Active uptake of inorganic carbon by Chlamydomonas reinhardtii: evidence for simultaneous transport of HCO3− and CO2 and characterization of active CO2 transport

1991 ◽  
Vol 69 (5) ◽  
pp. 995-1002 ◽  
Author(s):  
Dieter F. Sültemeyer ◽  
Heinrich P. Fock ◽  
David T. Canvin

Washed protoplasts of low CO2 grown cells of Chlamydomonas reinhardtii were used to further characterize the ability for active CO2 transport. The CO2 transport mechanism and the high affinity for dissolved inorganic carbon were completely induced within 4 h after transferring 5% CO2 grown cells to ambient air (0.035% CO2). Net O2 evolution and CO2 uptake were saturable processes showing saturation between 100 and 200 μM DIC (1.6–3.2 μM CO2) at pH 8.0. For both O2 evolution in whole cells and CO2 uptake in the protoplasts the concentration of dissolved inorganic carbon required for 50% of the maximal rates was about 12 μM (= 0.20 μM CO2). Studies with 3-(3,4-dichloro-phenyl)-1,1 dimethylurea, dibromo-thymoquinone, tetramethyl phenylenediamine and protoplasts of a cytochrome c oxidase deficient mutant of C. reinhardtii indicated the CO2 transport was driven by cyclic or pseudocyclic ATP formation and oxidative phosphorylation was not involved. These studies also show that CO2 transport and CO2 fixation are distinct mechanisms and that active CO2 uptake may occur in the absence of CO2 fixation. Key words: Chlamydomonas reinhardtii; CO2–HCO3− concentrating mechanism, CO2 transport, cyclic photophosphorylation, pseudocyclic photophosphorylation.

2014 ◽  
Vol 32 (3) ◽  
pp. 531
Author(s):  
Danilo R. Sá Teles ◽  
Antônio Expedito G. de Azevedo ◽  
Alexandre B. Costa ◽  
Maria R. Zucchi ◽  
Alexandre A. Ferreira

ABSTRACT. This paper presents a study of the physicochemical and isotopic characteristics of formation waters from the Castanhal, Siririzinho and Aguilhadafields in the Sergipe Basin, northeastern Brazil. In each of the samples, pH, conductivity, concentration of dissolved inorganic carbon (DIC), δ18O, δ2H, and δ13C weremeasured. These measurements are used to identify isotopic similarities among waters from local aquifers, which can be used as a proxy for groundwater connectivitywith formation water. Formation waters from the Castanhal and Siririzinho fields are enriched in deuterium, as evidenced by their δ2H values above the Global MeteoricWater Line (GMWL), which may be a result of significant isotopic exchange between water and H2S. These measurements are in accordance with the large enrichment in13C of DIC resulting from biodegradation, with the formation of methane depleted in 13C and CO2 enriched in 13C. These results indicate interaction between formationwater with rocks and other fluids.Keywords: stable isotopes, dissolved inorganic carbon, biodegradation. RESUMO. Este trabalho apresenta um estudo das características físico-químicas e isotópicas de águas de formação dos campos Castanhal, Siririzinho e Aguilhada, localizados na Bacia de Sergipe, nordeste do Brasil. Em cada uma das amostras foram medidos os parâmetros pH, condutividade elétrica, concentração de carbonoinorgânico dissolvido (CID), δ18O, δ2H e δ13C. A partir dos resultados obtidos, foi possível identificar a similaridade isotópica com as águas de aquíferos locais,indicando conexão de água subterrânea com água de formação. As águas de formação dos campos Castanhal e Siririzinho apresentaram um enriquecimento em deutério,resultado da troca isotópica entre a água e o H2S. Também foi verificado, um grande enriquecimento no δ13C do CID, resultado dos processos de biodegradação coma formação de metano empobrecido em 13C e CO2 enriquecido neste isótopo. Os resultados encontrados indicam interação entre a água de formação com as rochas ecom outros fluidos.Palavras-chave: isótopos estáveis, carbono inorgânico dissolvido, biodegradação.


2021 ◽  
Author(s):  
Sarah Schmid ◽  
Dale Chaput ◽  
Mya Breitbart ◽  
Rebecca Hines ◽  
Samantha Williams ◽  
...  

In nature, concentrations of dissolved inorganic carbon (DIC; = CO 2 + HCO 3 - + CO 3 2- ) can be low, and autotrophic organisms adapt with a variety of mechanisms to elevate intracellular DIC concentrations to enhance CO 2 fixation. Such mechanisms have been well-studied in Cyanobacteria , but much remains to be learned about their activity in other phyla. Novel multi-subunit membrane-spanning complexes capable of elevating intracellular DIC were recently described in three species of bacteria. Homologs of these complexes are distributed among 17 phyla in Bacteria and Archaea, and are predicted to consist of one, two, or three subunits. To determine whether DIC accumulation is a shared feature of these diverse complexes, seven of them, representative of organisms from four phyla, from a variety of habitats, and with three different subunit configurations were chosen for study. A high-CO 2 requiring, carbonic anhydrase-deficient ( yadF - cynT - ) strain of E. coli Lemo21(DE3), which could be rescued via elevated intracellular DIC concentrations, was created for heterologous expression and characterization of the complexes. Expression of all seven complexes rescued the ability of E. coli Lemo21(DE3) yadF - cynT - to grow under low CO 2 conditions, and six of the seven generated measurably elevated intracellular DIC concentrations when their expression was induced. For complexes consisting of two or three subunits, all subunits were necessary for DIC accumulation. Isotopic disequilibrium experiments clarified that CO 2 was the substrate for these complexes. In addition, the presence of an ionophore prevented the accumulation of intracellular DIC, suggesting that these complexes may couple proton potential to DIC accumulation. IMPORTANCE To facilitate the synthesis of biomass from CO 2 , autotrophic organisms use a variety of mechanisms to increase intracellular DIC concentrations. A novel type of multi-subunit complex has recently been described, which has been shown to generate measurably elevated intracellular DIC concentrations in three species of bacteria, begging the question of whether these complexes share this capability across the 17 phyla of Bacteria and Archaea where they are found. This study shows that DIC accumulation is a trait shared by complexes with varied subunit structures, from organisms with diverse physiologies and taxonomies, suggesting that this trait is universal among them. Successful expression in E. coli suggests the possibility of their expression in engineered organisms synthesizing compounds of industrial importance from CO 2 .


