Carbonic Anhydrase-Embedded ZIF-8 Electrospun PVA Fibers as an Excellent Biocatalyst Candidate
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PROVIDER: S-EPMC10210226 | biostudies-literature | 2023 May
REPOSITORIES: biostudies-literature
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Project description:Enzymes expressed by highly salt-tolerant organisms show many modifications compared with salt-affected counterparts including biased amino acid and lower α-helix content, lower solvent accessibility and negative surface charge. Here, we show that halotolerance can be generated in an enzyme solely by modifying surface residues. Rational design of carbonic anhydrase II is undertaken in three stages replacing 18 residues in total, crystal structures confirm changes are confined to surface residues. Catalytic activities and thermal unfolding temperatures of the designed enzymes increase at high salt concentrations demonstrating their shift to halotolerance, whereas the opposite response is found in the wild-type enzyme. Molecular dynamics calculations reveal a key role for sodium ions in increasing halotolerant enzyme stability largely through interactions with the highly ordered first Na(+) hydration shell. For the first time, an approach to generate extreme halotolerance, a trait with broad application in industrial biocatalysis, in a wild-type enzyme is demonstrated.
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Project description:Carbonic anhydrase is an enzyme that reversibly catalyzes the hydration of carbon dioxide (CO2). It has been suggested recently that this remarkably fast enzyme can be used for sequestration of CO2, a major greenhouse gas, making this a promising alternative for chemical CO2 mitigation. To promote the economical use of enzymes, we engineered the carbonic anhydrase from Neisseria gonorrhoeae (ngCA) in the periplasm of Escherichia coli, thereby creating a bacterial whole-cell catalyst. We then investigated the application of this system to CO2 sequestration by mineral carbonation, a process with the potential to store large quantities of CO2. ngCA was highly expressed in the periplasm of E. coli in a soluble form, and the recombinant bacterial cell displayed the distinct ability to hydrate CO2 compared with its cytoplasmic ngCA counterpart and previously reported whole-cell CA systems. The expression of ngCA in the periplasm of E. coli greatly accelerated the rate of calcium carbonate (CaCO3) formation and exerted a striking impact on the maximal amount of CaCO3 produced under conditions of relatively low pH. It was also shown that the thermal stability of the periplasmic enzyme was significantly improved. These results demonstrate that the engineered bacterial cell with periplasmic ngCA can successfully serve as an efficient biocatalyst for CO2 sequestration.
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Project description:Rare earth elements (REEs) are critical raw materials with a wide range of industrial applications. As a result, the recovery of REEs via adsorption from REE-rich matrices, such as water streams from processed electric and electronic waste, has gained increased attention for its simplicity, cost-effectiveness and high efficacy. In this work, the potential of nanometric cerium oxide-based materials as adsorbents for selected REEs is investigated. Ultra-small cerium oxide nanoparticles (CNPs, mean size diameter ≈ 3 nm) were produced via a precipitation-hydrothermal procedure and incorporated into woven-non-woven polyvinyl alcohol (PVA) nanofibres (d ≈ 280 nm) via electrospinning, to a final loading of ≈34 wt%. CNPs, CNP-PVA and the benchmark material CeO2 NM-212 (JRCNM02102, mean size diameter ≈ 28 nm) were tested as adsorbents for aqueous solutions of the REEs Eu3+, Gd3+ and Yb3+ at pH 5.8. Equilibrium adsorption data were interpreted by means of Langmuir and Freundlich data models. The maximum adsorption capacities ranged between 16 and 322 mgREE gCeO2 -1, with the larger value found for the adsorption of Yb3+ by CNP. The trend of maximum adsorption capacity was CNPs > NM-212 > CNP-PVA, which was ascribed to different agglomeration and surface area available for adsorption. Langmuir equilibrium constants K L were substantially larger for CNP-PVA, suggesting a potential higher affinity of REEs for CNPs due to a synergistic effect of PVA on adsorption. CNP-PVA were effectively used in repeated adsorption cycles under static and dynamic configurations and retained the vast majority of adsorptive material (>98% of CeO2 retained after 10 adsorption cycles). The small loss was attributed to partial solubilisation of fibre components with change in membrane morphology. The findings of this study pave the way for the application of CNP-PVA nanocomposites in the recovery of strategically important REEs from electrical and electronic waste.
