Copper Carbonate Nanoparticles Research Articles

Evaluation of Copper Nanoparticle Synthesis by Fusarium oxysporum Antibacterial Activity Against Staphylococcus aureus

The creation of effective, environmentally friendly antimicrobial alternatives is becoming more and more important in light of the development of antibiotic resistance. The objective of the present study was to illustrate the biosynthetic process of copper (II) carbonate nanoparticles, CuCo3 NPs, by using Fusarium oxysporum culture filtrate as an antibacterial activity against multidrug-resistant (MDR) clinical bacterial isolates, Staphylococcus aureus. Fusarium oxysporum isolate was diagnosed by PCR. One hundred and sixty specimens of pathogenic bacteria were collected from different sources (wounds, urine, sputum) then the bacterial isolates were diagnosed as Staphylococcus aureus by conventional morphological examination, biochemical tests, and the Vitek-2 system. Fusarium oxysporum culture filtrate was prepared by a culture of the fungus on Czapek Dox broth media modified by adding cornmeal and incubated for 14 days with shaking at 27±2°C and filtered in the last stages using a Millipore. The biosynthesis of CuCo3 NPs was done by adding 1g of CuCo3 Cu (OH)2 to 10 ml of Fusarium oxysporum culture filtrate. The NPs were diagnosed using modern methods, including UV-VIS, FT-IR, AFM, FE-SEM, and EDX techniques. Multidrug-resistant Staphylococcus aureus was tested on the produced Cu NPs. The results indicated that the prepared CuCo3 NPs inhibited pathogenic bacterial isolates of Staphylococcus aureus at concentration of 500, 250, 125, and 62.5 mg/ml.

Green Copper Carbonate Nanoparticles Produced by the Ureolytic Fungus Alternaria sp. Strain ccf7 and Their Antibacterial Activity

Background: Copper carbonate nanoparticles have several applications in the fields of pigments, insecticides, and fungicides. They are also used as catalysts in chemical processes and crude oil desulfurization. Fungi can biosynthesize metal nanoparticles due to their high tolerance, extracellular synthesis, simplicity of extraction, and large-scale exploitation. Objectives: This study aimed to investigate the potential of fungal isolates (which are resistant to copper chloride with urease activity) as biocatalysts for the synthesis of copper carbonate nanoparticles. This approach was considered due to the advantages of using fungal isolates in nanoparticle biosynthesis. Methods: In a PDA culture medium with 25 mM copper chloride, an enrichment culture was used to isolate copper-resistant fungal isolates. Fungal isolates’ urease enzyme was qualitatively assessed using 2% urea agar-based culture media. Studies on the synthesis of copper carbonate nanoparticles and the effect of different parameters on the synthesis of these nanoparticles were conducted using a mycelium-free supernatant strategy. Field emission scanning electron microscope (FESEM), dynamic light scattering (DLS), energy-dispersive X-ray spectroscopy (EDX), and Fourier transform infrared spectroscopy (FTIR) studies were used to determine the properties of calcium carbonate nanoparticles. The selected fungal isolate was identified using macroscopic and microscopic characteristics, as well as molecular analysis using amplification of the ITS1-5.8S-ITS2 gene sequences. Results: Alternaria sp. strain ccf7 (GenBank accession number OP242500) was chosen as the superior strain for copper carbonate nanoparticle synthesis tests based on the pattern of resistance to copper chloride salt and the qualitative assessment of urease activity. Based on the findings of the electron microscope studies, spherical copper carbonate nanoparticles with an average size of 66.7 nm were synthesized after 24 hours of incubation at the optimal concentration of 45 mM copper chloride, temperature of 25°C, and shaker speed of 100 rpm. The distribution of the produced nanoparticles was appropriate, as indicated by a polydispersity index (PDI) of 0.25. The strongest inhibitory impact of these copper carbonate nanoparticles was against Pseudomonas aeruginosa, with an average inhibition of 31 mm at a concentration of 50 mg/L, according to the results of their antibacterial activities. Conclusions: For the first time, the synthesis and development of a green approach for the fabrication of copper carbonate nanoparticles using the genus Alternaria have been proposed in this study.

