RESUMEN
Bacterial carbonic anhydrases (BCAs, EC 4.2.1.1) are indispensable enzymes in microbial physiology because they facilitate the hydration of carbon dioxide (CO2) to bicarbonate ions (HCO3-) and protons (H+), which are crucial for various metabolic processes and cellular homeostasis. Their involvement spans from metabolic pathways, such as photosynthesis, respiration, to organic compounds production, which are pivotal for bacterial growth and survival. This chapter elucidates the diversity of BCA genetic families, categorized into four distinct classes (α, ß, γ, and ι), which may reflect bacterial adaptation to environmental niches and their metabolic demands. The diversity of BCAs is essential not only for understanding their physiological roles but also for exploring their potential in biotechnology. Knowledge of their diversity enables researchers to develop innovative biocatalysts for industrial applications, including carbon capture technologies to convert CO2 emissions into valuable products. Additionally, BCAs are relevant to biomedical research and drug development because of their involvement in bacterial pathogenesis and microbial survival within the host. Understanding the diversity and function of BCAs can aid in designing targeted therapeutics that interfere with bacterial metabolism and potentially reduce the risk of infections.
Asunto(s)
Bacterias , Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/genética , Bacterias/enzimología , Bacterias/genética , Dióxido de Carbono/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismoRESUMEN
The increasing prevalence of antibiotic-resistant bacteria necessitates the exploration of novel therapeutic targets. Bacterial carbonic anhydrases (CAs) have been known for decades, but only in the past ten years they have garnered significant interest as drug targets to develop antibiotics having a diverse mechanism of action compared to the clinically used drugs. Significant progress has been made in the field in the past three years, with the validation in vivo of CAs from Neisseria gonorrhoeae, and vancomycin-resistant enterococci as antibiotic targets. This chapter compiles the state-of-the-art research on sulfonamide derivatives described as inhibitors of all known bacterial CAs. A section delves into the mechanisms of action of sulfonamide compounds with the CA classes identified in pathogenic bacteria, specifically α, ß, and γ classes. Therefore, the inhibitory profiling of the bacterial CAs with classical and clinically used sulfonamide compounds is reported and analyzed. Another section covers various other series of sulfonamide CA inhibitors studied for the development of new antibiotics. By synthesizing current research findings, this chapter highlights the potential of sulfonamide inhibitors as a novel class of antibacterial agents and paves the way for future drug design strategies.
Asunto(s)
Antibacterianos , Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Sulfonamidas , Sulfonamidas/farmacología , Sulfonamidas/química , Inhibidores de Anhidrasa Carbónica/farmacología , Inhibidores de Anhidrasa Carbónica/química , Anhidrasas Carbónicas/metabolismo , Antibacterianos/farmacología , Antibacterianos/química , Humanos , Bacterias/enzimología , Bacterias/efectos de los fármacos , Neisseria gonorrhoeae/enzimología , Neisseria gonorrhoeae/efectos de los fármacosRESUMEN
Non-sulfonamide chemical moieties able to inhibit the bacterial (b) expressed Carbonic Anhydrases (CAs; EC 4.2.1.1) constitute an important alternative to the prototypic modulators discussed in Chapter 6, as give access to large and variegate chemical classes, also of the natural origin. This contribution reports the main classes of compounds profiled in vitro on the bCAs and thus may be worth developing for the validation process of this class of enzymes.
Asunto(s)
Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Inhibidores de Anhidrasa Carbónica/farmacología , Inhibidores de Anhidrasa Carbónica/química , Anhidrasas Carbónicas/metabolismo , Humanos , Bacterias/efectos de los fármacos , Bacterias/enzimología , Sulfonamidas/farmacología , Sulfonamidas/químicaRESUMEN
Carbonic anhydrase metalloenzymes are encoded in genomes throughout all kingdoms of life with a conserved function catalyzing the reversible conversion of CO2 to bicarbonate. Carbonic anhydrases have been well-investigated in humans, but are still relatively understudied in bacterial organisms, including Enterococci. Studies over the past decade have presented bacterial carbonic anhydrases as potential drug targets, with some chemical scaffolds potently inhibiting the Enterococcus carbonic anhydrases in vitro and displaying antimicrobial efficacy against Enterococcus organisms. While carbonic anhydrases in Enterococci still have much to be explored, hypotheses may be drawn from similar Gram-positive organisms for which known information exists about carbonic anhydrase function and relevance. Within this chapter is reported information and rational hypotheses regarding the subcellar locations, potential physiological roles, essentiality, structures, and kinetics of carbonic anhydrases in Enterococci.
