The 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 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.展开更多
Carbonic anhydrase(CA)as a typical metalloenzyme in biological system can accelerate the hydration/dehydration of carbon dioxide(CO2,the major components of greenhouse gases),which performer with high selectivity,envi...Carbonic anhydrase(CA)as a typical metalloenzyme in biological system can accelerate the hydration/dehydration of carbon dioxide(CO2,the major components of greenhouse gases),which performer with high selectivity,environmental friendliness and superior efficiency.However,the free form of CA is quite expensive(~RMB 3000/100 mg),unstable,and non-reusable as the free form of CA is not easy for recovery from the reaction environment,which severely limits its large-scale industrial applications.The immobilization may solve these problems at the same time.In this context,many efforts have been devoted to improving the chemical and thermal stabilities of CA through immobilization strategy.Very recently,a wide range of available inorganic,organic and hybrid compounds have been explored as carrier materials for CA immobilization,which could not only improve the tolerance of CA in hazardous environments,but also improve the efficiency and recovery to reduce the cost of large-scale application of CA.Several excellent reviews about immobilization methods and application potential of CA have been published.By contrast,in our review,we stressed on the way to better retain the biocatalytic activity of immobilized CA system based on different carrier materials and to solve the problems facing in practical operations well.The concluding remarks are presented with a perspective on constructing efficient CO2 conversion systems through rational combining CA and advanced carrier materials.展开更多
A carbonic anhydrase( CA) transcript was obtained from the Contig library according to the published sequencing information of the buckwheat transcripts. The full length of the CA gene was amplified by reverse transcr...A carbonic anhydrase( CA) transcript was obtained from the Contig library according to the published sequencing information of the buckwheat transcripts. The full length of the CA gene was amplified by reverse transcription PCR( RT-PCR). The bioinformatics analysis showed that the full length of Fs CA1 gene was 1233 bp and open reading frame was 978 bp,and encoding 325 amino acids. The molecular weight was 35. 11 ku and the isoelectric point was 7. 59; there were 9 α helices,6 β folds,many randon coil and extension chain,containing one signal peptide and one transmembrane region,having a 2 amino acid conserved domains with typical beta-type carbonic anhydrase. Subcellular localization showed that the protein is most likely to appear in the chloroplast. The three-dimensional structure model of Fs CA1 was built by homologous modeling method,indicating that the homo-octamer of buckwheat CA and pea CA could match well,so it can be inferred that buckwheat CA is also homo-octamer. Real-time quantitative PCR was used to detect the expression of Fs CA1 in different organs of buckwheat.The results showed that Fs CA1 had the highest expression level in leaves,then in the stems,and the lowest in roots.展开更多
Many cancer cells in solid tumors are hypoxic or pseudohypoxic and create acidic environment for malignancy progression. Under low oxygen conditions (hypoxia), hypoxia-inducible factors (HIFs) play pathological roles ...Many cancer cells in solid tumors are hypoxic or pseudohypoxic and create acidic environment for malignancy progression. Under low oxygen conditions (hypoxia), hypoxia-inducible factors (HIFs) play pathological roles in cancer cell survival and spreading. HIF regulates several genes such as genes of glucose transporters that enhance anaerobic glycolysis, angiogenesis, erythropoiesis and carbonic anhydrase IX (CA-IX). CA-IX is a cell-surface glycoprotein that catalyzes the hydration of CO2 to protons and bicarbonate ions (respiratory acidification). This process is involved in adaptation to acidosis and implicated in cancer progression. Therefore, CA-IX inhibitors (such as sulfonamide-based compounds) showed hoping results in reduction malignancy progression. The article aims to reversal the malignant hypoxic environment in solid tumors to create a condition of weakness within the cancer for further focused cisplatin potency. This article suggests the use of modified synthesized HIF as a drug delivery molecule for both carbonic anhydrase IX inhibitor and glycosylated cisplatin that damages the DNA of malignant cell. HIF molecule has high affinity to bind with CA IX-expressing malignant cells, which is followed by cell entrance via endocytosis. Once the HIF-Cisplatin-CA-inhibitor complex enters the cell, the carbonic anhydrase inhibitor will improve the cellular pH that makes the environment unsuitable for HIF 1α function and it may be ubiquitinated. So, the raise in target genes transcription will be arrested. On the other hand, once the synthetized HIF is degraded, the cisplatin molecules will be released inside the malignant cell and start to damage its DNA. This approach may be a good solution for many solid tumors.展开更多
Carbonic anhydrase (CA) is a kind of zinc enzyme that reversibly catalyzes hydration of CO2. Sixteen CAisoenzymes have been described in mammals . These isozymes play physiological roles in erythrocytes, including C...Carbonic anhydrase (CA) is a kind of zinc enzyme that reversibly catalyzes hydration of CO2. Sixteen CAisoenzymes have been described in mammals . These isozymes play physiological roles in erythrocytes, including CO2 transport, ion secretion, pH regulation and so forth. CA1 and 2 are both cytosolic enzymes that are found in large quantities in erythrocytes. Excluding hemoglobin, CA I is the most abundant protein in erythrocytes展开更多
