scholarly journals Detection of urease and carbonic anhydrase activity using a rapid and economical field test to assess microbially-induced carbonate precipitation

2020 ◽  
Author(s):  
Fernando Medina Ferrer ◽  
Kathryn Hobart ◽  
Jake V. Bailey
Keyword(s):  
Carbon Sequestration ◽  
Carbonic Anhydrase ◽  
Marker Gene ◽  
Cost Effective ◽  
Carbonic Anhydrase Activity ◽  
Soil Restoration ◽  
Reaction Products ◽  
Calcareous Fens ◽  
Molecular Approaches

ABSTRACTMicrobial precipitation of calcium carbonate has diverse engineering applications, from building and soil restoration, to carbon sequestration. Urease-mediated ureolysis and CO2 (de)hydration by carbonic anhydrase (CA) are known for their potential to precipitate carbonate minerals, yet many microbial community studies rely on marker gene or metagenomic approaches that are unable to determine in situ activity. Here, we developed fast and cost-effective tests for the field detection of urease and CA activity using pH-sensitive strips inside microcentrifuge tubes that change color in response to the reaction products of urease (NH3) and CA (CO2). Samples from a saline lake, a series of calcareous fens, and ferrous springs were assayed in the field, finding relatively high urease activity in lake samples, whereas CA activity was only detected in a ferrous spring. Incubations of lake microbes with urea resulted in significantly higher CaCO3 precipitation compared to incubations with a urease inhibitor. Therefore, the rapid assay indicated an on-site active metabolism potentially mediating carbonate mineralization. Field urease and CA activity assays complement molecular approaches and facilitate the search for carbonate-precipitating microbes and their in situ activity, which could be applied toward agriculture, engineering and carbon sequestration technologies.

In Situ Photoregulation of Carbonic Anhydrase Activity Using Azobenzenesulfonamides

Biochemistry ◽  
2018 ◽  
Vol 58 (1) ◽  
pp. 48-53 ◽  
Author(s):  
Kanchan Aggarwal ◽  
Mandira Banik ◽  
Brenda Medellin ◽  
Emily L. Que
Keyword(s):  
Carbonic Anhydrase ◽  
Carbonic Anhydrase Activity

Changes in inorganic carbon uptake during the progression of a dinoflagellate bloom in a lake ecosystem

1998 ◽  
Vol 76 (6) ◽  
pp. 1043-1051 ◽  
Author(s):  
Ilana Berman-Frank ◽  
Jonathan Erez ◽  
Aaron Kaplan
Keyword(s):  
Carbonic Anhydrase ◽  
Growth Rates ◽  
Inorganic Carbon ◽  
Carbonic Anhydrase Activity ◽  
Quantum Yields ◽  
Carbon Uptake ◽  
In Situ Growth ◽  
Photosynthetic Rates ◽  
Inorganic Carbon Uptake

The physiological, biochemical, and genetic aspects of inorganic (Ci) carbon uptake in aquatic plants and algae have been studied extensively. Yet, to date, few studies examined these questions on dominant phytoplankton populations in their natural environment. Lake Kinneret, Israel, provides a good example of a system in which changes in CO2 availability play a vital role in the ecophysiology of inorganic carbon uptake and in the population dynamics during the annual bloom of the dinoflagellate Peridinium gatunense Nygaard. In this study we investigated whether the availability of CO2(aq) limited growth rates and primary productivity of in situ populations of P. gatunense and focused on the role of adaptive mechanisms for Ci uptake. At the onset of the bloom, when epilimnetic pH was low ( = 8) and Ci concentrations were high ( = 2.5 mM), carbonic anhydrase activity and cellular affinity to CO2(aq) were comparatively low. At this time photosynthetic rates, quantum yields, and in situ growth rates were high. As P. gatunense biomass increased, inorganic carbon decreased by 40%, while CO2(aq) concentrations declined 50-fold to values less than 2 µM. The algae adapted by acquiring a CO2-concentrating mechanism indicated by (i) intracellular Ci-concentrations higher by a factor of 5-70 relative to the ambient Ci; (ii) levels of carbonic anhydrase activity higher by 5- to 50-fold than those at the beginning of the bloom; and (iii) enhanced affinity for Ci and CO2(aq) 3- and 40-fold higher, respectively, than affinities at the start of the bloom. These mechanistic changes in carbon uptake were reflected in declining photosynthetic rates and quantum yields as well as in the carbon isotopic composition with lower fractionation (13C enrichment) of the algae as the bloom progressed. Finally, despite induction of adaptive uptake mechanisms to low CO2 availability; scarcity of other nutrients combined with low CO2 concentrations, increased temperatures, and increased turbulence cause a decline in in situ growth rates and the collapse of the dinoflagellate biomass.Key words: dinoflagellates, inorganic carbon uptake, CCM, carbonic anhydrase, Peridinium gatunense.