Asunto(s)
Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Enterococcus , Inhibidores de Anhidrasa Carbónica/farmacología , Anhidrasas Carbónicas/metabolismo , Enterococcus/efectos de los fármacos , Enterococcus/enzimología , Humanos , Antibacterianos/farmacologíaRESUMEN
Recent research has identified a novel class of carbonic anhydrases (CAs), designated ι-CA, predominantly found in marine diatoms, eukaryotic algae, cyanobacteria, bacteria, and archaea genomes. This class has garnered attention owing to its unique biochemical properties and evolutionary significance. Through bioinformatic analyses, LCIP63, a protein initially annotated with an unknown function, was identified as a potential ι-CA in the marine diatom Thalassiosira pseudonana. Subsequent biochemical characterization revealed that LCIP63 has CA activity and its preference for manganese ions over zinc, indicative of evolutionary adaptation to marine environments. Further exploration of bacterial ι-CAs, exemplified by Burkholderia territorii ι-CA (BteCAι), demonstrated catalytic efficiency and sensitivity to sulfonamide and inorganic anion inhibitors, the classical CA inhibitors (CAIs). The classification of ι-CAs into two variant types based on their sequences, distinguished by the COG4875 and COG4337 domains, marks a significant advancement in our understanding of these enzymes. Structural analyses of COG4337 ι-CAs from eukaryotic microalgae and cyanobacteria thereafter revealed a distinctive structural arrangement and a novel catalytic mechanism involving specific residues facilitating CO2 hydration in the absence of metal ion cofactors, deviating from canonical CA behavior. These findings underscore the biochemical diversity within the ι-CA class and highlight its potential as a target for novel antimicrobial agents. Overall, the elucidation of ι-CA properties and mechanisms advances our knowledge of carbon metabolism in diverse organisms and underscores the complexity of CA evolution and function.
Asunto(s)
Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Bacterias/efectos de los fármacos , Burkholderia , Diatomeas , Cianobacterias , Inhibidores de Anhidrasa Carbónica/farmacología , Inhibidores de Anhidrasa Carbónica/químicaRESUMEN
Carbonic anhydrases belonging to the α-class are widely distributed in bacterial species. These enzymes have been isolated from bacteria with completely different characteristics including both Gram-negative and Gram-positive strains. α-CAs show a considerable similarity when comparing the biochemical, kinetic and structural features, with only small differences which reflect the diverse role these enzymes play in Nature. In this chapter, we provide a comprehensive overview on bacterial α-CA data, with a highlight to their potential biomedical and biotechnological applications.
Asunto(s)
Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Bacterias/enzimología , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismoRESUMEN
Infections from Helicobacter pylori (Hp) are endangering Public Health safety worldwide, due to the associated high risk of developing severe diseases, such as peptic ulcer, gastric cancer, diabetes, and cardiovascular diseases. Current therapies are becoming less effective due to the rise of (multi)drug-resistant phenotypes and an urgent need for new antibacterial agents with innovative mechanisms of action is pressing. Among the most promising pharmacological targets, Carbonic Anhydrases (EC: 4.2.1.1) from Hp, namely HpαCA and HpßCA, emerged for their high druggability and crucial role in the survival of the pathogen in the host. Thereby, in the last decades, the two isoenzymes were isolated and characterized offering the opportunity to profile their kinetics and test different series of inhibitors.
Asunto(s)
Antibacterianos , Inhibidores de Anhidrasa Carbónica , Infecciones por Helicobacter , Helicobacter pylori , Helicobacter pylori/efectos de los fármacos , Helicobacter pylori/enzimología , Humanos , Inhibidores de Anhidrasa Carbónica/farmacología , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/microbiología , Antibacterianos/farmacología , Anhidrasas Carbónicas/metabolismo , Isoenzimas/antagonistas & inhibidoresRESUMEN
Carbonic anhydrases (CAs) are ubiquitous enzymes that are found in all kingdoms of life. Though different classes of CAs vary in their roles and structures, their primary function is to catalyze the reaction between carbon dioxide and water to produce bicarbonate and a proton. Neisseria gonorrhoeae encodes for three distinct CAs (NgCAs) from three different families: an α-, a ß-, and a γ-isoform. This chapter details the differences between the three NgCAs, summarizing their subcellular locations, roles, essentiality, structures, and enzyme kinetics. These bacterial enzymes have the potential to be drug targets; thus, previous studies have investigated the inhibition of NgCAs-primarily the α-isoform. Therefore, the classes of inhibitors that have been shown to bind to the NgCAs will be discussed as well. These classes include traditional CA inhibitors, such as sulfonamides, phenols, and coumarins, as well as non-traditional inhibitors including anions and thiocarbamates.
