Showing papers on "Substrate (chemistry) published in 2020"

Factors influencing volatile fatty acids production from food wastes via anaerobic digestion

Abstract: Volatile fatty acids (VFAs) are intermediate products in anaerobic digestion. The effect of substrate loading or inoculum to substrate ratio (ISR), the addition of methanogen inhibitor, O2 presence ...

Synthesis of magnetic Fe3O4@ZnO@graphene oxide nanocomposite for photodegradation of organic dye pollutant

Abstract: In this study graphene oxide (GO) is applied as a substrate for the synthesis of the magnetic ZnO nanocomposite (GO-Fe3O4-ZnO). The prepared samples are characterised using XRD and VSM. The...

Reduced Enzyme Dynamics upon Multipoint Covalent Immobilization Leads to Stability-Activity Trade-off.

Abstract: The successful incorporation of enzymes into materials through multipoint covalent immobilization (MPCI) has served as the foundation for numerous advances in diverse fields, including biocatalysis, biosensing, and chemical weapons defense. Despite this success, a mechanistic understanding of the impact of this approach on enzyme stability has remained elusive, which is critical for realizing the full potential of MPCI. Here, we showed that the stabilization of lipase upon MPCI to polymer brush surfaces resulted from the rigidification of the enzyme with an increase in the number of enzyme-brush attachments. This was evident by a 10-fold decrease in the rates of enzyme unfolding and refolding as well as a reduction of the intrinsic fluctuations of the folded and unfolded states, which was measured by single-molecule (SM) Forster Resonance Energy Transfer imaging. Moreover, our results illuminate an important trade-off between stability and activity as a function of this decrease in structural dynamics of the immobilized lipase. Notably, as the thermal stability of lipase increased, as indicated by the temperature optimum for activity of the enzyme, the specific activity of lipase decreased. This decrease in activity was attributed to a reduction in the essential motions of the folded state that are required for catalytic turnover of substrate. These results provide direct evidence of this effect, which has long been a matter of speculation. Furthermore, our findings suggest that the retention of activity and stabilization of an enzyme may be balanced by tuning the extent of enzyme attachment.

Ag-Nanoparticles@Bacterial Nanocellulose as a 3D Flexible and Robust Surface-Enhanced Raman Scattering Substrate.

Abstract: We present a well-designed, low-cost, and simple synthetic approach to realizing the hybrid composites of Ag nanoparticle-decorated bacterial nanocellulose (denoted as Ag-NPs@BNC) as a three-dimensional (3D) flexible surface-enhanced Raman scattering (SERS) substrate with ultrahigh SERS sensitivity, excellent signal reproducibility, and stability. The homogeneous Ag-NPs with high density were in situ grown on the networked BNC fibers by the controlled silver mirror reaction and volume shrinkage treatment, which created uniformly distributed SERS "hot spots" in the 3D networked hybrid substrate. Attributed to these unique 3D hot spots, the as-presented Ag-NPs@BNC substrates exhibited ultrahigh sensitivity and good spectral reproducibility. Moreover, the hydrophilic BNC exhibits good permeability and adsorption performances, which could capture the target molecules in the highly active hot spot areas to further improve the SERS sensitivity. As a result, not only dye molecules (rhodamine 6G) but also toxic organic pollutants such as 2-naphthalenethiol and thiram have been detected using the hybrid substrates as SERS substrates, with sensitivities of 1.6 × 10-8 and 3.8 × 10-9 M, respectively. The good linear response of the intensity and the logarithmic concentration revealed promising applications in the rapid and quantitative detection of toxic organic pollutants. Besides, this self-supported Ag-NPs@BNC substrate demonstrated good stability and flexibility for varied detection conditions. Therefore, the 3D networked, flexible, ultrasensitive, and stable Ag-NPs@BNC substrate shows potential as a versatile SERS substrate in the rapid identification of various organic molecules.