Background: Carbonic anhydrase (CA) is a ubiquitous enzyme catalyzing the reversible hydration of CO<sub>2</sub> to and H<sup>+</sup>. CA plays a crucial role in CO<sub>2</sub>...Background: Carbonic anhydrase (CA) is a ubiquitous enzyme catalyzing the reversible hydration of CO<sub>2</sub> to and H<sup>+</sup>. CA plays a crucial role in CO<sub>2</sub> transport, acid-base balance, and in linking local acidosis to O<sub>2</sub> unloading from hemoglobin and also facilitates lactate shuttling across the monocarboxylate transporters (MCT). The study aimed to investigate the influence of hyperglycemia on erythrocyte carbonic anhydrase activity and lactic acid in type II diabetic patients. Method: Red blood cell carbonic anhydrase activity was determined in washed lysed-hemolysate by the action of the enzyme on the substrate p-nitrophenyl acetate. The absorbance of released p-nitrophenol was recorded at 345 nm. Glycated Hemoglobin was determined by ion exchange method (Spectrum Diagnostic Kit). Blood glucose, lactate, cholesterol and triglyceride were determined using Accutrend GCT meter (Roche, Germany) with cobias<sup>®</sup> test strips. Results: The present study showed that hyperglycemia significantly (p < 0.05) increased both erythrocyte carbonic anhydrase activity and blood lactate level in type II diabetic patients. Conclusion: We may conclude that hyperglycemia may be responsible for the increased activity of carbonic anhydrase activity and blood lactate concentration.展开更多
Objective To evaluate the prognostic significance of carbonic anhydrase IX ( CA IX) expression in patients with clear cell renal cell carcinoma ( ccRCC) . Methods CA IX excression in a cohort of 120 patients with ccRC...Objective To evaluate the prognostic significance of carbonic anhydrase IX ( CA IX) expression in patients with clear cell renal cell carcinoma ( ccRCC) . Methods CA IX excression in a cohort of 120 patients with ccRCC was evaluated by P-V immunohistochemistry展开更多
Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it i...Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it is a classical protein model of greatly biocatalytic significance as well as a highly expressed biomarker with renal tumor. We herein proposed a single-molecule measurement on carbonic anhydrase using MspA nanopore, in [BMIM+] and asymmetric K^(+)/Ca^(2+) cationic coordinated environment, instead of usual symmetric KCl/NaCl electrolyte. Significantly, our empirical analysis showed that asymmetric K^(+)/Ca^(2+) cationic environment contributes to distinguishable current modulations, thus yielding better resolution for carbonic anhydrase measurement, which is independent of applied voltage and more importantly, is stable enough at varied pH conditions and for very low concentration test in urine sample. Our results provide a classical model for nanopore protein analysis, and may also permit biocatalytic measurement at single-molecule level.展开更多
To investigate the biochemical response of freshwater green algae to elevated CO2 concentrations, Chlorella pyrenoidosa Chick and Chlamydomonas reinhardtii Dang cells were cultured at different CO2 concentrations with...To investigate the biochemical response of freshwater green algae to elevated CO2 concentrations, Chlorella pyrenoidosa Chick and Chlamydomonas reinhardtii Dang cells were cultured at different CO2 concentrations within the range 3-186 ümol/L and the biochemical composition, carbonic anhydrase (CA), and nitrate reductase activities of the cells were investigated. Chlorophylls (Chl), carotenoids, carbonhydrate, and protein contents were enhanced to varying extents with increasing CO2 concentration from 3-186 ümol/L. The CO2 enrichment significantly increased the Chl a/Chl b ratio in Chlorella pyrenoidosa, but not in Chlamydomonas reinhardtii. The CO2 concentration had significant effects on CA and nitrate reductase activity. Elevating CO2 concentration to 186 ümol/L caused a decline in intracellular and extracellullar CA activity. Nitrate reductase activity, under either light or dark conditions, in C. reinhardtii and C. pyrenoidosa was also significantly decreased with CO2 enrichment. From this study, it can be concluded that CO2 enrichment can affect biochemical composition, CA, and nitrate reductase activity, and that the biochemical response was species dependent.展开更多
Carbonic anhydrases (CAs) are zinc metalloenzymes that catalyze the interconversion of CO2 and HCO3- and are ubiquitous in nature. Higher plants contain three evolutionarily distinct CA families, αCAs, 13CAs, and ...Carbonic anhydrases (CAs) are zinc metalloenzymes that catalyze the interconversion of CO2 and HCO3- and are ubiquitous in nature. Higher plants contain three evolutionarily distinct CA families, αCAs, 13CAs, and γCAs, where each family is represented by multiple isoforms in all species. Alternative splicing of CA transcripts appears common; consequently, the number of functional CA isoforms in a species may exceed the number of genes. CAs are expressed in numerous plant tissues and in different cellular locations. The most prevalent CAs are those in the chloroplast, cytosol, and mitochondria. This diversity in location is paralleled in the many physiological and biochemical roles that CAs play in plants. In this review, the number and types of CAs in C3, C4, and crassulacean acid metabolism (CAM) plants are considered, and the roles of the α and yCAs are briefly discussed. The remainder of the review focuses on plant βCAs and includes the identification of homologs between species using phylogenetic approaches, a consideration of the inter- and intracellular localization of the proteins, along with the evidence for alternative splice forms. Current understanding of βCA tissue-specific expression patterns and what controls them are reviewed, and the physiological roles for which βCAs have been implicated are presented.展开更多
The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that ...The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.展开更多
Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and ...Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and absolute quantitation(iTRAQ)-based quantitative proteomic strategy was used to study the secretomes in conditioned media from HCC cancerous tissues,surrounding noncancerous tissues,and distal noncancerous tissues to identify diagnostic and prognostic biomarkers for HCC.In total,22 and 49 dysregulated secretory proteins were identified in the cancerous and surrounding noncancerous tissues,respectively,compared with the distal noncancerous tissues.Among these proteins,carbonic anhydrase II(CA2)was identified to be significantly upregulated in the secretome of cancerous tissues;correspondingly,the serum concentrations of CA2 were remarkably increased in HCC patients compared with that in normal populations.Interestingly,a significant increase of serum CA2 in recurrent HCC patients after radical resection was also confirmed compared with HCC patients without recurrence,and the serum level of CA2 could act as an independent prognostic factor for time to recurrence and overall survival.Regarding the mechanism,the secreted CA2 enhances the migration and invasion of HCC cells by activating the epithelial mesenchymal transition pathway.Taken together,this study identified a novel biomarker for HCC diagnosis and prognosis,and provided a valuable resource of HCC secretome for investigating serological biomarkers.展开更多
This study provides the presence of carbonic anhydrase(CA) activity in waters of the Yangtze River basin, China, as well as the correlation of CA activity with HCO-3 concentration and CO2 sink flux. Different degree...This study provides the presence of carbonic anhydrase(CA) activity in waters of the Yangtze River basin, China, as well as the correlation of CA activity with HCO-3 concentration and CO2 sink flux. Different degrees of CA activity could be detected in almost all of the water samples from different geological eco-environments in all four seasons. The CA activity of water samples from karst areas was significantly higher than from non-karst areas(PP3-concentration(r = 0.672, P2 sink flux(r = 0.602, P = 0.076) in karst areas. This suggests that CA in waters might have a promoting effect on carbon sinks for atmospheric CO2 in karst river basins. In conditions of similar geological type, higher CA activity was generally detected in water samples taken from areas that exhibited better eco-environments, implying that the CA activity index of waters could be used as an indicator for monitoring ecological environments and protection of river basins. These findings suggest that the role of CA in waters in the karst carbon sink potential of river basins is worthy of further in-depth studies.展开更多
Mineralization catalyzed by carbonic anhydrase(CA)is one of the most promising technologies for capturing CO_(2).In this work,Escherichia coli BL21(DE3)was used as the host,and the N-terminus of ice nucleation protein...Mineralization catalyzed by carbonic anhydrase(CA)is one of the most promising technologies for capturing CO_(2).In this work,Escherichia coli BL21(DE3)was used as the host,and the N-terminus of ice nucleation protein(INPN)was used as the carrier protein.Different fusion patterns and vectors were used to construct CA surface display systems forα-carbonic anhydrase(HPCA)from Helicobacter pylori 26695 andα-carbonic anhydrase(SazCA)from Sulfurihydrogenibium azorense.The surface display system in which HPCA was fused with INPN via a flexible linker and intermediate repeat sequences showed higher whole-cell enzyme activity,while the enzyme activity of the SazCA expression system was significantly higher than that of the HPCA expression system.The pET22b vector with the signal peptide PelB was more suitable for the cell surface display of SazCA.Cell frac-tionation and western-blot analysis indicated that SazCA and INPN were successfully anchored on the cell’s outer membrane as a fusion protein.The enzyme activity of the surface display strain E-22b-I RL S(11.43 U⋅mL^(−1) OD 600−1)was significantly higher than that of the intracellular expression strain E-22b-S(8.355 U⋅mL^(−1) OD 600−1)under optimized induction conditions.Compared with free SazCA,E-22b-I RL S had higher thermal and pH stability.The long-term stability of SazCA was also significantly improved by surface display.When the engineered strain and free enzyme were used for CO_(2) mineralization,the amount of CaCO_(3) deposition catalyzed by the strain E-22b-I RL S on the surface(241 mg)was similar to that of the free SazCA and was significantly higher than the intracellular expression strain E-22b-S(173 mg).These results demonstrate that the SazCA surface display strain can serve as a whole-cell biocatalyst for CO_(2) capture and mineralization.展开更多
Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known...Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.展开更多
This study investigated the inhibitory effect of a high efficiency diuretic, furosemide, on carbonic anhydrase (CA). First, comparing the inhibitory effect of acetazolamide, a low efficiency diuretic, on CA, shows t...This study investigated the inhibitory effect of a high efficiency diuretic, furosemide, on carbonic anhydrase (CA). First, comparing the inhibitory effect of acetazolamide, a low efficiency diuretic, on CA, shows that furosemide or acetazolamide can quickly make CA inactive when its concentration is close to the enzyme concentration, different from the usual inhibitory kinetics in which the concentration of the inhibitor is far higher than the enzyme concentration. Secondly, the reaction of the enzyme indicates that the inhibitory effect of furosemide or acetazolamide on carbonic anhydrase is quickly reversible. Finally, the degree of the inhibitory effect of furosemide and of acetazolamide on CA are compared. The results show that furosemide inhibits CA less than acetazolamide.展开更多