Asunto(s)
Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Neisseria gonorrhoeae , Neisseria gonorrhoeae/enzimología , Neisseria gonorrhoeae/efectos de los fármacos , Inhibidores de Anhidrasa Carbónica/farmacología , Anhidrasas Carbónicas/metabolismo , Humanos , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/metabolismoRESUMEN
Acetazolamide, methazolamide, ethoxzolamide and dorzolamide, classical sulfonamide carbonic anhydrase (CA) inhibitors (CAIs) designed for targeting human enzymes, were also shown to effectively inhibit bacterial CAs and were proposed for repurposing as antibacterial agents against several infective agents. CAs belonging to the α-, ß- and/or γ-classes from pathogens such as Helicobacter pylori, Neisseria gonorrhoeae, vacomycin resistant enterococci (VRE), Vibrio cholerae, Mycobacterium tuberculosis, Pseudomonas aeruginosa and other bacteria were considered as drug targets for which several classes of potent inhibitors have been developed. Treatment of some of these pathogens with various classes of such CAIs led to an impairment of the bacterial growth, reduced virulence and for drug resistant bacteria, a resensitization to clinically used antibiotics. Here I will discuss the strategies and challenges for obtaining CAIs with enhanced selectivity for inhibiting bacterial versus human enzymes, which may constitute an important weapon for addressing the drug resistance to ß-lactams and other clinically used antibiotics.
Asunto(s)
Antibacterianos , Inhibidores de Anhidrasa Carbónica , Antibacterianos/farmacología , Humanos , Inhibidores de Anhidrasa Carbónica/farmacología , Bacterias/efectos de los fármacos , Anhidrasas Carbónicas/metabolismoRESUMEN
ß-Carbonic anhydrases (ß-CA; EC 4.2.1.1) are widespread zinc metalloenzymes which catalyze the interconversion of carbon dioxide and bicarbonate. They have been isolated in many pathogenic and non-pathogenic bacteria where they are involved in multiple roles, often related to their growth and survival. ß-CAs are structurally distant from the CAs of other classes. In the active site, located at the interface of a fundamental dimer, the zinc ion is coordinated to two cysteines and one histidine. ß-CAs have been divided in two subgroups depending on the nature of the fourth ligand on the zinc ion: class I have a zinc open configuration with a hydroxide ion completing the metal coordination, which is the catalytically active species in the mechanism proposed for the ß-CAs similar to the well-known of α-CAs, while in class II an Asp residue substitute the hydroxide. This latter active site configuration has been showed to be typical of an inactive form at pH below 8. An Asp-Arg dyad is thought to play a key role in the pH-induced catalytic switch regulating the opening and closing of the active site in class II ß-CAs, by displacing the zinc-bound solvent molecule. An allosteric site well-suited for bicarbonate stabilizes the inactive form. This bicarbonate binding site is composed by a triad of well conserved residues, strictly connected to the coordination state of the zinc ion. Moreover, the escort site is a promiscuous site for a variety of ligands, including bicarbonate, at the dimer interface, which may be the route for bicarbonate to the allosteric site.
