Promising approaches for the assembly of the catalytically active, recombinant Desulfomicrobium baculatum hydrogenase with substitutions at the active site.

BACKGROUND: Hydrogenases (H2ases) are metalloenzymes capable of the reversible conversion of protons and electrons to molecular hydrogen. Exploiting the unique enzymatic activity of H2ases can lead to advancements in the process of biohydrogen evolution and green energy production. RESULTS: Here we created of a functional, optimized operon for rapid and robust production of recombinant [NiFe] Desulfomicrobium baculatum hydrogenase (Dmb H2ase). The conversion of the [NiFeSe] Dmb H2ase to [NiFe] type was performed on genetic level by site-directed mutagenesis. The native dmb operon includes two structural H2ase genes, coding for large and small subunits, and an additional gene, encoding a specific maturase (protease) that is essential for the proper maturation of the enzyme. Dmb, like all H2ases, needs intricate bio-production machinery to incorporate its crucial inorganic ligands and cofactors. Strictly anaerobic, sulfate reducer D. baculatum bacteria are distinct, in terms of their biology, from E. coli. Thus, we introduced a series of alterations within the native dmb genes. As a result, more than 100 elements, further compiled into 32 operon variants, were constructed. The initial requirement for a specific maturase was omitted by the artificial truncation of the large Dmb subunit. The assembly of the produced H2ase subunit variants was investigated both, in vitro and in vivo. This approach resulted in 4 recombinant [NiFe] Dmb enzyme variants, capable of H2 evolution. The aim of this study was to overcome the gene expression, protein biosynthesis, maturation and ligand loading bottlenecks for the easy, fast, and cost-effective delivery of recombinant [NiFe] H2ase, using a commonly available E. coli strains. CONCLUSION: The optimized genetic constructs together with the developed growth and purification procedures appear to be a promising platform for further studies toward fully-active and O2 tolerant, recombinant [NiFeSe] Dmb H2ase, resembling the native Dmb enzyme. It could likely be achieved by selective cysteine to selenocysteine substitution within the active site of the [NiFe] Dmb variant.

Photocatalytic hydrogen evolution from glycerol-water mixture under visible light over zinc indium sulfide (ZnIn2S4) nanosheets grown on bismuth oxychloride (BiOCl) microplates.

ZnIn2S4 (ZIS) is one of the widely studied photocatalyst for photocatalytic hydrogen evolution applications due to its prominent visible light response and strong reduction ability. However, its photocatalytic glycerol reforming performance for hydrogen evolution has never been reported. Herein, the visible light driven BiOCl@ZnIn2S4 (BiOCl@ZIS) composite was synthesized by growth of ZIS nanosheets on a template-like hydrothermally pre-prepared wide-band-gap BiOCl microplates using simple oil-bath method to be used for the first time for photocatalytic glycerol reforming for photocatalytic hydrogen evolution (PHE) under visible light irradiation (λ > 420 nm). The optimum amount of BiOCl microplates in the composite was found 4 wt% (4% BiOCl@ZIS) in the presence of in-situ 1 wt% Pt deposition. Then, the in-situ Pt photodeposition optimization studies over 4% BiOCl@ZIS composite showed the highest PHE rate of 674 µmol g-1h-1 with the ultra-low platinum amount (0.0625 wt%). The possible mechanisms behind this improvement can be ascribed to the formation of Bi2S3 low-band-gap semiconductor during BiOCl@ZIS composite synthesis resulting in Z-scheme charge transfer mechanism between ZIS and Bi2S3 upon visible light irradiation. This work expresses not only the photocatalytic glycerol reforming over ZIS photocatalyst but also a solid proof of the contribution of wide-band-gap BiOCl photocatalysts to enhancement of ZIS PHE performance under visible light.

Catalytic Activity of New Oxovanadium(IV) Microclusters with 2-Phenylpyridine in Olefin Oligomerization.