Copper carbonate nanoparticles as an effective biomineralized carrier to load macromolecular drugs for multimodal therapy

Macromolecular drugs have attracted great interest as biotherapy to cure previously untreatable diseases. For clinical translation, biomacromolecules encounter several common druggability difficulties, such as in vivo instability and poor penetration to cross physiologic barriers, thus requiring sophisticated systems for drug delivery. Inspired by the natural biomineralization via interaction between inorganic ions and biomacromolecules, herein we rationally screened biocompatible transition metals to biomineralize with carbonate for macromolecules loading. Among the metal ions, Cu2+ was found to be the best candidate, and its superiority over the widely studied Ca2+ minerals was also demonstrated. Capitalized on this finding, copper carbonate nanoparticles were prepared via a simple mixing process to co-load glucose oxidase (GOx) and a HIF-α DNAzyme (DZ), achieving ultra-high loading capacity of 61%. Upon encapsulation into nanoparticles, enzymatic activity of both drugs was passivated to avoid potential side-effects during circulation, while the drugs could be rapidly released within 1 h in response to acidic pH to fully recover their activities. The nanoparticles could accumulate into tumor via intravenous injection, facilitate the cell membrane penetration, and release the payloads of GOx, DZ and Cu2+ inside cells to exert a series of anti-tumor effects. GOx caused tumor starvation by catalytic glucose consumption, and the concomitantly generated H2O2 byproduct boosted the Cu2+-mediated chemodynamic therapy (CDT). Meanwhile, the DZ silenced HIF-α expression to sensitize both starvation therapy and CDT. As a result, a synergistic tumor growth inhibition was achieved. This work provides a simple method to prepare biomineralized nanoparticles, and offers a general approach for macromolecular drugs delivery via Cu2+-based biomineralization.

Application of fungal copper carbonate nanoparticles as environmental catalysts: organic dye degradation and chromate removal

Biomineralization is a ubiquitous process in organisms to produce biominerals, and a wide range of metallic nanoscale minerals can be produced as a consequence of the interactions of micro-organisms with metals and minerals. Copper-bearing nanoparticles produced by biomineralization mechanisms have a variety of applications due to their remarkable catalytic efficiency, antibacterial properties and low production cost. In this study, we demonstrate the biotechnological potential of copper carbonate nanoparticles (CuNPs) synthesized using a carbonate-enriched biomass-free ureolytic fungal spent culture supernatant. The efficiency of the CuNPs in pollutant remediation was investigated using a dye (methyl red) and a toxic metal oxyanion, chromate Cr(VI). The biogenic CuNPs exhibited excellent catalytic properties in a Fenton-like reaction to degrade methyl red, and efficiently removed Cr(VI) from solution due to both adsorption and reduction of Cr(VI). X-ray photoelectron spectroscopy (XPS) identified the oxidation of reducing Cu species of the CuNPs during the reaction with Cr(VI). This work shows that urease-positive fungi can play an important role not only in the biorecovery of metals through the production of insoluble nanoscale carbonates, but also provides novel and simple strategies for the preparation of sustainable nanomineral products with catalytic properties applicable to the bioremediation of organic and metallic pollutants, solely and in mixtures.

The effect of the use of copper carbonate and copper nanoparticles in the diet of rats on the level of β-amyloid and acetylcholinesterase in selected organs

BackgroundCopper has an important role in nervous system function, as a cofactor of many enzymes and in the synthesis of neurotransmitters. Both the dose and the chemical form of copper can determine the impact of this element on metabolism, the neurological system and the immune system. AimsThe aim of the study was to determine whether and in what form the addition of copper changes the level of amyloid beta and acetylcholinesterase level in selected rat tissues. MethodsThirty, healthy, male, albino Wistar rats aged 7 weeks were randomly divided into 3 groups. Three experimental treatments were used to evaluate the effects of different levels and sources of Cu (6.5 mg kg of diet) in the diet: Cu0 – rats fed a diet without Cu supplementation; Cusalt – rats fed a diet with CuCO3 (6.5 mg kg of diet) during two months of feeding; CuNPs - rats fed a diet with Cu nanoparticles (6.5 mg kg of diet) during two months of feeding. In blood serum and tissue homogenates there rated the indicators proving the potential neurodegenerative effect and epigenetic DNA damage induced by chemical form of copper or lack of additional copper supplementation in diet were determined. There were analysed: level of acetylcholinesterase, β-amyloid, low-density lipoprotein receptor-related protein 1, apyrimidinic endonuclease, thymidine glycosidase, alkylpurine-DNA-N-glycosylase and glycosylated acetylcholinesterase. ResultsIrrespective of the form of copper added, it was found to increase acetylcholinesterase level in the brain, spleen and liver, as well as in the blood plasma of the rats. Copper in the form of CuCO3 was found to increase acetylcholinesterase level in the kidneys. The addition of both forms of copper caused a marked increase in the plasma concentration of β-amyloid in comparison with the diet with no added Cu. The addition of both forms of copper caused a marked increase in the plasma concentration of β-amyloid in comparison with the diet with no added Cu. ConclusionsA lack of added Cu in the diet of rats reduces the concentration of amyloid-β in the blood, whereas administration of copper, in the form of either CuNPs or CuCO3, increases the level of this peptide in the blood. The use of copper in the form of CuNPs in the diet of rats does not increase the level of β-amyloid more than the use of the carbonate form of this element. The use of CuNPs or CuCO3 in the diet of rats increases acetylcholinesterase level in the brain, spleen, liver, and blood. CuNPs in the diet of rats were not found to increase acetylcholinesterase level to a greater extent than Cu+2 carbonate.