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Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction
2012-03-25 | GSE36625 | GEO
Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction 6 dye-swap - gene knock out
2012-03-25 | E-GEOD-36625 | biostudies-arrayexpress
Project description:Ischemic stroke is a leading cause of death and disability worldwide. The only pharmacological treatment available to date for cerebral ischemia is tissue plasminogen activator (t-PA) and the search for successful therapeutic strategies still remains a major challenge. The loss of cerebral blood flow leads to reduced oxygen and glucose supply and a subsequent switch to the glycolytic pathway, which leads to tissue acidification. Carbonic anhydrase (CA, EC 4.2.1.1) is the enzyme responsible for converting carbon dioxide into a protons and bicarbonate, thus contributing to pH regulation and metabolism, with many CA isoforms present in the brain. Recently, numerous studies have shed light on several classes of carbonic anhydrase inhibitor (CAI) as possible new pharmacological agents for the management of brain ischemia. In the present review we summarized pharmacological, preclinical and clinical findings regarding the role of CAIs in strokes and we discuss their potential protective mechanisms.
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Project description:High temperature is required in carbon fiber synthesis in the carbonization step. However, direct high-temperature heating without the presence of additive materials would affect the yield and structure of carbon fibers produced. Thus, this study aims to synthesize carbon fibers from poly-vinyl alcohol (PVA), as the precursor and reducing agent, using silver nanoparticles (SNP) from silver nitrate (AgNO3) as additives. The pre-treatment of PVA was performed in three steps, i.e., mixing PVA/AgNO3, electrospinning, and iodination. The interaction of PVA and AgNO3 was assessed by FTIR, and SEM was used to characterize the electro-spun fibers prior and after iodination; Raman spectrophotometer was carried out to confirm the yield of carbon fibers. There was reduction in oxygen groups (3000-3800 cm-1) and emergence of -C=O (1100 cm-1) and -C=C- (1627 cm-1) functional groups, indicating formation of carbon layers. Based on the DT/GA results, the silver nanoparticles reduce the need of high temperature with optimum carbonization at 350 °C and lead to the formation of more regular graphene layers. Graphene layers with a size distribution of 0.438 nm and well-organized structures were successfully formed, and the Raman shifting showed higher intensities of G and G' bands in the presence of Ag. Based on DT/GA results, the yield of carbon fibers with iodinated PVA fibers and SNP as additive had higher rates around 800 µg/min, reaching 33% at 500 °C. Thus, it is demonstrated that iodinated PVA/AgNO3 samples can significantly improve carbon fiber yield at low temperatures.
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Project description:Enzyme prodrug therapy (EPT) enables localized conversion of inert prodrugs to active drugs by enzymes. Performance of EPT necessitates that the enzyme remains active throughout the time frame of the envisioned therapeutic application. β-glucuronidase is an enzyme with historically validated performance in EPT, however it retains its activity in biomaterials for an insufficiently long period of time, typically not exceeding 7 d. Herein, the encapsulation of β-glucuronidase in liposomal subcompartments within poly(vinyl alcohol) electrospun fibers is reported, leading to the assembly of biocatalytically active materials with activity of the enzyme sustained over at least seven weeks. It is further shown that liposomes provide the highly beneficial stabilization of the enzyme when incubated in cell culture media. The assembled biocatalytic materials successfully produce antiproliferative drugs (SN-38) using externally administered prodrugs (SN-38-glucuronide) and effectively suppress cell proliferation, with envisioned utility in the design of cardiovascular grafts.
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Project description:Specific isoforms from the carbonic anhydrase (CA) family of zinc metalloenzymes have been associated with a variety of diseases. Isoform-specific carbonic anhydrase inhibitors (CAIs) are therefore a major focus of attention for specific disease treatments. Classical CAIs, primarily sulfonamide-based compounds and their bioisosteres, are examined as antiglaucoma, antiepileptic, antiobesity, antineuropathic pain and anticancer compounds. However, many sulfonamide compounds inhibit all CA isoforms nonspecifically, diluting drug effectiveness and causing undesired side effects due to off-target inhibition. In addition, a small but significant percentage of the general population cannot be treated with sulfonamide-based compounds due to a sulfa allergy. Therefore, CAIs must be developed that are not only isoform specific, but also non-classical, i.e. not based on sulfonamides, sulfamates, or sulfamides. This review covers the classes of non-classical CAIs and the recent advances in the development of isoform-specific inhibitors based on phenols, polyamines, coumarins and their derivatives.
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