So far, few microclusters containing vanadium have been described in the literature. In this report, the synthesis protocol for the preparation of oxovanadium (IV) microclusters with 2-phenylpyridine is shown for the first time. Moreover, the crystal structure of these microclusters is also studied through the use of X-rays. The morphology of the prepared crystals is investigated using a field-emission Scanning Electron Microscope (SEM). The new compound, after activation by modified methylaluminoxane as the catalytic system, is investigated regarding the oligomerizations of 3-buten-1-ol, 2-chloro-2-propen-1-ol, allyl alcohol, and 2,3-dibromo-2-propen-1-ol. The products of oligomerization are tested by the TG-FTIR and MALDI-TOF-MS methods. Moreover, the values of catalytic activities for the new oxovanadium(IV) microclusters with 2-phenylpyridine are determined for the 3-buten-1-ol, 2-chloro-2-propen-1-ol, allyl alcohol, and 2,3-dibromo-2-propen-1-ol oligomerizations. Oxovanadium(IV) microclusters with 2-phenylpyridine are shown to be very highly active precatalysts for the oligomerization of allyl alcohol, 2,3-dibromo-2-propen-1-ol, and 3-buten-1-ol. However, in the case of 2-chloro-2-propen-1-ol oligomerization, the new microclusters are seen as highly active precatalysts.

A chemoinformatics approach for the characterization of hybrid nanomaterials: safer and efficient design perspective.

In this study, photocatalytic properties and in vitro cytotoxicity of 29 TiO2-based multi-component nanomaterials (i.e., hybrids of more than two composition types of nanoparticles) were evaluated using a combination of the experimental testing and supervised machine learning modeling. TiO2-based multi-component nanomaterials with metal clusters of silver, and their mixtures with gold, palladium, and platinum were successfully synthesized. Two activities, photocatalytic activity and cytotoxicity, were studied. A novel cheminformatic approach was developed and applied for the computational representation of the photocatalytic activity and cytotoxicity effect. In this approach, features of investigated TiO2-based hybrid nanomaterials were reflected by a series of novel additive descriptors for hybrid and hybrid nanostructures (denoted as "hybrid nanosctructure descriptors"). These descriptors are based on quantum chemical calculations and the Smoluchowski equation. The obtained experimental data and calculated hybrid-nanostructure descriptors were used to develop novel predictive Quantitative Structure-Activity Relationship computational models (called "nano-QSARmix"). The proposed modeling approach is an initial step in the understanding of the relationships between physicochemical properties of hybrid nanoparticles, their toxicity, and photochemical activity under UV-vis irradiation. Acquired knowledge supports the safe-by-design approaches relevant to the development of efficient hybrid nanomaterials with reduced hazardous effects.

Impact of gold nanoparticles shape on their cytotoxicity against human osteoblast and osteosarcoma in in vitro model. Evaluation of the safety of use and anti-cancer potential.

Due to development of nanotechnology and gold nanoparticles (AuNPs) increasing use in different areas of medicine, especially in oncology, better understanding of their potential cytotoxicity is necessary to protect patients safety. Shape and size of AuNPs is an important modulator of their cytotoxicity. Therefore, we investigated the cytotoxicity of AuNPs rods (≈39 nm length, 18 nm width), AuNPs stars (≈ 215 nm) and AuNPs spheres (≈ 6.3 nm) against human fetal osteoblast (hFOB 1.19), osteosarcoma (143B, MG63) and pancreatic duct cell (hTERT-HPNE) lines by MTT and neutral-red uptake assay. Moreover, influence of AuNPs on level of proapoptotic protein (Bax) and anti-apoptotic protein (Bcl-2) was measured by western blot. Cellular uptake of nanoparticles and ultrastructure changes were examined by transmission electron microscopy (TEM). In the present study we have proven that AuNPs stars are the most cytotoxic against human cells. We observed that cancer cells are more susceptible to AuNPs cytotoxic effect. Furthermore, AuNPs rods and AuNPs stars caused increased expression of Bax and decreased expression of Bcl-2 protein in osteosarcoma cells. We found that AuNPs penetrated through the cell membrane and caused ultrastructural changes. Our results clearly demonstrated that the cytotoxicity of AuNPs was shape-dependent. AuNPs stars with the highest anti-cancer potential were also the most cytotoxic type of tested NPs, whereas AuNPs spheres which appears to be the safest one had small anti-cancer potential.