Crossed beak is a complex mode of inheritance with prevalence ranging from 0.2 to 7.4% in at least 12 chicken strains worldwide.To reveal the intrinsic factors causing crossed beaks,genes expression patterns in bilate...Crossed beak is a complex mode of inheritance with prevalence ranging from 0.2 to 7.4% in at least 12 chicken strains worldwide.To reveal the intrinsic factors causing crossed beaks,genes expression patterns in bilateral mandibular condyle between affected and normal birds were characterized by RNA sequencing analysis in the present studies.Crossed beak was induced by short length of unilateral mandibular ramus,and a total of 110differentially expressed genes were up-or down-regulated in the affected(short)mandibular condyle side as compared to the normal side.Carbonic anhydrase 2(CA2)and Carbonic anhydrase 13(CA13)were enriched in the carbonate dehydratase activity,and high-expressed in mandibular condyle and osteoblasts(P<0.05).However,both were low-expressed in short mandibular condyle side of affected birds(P<0.05).The carbonate dehydratase inhibitor experiments confirmed that there is positive association between the calcification and carbonic anhydrase isoenzymes.Quantitative analysis with cetylpyridinium chloride showed a decrease in calcification when the cells were transfected with an anti-CA13 shRNA.Our research suggested that CA2 and CA13 are down-calcified in shortside mandibular condyle,and caused mandibular ramus to grow slowly.CA2 and CA13 have the critical role in crossed beaks by regulating calcification of mandibular condyle.展开更多
Vanadium and its derivatives are used in various industries,including steel,metallurgy,pharmaceuticals,and aerospace engineering.Although China has massive reserves of stone coal resources,these resources have low gra...Vanadium and its derivatives are used in various industries,including steel,metallurgy,pharmaceuticals,and aerospace engineering.Although China has massive reserves of stone coal resources,these resources have low grades.Therefore,the effective extraction and recovery of metallic vanadium from stone coal is an important way to realize the efficient resource utilization of stone coal vanadium ore.Herein,Bacillus mucilaginosus was selected as the leaching strain.The vanadium leaching rate reached 35.5%after 20 d of bioleaching under optimal operating conditions.The cumulative vanadium leaching rate in the contact group reached 35.5%,which was higher than that in the noncontact group(9.3%).The metabolites of B.mucilaginosus,such as oxalic,tartaric,citric,and malic acids,dominated in bioleaching,accounting for 73.8%of the vanadium leaching rate.Interestingly,during leaching,the presence of stone coal stimulated the expression of carbonic anhydrase in bacterial cells,and enzyme activity increased by 1.335-1.905 U.Enzyme activity positively promoted the production of metabolite organic acids,and total organic acid content increased by 39.31 mg·L^(-1),resulting in a reduction of 2.51 in the pH of the leaching system with stone coal.This effect favored the leaching of vanadium from stone coal.Atomic force microscopy illustrated that bacterial leaching exacerbated corrosion on the surface of stone coal beyond 10 nm.Our study provides a clear and promising strategy for exploring the bioleaching mechanism from the perspective of microbial enzyme activity and metabolites.展开更多
The study investigates the potential of the serpulid tubeworm Ficopomatus sp.,sourced from Jesolo Lido(Venice,Italy),for CCS(Carbon Capture and Storage).Specimens were processed to extract bioactive compounds using a ...The study investigates the potential of the serpulid tubeworm Ficopomatus sp.,sourced from Jesolo Lido(Venice,Italy),for CCS(Carbon Capture and Storage).Specimens were processed to extract bioactive compounds using a 50%polypropylene glycol-water solvent system.Extracts were analysed and purified using column gel filtration chromatography,with fractions identified by TLC(Thin Layer Chromatography)and further characterized for antioxidant and antibacterial activities.Antioxidant activity was detected via DPPH(2,2-Diphenyl-1-Picrylhydrazyl)spraying on TLC plates,while antibacterial activity was evaluated using the antibiogram paper disk diffusion method.Enzyme activity in the fractions was previously confirmed through a bromothymol blue test followed by spectrophotometric analysis.The primary goal was to explore the CCS potential,using an experimental module involving Arduino Uno embedded microprocessor for the CO2 measurement and to confirm the conversion into insoluble carbonates(storage).The most active fraction,identified as S1,showed significant CCS action,confirmed by microscopic observation of calcareous deposits on treated sponges.These findings suggest that Ficopomatus sp.can be used in CCS research highlighting its potential for biotechnological applications in mitigating climate change.The paper underscores the importance of marine organisms in CCS and offers insights into innovative strategies for environmental conservation and carbon management.展开更多
This work demonstrated quantitatively that DSA (dansylamide) and apo CA (dezincified carbon anhydrase) interacted with each other in the presence of Zn2+ (zinc ion). The fluorescence emission of DSA in the pres...This work demonstrated quantitatively that DSA (dansylamide) and apo CA (dezincified carbon anhydrase) interacted with each other in the presence of Zn2+ (zinc ion). The fluorescence emission of DSA in the presence of Zn2+ and the intensity of fluorescence, which was proportional to the concentration of Zn2+, could be used in the measurement of Zn2+ concentration. Considering that Zn2+ was the active center of enzymes like CA (carbon anhydrase), two dezincification reagents were compared to ensure the validity of our method. A certain range concentration of Zn2+ could be measured by the reaction product of apo CA, DSA and Zn2+, where the measurement limitation was about 60 nmol/L. Interaction of CA with DSA was also studied with infrared spectrometry.展开更多
基金Financial supports for this research was provided by the National Nature Science Foundation of China(Grant 40073026)Ministry of Science and Technology of China(Grant 164)+1 种基金Natural Science Foundation of Guangxi(Grant 9824021)Ministry of Land and Resources of China(Grant 9806)and Bremen University of Germany.
