Addition Of Carbonic Anhydrase Research Articles

Periplasmic carbonic anhydrase CAH1 contributes to high inorganic carbon affinity in Chlamydomonas reinhardtii.

Carbonic anhydrase (CA), an enzyme conserved across species, is pivotal in the interconversion of inorganic carbon (Ci; CO2, and HCO3-). Compared to the well-studied intracellular CA, the specific role of extracellular CA in photosynthetic organisms is still not well understood. In the green alga Chlamydomonas (Chlamydomonas reinhardtii), carbonic anhydrase 1 (CAH1), located at the periplasmic space, is strongly induced under CO2-limiting conditions by the Myb transcription factor LCR1. While the lcr1 mutant shows decreased Ci-affinity, the detailed mechanisms behind this phenomenon are yet to be elucidated. In this study, we aimed to unravel the LCR1-dependent genes essential for maintaining high Ci-affinity. To achieve this, we identified a total of 12 LCR1-dependent inducible genes under CO2-limiting conditions, focusing specifically on the most prominent ones-CAH1, LCI1, LCI6, and Cre10.g426800. We then created mutants of these genes using the CRISPR-Cas9 system, all from the same parental strain, and compared their Ci-affinity. Contrary to earlier findings that reported no reduction in Ci-affinity in the cah1 mutant, our cah1-1 mutant exhibited a decrease in Ci-affinity under high HCO3-/CO2-ratio conditions. Additionally, when we treated wild-type cells with a CA inhibitor with low membrane permeability, a similar reduction in Ci-affinity was observed. Moreover, the addition of exogenous CA to the cah1 mutant rescued the decreased Ci-affinity. These results, highlighting the crucial function of the periplasmic CAH1 in maintaining high Ci-affinity in Chlamydomonas cells, provide insights into the functions of periplasmic CA in algal carbon assimilation.

The Kinetics Investigation of CO2 Absorption into TEA and DEEA Amine Solutions Containing Carbonic Anhydrase

Tertiary amines have been used as alternative absorbents for traditional primary and secondary amines in the process of carbon capture. However, the carbon dioxide (CO2) absorption rates in these kinds of amine are relatively slow, which implies greater investment and construction costs and limits the large-scale application of carbon capture. Carbonic anhydrase (CA) is considered to be an ideal homogeneous catalyst for accelerating the rate of CO2 into aqueous amine solution. In this work, CO2 absorption combining CA with two single aqueous tertiary amines, namely triethanolamine (TEA) and 2-(diethylamino)ethanol (DEEA), was studied by use of the stopped-flow apparatus over temperature ranging from 293 to 313 K. The concentrations of selected aqueous amine solution and CA used in the experiment were ranging among 0.1–0.5 kmol/m3 and 0–50 g/m3 , respectively. Compared to the solution without the addition of CA, the pseudo first-order reaction rate in the presence of CA (k0,withCA) is significantly increased. The values of k0,withCA have been calculated by a new kinetics model. The results of experimental and calculated k0,amine and k0,withCA in CO2-amine-H2O solutions were also investigated,respectively.

Pore water conditions driving calcium carbonate dissolution in reef sands

Pore water conditions driving calcium carbonate dissolution in reef sands

Addition of carbonic anhydrase 9 inhibitor SLC-0111 to temozolomide treatment delays glioblastoma growth in vivo.

Tumor microenvironments can promote stem cell maintenance, tumor growth, and therapeutic resistance, findings linked by the tumor-initiating cell hypothesis. Standard of care for glioblastoma (GBM) includes temozolomide chemotherapy, which is not curative, due, in part, to residual therapy-resistant brain tumor-initiating cells (BTICs). Temozolomide efficacy may be increased by targeting carbonic anhydrase 9 (CA9), a hypoxia-responsive gene important for maintaining the altered pH gradient of tumor cells. Using patient-derived GBM xenograft cells, we explored whether CA9 and CA12 inhibitor SLC-0111 could decrease GBM growth in combination with temozolomide or influence percentages of BTICs after chemotherapy. In multiple GBMs, SLC-0111 used concurrently with temozolomide reduced cell growth and induced cell cycle arrest via DNA damage in vitro. In addition, this treatment shifted tumor metabolism to a suppressed bioenergetic state in vivo. SLC-0111 also inhibited the enrichment of BTICs after temozolomide treatment determined via CD133 expression and neurosphere formation capacity. GBM xenografts treated with SLC-0111 in combination with temozolomide regressed significantly, and this effect was greater than that of temozolomide or SLC-0111 alone. We determined that SLC-0111 improves the efficacy of temozolomide to extend survival of GBM-bearing mice and should be explored as a treatment strategy in combination with current standard of care.