Design, Synthesis, and Enzymatic Evaluation of Novel ZnO Quantum Dot-Based Assay for Detection of Proteinase 3 Activity.

Herein, the synthesis and application of functionalized quantum dot-based protease probes is described. Such probes are composed of nontoxic ZnO nanocrystals decorated by amino groups followed by linker and labeled peptide attachment. Spherical NH2-terminated ZnO quantum dots (QDs) with the average size ranging from 4 to 8 nm and strong emission centered at 530 nm were prepared using the sol-gel method. The fluorescence of ZnO QDs was quenched by the BHQ1 moiety present on the N-terminal amino group of the peptide. The enzymatic cleavage of the peptide mediated by the proteinase 3 (PR3) bond resulted in an increase in the QD probe fluorescence. This observation was verified using both model and biological systems; and the picomolar detection limit was found to be more than 30 times lower than that of the previously reported internally quenched peptide (a decrease in detection limit from 43 to 1.3 pmol was observed).

Quantum dot-decorated semiconductor micro- and nanoparticles: A review of their synthesis, characterization and application in photocatalysis.

Quantum dot (QD)-decorated semiconductor micro- and nanoparticles are a new class of functional nanomaterials that have attracted considerable interest for their unique structural, optical and electronic properties that result from the large surface-to-volume ratio and the quantum confinement effect. In addition, because of QDs' excellent light-harvesting capacity, unique photoinduced electron transfer, and up-conversion behaviour, semiconductor nanoparticles decorated with quantum dots have been used widely in photocatalytic applications for the degradation of organic pollutants in both the gas and aqueous phases. This review is a comprehensive overview of the recent progress in synthesis methods for quantum dots and quantum dot-decorated semiconductor composites with an emphasis on their composition, morphology and optical behaviour. Furthermore, various approaches used for the preparation of QD-based composites are discussed in detail with respect to visible and UV light-induced photoactivity. Finally, an outlook on future development is proposed with the goal of overcoming challenges and stimulating further research into this promising field.

Evaluating the toxicity of TiO2-based nanoparticles to Chinese hamster ovary cells and Escherichia coli: a complementary experimental and computational approach.

Titania-supported palladium, gold and bimetallic nanoparticles (second-generation nanoparticles) demonstrate promising photocatalytic properties. However, due to unusual reactivity, second-generation nanoparticles can be hazardous for living organisms. Considering the ever-growing number of new types of nanoparticles that can potentially contaminate the environment, a determination of their toxicity is extremely important. The main aim of presented study was to investigate the cytotoxic effect of surface modified TiO2-based nanoparticles, to model their quantitative nanostructure-toxicity relationships and to reveal the toxicity mechanism. In this context, toxicity tests for surface-modified TiO2-based nanoparticles were performed in vitro, using Gram-negative bacteria Escherichia coli and Chinese hamster ovary (CHO-K1) cells. The obtained cytotoxicity data were analyzed by means of computational methods (quantitative structure-activity relationships, QSAR approach). Based on a combined experimental and computational approach, predictive models were developed, and relationships between cytotoxicity, size, and specific surface area (Brunauer-Emmett-Teller surface, BET) of nanoparticles were discussed.

Size and shape-dependent cytotoxicity profile of gold nanoparticles for biomedical applications.