Copper–Oxygen Dynamics in the Tyrosinase Mechanism

Abstract: The dinuclear copper enzyme, tyrosinase, activates O2 to form a (μ-η2 :η2 -peroxido)dicopper(II) species, which hydroxylates phenols to catechols. However, the exact mechanism of phenolase reaction in the catalytic site of tyrosinase is still under debate. We herein report the near atomic resolution X-ray crystal structures of the active tyrosinases with substrate l-tyrosine. At their catalytic sites, CuA moved toward l-tyrosine (CuA1 → CuA2), whose phenol oxygen directly coordinates to CuA2, involving the movement of CuB (CuB1 → CuB2). The crystal structures and spectroscopic analyses of the dioxygen-bound tyrosinases demonstrated that the peroxide ligand rotated, spontaneously weakening its O-O bond. Thus, the copper migration induced by the substrate-binding is accompanied by rearrangement of the bound peroxide species so as to provide one of the peroxide oxygen atoms with access to the phenol substrate's ϵ carbon atom.

Improving Artificial Photosynthesis over Carbon Nitride by Gas-Liquid-Solid Interface Management for Full Light-Induced CO2 Reduction to C1 and C2 Fuels and O2.

Abstract: The activity and selectivity of simple photocatalysts for CO2 reduction remain limited by the insufficient photophysics of the catalysts, as well as the low solubility and slow mass transport of gas molecules in/through aqueous solution. In this study, these limitations are overcome by constructing a triphasic photocatalytic system, in which polymeric carbon nitride (CN) is immobilized onto a hydrophobic substrate, and the photocatalytic reduction reaction occurs at a gas-liquid-solid (CO2 -water-catalyst) triple interface. CN anchored onto the surface of a hydrophobic substrate exhibits an approximately 7.2-fold enhancement in total CO2 conversion, with a rate of 415.50 μmol m-2 h-1 under simulated solar light irradiation. This value corresponds to an overall photosynthetic efficiency for full water-CO2 conversion of 0.33 %, which is very close to biological systems. A remarkable enhancement of direct C2 hydrocarbon production and a high CO2 conversion selectivity of 97.7 % are observed. Going from water oxidation to phosphate oxidation, the quantum yield is increased to 1.28 %.

GaN Nanotowers Grown on Si (111) and Functionalized with Au Nanoparticles and ZnO Nanorods for Highly Responsive UV Photodetectors

Abstract: Vertically aligned GaN Nanotowers (NTs) were grown on Si (111) substrate by plasma-assisted molecular beam epitaxy to design a highly responsive ultraviolet (UV) photodetector. The UV detector fabr...

Enzyme Immobilization on Graphite Oxide (GO) Surface via One-pot Synthesis of GO/Metal-Organic Framework Composites for Large-Substrate Biocatalysis

Abstract: Although enzyme immobilization has improved many areas, biocatalysis involving large-size substrates is still challenging for immobilization platform design because of the protein damage under the often "harsh" reaction conditions required for these reactions. Our recent efforts indicate the potential of using Metal-Organic Frameworks (MOFs) to partially confine enzymes on the surface of MOF-based composites while offering sufficient substrate contact. Still, improvements are required to expand the feasible pH range and the efficiency of contacting substrates. In this contribution, we discovered that Zeolitic Imidazolate Framework (ZIF) and a new calcium-carboxylate based MOF (CaBDC) can both be coprecipitated with a model large-substrate enzyme, lysozyme (lys), to anchor the enzyme on the surface of graphite oxide (GO). We observed lys activity against its native substrate, bacterial cell walls, indicating lys was confined on composite surface. Remarkably, lys@GO/CaBDC displayed a stronger catalytic efficiency at pH 6.2 as compared to pH 7.4, indicating CaBDC is a good candidate for biocatalysis under acidic conditions as compared to ZIFs which disassemble under pH < 7. Furthermore, to understand the regions of lys being exposed to the reaction medium, we carried out a site-directed spin labeling (SDSL) electron paramagnetic resonance (EPR) spectroscopy study. Our data showed a preferential orientation of lys in GO/ZIF composite, whereas a random orientation in GO/CaBDC. This is the first report on immobilizing solution-state large-substrate enzymes on GO surface using two different MOFs via one-pot synthesis. These platforms can be generalized to other large-substrate enzymes to carry out catalysis under the optimal buffer/pH conditions. The orientation of enzyme at the molecular level on composite surfaces is critical for guiding the rational design of new composites.