Pilot Absorption Experiments with Carbonic Anhydrase Enhanced MDEA

Pilot Absorption Experiments with Carbonic Anhydrase Enhanced MDEA

Enzymatic Electrosynthesis of Formic Acid through Carbon Dioxide Reduction in a Bioelectrochemical System: Effect of Immobilization and Carbonic Anhydrase Addition.

The enzymatic electrosynthesis of formic acid from the reduction of carbon dioxide (CO2 ) by using formate dehydrogenase (FDH) as a catalyst at the cathode in both its free and immobilized forms was studied in detail in a bioelectrochemical system (BES). The essential role of solubilizing CO2 for its conversion was also studied by adding carbonic anhydrase (CA) to the FDH enzyme in both its free and immobilized forms. FDH alone in the free form showed large variation in the reduction current [(-6.2±3.9) A m-2 ], whereas the immobilized form showed less variation [(-3.8±0.5) A m-2 ] due to increased enzyme stability. The addition of CA with FDH increased the consumption of the current in both forms due to the fact that it allowed rapid dissolution of CO2 , which made it available for the catalytic reaction with FDH. Remarkably, stable consumption of the current was observed throughout the operation if both CA and FDH were immobilized onto the electrode [(-3.9±0.2) A m-2 ]. Product formation by the immobilized enzyme was also continued for three repetitive cycles, which revealed the longevity of the enzyme after immobilization. The recyclability of NADH (NAD=nicotinamide adenine dinucleotide) was also clearly evidenced on the derivative voltammetric signature. Extension of this study for continuous and long-term operation may reveal more possibilities for the rapid capture and conversion of CO2 .

Role of bacterial carbonic anhydrase during CO2 capture in the CO2-H2O-carbonate system

Role of bacterial carbonic anhydrase during CO2 capture in the CO2-H2O-carbonate system

Effect of water activity on carbon dioxide transport in cholinium-based ionic liquids with carbonic anhydrase

Effect of water activity on carbon dioxide transport in cholinium-based ionic liquids with carbonic anhydrase

Reactive absorption of CO2 into enzyme accelerated solvents: From laboratory to pilot scale

Reactive absorption of CO2 into enzyme accelerated solvents: From laboratory to pilot scale

Effect of carbonic anhydrase on enzymatic conversion of CO2 to formic acid and optimization of reaction conditions

Effect of carbonic anhydrase on enzymatic conversion of CO2 to formic acid and optimization of reaction conditions

재조합 탄산무수화 효소 첨가 생산배지를 이용한 Actinobacillus succinogenes 유래의 숙신산 생산성 향상

Succinic acid, a representative biomass-derived platform chemical, is a major fermentation product of Actinobacillus succinogenes. It is well known that carbon dioxide is consumed during the succinate fermentation, but the biochemical mechanism behind this phenomenon is not yet understood well. In this study, it was found that the addition of carbonic anhydrase (CA)s into media significantly enhances the succinic acid production by A. succinogenes during the fermentation supplied with carbon dioxide. It is likely that the (bi) carbonate produced by the CA activity from gaseous carbon dioxide is favoured by A. succinogenes for consumption and utilization. Therefore, the MgCO₃ requirement could be significantly reduced without compromising the succinate productivity. Furthermore, because of too high price of the commercial carbonic anhydrase, it was undertaken to economically overproduce a cyanobacterial carbonic anhydrase by the use of a recombinant Pichia pastoris. An expression vector system was constructed with the carbonic anhydrase gene PCR-cloned from Cyanobacterium Synechocystis sp., and introduced into P. pastoris for fermentation studies. About 95.9 g/L of succinic acid was produced in the production medium with 30 ppm of carbonic anhydrase, approximately 2 fold higher productivity compared to the parallel process with no supplementation of the enzyme. It is expected that this method can provide a valuable way of overcoming inefficiencies inherent in gas supply during CO₂-based bioprocesses like succinic acid fermentation.

The effect of carbonic anhydrase on the kinetics and equilibrium of the oxygen isotope exchange in the CO2–H2O system: Implications for δ18O vital effects in biogenic carbonates

The effect of carbonic anhydrase on the kinetics and equilibrium of the oxygen isotope exchange in the CO2–H2O system: Implications for δ18O vital effects in biogenic carbonates