文摘The 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.
基金the National Natural Science Fundation of China(21776213)Natural Science Fund of Tianjin(19JCYBJC19700)for financial support。
文摘Carbonic anhydrase(CA)as a typical metalloenzyme in biological system can accelerate the hydration/dehydration of carbon dioxide(CO2,the major components of greenhouse gases),which performer with high selectivity,environmental friendliness and superior efficiency.However,the free form of CA is quite expensive(~RMB 3000/100 mg),unstable,and non-reusable as the free form of CA is not easy for recovery from the reaction environment,which severely limits its large-scale industrial applications.The immobilization may solve these problems at the same time.In this context,many efforts have been devoted to improving the chemical and thermal stabilities of CA through immobilization strategy.Very recently,a wide range of available inorganic,organic and hybrid compounds have been explored as carrier materials for CA immobilization,which could not only improve the tolerance of CA in hazardous environments,but also improve the efficiency and recovery to reduce the cost of large-scale application of CA.Several excellent reviews about immobilization methods and application potential of CA have been published.By contrast,in our review,we stressed on the way to better retain the biocatalytic activity of immobilized CA system based on different carrier materials and to solve the problems facing in practical operations well.The concluding remarks are presented with a perspective on constructing efficient CO2 conversion systems through rational combining CA and advanced carrier materials.
基金Supported by Project of National Natural Science Foundation(31360300&31560362)Key Project of the Tibet Autonomous Region(XZXTCX-2016)
文摘A carbonic anhydrase( CA) transcript was obtained from the Contig library according to the published sequencing information of the buckwheat transcripts. The full length of the CA gene was amplified by reverse transcription PCR( RT-PCR). The bioinformatics analysis showed that the full length of Fs CA1 gene was 1233 bp and open reading frame was 978 bp,and encoding 325 amino acids. The molecular weight was 35. 11 ku and the isoelectric point was 7. 59; there were 9 α helices,6 β folds,many randon coil and extension chain,containing one signal peptide and one transmembrane region,having a 2 amino acid conserved domains with typical beta-type carbonic anhydrase. Subcellular localization showed that the protein is most likely to appear in the chloroplast. The three-dimensional structure model of Fs CA1 was built by homologous modeling method,indicating that the homo-octamer of buckwheat CA and pea CA could match well,so it can be inferred that buckwheat CA is also homo-octamer. Real-time quantitative PCR was used to detect the expression of Fs CA1 in different organs of buckwheat.The results showed that Fs CA1 had the highest expression level in leaves,then in the stems,and the lowest in roots.
文摘Many cancer cells in solid tumors are hypoxic or pseudohypoxic and create acidic environment for malignancy progression. Under low oxygen conditions (hypoxia), hypoxia-inducible factors (HIFs) play pathological roles in cancer cell survival and spreading. HIF regulates several genes such as genes of glucose transporters that enhance anaerobic glycolysis, angiogenesis, erythropoiesis and carbonic anhydrase IX (CA-IX). CA-IX is a cell-surface glycoprotein that catalyzes the hydration of CO2 to protons and bicarbonate ions (respiratory acidification). This process is involved in adaptation to acidosis and implicated in cancer progression. Therefore, CA-IX inhibitors (such as sulfonamide-based compounds) showed hoping results in reduction malignancy progression. The article aims to reversal the malignant hypoxic environment in solid tumors to create a condition of weakness within the cancer for further focused cisplatin potency. This article suggests the use of modified synthesized HIF as a drug delivery molecule for both carbonic anhydrase IX inhibitor and glycosylated cisplatin that damages the DNA of malignant cell. HIF molecule has high affinity to bind with CA IX-expressing malignant cells, which is followed by cell entrance via endocytosis. Once the HIF-Cisplatin-CA-inhibitor complex enters the cell, the carbonic anhydrase inhibitor will improve the cellular pH that makes the environment unsuitable for HIF 1α function and it may be ubiquitinated. So, the raise in target genes transcription will be arrested. On the other hand, once the synthetized HIF is degraded, the cisplatin molecules will be released inside the malignant cell and start to damage its DNA. This approach may be a good solution for many solid tumors.