Immobilization of β-Glucosidase from Thermatoga maritima on Chitin-functionalized Magnetic Nanoparticle via a Novel Thermostable Chitin-binding Domain.

Abstract: Enzyme immobilization is a powerful tool not only as a protective agent against harsh reaction conditions but also for the enhancement of enzyme activity, stability, reusability, and for the improvement of enzyme properties as well. Herein, immobilization of β-glucosidase from Thermotoga maritima (Tm-β-Glu) on magnetic nanoparticles (MNPs) functionalized with chitin (Ch) was investigated. This technology showed a novel thermostable chitin-binding domain (Tt-ChBD), which is more desirable in a wide range of large-scale applications. This exclusive approach was fabricated to improve the Galacto-oligosaccharide (GOS) production from a cheap and abundant by-product such as lactose through a novel green synthesis route. Additionally, SDS-PAGE, enzyme activity kinetics, transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) revealed that among the immobilization strategies for Thermotoga maritime-β-Glucosidase thermostable chitin-binding domain (Tm-β-Glu-Tt-ChBD) on the attractive substrate; Ch-MNPs had the highest enzyme binding capacity and GOS production ratio when compared to the native enzyme. More interestingly, a magnetic separation technique was successfully employed in recycling the immobilized Tm-β-Glu for repetitive batch-wise GOS without significant loss or reduction of enzyme activity. This immobilization system displayed an operative stability status under various parameters, for instance, temperature, pH, thermal conditions, storage stabilities, and enzyme kinetics when compared with the native enzyme. Conclusively, the GOS yield and residual activity of the immobilized enzyme after the 10th cycles were 31.23% and 66%, respectively. Whereas the GOS yield from native enzyme synthesis was just 25% after 12 h in the first batch. This study recommends applying Tt-ChBD in the immobilization process of Tm-β-Glu on Ch-MNPs to produce a low-cost GOS as a new eco-friendly process besides increasing the biostability and efficiency of the immobilized enzyme.

Size-Tunable Metal–Organic Framework-Coated Magnetic Nanoparticles for Enzyme Encapsulation and Large-Substrate Biocatalysis

Abstract: Immobilizing enzymes on nanoparticles (NPs) enhances the cost-efficiency of biocatalysis; however, when the substrates are large, it becomes difficult to separate the enzyme@NP from the products wh...

Solvent-Free Photobiocatalytic Hydroxylation of Cyclohexane

Abstract: The use of neat reaction media, that is the avoidance of additional solvents, is the simplest and the most efficient approach to follow in biocatalysis. Here, we show that unspecific peroxygenase from Agrocybe aegerita (AaeUPO) can hydroxylate the neat model substrate cyclohexane. H2O2 was photocatalytically generated in situ by nitrogen-doped carbon nanodots (N−CNDs) and UV LED illumination. AaeUPO entrapment in alginate beads increased enzyme stability and facilitated the reaction in neat cyclohexane. N−CNDs absorption in beads containing AaeUPO created a 2-in-1 heterogeneous photobiocatalyst that was active for up to seven days under reaction conditions and produced cyclohexanol, 2.5 mM. To increase productivity, the bead size and the photocatalyst-to-enzyme ratio have been identified as promising targets for optimisation.

Tuning Triplet Energy Transfer of Hydroxamates as the Nitrene Precursor for Intramolecular C(sp3)-H Amidation.

Abstract: Reported herein is the design of a photosensitization strategy to generate triplet nitrenes and its applicability for the intramolecular C–H amidation reactions. Substrate optimization by tuning ph...

The Effect of pH on the Size of Silver Nanoparticles Obtained in the Reduction Reaction with Citric and Malic Acids.

