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Functional convergence of rTCA-related carbon-fixation potential and biochemical residue accumulation in seagrass sediments.

Seagrass meadows are globally significant blue carbon ecosystems, yet the microbial and biochemical mechanisms driving sediment organic carbon (SOC) accumulation remain poorly understood. To address this, we employed an integrated approach combining metagenomic sequencing, biochemical assays, and structural equation modeling to investigate carbon cycling in the seagrass and adjacent unvegetated sediments of Swan Lake, China. A total of 115,179 carbon fixation genes and 119,615 decomposition genes were identified, revealing distinct microbial community structures among the habitats. Seagrass sediments harbored more diverse carbon-fixing (CFMs) and decomposing microorganisms (CDMs), with 83 medium-to high-quality metagenome-assembled genomes (MAGs) recovered. While neutral community model analysis indicated that stochastic processes predominantly governed community assembly, functional analyses highlighted specific drivers of sequestration. The reductive tricarboxylic acid (rTCA) cycle emerged as the dominant carbon fixation pathway, with key genes (e.g., aclA, korA) showing strong positive correlations with SOC. Conversely, decomposition pathways for starch and lignin were negatively associated with SOC. Furthermore, seagrass sediments exhibited elevated concentrations of total amino sugars (TAS) and lignin phenols (TLP), which linked significantly to carbon fixation rather than decomposition. PLS-SEM revealed statistically significant associations among seagrass traits, environmental variables, microbial carbon-fixation potential, biochemical residue pools, and SOC, supporting a mechanistic pathway in which enhanced microbial functional potential drives the accumulation of recalcitrant biochemical residues, thereby facilitating long-term carbon retention in sediments. These findings emphasize the pivotal role of microbial anabolism and the accumulation of biosynthetic residues in sediment carbon storage, suggesting a functional convergence in seagrass-driven carbon sinks.

Metagenomics

Variations in short term products of inorganic carbon fixation in exponential and stationary phase cultures of Aphanocapsa 6308.

Aphanocapsa 6308 metabolizes both NaHCO3 and Na2CO3. The short term incorporation (5-s) metabolic pattern and the patterns of incorporation of bicarbonate for exponential versus stationary phase cultures differ, however. Cells were equilibrated for 10 min in air and distilled water prior to injection of either NaH14CO3 at pH 8.0, or Na214CO3 at pH 11.0. Hot ethanol extracts were analyzed via paper chromatography and autoradiography for products of CO2 fixation. At 5 s, malate (51.5%) predominates slightly as a primary bicarbonate fixation product over 3-phosphoglycerate (40.3%); 3-phosphoglycerate is the primary product of carbonate fixation. At 60 s, the carbonate and bicarbonate labelling patterns are similar. Cells in stationary phase fix in 5 s a greater proportion of bicarbonate into malate (36% vs. 14% for 3-phosphoglycerate) than do cells in exponential growth. Likewise, 60 s incorporations show a large amount of bicarbonate fixed into aspartate (30.9%) in stationary phase cells over that of exponential phase (11.6%). These data suggest an operative C4 pathway for purposes not related to carbohydrate synthesis but rather as compensation for the incomplete tricarboxylic acid cycle in cyanobacteria. The enhancement of both aspartate fixation and CO2 fixation into citrulline in stationary phase correlates with an increase in cyanophycin granule production which requires both aspartate and arginine.

Aspartic Acid

Toxicity of polychlorinated biphenyls (PCBs) to Euglena gracilis: cell population growth, carbon fixation, chlorophyll level, oxygen consumption, and protein and nucleic acid synthesis.

Populations of Euglena gracilis in exponential growth under light were exposed to 2.5, 5.0, 7.5, and 10 ppm of Aroclor 1221. The ID50/48 of Aroclor 1221 was estimated to be 4.4 ppm, while Aroclor 1232 tested at 20, 35, 50, and 100 ppm resulted in an id50/48 of 55 ppm. With Aroclor 1242, no inhibition of growth was observed with up to 100 ppm exposure. Cell cultures exposed to 4.4 ppm of Aroclor 1221 for 48 hrs had a significantly reduced rate of carbon fixation and reduced levels of chlorophyll after correction for cell density. Oxygen consumption was not affected at the ID50 level of the Aroclor. Uptake of [3H]-leucine in treated cultures was twice that of controls, and [3H]-uridine uptake was significantly lower. Uptake of [3H]-thymidine, and incorporation of [3H]-leucine, [3H]-thymidine, and [3H]-uridine were not significantly different in treated and control cultures. Thes results suggest that at the ID50 level, polychlorinated biphenyls (PCBs) reduce cell population growth in Euglena gracilis by inhibition of photosynthesis and/or chlorophyll production.