Asunto(s)
Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Dominio Catalítico , Bacterias/enzimología , Zinc/química , Zinc/metabolismo , Bicarbonatos/metabolismo , Bicarbonatos/química , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Modelos MolecularesRESUMEN
Carbonic anhydrases (CAs) are a ubiquitous family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate and protons, playing pivotal roles in a variety of biological processes including respiration, calcification, acid-base balance, and CO2 fixation. Recent studies have expanded the understanding of CAs, particularly the γ-class from diverse biological sources such as pathogenic bacteria, extremophiles, and halophiles, revealing their unique structural adaptations and functional mechanisms that enable operation under extreme environmental conditions. This chapter discusses the comprehensive catalytic mechanism and structural insights from X-ray crystallography studies, highlighting the molecular adaptations that confer stability and activity to these enzymes in harsh environments. It also explores the modulation mechanism of these enzymes, detailing how different modulators interact with the active site of γ-CAs. Comparative analyzes with other CA classes elucidate the evolutionary trajectories and functional diversifications of these enzymes. The synthesis of this knowledge not only sheds light on the fundamental aspects of CA biology but also opens new avenues for therapeutic and industrial applications, particularly in designing targeted inhibitors for pathogenic bacteria and developing biocatalysts for industrial processes under extreme conditions. The continuous advancement in structural biology promises further insights into this enzyme family, potentially leading to novel applications in medical and environmental biotechnology.
Asunto(s)
Bacterias , Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Bacterias/enzimología , Bacterias/efectos de los fármacos , Dominio Catalítico , Cristalografía por Rayos X , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Dióxido de Carbono/metabolismo , Dióxido de Carbono/químicaRESUMEN
Carbonic anhydrases (CAs) catalyze the reversable hydration of carbon dioxide to bicarbonate placing them into the core of the biochemical carbon cycle. Due to the fundamental importance of their function, they evolved independently into eight classes, three of which have been recently discovered. Most research on CAs has focused on their representatives in eukaryotic organisms, while prokaryotic CAs received significantly less attention. Nevertheless, prokaryotic CAs play a key role in the fundamental ability of the biosphere to acquire CO2 for photosynthesis and to decompose the organic matter back to CO2. They also contribute to a broad spectrum of processes in pathogenic bacteria, enhancing their ability to survive in a host and, therefore, present a promising target for developing antimicrobials. This review focuses on the distribution of CAs among bacterial pathogens and their importance in bacterial virulence and host-pathogen interactions.
Asunto(s)
Bacterias , Anhidrasas Carbónicas , Anhidrasas Carbónicas/metabolismo , Bacterias/enzimología , Bacterias/patogenicidad , Interacciones Huésped-Patógeno , Humanos , Dióxido de Carbono/metabolismo , VirulenciaRESUMEN
Mycobacterium tuberculosis (Mtb), which causes tuberculosis (TB), is still a major global health problem. According to the World Health Organization (WHO), TB still causes more deaths worldwide than any other infectious agent. Drug-sensitive TB is treatable using first-line drugs; treatment of multidrug-resistant (MDR) and extensively drug-resistant (XDR) TB requires second- and third-line drugs. However, due to the long duration of treatment, the noncompliance of patients with different levels of resistance of Mtb to these drugs has worsened the situation. Previously developed anti-TB drugs targeted the replication machinery, protein synthesis, and cell wall biosynthesis pathways of Mtb. Therefore, novel drugs targeting alternate pathways crucial for the survival and pathogenesis of Mtb in the human host are needed. The genome of Mtb encodes three ß-carbonic anhydrases (CAs) that are fundamental for pH homeostasis, hypoxia, survival, and pathogenesis. Recently, several studies have shown that the ß-CAs of Mtb could be inhibited both in vitro and in vivo using small chemical molecules, suggesting that these enzymes could be novel targets for developing anti-TB compounds that are devoid of resistance by Mtb. In addition, homologs of ß-CAs are absent in humans; therefore, drugs developed to target these enzymes might have minimal off-target effects. In this work, we describe the roles of ß-CAs in Mtb and discuss bioinformatics and cheminformatics tools used in development and discovery of novel inhibitors of these enzymes. In addition, we summarize the in vitro and in vivo studies demonstrating that the ß-CAs of Mtb are indeed druggable targets.
