y _ATP, markedly decreased, while the butyrate and H₂ production increased. The latter reached a value of 2·21 mol H₂ per mol mannitol consumed.     

Diurnal variations in the kinetics of delayed luminescence from Scenedesmus obtusiusculus after exposure to various light and CO2 regimes

Delayed luminescence was measured from samples of a synchronously growing cell culture of the unicellular green alga, Scenedesmus obtusiusculus Chod., every second hour during the 24 h cell cycle under a 15/9 h lighi/dark regime. Both high (air + 2.5% CO₂) and low (0.03% CO₂) CO₂ conditions were used. Under high CO₂ conditions, while light excitation induces formation of a late (maximum reached after 10–60 s) transient peak in delayed luminescence in cells sampled after 10–16 h in the cell cycle. During most of the cell cycle low CO₂ conditions stimulate a late transient peak formation. Excitation with 700 nm light, but not with 680 nm light, induces a late transient peak in delayed luminescence under high CO₂ conditions. The transient peak is more or less pronounced depending on the cell stage. The variations might be due to a changing capacity for light-induced state I/stale II transitions during the cell cycle. It is assumed that the formation of a late transient peak in delayed luminescence is due to ATP hydrolyzation and is thus favoured by a high ATP/NADPH ratio. Hydrolyzation of ATP affects the transthylakoidal ΔpH, which regulates the reverse electron flow to the plastoquinone-pool and Q_A/Q_B, thus affecting the decay kinetics of the delayed luminescence.     

A model for photosynthesis and photorespiration in C3 plants based on the biochemistry and stoichiometry of the pathways

Abstract. A model is developed for photosynthesis and photorespiration in C₃ plants, using an equation for the multisubslrate ordered reaction of ribulose 1,5-bisphosphalc carboxylase-oxygenase (Farazdaghi & Edwards, 1988). The model examines net CO₂ fixation with O₂ inhibition, and mutual inhibition when equilibrium exists between carboxylation and oxygenation (at the CO₂ compensation point). It is based on the stoichiometry of energy requirements and O₂, and CO₂ exchange in the cycles, the quantum efficiency for RuBP generation, the maximum capacity for RuBP generation, the carboxylation efficiency with respect to [CO₂], and the oxygenation efficiency with respect to [O₂]. With increasing concentrations of CO₂ above the CO₂ compensation point, decreasing quantum flux density, or decreasing O₂, simulations show that the rate of photorespiration progressively decreases. The two components of O₂ inhibition of photosynthesis change disproportionately with increasing CO₂ concentration. According to the model, the energy utilized during photosynthesis at the CO₂ compensation point is about half that under atmospheric conditions.     

The regulation of photosynthetic rate and activation of extracellular carbonic anhydrase under CO2-limiting conditions in the marine diatom Skeletonema costatum

In the marine diatom Skeletonema costatum , carbonic anhydrase activity exterior to the plasma membrane (CA_ext) was detected only when the available CO₂ concentration was less than 5·0 mmol m^–3, this activity being unaffected by the total dissolved inorganic carbon concentration. The inhibition of CA_ext by dextran bound sulphonamide (DBS) demonstrated the key role of this enzyme in maintaining photosynthetic rate under CO₂-limited conditions. Treatment with trypsin followed by affinity chromatography on p-aminomethylbenzene-sulphamide agarose and subsequent SDS-PAGE analysis revealed a polypeptide from carbon-replete cells of identical molecular mass to the CA_ext released by trypsin from CO₂-limited cells. Redox activity in the plasma membrane of intact cells was measured by following the light-dependent reduction of ferricyanide or NADP, the greatest activity being shown by CO₂-limited cells. Overall the results suggest that high rates of redox activity under conditions of CO₂-limitation were required for the activation of CA_ext.     