Animals

The mechanism of the control of carbon fixation by the pH in the chloroplast stroma. Studies with nitrite-mediated proton transfer across the envelope.

1. CO2 fixation of intact spinach chloroplasts is inhibited by nitrite in a pH-dependent mode. At pH 7.3 in the medium 1 mM NaNO2 and at pH 7.9 5 mM NaNO2 were required for 50% inhibition. 2. The addition of nitrite leads to an acidificiation in the stroma. It appears that nitrite renders the envelope permeable for protons resulting in a breakdown of the pH gradient between the external space and the stroma. 3. In view of earlier results on the pH sensitivity of C02 fixation it is concluded that this pH shift in the stroma is responsible for the observed inhibition of CO2 fixation by nitrite. 4. Octanoate and to some extent also high concentrations of bicarbonate and acetate have a similar effect as nitrite in inhibiting CO2 fixation through an acidification in the stroma. 5. The levels of the intermediates of the CO2 fixation cycle were measured. A strong rise of the levels of fructose- and sedoheptulose biphosphates and a concomitant decrease of the corresponding monophosphates was observed during inhibition of CO2 fixation. It appears that the enzymatic steps of the CO2 fixation cycle responsible for the overall inhibition of CO2 fixation caused by lowering of the H+ concentration in the stroma are fructose- and sedopheptulose bisphosphatase. These two enzymes have an important function in the light regulation of CO2 fixation.

Ammonium Chloride

The role of pH in the regulation of carbon fixation in the chloroplast stroma. Studies on CO2 fixation in the light and dark.

1. The pH in the stroma and in the thylakoid space has been measured in a number of chloroplast preparations in the dark and in the light at 20 degrees C. Illumination causes a decrease of the pH in the thylakoid space by 1.5 and an increase of the pH in the stroma by almost 1 pH unit. 2. CO2 fixation is shown to be strongly dependent on the pH in the stroma. The pH optimum was 8.1, with almost zero activity below pH 7.3.Phosphoglycerate reduction, which is a partial reaction of CO2 fixation, shows very little pH dependency. 3. Low concentrations of the uncoupler m-chlorocarbonylcyanide phenylhydrazone (CCCP) inhibit CO2 fixation without affecting phosphoglycerate reduction. This inhibition of CO2 fixation appears to be caused by reversal of light induced alkalisation in the stroma by CCCP. 4. Methylamine has a very different effect compared to CCCP. Increasing concentrations of methylamine inhibit CO2 fixation and phosphoglycerate reduction to the same extent. The light induced alkalisation of the stroma appears not to be significantly inhibited by methylamine, but the protons in the thylakoid space are neutralized. The inhibition of CO2 fixation by higher concentrations of methylamine is explained by an inhibition of photophosphorylation. It appears that methylamine does not abolish proton transport. 5. It is shown that intact chloroplasts are able to fix CO2 in the dark, yielding 3-phosphoglycerate. This requires the addition of dihydroxyacetone phosphate as precursor of ribulosemonophosphate and also to supply ATP, and the addition of oxaloacetate for reoxidation of the NADPH in the stroma. 6. Dark CO2 fixation in the presence of dihydroxyacetone phosphate and oxaloacetate has the same pH dependency as CO2 fixation in the light. This demonstrates that CO2 fixation in the dark is not possible, unless the pH in the medium is artificially raised to pH 8.8.

Adenosine Triphosphate

Polyphosphate acts as an architectural regulator of carbon fixation and nucleoid structure in cyanobacteria.