Mycology: Protein Control of Fungal Nanoparticle Formation

The ability of fungi to immobilize metals is an extraordinary phenomenon. Now, a protein involved in the formation of copper carbonate nanoparticles has been identified, and suggests that the process is under the control of the fungus, opening new avenues for the synthesis of nanomaterials.

Role of Protein in Fungal Biomineralization of Copper Carbonate Nanoparticles

Biomineralization processes are of key importance in the biogeochemical cycling of metals and other elements by microorganisms, and several studies have highlighted the potential applications of nanoparticle synthesis via biomineralization. The roles played by proteins in the transformation and biologically induced biomineralization of metals by microorganisms is not well understood, despite the interactions of protein and nanoparticles at mineral interfaces attracting much interest in various emerging fields for novel biomaterial synthesis. Here, we have elucidated the association and involvement of fungal proteins in the formation of biogenic copper carbonate nanoparticles (CuNPs) using a carbonate-enriched biomass-free ureolytic fungal culture supernatant. Proteomic analysis was conducted that identified the major proteins present in the culture supernatant. Of the proteins identified, triosephosphate isomerase (TPI) exhibited a strong affinity to the CuNPs, and the impact of purified TPI on CuNP formation was studied in detail. The combined use of scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) confirmed that TPI played an important role in controlling the morphology and structure of the nanomaterials. Fourier transform infrared spectroscopy (FTIR) was applied to examine conformational changes of the proteins to further clarity the interaction mechanisms with CuNPs during biomineralization. Such analyses revealed unfolding of proteins on the mineral surface and an increase in β sheets within the protein structure. These results extend understanding of how microbial systems can influence biomineral formation through protein secretion, the mechanisms involved in formation of complex protein/inorganic systems, and provide useful guidelines for the synthesis of inorganic-protein based nanomaterials.

Amino acid secretion influences the size and composition of copper carbonate nanoparticles synthesized by ureolytic fungi

The ureolytic activity of Neurospora crassa results in an alkaline carbonate-rich culture medium which can precipitate soluble metals as insoluble carbonates. Such carbonates are smaller, often of nanoscale dimensions, than metal carbonates synthesized abiotically which infers that fungal excreted products can markedly affect particle size. In this work, it was found that amino acid excretion was a significant factor in affecting the particle size of copper carbonate. Eleven different amino acids were found to be secreted by Neurospora crassa, and l-glutamic acid, l-aspartic acid and l-cysteine were chosen to examine the impact of amino acids on the morphology and chemical composition of copper carbonate minerals. X-ray powder diffraction (XRPD), scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA) and X-ray photoelectron spectroscopy (XPS) were used to characterize the obtained copper carbonate samples. Copper carbonate nanoparticles with a diameter of 100–200 nm were produced with l-glutamic acid, and the presence of l-glutamic acid was found to stabilize these particles in the early phase of crystal growth and prevent them from aggregation. FTIR and TG analysis revealed that the amino acid moieties were intimately associated with the copper mineral particles. Component analysis of the final products of TG analysis of the copper minerals synthesized under various conditions showed the ultimate formation of Cu, Cu2O and Cu2S, suggesting a novel synthesis method for producing these useful Cu-containing materials.

Toxicity of copper oxide and basic copper carbonate nanoparticles after short-term oral exposure in rats

Copper oxide (CuO) nanoparticles (NPs) and copper carbonate nanoparticles (Cu2CO3(OH)2 NPs have applications as antimicrobial agents and wood preservatives: an application that may lead to oral ingestion via hand to mouth transfer. Rats were exposed by oral gavage to CuO NPs and Cu2CO3(OH)2 NPs for five consecutive days with doses from 1 to 512 mg/kg and 4 to 128 mg/kg per day, respectively, and toxicity was evaluated at days 6 and 26. Both CuO NPs and Cu2CO3(OH)2 NPs induced changes in hematology parameters, as well as clinical chemistry markers (e.g. increased alanine aminotransferase, ALT) indicative of liver damage For CuO NPs histopathological alterations were observed in bone marrow, stomach and liver mainly consisting of an inflammatory response, ulceration, and degeneration. Cu2CO3(OH)2 NPs induced morphological alterations in the stomach, liver, intestines, spleen, thymus, kidneys, and bone marrow. In spleen and thymus lymphoid, depletion was noted that warrants further immunotoxicological evaluation. The NPs showed partial dissolution in artificial simulated stomach fluids, while in intestinal conditions, the primary particles simultaneously shrank and agglomerated into large structures. This means that both copper ions and the particulate nanoforms should be considered as potential causal agents for the observed toxicity. For risk assessment, the lowest bench mark dose (BMD) was similar for both NPs for the serum liver enzyme AST (an indication of liver toxicity), being 26.2 mg/kg for CuO NPs and 30.8 mg/kg for Cu2CO3(OH)2 NPs. This was surprising since the histopathology evidence demonstrates more severe organ damage for Cu2CO3(OH)2 NPs than for CuO NPs.