THE EFFECT OF MODERATE AND MARKED HYPERCAPNIA UPON THE ENERGY STATE AND UPON THE CYTOPLASMIC NADH/NAD+ RATIO OF THE RAT BRAIN

Abstract— The energy state of brain tissue was evaluated from the tissue concentrations of ATP, ADP and AMP and the cytoplasmic NADH/NAD⁺ ratio from the tissue, CSF and blood concentrations of lactate and pyruvate, and from the intracellular pH', in rats exposed to carbon dioxide concentrations of 640 per cent. The hypercapnia had no significant effect on the energy state of the tissue. Hypercapnia of increasing severity gave rise to a progressive decrease in the pyruvate concentration; the lactate concentration fell at low CO₂ concentrations, but no further decrease was observed at CO₂ concentrations greater than 20 per cent. There was a progressive rise in the intracellular lactate/pyruvate ratio at increasing CO₂ concentrations, corresponding to the fall in intracellular pH, i.e. the calculated NADH/NAD⁺ ratios remained normal. It is therefore concluded that hypercapnia does not affect the cytoplasmic redox state.     

The effects of elevated atmospheric CO2 on lipid metabolism in leaves from mature wheat (Triticum aestivum cv. Hereward) plants

Winter wheat (Triticum aestivum L. cv. Hereward) plants were grown for 35 d either at 350 μ mol mol^–1 CO₂ or at 650 μ mol mol^–1 CO₂. Lipid synthesis was studied in these plants by incubating the 5th leaf on the main stem with [1–¹⁴C]acetate. Increased CO₂ concentrations did not significantly affect the total incorporation of radiolabel into lipids of whole leaf tissue, but altered the distribution for individual lipid classes. Most noticeable amongst acyl lipids was the reduction in labelling of diacylglycerol and a corresponding increase in the proportion of phosphatidylcholine labelling. In the basal regions, there were similar changes and, in addition, phosphatidylglycerol labelling was particularly increased following growth in an enriched CO₂ atmosphere. The stimulation of labelling of the mitochondrial-specific lipid, diphosphatidylglycerol, prompted an examination of the mitochondrial population in wheat plants. Mitochondria were localized in intact wheat sections by immunolabelling for the mitochondrial-specific chaperonin probe. Growth in elevated CO₂ doubled the number of mitochondria compared to growth in ambient CO₂. Fatty acid labelling was also significantly influenced following growth at elevated CO₂ concentrations. Most noticeable were the changes in 16C:18C ratios for the membrane lipids, phosphatidylcholine, phosphatidylglycerol and monogalactosyldiacylglycerol. These data imply a change in the apportioning of newly synthesized fatty acids between the 'eukaryotic' and 'prokaryotic' pathways of metabolism under elevated CO₂.     

The acetyl-CoA pathway of autotrophic growth

Abstract The most direct conceivable route for synthesis of multicarbon compounds from CO₂ is to join two molecules of CO₂ together to make a 2-carbon compound and then polymerize the 2-carbon compound or add CO₂ successively to the 2-carbon compound to make multicarbon compounds. Recently, it has been demonstrated that the bacterium, Clostridium thermoaceticum , grows autotrophically by such a process. The mechanism involves the reduction of one molecule of CO₂ to a methyl group and then its combination with a second molecule of CO₂ and CoA to form acetyl-CoA. We have designated this autotrophic pathway the acetyl-CoA pathway [1]. Evidence is accumulating that this pathway is utilized by other bacteria that grow with CO₂ and H₂ as the source of carbon and energy. This group includes bacteria which, like C. thermoaceticum , produce acetate as a major end product and are called acetogens or acetogenic bacteria. It also includes the methane-producing bacteria and sulfate-reducing bacteria.
The purpose of this review is to examine critically the evidence that the acetyl-CoA pathway occurs in other bacteria by a mechanism that is the same or similar to that found in C. thermoaceticum . For this purpose, the mechanism of the acetyl-CoA pathway, as found in C. thermoaceticum , is described and hypothetical mechanisms for other organisms are presented based on the acetyl-CoA pathway of C. thermoaceticum . The available data have been reviewed to determine if the hypothetical schemes are in accord with presently known facts. We conclude that the formation of acetyl-CoA by other acetogens, the methanogens and sulphate-reducing bacteria occurs by a mechanism very similar to that of C. thermoaceticum .     