Metallic nanoparticles, in particular gold nanoparticles (AuNPs), offer a wide spectrum of applications in biomedicine. A crucial issue is their cytotoxicity, which depends greatly on various factors, including morphology of nanoparticles. Because metallic nanoparticles have an effect on cell membrane integrity, their shape and size may affect the viability of cells, due to their different geometries as well as physical and chemical interactions with cell membranes. Variations in the size and shape of gold nanoparticles may indicate particular nanoparticle morphologies that provide strong cytotoxicity effects. Synthesis of different sized and shaped bare AuNPs was performed with spherical (~ 10 nm), nanoflowers (~ 370 nm), nanorods (~ 41 nm), nanoprisms (~ 160 nm) and nanostars (~ 240 nm) morphologies. These nanostructures were characterized and interacting with cancer (HeLa) and normal (HEK293T) cell lines and cell viability tests were performed by WST-1 tests and fluorescent live/dead cell imaging experiments. It was shown that various shapes and sizes of gold nanostructures may affect the viability of the cells. Gold nanospheres and nanorods proved to be more toxic than star, flower and prism gold nanostructures. This may be attributed to their small size and aggregation process. This is the first report concerning a comparison of cytotoxic profile in vitro with a wide spectrum of bare AuNPs morphology. The findings show their possible use in biomedical applications.

Self-Organized TiO2-MnO2 Nanotube Arrays for Efficient Photocatalytic Degradation of Toluene.

Vertically oriented, self-organized TiO2-MnO2 nanotube arrays were successfully obtained by one-step anodic oxidation of Ti-Mn alloys in an ethylene glycol-based electrolyte. The as-prepared samples were characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), UV-Vis absorption, photoluminescence spectroscopy, X-ray diffraction (XRD), and micro-Raman spectroscopy. The effect of the applied potential (30-50 V), manganese content in the alloy (5-15 wt. %) and water content in the electrolyte (2-10 vol. %) on the morphology and photocatalytic properties was investigated for the first time. The photoactivity was assessed in the toluene removal reaction under visible light, using low-powered LEDs as an irradiation source (λmax = 465 nm). Morphology analysis showed that samples consisted of auto-aligned nanotubes over the surface of the alloy, their dimensions were: diameter = 76-118 nm, length = 1.0-3.4 µm and wall thickness = 8-11 nm. It was found that the increase in the applied potential led to increase the dimensions while the increase in the content of manganese in the alloy brought to shorter nanotubes. Notably, all samples were photoactive under the influence of visible light and the highest degradation achieved after 60 min of irradiation was 43%. The excitation mechanism of TiO2-MnO2 NTs under visible light was presented, pointing out the importance of MnO2 species for the generation of e- and h⁺.

Growth, Structure, and Photocatalytic Properties of Hierarchical V2O5-TiO2 Nanotube Arrays Obtained from the One-step Anodic Oxidation of Ti-V Alloys.

V2O5-TiO2 mixed oxide nanotube (NT) layers were successfully prepared via the one-step anodization of Ti-V alloys. The obtained samples were characterized by scanning electron microscopy (SEM), UV-Vis absorption, photoluminescence spectroscopy, energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (DRX), and micro-Raman spectroscopy. The effect of the applied voltage (30-50 V), vanadium content (5-15 wt %) in the alloy, and water content (2-10 vol %) in an ethylene glycol-based electrolyte was studied systematically to determine their influence on the morphology, and for the first-time, on the photocatalytic properties of these nanomaterials. The morphology of the samples varied from sponge-like to highly-organized nanotubular structures. The vanadium content in the alloy was found to have the highest influence on the morphology and the sample with the lowest vanadium content (5 wt %) exhibited the best auto-alignment and self-organization (length = 1 µm, diameter = 86 nm and wall thickness = 11 nm). Additionally, a probable growth mechanism of V2O5-TiO2 nanotubes (NTs) over the Ti-V alloys was presented. Toluene, in the gas phase, was effectively removed through photodegradation under visible light (LEDs, λmax = 465 nm) in the presence of the modified TiO2 nanostructures. The highest degradation value was 35% after 60 min of irradiation. V2O5 species were ascribed as the main structures responsible for the generation of photoactive e- and h⁺ under Vis light and a possible excitation mechanism was proposed.