Polyphosphate (polyP) is a conserved inorganic polymer traditionally viewed as a stress-induced phosphate and energy reserve. In cyanobacteria, however, polyP granules are constitutively present and are frequently observed in close proximity to carboxysomes, the bacterial microcompartments that mediate CO2 fixation. Here, we show that polyP functions as a spatially organized regulator of the photosynthetic cytoplasm in Synechococcus elongatus. PolyP granules localize to the nucleoid and are periodically arranged along the cell axis, independent of the McdAB carboxysome positioning system. Despite this independence, polyP and carboxysomes associate non-randomly, and this association is enhanced when active carboxysome positioning by the McdAB system is disrupted. Loss of polyP synthesis leads to nucleoid expansion, an increased number of smaller carboxysomes with high mobility, and severe defects in growth under ambient CO2. Perturbation of polyP turnover further reveals structural connections to both carboxysomes and thylakoid membranes. Together, these findings identify polyP as an architectural integrator that couples chromosome organization, metabolic compartmentalization, and photosynthetic fitness.IMPORTANCEPolyphosphate (polyP) is a ubiquitous storage polymer found across all three domains of life. In bacteria, polyP is involved in virulence, energy metabolism, DNA structure, and stress. Here, we investigate the physical and functional relationships between polyP and the carboxysome-a carbon-fixing organelle that encapsulates the most abundant enzyme on Earth and is responsible for a significant amount of atmospheric CO2 sequestration. We discover that polyP is important for proper carboxysome assembly and organization in the cell and is also required for robust cell growth under ambient CO2 conditions. As carboxysomes are one class of the broader group of bacterial microcompartments, these findings have broader implications for polyP in the function and organization of the bacterial cytoplasm.

Polyphosphates

Polyphosphate acts as an architectural regulator of carbon fixation and nucleoid structure in cyanobacteria.

Polyphosphate (polyP) is a conserved inorganic polymer traditionally viewed as a stress-induced phosphate and energy reserve. In cyanobacteria, however, polyP granules are constitutively present and frequently observed in proximity to carboxysomes, the bacterial microcompartments that mediate CO2 fixation. Here we show that polyP functions as a spatially organized regulator of the photosynthetic cytoplasm in Synechococcus elongatus. PolyP granules localize to the nucleoid and are periodically arranged along the cell axis, independently of the McdAB carboxysome positioning system. Despite this independence, polyP and carboxysomes associate non-randomly, and this association is enhanced when active carboxysome positioning by the McdAB system is disrupted. Loss of polyP synthesis leads to nucleoid expansion, an increased number of smaller carboxysomes with high mobility, and severe defects in growth under ambient CO2. Perturbation of polyP turnover further reveals structural connections to both carboxysomes and thylakoid membranes. Together, these findings identify polyP as an architectural integrator that couples chromosome organization, metabolic compartmentalization, and photosynthetic fitness.

Journal Article

Pyruvate inhibition of the carbon dioxide fixation of the strict chemolithotroph Thiobacillus thiooxidans.

A flow-through dialysis system used to decrease the concentrations of toxic organic materials excreted by Thiobacillus thiooxidans permitted an improved efficiency of carbon dioxide fixation when compared with cells taken from the usual shaken culture. The additions of various concentrations of pyruvic acid and succinic acid inhibited growth significantly. Pyruvate at a concentration of 5 X 10(-3) M completely inhibited the respiration of resting cells oxidizing sulfur. The toxicity of pyruvic acid was found to be permanent as evidenced by the inability to obtain satisfactory oxidation rates after washing the exposed cells twice in buffer. Both pyruvate (10(-3) M) and succinate (10(-3) M) inhibited carbon dioxide fixation by 84%.

Acidithiobacillus thiooxidans

[New data on carbonic acid fixation in animals and man].

Recently a new enzymic carboxylation reaction has been discovered at the final-step of prothrombin and other proteins synthesis. The presence of the unique amino acid, gamma-carboxyglutaminic acid, is established and its structure is determined. The role of the acid is shown in Ca2+ fixation in the known proteins, playing an important role in the blood coagulation processes, and in some new proteins participating in the tissue calcification. Data are systematized on the necessity of vitamin K in the new reaction of the carbonic acid fixation under conditions of its in vitro simulation.

1-Carboxyglutamic Acid

Carbon dioxide fixation in the brain: its relation to glucose synthesis.