The long-term responses of the photosynthetic proton circuit to drought

Proton motive force (pmf) across thylakoid membranes is not only for harnessing solar energy for photosynthetic CO₂ fixation, but also for triggering feedback regulation of photosystem II antenna. The mechanisms for balancing these two roles of the proton circuit under the long-term environmental stress, such as prolonged drought, have been poorly understood. In this study, we report on the response of wild watermelon thylakoid 'proton circuit' to drought stress using both in vivo spectroscopy and molecular analyses of the representative photosynthetic components. Although drought stress led to enhanced proton flux via a ∼34% increase in cyclic electron flow around photosystem I (PS I), an observed ∼fivefold decrease in proton conductivity, g_H⁺, across thylakoid membranes suggested that decreased ATP synthase activity was the major factor for sustaining elevated q_E. Western blotting analyses revealed that ATP synthase content decreased significantly, suggesting that quantitative control of the complex plays a pivotal role in down-regulation of g_H⁺. The expression level of cytochrome b ₆ f complex – another key control point in photosynthesis – also declined, probably to prevent excess-reduction of PS I electron acceptors. We conclude that plant acclimation to long-term environmental stress involves global changes in the photosynthetic proton circuit, in which ATP synthase represents the key control point for regulating the relationship between electron transfer and pmf.     

The interaction of high temperature and elevated CO2 on photosynthetic acclimation of single leaves of rice in situ

Rice ( Oryza sativa L. cv. IR72) was grown at three different CO₂ concentrations (ambient, ambient + 200 μmol mol⁻¹, ambient + 300 μmol mol⁻¹) at two different growth temperatures (ambient, ambient + 4°C) from sowing to maturity to determine longterm photosynthetic acclimation to elevated CO₂ with and without increasing temperature. Single leaves of rice showed a cooperative enhancement of photosynthetic rate with elevated CO₂ and temperature during tillering, relative to the elevated CO₂ condition alone. However, after flowering, the degree of photosynthetic stimulation by elevated CO₂ was reduced for the ambient + 4°C treatment. This increasing insensitivity to CO₂ appeared to be accompanied by a reduction in ribulose-1.5-bisphosphate carboxylase/oxygenase (Rubisco) activity and/or concentration as evidenced by the reduction in the assimilation (A) to internal CO₂ (C₁) response curve. The reproductive response (e.g. percent filled grains, panicle weight) was reduced at the higher growth temperature and presumably reflects a greater increase in floral sterility. Results indicate that while CO₂ and temperature could act synergistically at the biochemical level, the direct effect of temperature on floral development with a subsequent reduction in carbon utilization may change sink strength so as to limit photosynthetic stimulation by elevated CO₂ concentration.     

Growth in elevated CO2 enhances temperature response of photosynthesis in wheat

The temperature dependence of C₃ photosynthesis may be altered by the growth environment. The effects of long-term growth in elevated CO₂ on photosynthesis temperature response have been investigated in wheat ( Triticum aestivum L.) grown in controlled chambers with 370 or 700 μmol mol⁻¹ CO₂ from sowing through to anthesis. Gas exchange was measured in flag leaves at ear emergence, and the parameters of a biochemical photosynthesis model were determined along with their temperature responses. Elevated CO₂ slightly decreased the CO₂ compensation point and increased the rate of respiration in the light and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) V_cmax, although the latter effect was reversed at 15°C. With elevated CO₂, J_max decreased in the 15–25°C temperature range and increased at 30 and 35°C. The temperature response (activation energy) of V_cmax and J_max increased with growth in elevated CO₂. CO₂ enrichment decreased the ribulose 1,5-bisphosphate (RuBP)-limited photosynthesis rates at lower temperatures and increased Rubisco- and RuBP-limited rates at higher temperatures. The results show that the photosynthesis temperature response is enhanced by growth in elevated CO₂. We conclude that if temperature acclimation and factors such as nutrients or water availability do not modify or negate this enhancement, the effects of future increases in air CO₂ on photosynthetic electron transport and Rubisco kinetics may improve the photosynthetic response of wheat to global warming.     