Inorganic carbon acquisition in two green marine Stichococcus species

The mechanism of inorganic carbon (C(i)) uptake was examined in the marine green microalgae Stichococcus cylindricus and Stichococcus minor. External carbonic anhydrase (CA) activity was not detected in either species, by potentiometric assay or by mass spectrometry. Photosynthetic characteristics of C(i) uptake indicate that both species have high apparent affinity for CO(2) with a low K(1/2) (CO(2)) of about 10 µm. The O(2) evolution rates in light exceeded the spontaneous CO(2) formation rate by 2.5-fold in both species, which thus have active bicarbonate uptake. Mass spectrometric monitoring of CO(2) and O(2) fluxes showed that rates of O(2) evolution exceeded those of CO(2) depletion by about three- and twofold in S. minor and S. cylindricus, respectively, and also showed, in cells photosynthesizing at pH 8.2, a rapid depletion of CO(2) upon illumination to a CO(2) compensation concentration of 15.42 and 12.03 µm in S. minor and S. cylindricus, respectively. Both species also exhibit active CO(2) uptake: addition of bovine CA at CO(2) compensation concentration caused a rapid rise in CO(2) as the CO(2) -HCO(3) (-) equilibrium was restored. Accumulation of unfixed C(i) by cells at pH 8.2 was calculated to be 84.33 mm in S. cylindricus, and 30.37 mm in S. minor to give internal accumulations of 23- and 8-fold, respectively, compared to the external C(i) concentration.

Plasma-accessible carbonic anhydrase at the tissue of a teleost fish may greatly enhance oxygen delivery:in vitroevidence in rainbow trout,Oncorhynchus mykiss

During a generalized acidosis in rainbow trout, catecholamines are released into the blood, activating red blood cell (RBC) Na(+)/H(+) exchange (βNHE), thus protecting RBC intracellular pH (pH(i)) and subsequent O(2) binding at the gill. Because of the presence of a Root effect (a reduction in oxygen carrying capacity of the blood with a reduction in pH), the latter could otherwise be impaired. However, plasma-accessible carbonic anhydrase (CA) at the tissues (and absence at the gills) may result in selective short-circuiting of RBC βNHE pH regulation. This would acidify the RBCs and greatly enhance O(2) delivery by exploitation of the combined Bohr-Root effect, a mechanism not previously proposed. As proof-of-principle, an in vitro closed system was developed to continuously monitor extracellular pH (pH(e)) and O(2) tension (P(O(2))) of rainbow trout blood. In this closed system, adding CA to acidified, adrenergically stimulated RBCs short-circuited βNHE pH regulation, resulting in an increase in P(O(2)) by >30 mmHg, depending on the starting Hb-O(2) saturation and degree of initial acidification. Interestingly, in the absence of adrenergic stimulation, addition of CA still elevated P(O(2)), albeit to a lesser extent, a response that was absent during general NHE inhibition. If plasma-accessible CA-mediated short-circuiting is operational in vivo, the combined Bohr-Root effect system unique to teleost fishes could markedly enhance tissue O(2) delivery far in excess of that in vertebrates possessing a Bohr effect alone and may lead to insights about the early evolution of the Root effect.

Methanol Production via Bioelectrocatalytic Reduction of Carbon Dioxide: Role of Carbonic Anhydrase in Improving Electrode Performance

Electrocatalytic production of methanol from CO2 has recently been studied. This paper focuses on understanding the role of carbonic anhydrase to efficiently facilitate uptake of CO2, which can be the rate determining step. The three oxidoreductase enzymes responsible for CO2 reduction to methanol are formate, aldehyde, and alcohol dehydrogenase. This enzyme cascade was coupled to a poly(neutral red) modified electrode to regenerate NADH. We have found that the dehydrogenases alone can achieve reduction of CO2, but the process is accelerated by the addition of carbonic anhydrase. As researchers focus on electrofuels, carbonic anhydrase will likely improve performance.

Carbon Dioxide Transport through Membranes

Several membrane channels, like aquaporin-1 (AQP1) and the RhAG protein of the rhesus complex, were hypothesized to be of physiological relevance for CO(2) transport. However, the underlying assumption that the lipid matrix imposes a significant barrier to CO(2) diffusion was never confirmed experimentally. Here we have monitored transmembrane CO(2) flux (J(CO2)) by imposing a CO(2) concentration gradient across planar lipid bilayers and detecting the resulting small pH shift in the immediate membrane vicinity. An analytical model, which accounts for the presence of both carbonic anhydrase and buffer molecules, was fitted to the experimental pH profiles using inverse problems techniques. At pH 7.4, the model revealed that J(CO2) was entirely rate-limited by near-membrane unstirred layers (USL), which act as diffusional barriers in series with the membrane. Membrane tightening by sphingomyelin and cholesterol did not alter J(CO2) confirming that membrane resistance was comparatively small. In contrast, a pH-induced shift of the CO(2) hydration-dehydration equilibrium resulted in a relative membrane contribution of about 15% to the total resistance (pH 9.6). Under these conditions, a membrane CO(2) permeability (3.2 +/- 1.6 cm/s) was estimated. It indicates that cellular CO(2) uptake (pH 7.4) is always USL-limited, because the USL size always exceeds 1 mum. Consequently, facilitation of CO(2) transport by AQP1, RhAG, or any other protein is highly unlikely. The conclusion was confirmed by the observation that CO(2) permeability of epithelial cell monolayers was always the same whether AQP1 was overexpressed in both the apical and basolateral membranes or not.