The incorporation in vivo of radiocarbon from 14C-bicarbonate in blood into relevant metabolites in rat brain is described. The animals, partially hepatectomized and nephrectomized, received the tracer bicarbonate via the intravenous route. The time course of label was followed in CO2 of blood and brain, in the anionic and cationic fractions of brain extract, in aspartate, glutamate, glutamine and in free glucose and in glycogen. From the tracer kinetic data a flux of 0.08 microgram atom fixed carbon min-1.g-1 brain tissue was calculated. Substantial amounts of 14C were found in free glucose, only a few percent in glycogen. The flux of newly synthetized glucose was approximated to 0.5--1.0 percent of the steady state level of glucose in brain tissue. In special experiments the localization of 14C in the carbon chain of aspartate and glucose was examined. 5 min following the tracer injection a practically total randomization of 14C between C-1 and C-4 aspartate was seen. From the radioactivity in glucose 94 percent were found in C-3 and C-4, only 6 percent in residual carbon. This 14C-pattern is typical for the labelling of glucose by CO2 fixation and retrograde Embden-Meyerhof pathway.

Animals

[Carbon dioxide fixation in aquatic animals and its metabolic significance].

In experiments on the carp Cyprinus carpio and freshwater lamellibranch mollusc Anodonta cygnea, tissue and organ peculiarities of carboxylic reactions have been revealed together with their relationship to the temperatute and ionic composition of the incubation medium. It was shown that in fish the highest intensity of fixation of CO2 is exhibited by glandular organs with biosynthetic profile of the metabolism (liver), whereas in molluscs it is exhibited in the mantle which plays the key role in the formation of the shell, containing carbonate compounds of calcium.

Animals

Inhibition of carbon dioxide fixation by lead acetate in rat liver mitochondria.

These studies were undertaken to determine the mechanism by which intravenously administered lead salts inhibit hepatic gluconeogenesis. Within 1 h after the intravenous administration of lead acetate (10 mg), there is 97% inhibition of CO2 fixation in isolated rat liver mitochondria. This effect is concentration-dependent. The induction of phosphoenolpyruvate carboxykinase activity observed with starvation was also inhibited by intravenously administered lead acetate, but the activities of pyruvate kinase, glucose 6-phosphate dehydrogenase and pyruvate carboxylase were unaffected, as was the oxidation of palmitate and palmitoyl-CoA by mitochondria from Pb2+-treated animals. The addition of reduced glutathione to mitochondria from Pb2+-treated animals had no effect on the inhibited CO2 fixation. ATP concentrations in mitochondria from Pb2+-treated animals are decreased and the dose-response relationships for the effect of Pb2+ on CO2 fixation and ATP concentrations correspond. We conclude that the decrease in mitochondrial ATP in Pb2+-treated animals is probably responsible for the marked inhibition ov CO2 fixation, and hence the impairment of gluconeogenesis from alanine, lactate and pyruvate observed by others.

Adenosine Triphosphate

Carbon dioxide fixation in Helminthosporium cynodontis.

A mycelial suspension of Helminthosporium cynodontis (ATCC24938), grown on glucose-peptone-yeast extract broth and exposed to NaH14CO3 for 5 h, fixed significant quantities of 14C into the following fractions (%): small molecular weight components, 7-4; lipid and lipoproteins, 3-9; nucleic acids, 59; the residual protein and cell wall fragments, 29-2. The labelled protein components were (%): aspartate, 39; glutamate, 18; cystine, 15; threonine, 9. Radioactive nucleic acid components were (%): adenine, 18; guanine, 18; cytidylate, 34; uridylate, 30. When the mycelium was grown in Czapek-Dox glucose medium and incubated in this medium plus NaH14CO3, the nucleic acid fraction contained 29-9% and the residual protein 49-5% of the cellular radioactivity. The removal of CO2 from the atmosphere did not reduce growth. Pyruvate carboxylase (PC) and phosphoenolypyruvate carboxykinase (PEPCK) activities were demonstrated in extracts of H. cynodontis. Synthesis of PEPCK was stimulated under conditions promoting gluconeogenesis and was reduced under conditions promoting glycolysis, while PC synthesis was similar under both conditions.

Carbon Dioxide