Asunto(s)
Antituberculosos , Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Mycobacterium tuberculosis , Tuberculosis , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/enzimología , Humanos , Anhidrasas Carbónicas/metabolismo , Antituberculosos/farmacología , Inhibidores de Anhidrasa Carbónica/farmacología , Inhibidores de Anhidrasa Carbónica/uso terapéutico , Tuberculosis/tratamiento farmacológico , Tuberculosis/microbiología , Animales , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/genéticaRESUMEN
Carbonic anhydrase (CA) plays a crucial role in the CO2 capture processes by catalyzing the hydration of CO2. In this study, we synthesized a bioinspired carbonic anhydrase Zn-MOF (metal-organic framework) incorporating 2-aminoimidazole and Zn2+ as initial constituents. The synthesized Zn-MOF exhibited promising potential for efficiently catalyzing the CO2 hydration. Structural analyses such as SEM, XRD, and BET confirmed that the Zn-MOF crystal consisted of stacked grains with an average size of approximately 36 nm, forming a micron-sized spherical structure. Functionally, Zn-MOF exhibited effective catalytic activity toward both CO2 hydration and ester hydrolysis. The introduction of amino groups significantly enhanced the esterase activity of Zn-MOF to 0.28 U/mg at ambient temperature, which was twice that of ZIF-8. Furthermore, the introduction of amino groups resulted in remarkable hydrothermal stability, with the esterase activity reaching 0.72 U/mg after undergoing hydrothermal treatment at 80 °C for 12 h. Additionally, Zn-MOF exhibited enhanced capability in CO2 hydration at a pH value exceeding 8.5. After six repeated uses, ZIF-8 and Zn-MOF retained approximately 68 and 65% of their initial enzyme activity, respectively, underscoring the potential practical applicability of Zn-MOF in industrial CO2 capture processes. This work showcases the development of a novel Zn-MOF crystal as an efficient CA mimic, effectively emulating the active sites of natural CA using 2-aminoimidazole as a coordinating ligand for Zn2+ coordination. These findings not only advance the field of innovative enzyme mimics but also pave the way for further exploration of industrial CO2 capture catalysts.
Asunto(s)
Dióxido de Carbono , Anhidrasas Carbónicas , Imidazoles , Zinc , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Imidazoles/química , Zinc/química , Dióxido de Carbono/química , Estructuras Metalorgánicas/química , Estructuras Metalorgánicas/síntesis química , Materiales Biomiméticos/química , Materiales Biomiméticos/síntesis química , Catálisis , Esterasas/química , Esterasas/metabolismoRESUMEN
Microplastics (MPs) and nanoplastics (NPs) have emerged as significant environmental pollutants with potential detrimental effects on ecosystems and human health. Several studies indicate their interaction with enzymes; this topic represents a multifaceted research field encompassing several areas of interest from the toxicological and ecotoxicological impact of MPs and NPs on humans and wildlife to the biodegradation of plastics by microbial enzymes. This review aims to provide a critical analysis of the state-of-the-art knowledge of the interaction of MPs and NPs on the enzyme carbonic anhydrase (CA), providing recent insights, analyzing the knowledge gaps in the field, and drawing future perspectives of the research and its application. CA is a widespread and crucial enzyme in various organisms; it is critical for various physiological processes in animals, plants, and bacteria. It catalyzes the reversible hydration of CO2, which is essential for respiration, acid-base balance, pH homeostasis, ion transport, calcification, and photosynthesis. Studies demonstrate that MPs and NPs can inhibit CA activity with mechanisms including adsorption to the enzyme surface and subsequent conformational changes. In vitro and in silico studies highlight the role of electrostatic and hydrophobic interactions in these processes. In vivo studies present mixed results, which are influenced by factors like particle type, size, concentration, and organism type. Moreover, the potentiality of the esterase activity of CA for plastic degradation is discussed. The complexity of the interaction between CA and MPs/NPs underscores the need for further research to fully understand the ecological and health impacts of MPs and NPs on CA activity and expression and glimpses of the potentiality and perspectives in this field.
Asunto(s)
Anhidrasas Carbónicas , Microplásticos , Nanopartículas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Microplásticos/toxicidad , Humanos , Animales , Nanopartículas/química , Contaminantes Ambientales/metabolismo , Contaminantes Ambientales/toxicidadRESUMEN
Utilizing carbonic anhydrase (CA) to catalyze CO2 hydration offers a sustainable and potent approach for carbon capture and utilization. To enhance CA's reusability and stability for successful industrial applications, enzyme immobilization is essential. In this study, delignified bamboo cellulose served as a renewable porous scaffold for immobilizing CA through oxidation-induced cellulose aldehydation followed by Schiff base linkage. The catalytic performance of the resulting immobilized CA was evaluated using both p-NPA hydrolysis and CO2 hydration models. Compared to free CA, immobilization onto the bamboo scaffold increased CA's optimal temperature and pH to approximately 45 °C and 9.0, respectively. Post-immobilization, CA activity demonstrated effective retention (>60 %), with larger scaffold sizes (i.e., 8 mm diameter and 5 mm height) positively impacting this aspect, even surpassing the activity of free CA. Furthermore, immobilized CA exhibited sustained reusability and high stability under thermal treatment and pH fluctuation, retaining >80 % activity even after 5 catalytic cycles. When introduced to microalgae culture, the immobilized CA improved biomass production by â¼16 %, accompanied by enhanced synthesis of essential biomolecules in microalgae. Collectively, the facile and green construction of immobilized CA onto bamboo cellulose block demonstrates great potential for the development of various CA-catalyzed CO2 conversion and utilization technologies.
