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
2020 ◽  
Author(s):  
Fernando Medina Ferrer ◽  
Kathryn Hobart ◽  
Jake V. Bailey

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.


1998 ◽  
Vol 76 (6) ◽  
pp. 1043-1051 ◽  
Author(s):  
Ilana Berman-Frank ◽  
Jonathan Erez ◽  
Aaron Kaplan

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.


1980 ◽  
Vol 49 (4) ◽  
pp. 589-600 ◽  
Author(s):  
R. M. Effros ◽  
L. Shapiro ◽  
P. Silverman

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.


1990 ◽  
Vol 69 (6) ◽  
pp. 2155-2162 ◽  
Author(s):  
T. A. Heming ◽  
A. Bidani

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.


2004 ◽  
Vol 171 (4S) ◽  
pp. 296-296
Author(s):  
Michael Straub ◽  
Joséphine Befolo-Elo ◽  
Richard E Hautmann ◽  
Edgar Braendle

PEDIATRICS ◽  
1951 ◽  
Vol 7 (2) ◽  
pp. 182-185
Author(s):  
RICHARD DAY ◽  
JANE FRANKLIN

The carbonic anhydrase activity in the kidneys of premature infants was studied because it was thought that if the renal enzyme is as deficient as that in the blood, inefficiency in acidification of urine might result. In contrast with the blood, postmortem specimens of kidneys of premature infants were found to exhibit carbonic anhydrase activity similar to that found in the case of kidneys from older infants and adults.


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