Comparative study of dissolution rate-determining mechanisms of limestone and dolomite

The dissolution rate-determining processes of carbonate rocks include: (1) heterogeneous reactions on rock surfaces; (2) mass transport of ions into solution from rock surfaces via diffusion; and (3) the conversion reaction of CO2 into H+ and HCO3−. Generally, it is the slowest of these three processes that limits the dissolution rate of carbonate rock. However, from experiment and theoretical analysis under similar conditions not only were the initial dissolution rates of dolomite lower by a factor of 3–60 than those of limestone, but also there are different dissolution rate-determining mechanisms between limestone and dolomite. For example, for limestone under the condition of CO2 partial pressures \( {\left( {P_{{{\text{CO}}_{{\text{2}}} }} } \right)} > 100\,{\text{Pa}} \) dissolution rates increased significantly by a factor of about ten after addition of carbonic anhydrase (CA) into solution, which catalysed the conversation reaction of CO2, whereas CA had little influence on dolomite dissolution. For dolomite, the increase of dissolution rate after addition of CA into solution appeared at \( P_{{{\text{CO}}_{{\text{2}}} }} < 10,000\,{\text{Pa}}{\text{.}} \) Moreover, the enhancement factor of CA on dolomite dissolution rate was much lower (by a factor of about 3). In addition, when dissolution of both limestone and dolomite was determined by hydrodynamics (rotation speed or flow speed), especially under \( P_{{{\text{CO}}_{{\text{2}}} }} {\text{ $<$ 1,000}}\,{\text{Pa,}} \) the dissolution of limestone was more sensitive to hydrodynamic change than that of dolomite. These findings are of significance in understanding the differences in karstification and relevant problems of resource and environment in dolomite and limestone areas.

Buffering limits plasma HCO3− dehydration when red blood cell anion exchange is inhibited

Buffering limits plasma HCO3− dehydration when red blood cell anion exchange is inhibited

ACQUISITION OF INORGANIC CARBON BY THE MARINE HAPTOPHYTE ISOCHRYSIS GALBANA (PRYMNESIOPHYCEAE)1

Inorganic carbon acquisition has been investigated in the marine haptophyte Isochrysis galbana. External carbonic anhydrase (CA) was present in air‐grown (0.034% CO2) cells but completely repressed in high (3%) CO2‐grown cells. External CA was not inhibited by 1.0 mM acetazolamide. The capacity of cells to take up bicarbonate was examined by comparing the rate of photosynthetic O2 evolution with the calculated rate of spontaneous CO2 supply; at pH 8.2 the rates of O2 evolution exceeded the CO2 supply rate 14‐fold, indicating that this alga was able to take up HCO3−. Monitoring CO2 concentrations by mass spectrometry showed that suspensions of high CO2‐grown cells caused a rapid drop in the extracellular CO2 in the light and addition of bovine CA raised the CO2 concentration by restoring the HCO3−‐CO2 equilibrium, indicating that cells were maintaining the CO2 in the medium below its equilibrium value during photosynthesis. A rapid increase in extracellular CO2 concentration occurred on darkening the cells, indicating that the cells had accumulated an internal pool of unfixed inorganic carbon. Active CO2 uptake was blocked by the photosynthetic electron transport inhibitor 3‐(3′,4′‐dichlorphenyl)‐1,1‐dimethylurea, indicating that CO2 transport was supported by photosynthetic reactions. These results demonstrate that this species has the capacity to take up HCO3− and CO2 actively as sources of substrate for photosynthesis and that inorganic carbon transport is not repressed by growth on high CO2, although external CA expression is regulated by CO2 concentration.

Role of Carbonic Anhydrase as an Activator in Carbonate Rock Dissolution and Its Implication for Atmospheric CO2 Sink

AbstractThe conversion of CO2 into H+ and is a relatively slow reaction. Hence, its kinetics may be rate determining in carbonate rock dissolution. Carbonic anhydrase (CA), which is widespread in nature, was used to catalyze the CO2 conversion process in dissolution experiments of limestone and dolomite. It was found that the rate of dissolution increases by a factor of about 10 after the addition of CA at a high CO2 partial pressure (Pco2) for limestone and about 3 at low Pco2 for dolomite. This shows that reappraisal is necessary for the importance of chemical weathering (including carbonate rock dissolution and silicate weathering) in the atmospheric CO2 sink and the mysterious missing sink in carbon cycling. It is doubtless that previous studies of weathering underestimated weathering rates due to the ignorance of CA as an activator in weathering, thus the contribution of weathering to the atmospheric CO2 sink is also underestimated. This finding also shows the need to examine the situ distribution and activity of CA in different waters and to investigate the role of CA in weathering.