2002 ◽  
Vol 29 (3) ◽  
pp. 243 ◽  
Author(s):  
Eddy van Hunnik ◽  
Dieter Sültemeyer

In order to understand the function of the lumen carbonic anhydrase (CA) which is bound to PSII at the lumenal side of the thylakoids in chloroplasts of eukaryotic algae, thylakoids were isolated from chloroplasts of Tetraedron minimum, Chlamydomonas noctigama, the cell wall-less mutant Chlamydomonas reinhardtii CW15, and a C. reinhardtii CW15/CIA3 mutant which lacks the lumen CA. The isolated thylakoids produced O2 on illumination and exhibited electron flow between PSII and PSI, indicating that the thylakoids were intact and the photosynthetic apparatus were functional. We could not detect any uptake of HCO3–,nor efflux of CO2, from the thylakoids upon illumination, making it improbable that the CA present in the lumen of the thylakoids would play a role in furnishing CO2 for Rubisco. We were able to determine ATP production upon illumination in isolated thylakoids. Under high inorganic carbon (Ci; 5 mM), all species showed significant amounts of ATP being produced. Under low Ci (200 M), we could not detect ATP formation from C. reinhardtii CW15/CIA3 upon illumination. This mutant was not able to survive more then 4 h of low Ci in culture. We therefore suggest that the lumen CA is not involved in the CO2 concentrating mechanism, but might play a role in the formation of a proton gradient across the thylakoid membranes.


2017 ◽  
Vol 68 (14) ◽  
pp. 3879-3890 ◽  
Author(s):  
Marylou C Machingura ◽  
Joanna Bajsa-Hirschel ◽  
Susan M Laborde ◽  
Joshua B Schwartzenburg ◽  
Bratati Mukherjee ◽  
...  

1990 ◽  
Vol 63 (02) ◽  
pp. 193-203 ◽  
Author(s):  
John R Shainoff ◽  
Deborah J Stearns ◽  
Patricia M DiBello ◽  
Youko Hishikawa-Itoh

SummaryThe studies reported here probe the existence of a receptor-mediated mode of fibrin-binding by macrophages that is associated with the chemical change underlying the fibrinogen-fibrin conversion (the release of fibrinopeptides from the amino-terminal domain) without depending on fibrin-aggregation. The question is pursued by 1) characterization of binding in relation to fibrinopeptide content of both the intact protein and the CNBr-fragment comprising the amino-terminal domain known as the NDSK of the protein, 2) tests of competition for binding sites, and 3) photo-affinity labeling of macrophage surface proteins. The binding of intact monomers of types lacking either fibrinopeptide A alone (α-fibrin) or both fibrinopeptides A and B (αβ-fibrin) by peritoneal macrophages is characterized as proceeding through both a fibrin-specific low density/high affinity (BMAX ≃ 200–800 molecules/cell, KD ≃ 10−12 M) interaction that is not duplicated with fibrinogen, and a non-specific high density/low affinity (BMAX ≥ 105 molecules/cell, KD ≥ 10−6 M) interaction equivalent to the weak binding of fibrinogen. Similar binding characteristics are displayed by monocyte/macrophage cell lines (J774A.1 and U937) as well as peritoneal macrophages towards the NDSK preparations of these proteins, except for a slightly weaker (KD ≃ 10−10 M) high-affinity binding. The high affinity binding of intact monomer is inhibitable by fibrin-NDSK, but not fibrinogen-NDSK. This binding appears principally dependent on release of fibrinopeptide-A, because a species of fibrin (β-fibrin) lacking fibrinopeptide-B alone undergoes only weak binding similar to that of fibrinogen. Synthetic Gly-Pro-Arg and Gly-His-Arg-Pro corresponding to the N-termini of to the α- and the β-chains of fibrin both inhibit the high affinity binding of the fibrin-NDSKs, and the cell-adhesion peptide Arg-Gly-Asp does not. Photoaffinity-labeling experiments indicate that polypeptides with elec-trophoretically estimated masses of 124 and 187 kDa are the principal membrane components associated with specifically bound fibrin-NDSK. The binding could not be up-regulated with either phorbol myristyl acetate, interferon gamma or ADP, but was abolished by EDTA and by lipopolysaccharide. Because of the low BMAX, it is suggested that the high-affinity mode of binding characterized here would be too limited to function by itself in scavenging much fibrin, but may act cooperatively with other, less limited modes of fibrin binding.


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