Asunto(s)
Dióxido de Carbono , Anhidrasas Carbónicas , Celulosa , Enzimas Inmovilizadas , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Dióxido de Carbono/química , Dióxido de Carbono/metabolismo , Celulosa/química , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Concentración de Iones de Hidrógeno , Temperatura , Sasa/química , Hidrólisis , Porosidad , BiomasaRESUMEN
Despite its prominence, the ability to engineer Cupriavidus necator H16 for inorganic carbon uptake and fixation is underexplored. We tested the roles of endogenous and heterologous genes on C. necator inorganic carbon metabolism. Deletion of ß-carbonic anhydrase can had the most deleterious effect on C. necator autotrophic growth. Replacement of this native uptake system with several classes of dissolved inorganic carbon (DIC) transporters from Cyanobacteria and chemolithoautotrophic bacteria recovered autotrophic growth and supported higher cell densities compared to wild-type (WT) C. necator in batch culture. Strains expressing Halothiobacillus neopolitanus DAB2 (hnDAB2) and diverse rubisco homologs grew in CO2 similarly to the wild-type strain. Our experiments suggest that the primary role of carbonic anhydrase during autotrophic growth is to support anaplerotic metabolism, and an array of DIC transporters can complement this function. This work demonstrates flexibility in HCO3- uptake and CO2 fixation in C. necator, providing new pathways for CO2-based biomanufacturing.
Asunto(s)
Dióxido de Carbono , Cupriavidus necator , Dióxido de Carbono/metabolismo , Cupriavidus necator/metabolismo , Cupriavidus necator/genética , Bicarbonatos/metabolismo , Ciclo del Carbono/fisiología , Anhidrasas Carbónicas/metabolismo , Procesos Autotróficos , Halothiobacillus/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Ribulosa-Bifosfato Carboxilasa/metabolismoRESUMEN
INTRODUCTION: With rapid elevation in population, urbanization and industrialization, the environment is exposed to uncontrolled discharge of effluents filled with broad-spectrum toxicity, persistence and long-distance transmission anthropogenic compounds, among them heavy metals. That put our ecosystem on the verge or at a stake of drastic ecological deterioration, which eventually adversely influence on public health. Therefore, this study employed marine fungal strain Rhodotorula sp. MZ312369 for Zn2+ and Cr6+ remediation using the promising calcium carbonate (CaCO3) bioprecipitation technique, for the first time. RESULTS: Initially, Plackett-Burman design followed by central composite design were applied to optimize carbonic anhydrase enzyme (CA), which succeeded in enhancing its activity to 154 U/mL with 1.8-fold increase comparing to the basal conditions. The potentiality of our biofactory in remediating Zn2+ (50 ppm) and Cr6+ (400 ppm) was monitored through dynamic study of several parameters including microbial count, CA activity, CaCO3 weight, pH fluctuation, changing the soluble concentrations of Ca2+ along with Zn2+ and Cr6+. The results revealed that 9.23 × 107 ± 2.1 × 106 CFU/mL and 10.88 × 107 ± 2.5 × 106 CFU/mL of cells exhibited their maximum CA activity by 124.84 ± 1.24 and 140 ± 2.5 U/mL at 132 h for Zn2+ and Cr6+, respectively. Simultaneously, with pH increase to 9.5 ± 0.2, a complete removal for both metals was observed at 168 h; Ca2+ removal percentages recorded 78.99% and 85.06% for Zn2+ and Cr6+ remediating experiments, respectively. Further, the identity, elemental composition, functional structure and morphology of bioremediated precipitates were also examined via mineralogical analysis. EDX pattern showed the typical signals of C, O and Ca accompanying with Zn2+ and Cr6+ peaks. SEM micrographs depicted spindle, spherical and cubic shape bioliths with size range of 1.3 ± 0.5-23.7 ± 3.1 µm. Meanwhile, XRD difractigrams unveiled the prevalence of vaterite phase in remediated samples. Besides, FTIR profiles emphasized the presence of vaterite spectral peaks along with metals wavenumbers. CONCLUSION: CA enzyme mediated Zn2+ and Cr6+ immobilization and encapsulation inside potent vaterite trap through microbial biomineralization process, which deemed as surrogate ecofriendly solution to mitigate heavy metals toxicity and restrict their mobility in soil and wastewater.