基金Supported by the National Natural Science Foundation of China (No.81170827)Fundamental Research Funds of State Key Lab
文摘Carbonic anhydrase (CA) is a kind of zinc enzyme that reversibly catalyzes hydration of CO2. Sixteen CAisoenzymes have been described in mammals . These isozymes play physiological roles in erythrocytes, including CO2 transport, ion secretion, pH regulation and so forth. CA1 and 2 are both cytosolic enzymes that are found in large quantities in erythrocytes. Excluding hemoglobin, CA I is the most abundant protein in erythrocytes
文摘Background: Carbonic anhydrase (CA) is a ubiquitous enzyme catalyzing the reversible hydration of CO<sub>2</sub> to and H<sup>+</sup>. CA plays a crucial role in CO<sub>2</sub> transport, acid-base balance, and in linking local acidosis to O<sub>2</sub> unloading from hemoglobin and also facilitates lactate shuttling across the monocarboxylate transporters (MCT). The study aimed to investigate the influence of hyperglycemia on erythrocyte carbonic anhydrase activity and lactic acid in type II diabetic patients. Method: Red blood cell carbonic anhydrase activity was determined in washed lysed-hemolysate by the action of the enzyme on the substrate p-nitrophenyl acetate. The absorbance of released p-nitrophenol was recorded at 345 nm. Glycated Hemoglobin was determined by ion exchange method (Spectrum Diagnostic Kit). Blood glucose, lactate, cholesterol and triglyceride were determined using Accutrend GCT meter (Roche, Germany) with cobias<sup>®</sup> test strips. Results: The present study showed that hyperglycemia significantly (p < 0.05) increased both erythrocyte carbonic anhydrase activity and blood lactate level in type II diabetic patients. Conclusion: We may conclude that hyperglycemia may be responsible for the increased activity of carbonic anhydrase activity and blood lactate concentration.
文摘Objective To evaluate the prognostic significance of carbonic anhydrase IX ( CA IX) expression in patients with clear cell renal cell carcinoma ( ccRCC) . Methods CA IX excression in a cohort of 120 patients with ccRCC was evaluated by P-V immunohistochemistry
基金National Key Research and Development Program of China(2022YFB3205600)Chongqing Talents:Exceptional Young Talents Project(cstc2021ycjh-bgzxm0016)+1 种基金Beibei Technology Talents and Independent Innovation Project(No.2022-34)the Natural Science Foundation of Chongqing,China(cstc2021jcyj-jqx0030).
文摘Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it is a classical protein model of greatly biocatalytic significance as well as a highly expressed biomarker with renal tumor. We herein proposed a single-molecule measurement on carbonic anhydrase using MspA nanopore, in [BMIM+] and asymmetric K^(+)/Ca^(2+) cationic coordinated environment, instead of usual symmetric KCl/NaCl electrolyte. Significantly, our empirical analysis showed that asymmetric K^(+)/Ca^(2+) cationic environment contributes to distinguishable current modulations, thus yielding better resolution for carbonic anhydrase measurement, which is independent of applied voltage and more importantly, is stable enough at varied pH conditions and for very low concentration test in urine sample. Our results provide a classical model for nanopore protein analysis, and may also permit biocatalytic measurement at single-molecule level.
文摘To investigate the biochemical response of freshwater green algae to elevated CO2 concentrations, Chlorella pyrenoidosa Chick and Chlamydomonas reinhardtii Dang cells were cultured at different CO2 concentrations within the range 3-186 ümol/L and the biochemical composition, carbonic anhydrase (CA), and nitrate reductase activities of the cells were investigated. Chlorophylls (Chl), carotenoids, carbonhydrate, and protein contents were enhanced to varying extents with increasing CO2 concentration from 3-186 ümol/L. The CO2 enrichment significantly increased the Chl a/Chl b ratio in Chlorella pyrenoidosa, but not in Chlamydomonas reinhardtii. The CO2 concentration had significant effects on CA and nitrate reductase activity. Elevating CO2 concentration to 186 ümol/L caused a decline in intracellular and extracellullar CA activity. Nitrate reductase activity, under either light or dark conditions, in C. reinhardtii and C. pyrenoidosa was also significantly decreased with CO2 enrichment. From this study, it can be concluded that CO2 enrichment can affect biochemical composition, CA, and nitrate reductase activity, and that the biochemical response was species dependent.
文摘Carbonic anhydrases (CAs) are zinc metalloenzymes that catalyze the interconversion of CO2 and HCO3- and are ubiquitous in nature. Higher plants contain three evolutionarily distinct CA families, αCAs, 13CAs, and γCAs, where each family is represented by multiple isoforms in all species. Alternative splicing of CA transcripts appears common; consequently, the number of functional CA isoforms in a species may exceed the number of genes. CAs are expressed in numerous plant tissues and in different cellular locations. The most prevalent CAs are those in the chloroplast, cytosol, and mitochondria. This diversity in location is paralleled in the many physiological and biochemical roles that CAs play in plants. In this review, the number and types of CAs in C3, C4, and crassulacean acid metabolism (CAM) plants are considered, and the roles of the α and yCAs are briefly discussed. The remainder of the review focuses on plant βCAs and includes the identification of homologs between species using phylogenetic approaches, a consideration of the inter- and intracellular localization of the proteins, along with the evidence for alternative splice forms. Current understanding of βCA tissue-specific expression patterns and what controls them are reviewed, and the physiological roles for which βCAs have been implicated are presented.
基金This research was supported by Shanghai Universities Peak Discipline Project of Aquaculture and“Double First-Class Initiative”program for its First-Class Discipline of aquaculture.
文摘The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.