Biosynthesis of copper carbonate nanoparticles by ureolytic fungi

In this research, the ureolytic fungi Neurospora crassa, Pestalotiopsis sp. and Myrothecium gramineum were investigated for the preparation of nanoscale copper carbonate and the role of fungal extracellular protein in such mineral formation. After incubation in urea-modified media, carbonate-laden fungal supernatants were used for the precipitation of copper carbonate, with experimental results agreeing closely with those obtained using geochemical modelling (Geochemist’s Workbench). Compared with commercial and chemically synthesized copper carbonate, the minerals obtained using fungal supernatants were nanoscale and showed varying morphologies. It was found that extracellular protein played an important role in determining the size and morphology of the carbonate minerals precipitated, and after mixture with CuCl2 and resultant copper carbonate precipitation, more than 80% protein was removed from the N. crassa supernatant. Moreover, with addition of extracellular protein extracted from different fungal supernatants or standard bovine serum albumin, more than 96% of protein was removed by carbonate mineral precipitation. These results provide direct experimental evidence for the preparation of copper carbonate nanoparticles utilizing fungal ureolytic activity and show that fungal extracellular protein plays an important role in the formation and size of specific nano metal carbonates. Such a process provides opportunities for production of specific and/or novel metal carbonate nanoparticles of applied relevance, and as precursors of other useful biomineral products such as oxides.

Release of copper-amended particles from micronized copper-pressure-treated wood during mechanical abrasion.

BackgroundWe investigated the particles released due to abrasion of wood surfaces pressure-treated with micronized copper azole (MCA) wood preservative and we gathered preliminary data on its in vitro cytotoxicity for lung cells. The data were compared with particles released after abrasion of untreated, water (0% MCA)-pressure-treated, chromated copper (CC)-pressure-treated wood, and varnished wood. Size, morphology, and composition of the released particles were analyzed.ResultsOur results indicate that the abrasion of MCA-pressure-treated wood does not cause an additional release of nanoparticles from the unreacted copper (Cu) carbonate nanoparticles from of the MCA formulation. However, a small amount of released Cu was detected in the nanosized fraction of wood dust, which could penetrate the deep lungs. The acute cytotoxicity studies were performed on a human lung epithelial cell line and human macrophages derived from a monocytic cell line. These cell types are likely to encounter the released wood particles after inhalation.ConclusionsOur findings indicate that under the experimental conditions chosen, MCA does not pose a specific additional nano-risk, i.e. there is no additional release of nanoparticles and no specific nano-toxicity for lung epithelial cells and macrophages.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-016-0232-7) contains supplementary material, which is available to authorized users.

Electrosynthesis and thermal characterization of basic copper carbonate nanoparticles

AbstractThe present study concerns the electrochemical synthesis of basic copper carbonate nanoparticles by oxidation of metallic copper on the anode in an aqueous bicarbonate solution. This simple and one-step preparation can be considered as green synthesis. The scanning electron microscopy (SEM) analysis indicates that average particle size of the product is in the range of about 70 nm. On the other hand, basic copper carbonate micro-powder has been prepared, by mixing solutions of copper(II) sulphate and sodiu bicarbonate. The SEM analysis showed that the size of particles prepared in the same way is in the range of about 1 µm. In another part of this study, the thermal decomposition of micro and nanoparticles of copper carbonate produced by various methods was studied in air using TG-DTA techniques. The results of thermal study show that the decomposition of both samples occurs in single step. Also, the TG-DTA analysis of the nanoparticles indicates that the main thermal degradation occurs in the temperature range of 245–315°C. However, microparticles of Cu(OH)2 · CuCO3 decomposed endothermally in the temperature range of 230–330°C.

Microdistribution of copper-carbonate and iron oxide nanoparticles in treated wood

Aqueous dispersions of copper-carbonate nanoparticles and microparticles have just begun to be exploited commercially for the preservative treatment of wood. The success of the new systems will depend, in part, on the uniform distribution of the preservative in wood and the ability of copper to penetrate cell walls. We examined the distribution of copper in wood treated with a nano-Cu preservative. Copper particles are not uniformly distributed in treated wood, but they accumulate in voids that act as the flow paths for liquids in wood. Particles are deposited on, but not within cell walls. Nevertheless, elemental copper is present within cell walls, but at a lower level than that in wood treated with a conventional wood preservative. These findings suggest that nano-Cu preservatives are able to deliver bioactive components into wood cell walls even though the majority of copper particles are too large to penetrate the cell wall’s nanocapillary network.