Nodulation and nitrogenase activity in nitrogen-fixing woody plants stimulated by CO2 enrichment of the atmosphere

The responses of three species of nitrogen-fixing trees to CO₂ enrichment of the atmosphere were investigated under nutrient-poor conditions. Seedlings of the legume, Robinia pseudoacacia L. and the actinorhizal species, Alnus glutinosa (L.) Gaertn. and Elaeagnus angustifolia L. were grown in an infertile forest soil in controlled-environment chambers with atmospheric CO₂ concentrations of 350 μl ⁻¹ (ambient) or 700 μl ⁻¹. In R. pseudoacacia and A. glutinosa , total nitrogenase (N₂ reduction) activity per plant, assayed by the acetylene reduction method, was significantly higher in elevated CO₂, because the plants were larger and had more nodule mass than did plants in ambient CO₂. The specific nitrogenase activity of the nodules, however, was not consistently or significantly affected by CO₂ enrichment. Substantial increases in plant growth occurred with CO₂ enrichment despite probable nitrogen and phosphorus deficiencies. These results support the premises that nutrient limitations will not preclude growth responses of woody plants to elevated CO₂ and that stimulation of symbiotic activity by CO₂ enrichment of the atmosphere could increase nutrient availability in infertile habitats.     

The response of nodulated alfalfa to water supply, temperature and elevated CO2: photosynthetic downregulation

Plants grown in an environment of elevated CO₂ and temperature often show reduced CO₂ assimilation capacity, providing evidence of photosynthetic downregulation. The aim of this study was to analyse the downregulation of photosynthesis in elevated CO₂ (700 µmol mol⁻¹) in nodulated alfalfa plants grown at different temperatures (ambient and ambient + 4°C) and water availability regimes in temperature gradient tunnels. When the measurements were taken in growth conditions, a combination of elevated CO₂ and temperature enhanced the photosynthetic rate; however, when they were carried out at the same CO₂ concentration (350 and 700 µmol mol⁻¹), elevated CO₂ induced photosynthetic downregulation, regardless of temperature and drought. Intercellular CO₂ concentration measurements revealed that photosynthetic acclimation could not be accounted for by stomatal limitations. Downregulation of plants grown in elevated CO₂ was a consequence of decreased carboxylation efficiency as a result of reduced rubisco activity and protein content; in plants grown at ambient temperature, downregulation was also induced by decreased quantum efficiency. The decrease in rubisco activity was associated with carbohydrate accumulation and depleted nitrogen availability. The root nodules were not sufficiently effective to balance the source–sink relation in elevated CO₂ treatments and to provide the required nitrogen to counteract photosynthetic acclimation.     

Wood composition and energy content in a poplar short rotation plantation on fertilized agricultural land in a future CO2 atmosphere

To study the influence of elevated CO₂ and nitrogen (N) fertilization on wood properties and energy, Populus × euramericana trees were exposed to ambient CO₂ (about 370 μmol mol⁻¹ CO₂) or elevated CO₂ (about 550 μmol mol⁻¹ CO₂) using Free Air CO₂ Enrichment (FACE) technology in combination with two N levels. Elevated CO₂ was maintained for 5 years. After three growing seasons, the plantation was coppiced, one half of each experimental plot was fertilized and secondary sprouts were harvested after two growing seasons. Fourier transform infrared (FT-IR) spectra of wood revealed significant effects of both elevated CO₂ and N fertilization on wood chemistry, in particular, significant increases in lignin and decreases in N content. These results were corroborated by chemical analysis. Neither elevated CO₂ nor N fertilization affected the calorific value of wood, which was 19.3 MJ kg⁻¹. N fertilization enhanced the energy production per land area by 16–69% because of higher aboveground woody biomass production than on nonfertilized land. Estimates indicate that high yielding poplar short rotation cultivation may significantly contribute as an alternative feedstock for energy production.