基金supported by the National Natural Science Foundation of China(Grant Nos.81702910 and 81672376)the Educational Commission of Fujian Province,China(Grant No.2018B013)the Natural Science Foundation of Fujian Province,China(Grant Nos.2019J01298,2017J01159,and 2016J01417)
文摘Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and absolute quantitation(iTRAQ)-based quantitative proteomic strategy was used to study the secretomes in conditioned media from HCC cancerous tissues,surrounding noncancerous tissues,and distal noncancerous tissues to identify diagnostic and prognostic biomarkers for HCC.In total,22 and 49 dysregulated secretory proteins were identified in the cancerous and surrounding noncancerous tissues,respectively,compared with the distal noncancerous tissues.Among these proteins,carbonic anhydrase II(CA2)was identified to be significantly upregulated in the secretome of cancerous tissues;correspondingly,the serum concentrations of CA2 were remarkably increased in HCC patients compared with that in normal populations.Interestingly,a significant increase of serum CA2 in recurrent HCC patients after radical resection was also confirmed compared with HCC patients without recurrence,and the serum level of CA2 could act as an independent prognostic factor for time to recurrence and overall survival.Regarding the mechanism,the secreted CA2 enhances the migration and invasion of HCC cells by activating the epithelial mesenchymal transition pathway.Taken together,this study identified a novel biomarker for HCC diagnosis and prognosis,and provided a valuable resource of HCC secretome for investigating serological biomarkers.
基金supported by the Sub-project of Geological Survey Work Item of China Geological Survey Bureau (No. 12120113005200)the Fundamental Research Funds for the Central Universities (No. 2015QN102)
文摘This study provides the presence of carbonic anhydrase(CA) activity in waters of the Yangtze River basin, China, as well as the correlation of CA activity with HCO-3 concentration and CO2 sink flux. Different degrees of CA activity could be detected in almost all of the water samples from different geological eco-environments in all four seasons. The CA activity of water samples from karst areas was significantly higher than from non-karst areas(PP3-concentration(r = 0.672, P2 sink flux(r = 0.602, P = 0.076) in karst areas. This suggests that CA in waters might have a promoting effect on carbon sinks for atmospheric CO2 in karst river basins. In conditions of similar geological type, higher CA activity was generally detected in water samples taken from areas that exhibited better eco-environments, implying that the CA activity index of waters could be used as an indicator for monitoring ecological environments and protection of river basins. These findings suggest that the role of CA in waters in the karst carbon sink potential of river basins is worthy of further in-depth studies.
基金the financial support provided by the National Key Research and Development Program of China(Project No.2018YFA0902100)the National Natural Science Foundation of China(No.22178262,No.21576197).
文摘Mineralization catalyzed by carbonic anhydrase(CA)is one of the most promising technologies for capturing CO_(2).In this work,Escherichia coli BL21(DE3)was used as the host,and the N-terminus of ice nucleation protein(INPN)was used as the carrier protein.Different fusion patterns and vectors were used to construct CA surface display systems forα-carbonic anhydrase(HPCA)from Helicobacter pylori 26695 andα-carbonic anhydrase(SazCA)from Sulfurihydrogenibium azorense.The surface display system in which HPCA was fused with INPN via a flexible linker and intermediate repeat sequences showed higher whole-cell enzyme activity,while the enzyme activity of the SazCA expression system was significantly higher than that of the HPCA expression system.The pET22b vector with the signal peptide PelB was more suitable for the cell surface display of SazCA.Cell frac-tionation and western-blot analysis indicated that SazCA and INPN were successfully anchored on the cell’s outer membrane as a fusion protein.The enzyme activity of the surface display strain E-22b-I RL S(11.43 U⋅mL^(−1) OD 600−1)was significantly higher than that of the intracellular expression strain E-22b-S(8.355 U⋅mL^(−1) OD 600−1)under optimized induction conditions.Compared with free SazCA,E-22b-I RL S had higher thermal and pH stability.The long-term stability of SazCA was also significantly improved by surface display.When the engineered strain and free enzyme were used for CO_(2) mineralization,the amount of CaCO_(3) deposition catalyzed by the strain E-22b-I RL S on the surface(241 mg)was similar to that of the free SazCA and was significantly higher than the intracellular expression strain E-22b-S(173 mg).These results demonstrate that the SazCA surface display strain can serve as a whole-cell biocatalyst for CO_(2) capture and mineralization.
基金This research was supported by the National Key R&D Program of China(Grant No.2018YFD0901500)the National Natural Science Foundation of China(Grant No.41376136)the Double First-Class Discipline of Fisheries Science of China.
文摘Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.
文摘This study investigated the inhibitory effect of a high efficiency diuretic, furosemide, on carbonic anhydrase (CA). First, comparing the inhibitory effect of acetazolamide, a low efficiency diuretic, on CA, shows that furosemide or acetazolamide can quickly make CA inactive when its concentration is close to the enzyme concentration, different from the usual inhibitory kinetics in which the concentration of the inhibitor is far higher than the enzyme concentration. Secondly, the reaction of the enzyme indicates that the inhibitory effect of furosemide or acetazolamide on carbonic anhydrase is quickly reversible. Finally, the degree of the inhibitory effect of furosemide and of acetazolamide on CA are compared. The results show that furosemide inhibits CA less than acetazolamide.