Asunto(s)
Biodegradación Ambiental , Carbonato de Calcio , Anhidrasas Carbónicas , Cromo , Rhodotorula , Zinc , Zinc/metabolismo , Anhidrasas Carbónicas/metabolismo , Cromo/metabolismo , Carbonato de Calcio/metabolismo , Carbonato de Calcio/química , Rhodotorula/enzimología , Concentración de Iones de Hidrógeno , Contaminantes Químicos del Agua/metabolismoRESUMEN
In the search of new cymantrenyl- and ferrocenyl-sulfonamides as potencial inhibitors of human carbonic anhydrases (hCAs), four compounds based on N-ethyl or N-methyl benzenesulfonamide units have been obtained. These cymantrenyl (1a-b) and ferrocenyl (2a-b) derivatives were prepared by the reaction between aminobenzene sulfonamides ([NH2-(CH2)n-(C6H4)-SO2-NH2)], where n = 1, 2) with cymantrenyl sulfonyl chloride (P1) or ferrocenyl sulfonyl chloride (P2), respectively. All compounds were characterized by conventional spectroscopic techniques and cyclic voltammetry. In the solid state, the molecular structures of compounds 1a, 1b, and 2b were determined by single-crystal X-ray diffraction. Biological evaluation as carbonic anhydrases inhibitors were carried out and showed derivatives 1b y 2b present a higher inhibition than the drug control for the Human Carbonic Anhydrase (hCA) II and IX isoforms (KI = 7.3 nM and 5.8 nM, respectively) and behave as selective inhibition for hCA II isoform. Finally, the docking studies confirmed they share the same binding site and interactions as the known inhibitors acetazolamide (AAZ) and agree with biological studies.
Asunto(s)
Inhibidores de Anhidrasa Carbónica , Anhidrasas Carbónicas , Simulación del Acoplamiento Molecular , Sulfonamidas , Humanos , Inhibidores de Anhidrasa Carbónica/química , Inhibidores de Anhidrasa Carbónica/síntesis química , Inhibidores de Anhidrasa Carbónica/farmacología , Sulfonamidas/química , Sulfonamidas/farmacología , Sulfonamidas/síntesis química , Anhidrasas Carbónicas/metabolismo , Anhidrasas Carbónicas/química , Anhidrasa Carbónica II/antagonistas & inhibidores , Anhidrasa Carbónica II/metabolismo , Anhidrasa Carbónica II/química , Anhidrasa Carbónica IX/antagonistas & inhibidores , Anhidrasa Carbónica IX/metabolismo , Antígenos de Neoplasias/metabolismo , Antígenos de Neoplasias/química , Anhidrasa Carbónica I/antagonistas & inhibidores , Anhidrasa Carbónica I/metabolismo , Bencenosulfonamidas , Compuestos Organometálicos/química , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/farmacología , Cristalografía por Rayos XRESUMEN
Carbonic anhydrases (CAs) are a family of enzymes that play an important pH regulatory role in health and disease. While different CA isozymes have a high degree of structural similarity, they have variable enzymatic activity, with CA III being the least active and having less than 1% of the activity of CA II, the most active. Furthermore, ligand binding studies for CA III are limited, and a resulting lack of chemical probes impedes understanding of this CA isozyme in comparison to other CA family members where studies are abundant. Therefore, we employed native mass spectrometry (nMS), also known as intact mass spectrometry, to assess ligand binding to CA II and CA III and discovered two novel compounds that for the first time display strong binding to CA III. We present a new data visualization and quantification tool developed to display native mass spectra as an intuitive stacked heat map representation for rapidly interpreting the results of ligand-protein binding from nMS screening.