Abstract: In colloidal methods, the morphology of nanoparticles (size and shape) as well as their stability can be controlled by changing the concentration of the substrate, stabilizer, adding inorganic salts, changing the reducer/substrate molar ratio, and changing the pH and reaction time. The synthesis of silver nanoparticles was carried out according to the modified Lee and Meisel method in a wide pH range (from 2.0 to 11.0) using citric acid and malic acid, without adding any additives or stabilizers. Keeping the same reaction conditions as the concentration of acid and silver ions, temperature, and heating time, it was possible to determine the relationship between the reaction pH, the type of acid, and the size of the silver nanoparticles formed. Obtained colloids were analyzed by UV-Vis spectroscopy and investigated by means of Transmission Electron Microscope (TEM). The study showed that the colloids reduced with citric acid and malic acid are stable over time for a minimum of seven weeks. We observed that reactions occurred for citric acid from pH 6.0 to 11.0 and for malic acid from pH 7.0 to 11.0. The average size of the quasi-spherical nanoparticles changed with pH due to the increase of reaction rate.

Pre‐targeted Imaging of Protease Activity through In Situ Assembly of Nanoparticles

Abstract: The pre-targeted imaging of enzyme activity has not been reported, likely owing to the lack of a mechanism to retain the injected substrate in the first step for subsequent labeling. Herein, we report the use of two bioorthogonal reactions-the condensation reaction of aromatic nitriles and aminothiols and the inverse-electron demand Diels-Alder reaction between tetrazine and trans-cyclooctene (TCO)-to develop a novel strategy for pre-targeted imaging of the activity of proteases. The substrate probe (TCO-C-SNAT4) can be selectively activated by an enzyme target (e.g. caspase-3/7), which triggers macrocyclization and subsequent in situ self-assembly into nanoaggregates retained at the target site. The tetrazine-imaging tag conjugate labels TCO in the nanoaggregates to generate selective signal retention for imaging in vitro, in cells, and in mice. Owing to the decoupling of enzyme activation and imaging tag immobilization, TCO-C-SNAT4 can be repeatedly injected to generate and accumulate more TCO-nanoaggregates for click labeling.

Modulated Sn Oxidation States over a Cu2O-Derived Substrate for Selective Electrochemical CO2 Reduction

Abstract: Pursuing high catalytic selectivity is challenging but paramount for an efficient and low-cost CO2 electrochemical reduction (CO2R). In this work, we demonstrate a significant correlation between the selectivity of CO2R to formate and the duration of tin (Sn) electrodeposition over a cuprous oxide (Cu2O)-derived substrate. A Sn electrodeposition time of 120 s led to a cathode with a formate Faradaic efficiency of around 81% at -1.1 V vs reversible hydrogen electrode (RHE), which was more than 37% higher than those of the Sn foil and the sample treated for 684 s. This result highlights the significant role of the interface between deposited Sn and the cuprous-derived substrate in determining the selectivity of CO2R. High-resolution X-ray photoelectron spectra revealed that the residual cuprous species at the Cu/Sn interfaces could stabilize Sn species in oxidation states of 2+ and 4+, a mixture of which is essential for a selective formate conversion. Such modulation effects likely arise from the moderate electronegativity of the cuprous species that is lower than that of Sn2+ but higher than that of Sn4+. Our work highlights the significant role of the substrate in the selectivity of the deposited catalyst and provides a new avenue to advance selective electrodes for CO2 electrochemical reduction.

Game Change from Reagent- to Substrate-Controlled Peptide Synthesis

Abstract: An account of the development of Lewis-acid-catalyzed methods for racemization-free peptide synthesis is presented. These methods are based on the substrate control concept that has been exploited ...

Robust single-molecule enzyme nanocapsules for biosensing with significantly improved biosensor stability.