Carbonic anhydrase activity of rabbit lungs

1980 ◽  
Vol 49 (4) ◽  
pp. 589-600 ◽  
Author(s):  
R. M. Effros ◽  
L. Shapiro ◽  
P. Silverman
Keyword(s):  
Carbonic Anhydrase ◽  
Pulmonary Artery ◽  
Extravascular Lung Water ◽  
Carbonic Anhydrase Activity ◽  
Co2 Exchange ◽  
Tissue Binding ◽  
Lung Water ◽  
Expired Gas

Pulmonary carbonic anhydrase (CA) activity was studied in rabbit lungs perfused with solutions containing no CA. Measurements were made of the amount of 14CO2 appearing in the expired gas following injections of H14CO3(-), 14CO2, or a 20:1 mixture of each into the pulmonary artery. The fraction of the injected label in the expired gas was only 17% greater for 14CO2 than for the mixture, suggesting that equilibration between H14CO3(-) and 14CO2 was nearly complete during the capillary transit time. Inhibition of pulmonary CA decreased excretion of H14CO3(-) and the mixture by 40 and 49% and increased the excretion of 14CO2 by 96%. Addition of CA to the perfusate had no effect. Thus, CO2 exchange is not significantly limited by pulmonary CA if inhibitors are absent. Tissue binding of [3H]acetazolamide injected into the pulmonary artery was diminished by 50% when acetazolamide concentrations reached 0.13 x 10(-6) M. Each liter of extravascular lung water contained 1.25 x 10(-6) mol of receptors for acetazolamide that were accessible to plasma during a single circulation. Binding of [3H]acetazolamide was also observed in lungs of anesthetized rabbits, suggesting that pulmonary CA is accessible to plasma in vivo as well as in situ.

In situ characterization of carbonic anhydrase activity in isolated rat lungs

1990 ◽  
Vol 69 (6) ◽  
pp. 2155-2162 ◽  
Author(s):  
T. A. Heming ◽  
A. Bidani
Keyword(s):  
Carbonic Anhydrase ◽  
Time Course ◽  
Carbonic Anhydrase Activity ◽  
In Situ Characterization ◽  
Rat Lungs ◽  
Isolated Lungs ◽  
Menten Constant ◽  
Co2 Excretion

Lung carbonic anhydrase (CA) participates directly in plasma CO2-HCO3(-)-H+ reactions. To characterize pulmonary CA activity in situ, CO2 excretion and capillary pH equilibration were examined in isolated saline-perfused rat lungs. Isolated lungs were perfused at 25, 30, and 37 degrees C with solutions containing various concentrations of HCO3- and a CA inhibitor, acetazolamide (ACTZ). Total CO2 excretion was partitioned into those fractions attributable to dissolved CO2, uncatalyzed HCO3- dehydration, and catalyzed HCO3- dehydration. Approximately 60% of the total CO2 excretion at each temperature was attributable to CA-catalyzed HCO3- dehydration. Inhibition of pulmonary CA diminished CO2 excretion and produced significant postcapillary perfusate pH disequilibria, the magnitude and time course of which were dependent on temperature and the extent of CA inhibition. The half time for pH equilibration increased from approximately 5 s at 37 degrees C to 14 s at 25 degrees C. For the HCO3- dehydration reaction, pulmonary CA in situ displayed an apparent inhibition constant for ACTZ of 0.9-2.2 microM, a Michaelis-Menten constant of 90 mM, a maximal reaction velocity of 9 mM/s, and an apparent activation energy of 3.0 kcal/mol.

1123: Renal Oxalate Transport Depends on Local Carbonic Anhydrase Activity in the Proximal Tubule

2004 ◽  
Vol 171 (4S) ◽  
pp. 296-296
Author(s):  
Michael Straub ◽  
Joséphine Befolo-Elo ◽  
Richard E Hautmann ◽  
Edgar Braendle
Keyword(s):  
Carbonic Anhydrase ◽  
Proximal Tubule ◽  
Carbonic Anhydrase Activity ◽  
Oxalate Transport

scholarly journals Fabrication of a liquid cell for in situ transmission electron microscopy

Microscopy ◽  
2020 ◽  
Author(s):  
Xiaoguang Li ◽  
Kazutaka Mitsuishi ◽  
Masaki Takeguchi
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cost Effective ◽  
Production Method ◽  
Microscopy Imaging ◽  
Transmission Electron ◽  
In Situ Electron Microscopy ◽  
Liquid Cell ◽  
Si Wafer

Abstract Liquid cell transmission electron microscopy (LCTEM) enables imaging of dynamic processes in liquid with high spatial and temporal resolution. The widely used liquid cell (LC) consists of two stacking microchips with a thin wet sample sandwiched between them. The vertically overlapped electron-transparent membrane windows on the microchips provide passage for the electron beam. However, microchips with imprecise dimensions usually cause poor alignment of the windows and difficulty in acquiring high-quality images. In this study, we developed a new and efficient microchip fabrication process for LCTEM with a large viewing area (180 µm × 40 µm) and evaluated the resultant LC. The new positioning reference marks on the surface of the Si wafer dramatically improve the precision of dicing the wafer, making it possible to accurately align the windows on two stacking microchips. The precise alignment led to a liquid thickness of 125.6 nm close to the edge of the viewing area. The performance of our LC was demonstrated by in situ transmission electron microscopy imaging of the dynamic motions of 2-nm Pt particles. This versatile and cost-effective microchip production method can be used to fabricate other types of microchips for in situ electron microscopy.

Cost effective in-situ methane enrichment for chicken farm biogas system

2021 ◽  
pp. 100773
Author(s):  
Kamoldara Reansuwan ◽  
Rotsukon Jawana ◽  
Saoharit Nitayavardhana ◽  
Sirichai Koonaphapdeelert
Keyword(s):  
Cost Effective ◽  
Chicken Farm
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