基金supported by the Beijing Featured Livestock and Poultry Genetic Resources Preservation Project,China(202203310002)China Agriculture Research System of MOF and MARA(CARS40)+1 种基金the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(ASTIPIAS04)the Central Guidance on Local Science and Technology Development Fund of Hebei Province,China(236Z6602G)。
文摘Crossed beak is a complex mode of inheritance with prevalence ranging from 0.2 to 7.4% in at least 12 chicken strains worldwide.To reveal the intrinsic factors causing crossed beaks,genes expression patterns in bilateral mandibular condyle between affected and normal birds were characterized by RNA sequencing analysis in the present studies.Crossed beak was induced by short length of unilateral mandibular ramus,and a total of 110differentially expressed genes were up-or down-regulated in the affected(short)mandibular condyle side as compared to the normal side.Carbonic anhydrase 2(CA2)and Carbonic anhydrase 13(CA13)were enriched in the carbonate dehydratase activity,and high-expressed in mandibular condyle and osteoblasts(P<0.05).However,both were low-expressed in short mandibular condyle side of affected birds(P<0.05).The carbonate dehydratase inhibitor experiments confirmed that there is positive association between the calcification and carbonic anhydrase isoenzymes.Quantitative analysis with cetylpyridinium chloride showed a decrease in calcification when the cells were transfected with an anti-CA13 shRNA.Our research suggested that CA2 and CA13 are down-calcified in shortside mandibular condyle,and caused mandibular ramus to grow slowly.CA2 and CA13 have the critical role in crossed beaks by regulating calcification of mandibular condyle.
基金This work was financially supported by the National Natural Science Foundation of China(No.51874018)the Open Foundation of State Key Laboratory of Mineral Processing(No.BGRIMM-KJSKL-2022-07).
文摘Vanadium and its derivatives are used in various industries,including steel,metallurgy,pharmaceuticals,and aerospace engineering.Although China has massive reserves of stone coal resources,these resources have low grades.Therefore,the effective extraction and recovery of metallic vanadium from stone coal is an important way to realize the efficient resource utilization of stone coal vanadium ore.Herein,Bacillus mucilaginosus was selected as the leaching strain.The vanadium leaching rate reached 35.5%after 20 d of bioleaching under optimal operating conditions.The cumulative vanadium leaching rate in the contact group reached 35.5%,which was higher than that in the noncontact group(9.3%).The metabolites of B.mucilaginosus,such as oxalic,tartaric,citric,and malic acids,dominated in bioleaching,accounting for 73.8%of the vanadium leaching rate.Interestingly,during leaching,the presence of stone coal stimulated the expression of carbonic anhydrase in bacterial cells,and enzyme activity increased by 1.335-1.905 U.Enzyme activity positively promoted the production of metabolite organic acids,and total organic acid content increased by 39.31 mg·L^(-1),resulting in a reduction of 2.51 in the pH of the leaching system with stone coal.This effect favored the leaching of vanadium from stone coal.Atomic force microscopy illustrated that bacterial leaching exacerbated corrosion on the surface of stone coal beyond 10 nm.Our study provides a clear and promising strategy for exploring the bioleaching mechanism from the perspective of microbial enzyme activity and metabolites.
文摘The study investigates the potential of the serpulid tubeworm Ficopomatus sp.,sourced from Jesolo Lido(Venice,Italy),for CCS(Carbon Capture and Storage).Specimens were processed to extract bioactive compounds using a 50%polypropylene glycol-water solvent system.Extracts were analysed and purified using column gel filtration chromatography,with fractions identified by TLC(Thin Layer Chromatography)and further characterized for antioxidant and antibacterial activities.Antioxidant activity was detected via DPPH(2,2-Diphenyl-1-Picrylhydrazyl)spraying on TLC plates,while antibacterial activity was evaluated using the antibiogram paper disk diffusion method.Enzyme activity in the fractions was previously confirmed through a bromothymol blue test followed by spectrophotometric analysis.The primary goal was to explore the CCS potential,using an experimental module involving Arduino Uno embedded microprocessor for the CO2 measurement and to confirm the conversion into insoluble carbonates(storage).The most active fraction,identified as S1,showed significant CCS action,confirmed by microscopic observation of calcareous deposits on treated sponges.These findings suggest that Ficopomatus sp.can be used in CCS research highlighting its potential for biotechnological applications in mitigating climate change.The paper underscores the importance of marine organisms in CCS and offers insights into innovative strategies for environmental conservation and carbon management.
基金Supported by National Natural Science Foundation of China(NSFC)(No.61177076)the Fundamental Research Funds for the Central Universities of WUT(Nos.2012-Ia-020 and 2013-Ia-038)
文摘This work demonstrated quantitatively that DSA (dansylamide) and apo CA (dezincified carbon anhydrase) interacted with each other in the presence of Zn2+ (zinc ion). The fluorescence emission of DSA in the presence of Zn2+ and the intensity of fluorescence, which was proportional to the concentration of Zn2+, could be used in the measurement of Zn2+ concentration. Considering that Zn2+ was the active center of enzymes like CA (carbon anhydrase), two dezincification reagents were compared to ensure the validity of our method. A certain range concentration of Zn2+ could be measured by the reaction product of apo CA, DSA and Zn2+, where the measurement limitation was about 60 nmol/L. Interaction of CA with DSA was also studied with infrared spectrometry.