Abstract: The present study demonstrates the use of highly stable single-molecule enzyme nanocapsules (SMENs) instead of traditional native enzyme as biorecognition element in enzyme-based biosensors. The main purpose of this study is to resolve the major obstacle and challenge in the biosensor field, i.e., the poor stability of enzyme-based biosensors, including thermal stability, organic solvent tolerance, long-term operational stability, etc. Highly active and robust SMENs of glucose oxidase (GOx, as a model enzyme) were synthesized (nGOx) using an in situ polymerization strategy in an aqueous environment. The particle-size distribution, transmission electron microscopic (TEM) images, and UV-vis spectral characterization revealed the formation of a thin polymer layer around each enzyme molecule. The polymer shell effectively stabilized the GOx enzyme core while enabling rapid substrate transportation, resulting in a new class of biocatalytic nanocapsules. Multiple covalent attachments between a thin polymer layer and an enzyme molecule strengthened the encapsulated GOx molecule. Encapsulation created a favorable microenvironment to avoid any structural dissociation at high temperature and helped to retain essential water during the organic solvent operation. The present work reports a study implementing nGOx SMENs as highly stable nano(bio)sensors for point-of-care diagnostic applications. Prepared nGOx SMENs manifested significantly improved thermal stability (even at 65 °C) and organic solvent tolerance without any compromise in biocatalytic activity. For example, the native GOx-based biosensor lost its catalytic activity for glucose after 4 h of incubation at high temperature (65 °C), while the nGOx/N-CNTs-Chi/GCE nano(bio)sensor maintained ∼56% of its original catalytic activity for glucose oxidation. The proposed SMENs-based nano(bio)sensors with robust stability in variable working environment could promote the development and applications of biosensors in point-of care diagnostics, biomedical detection, wearable devices, implantable equipment, and biofuel cells.

CeO2/C nanowire derived from a cerium(III) based organic framework as a peroxidase mimic for colorimetric sensing of hydrogen peroxide and for enzymatic sensing of glucose.

Abstract: A metal organic framework obtained from cerium(III) and trimesic acid was pyrolyzed to obtain a novel nanostructure referred to as CeO2/C nanowires. The experimental parameters temperature, precursor and gas atmosphere were optimized. The nanowires show good dispersion and a large number of oxygen vacancies, and this leads to excellent peroxidase-like activity. The nanowires are stable at pH values between 2 and 10, and in the 4–80 °C temperature range. The peroxidase-mimicking activity was exploited in a sensitive colorimetric method for determination of H2O2 by using 3,3′,5,5′-tetramethylbenzidine as the chromogenic substrate. The absorbance at 652 nm increases linearly in the 0.5 to 100 μM H2O2 concentration range. If glucose oxidase is added to a solution containing glucose, H2O2 will be enzymatically produced. This was exploited to design a new method for determination of glucose. The optical response is linear in the 1–100 μM glucose concentration range, and the detection limit is 0.69 μM (at S/N = 3). The method was successfully applied to the determination of glucose in serum samples.

Rhodium-Catalyzed Electrooxidative C-H Olefination of Benzamides.

Abstract: Metal-catalyzed chelation-assisted C-H olefinations have emerged as powerful tools for the construction of functionalized alkenes. Herein, we describe the rhoda-electrocatalyzed C-H activation/alkenylation of arenes. The olefinations of challenging electron-poor benzamides were thus accomplished in a fully dehydrogenative fashion under electrochemical conditions, avoiding stoichiometric chemical oxidants, and with H2 as the only byproduct. This versatile alkenylation reaction also features broad substrate scope and used electricity as a green oxidant.

Film forming method for SiC film

Abstract: A method for forming a SiC film on a target substrate by ALD, comprises: activating a surface of the target substrate by activation gas plasma which is plasmatized an activation gas; and forming a SiC film by supplying a source gas containing a precursor represented by a chemical formula RSiX13 or RSiHClX2 onto the target substrate whose the surface is activated by activating the surface of the target substrate, where, R is an organic group having an unsaturated bond, X1 is selected from a group consisting of H, F, Cl, Br and I, and X2 is one selected from a group consisting of Cl, Br and I.