Ionic liquids for nano- and microstructures preparation. Part 2: Application in synthesis.

Ionic liquids (ILs) are widely applied to prepare metal nanoparticles and 3D semiconductor microparticles. Generally, they serve as a structuring agent or reaction medium (solvent), however it was also demonstrated that ILs can play a role of a co-solvent, metal precursor, reducing as well as surface modifying agent. The crucial role and possible types of interactions between ILs and growing particles have been presented in the Part 1 of this review paper. Part 2 of the paper gives a comprehensive overview of recent experimental studies dealing with application of ionic liquids for preparation of metal and semiconductor based nano- and microparticles. A wide spectrum of preparation routes using ionic liquids is presented, including precipitation, sol-gel technique, hydrothermal method, nanocasting and ray-mediated methods (microwave, ultrasound, UV-radiation and γ-radiation). It was found that ionic liquids formed of a 1-butyl-3-methylimidazolium [BMIM] combined with tetrafluoroborate [BF4], hexafluorophosphate [PF6], and bis(trifluoromethanesulfonyl)imide [Tf2N] are the most often used ILs in the synthesis of nano- and microparticles, due to their low melting temperature, low viscosity and good transportation properties. Nevertheless, examples of other IL classes with intrinsic nanoparticles stabilizing abilities such as phosphonium and ammonium derivatives are also presented. Experimental data revealed that structure of ILs (both anion and cation type) affects the size and shape of formed metal particles, and in some cases may even determine possibility of particles formation. The nature of the metal precursor determines its affinity to polar or nonpolar domains of ionic liquid, and therefore, the size of the nanoparticles depends on the size of these regions. Ability of ionic liquids to form varied extended interactions with particle precursor as well as other compounds presented in the reaction media (water, organic solvents etc.) provides nano- and microstructures with different morphologies (0D nanoparticles, 1D nanowires, rods, 2D layers, sheets, and 3D features of molecules). ILs interact efficiently with microwave irradiation, thus even small amount of IL can be employed to increase the dielectric constant of nonpolar solvents used in the synthesis. Thus, combining the advantages of ionic liquids and ray-mediated methods resulted in the development of new ionic liquid-assisted synthesis routes. One of the recently proposed approaches of semiconductor particles preparation is based on the adsorption of semiconductor precursor molecules at the surface of micelles built of ionic liquid molecules playing a role of a soft template for growing microparticles.

Ionic liquids for nano- and microstructures preparation. Part 1: Properties and multifunctional role.

Ionic liquids (ILs) are a broad group of organic salts of varying structure and properties, used in energy conversion and storage, chemical analysis, separation processes, as well as in the preparation of particles in nano- and microscale. In material engineering, ionic liquids are applied to synthesize mainly metal nanoparticles and 3D semiconductor microparticles. They could generally serve as a structuring agent or as a reaction medium (solvent). This review deals with the resent progress in general understanding of the ILs role in particle growth and stabilization and the application of ionic liquids for nano- and microparticles synthesis. The first part of the paper is focused on the interactions between ionic liquids and growing particles. The stabilization of growing particles by steric hindrance, electrostatic interaction, solvation forces, viscous stabilization, and ability of ILs to serve as a soft template is detailed discussed. For the first time, the miscellaneous role of the ILs in nano- and microparticle preparation composed of metals as well as semiconductors is collected, and the formation mechanisms are graphically presented and discussed based on their structure and selected properties. The second part of the paper gives a comprehensive overview of recent experimental studies dealing with the applications of ionic liquids for preparation of metal and semiconductor-based nano- and microparticles. A wide spectrum of preparation routes using ionic liquids are presented, including precipitation, sol-gel technique, hydrothermal method, nanocasting, and microwave or ultrasound-mediated methods.