Covalent Linkage of an R-ω-Transaminase to a d-Amino Acid Oxidase through Protein Splicing to Enhance Enzymatic Catalysis of Transamination

R-ω-Transaminases (RTAs) catalyse the conversion of R-configured amines [e.g., (R)-1-phenylethylamine] into the corresponding ketones (e.g., acetophenone), by transferring an amino group from an amino donor [e.g., (R)-1-phenylethylamine] onto an amino acceptor (e.g., pyruvate), resulting in a co-product (e.g., d -alanine). d -Alanine can be deaminated back to pyruvate by d -amino acid oxidase (DAAOs). Here, through in vivo subunit splicing, the N terminus of an RTA subunit (RTA^S) was specifically ligated to the C terminus of a DAAO subunit (DAAO^S) through native peptide bonds (RTA&DAAO). RTA^S is in close proximity to DAAO^S, at a molecular-scale distance. Thus the transfer of pyruvate and d -alanine between RTA and DAAO can be directional and efficient. Pyruvate→d -alanine→pyruvate cycles are efficiently formed, thus promoting the forward transamination reaction. In a different, in vitro noncovalent approach, based on coiled-coil association, the RTA^S N terminus was specifically associated with the DAAO^S C terminus (RTA#DAAO). In addition, the two mixed individual enzymes (RTA+DAAO) were also studied. RTA&DAAO has a shorter distance between the paired subunits (RTA^S–DAAO^S) than RTA#DAAO, and the number of the paired subunits is higher than in the case of RTA#DAAO, whereas RTA+DAAO cannot form the paired subunits. RTA&DAAO exhibited a transamination catalysis efficiency higher than that of RTA#DAAO and much higher than that of RTA+DAAO.     

Dual pH‐ and thermal‐responsive nanocomposite hydrogels for controllable delivery of hydrophobic drug baicalein

Hydrogels have been widely used as mild biomaterials due to their bio‐affinity, high drug loading capability and controllable release profiles. However, hydrogel‐based carriers are greatly limited for the delivery of hydrophobic payloads due to the lack of hydrophobic binding sites. Herein, nano‐liposome micelles were embedded in semi‐interpenetrating poly[(N‐isopropylacrylamide)‐co‐chitosan] (PNIPAAm‐co‐CS) and poly[(N‐isopropylacrylamide)‐co‐(sodium alginate)] (PNIPAAm‐co‐SA) hydrogels which were responsive to both temperature and pH, thereby establishing tunable nanocomposite hydrogel delivery systems. Nano‐micelles formed via the self‐assembly of phospholipid could serve as the link between hydrophobic drug and hydrophilic hydrogel due to their special amphiphilic structure. The results of transmission and scanning electron microscopies and infrared spectroscopy showed that the porous hydrogels were successfully fabricated and the liposomes encapsulated with baicalein could be well contained in the network. In addition, the experimental results of response release in vitro revealed that the smart hydrogels showed different degree of sensitiveness under different pH and temperature stimuli. The results of the study demonstrate that combining PNIPAAm‐co‐SA and PNIPAAm‐co‐CS hydrogels with liposomes encapsulated with hydrophobic drugs is a feasible method for hydrophobic drug delivery and have potential application prospects in the medical field. © 2018 Society of Chemical Industry     

Fully transient analytical solution for degradable organic contaminant transport through GMB/GCL/AL composite liners

In this study, analytical solution for degradable organic contaminant transport through a composite liner consisting of a geomembrane (GMB) layer, a geosynthetic clay liner (GCL) and an attenuation layer (AL) is derived by the separation of variables method. The transient contaminant transport in the whole composite liner can be well described avoiding some weird phenomena in existing analytical solutions. The results of parametric study show that GCL has significant effect on improving the barrier efficiency especially for scenarios with high leachate head. The biodegradation and adsorption in GCL have significant influence on the contaminant transport through the composite liner when the half-life of contaminant in GCL is less than 5 years. Otherwise, the effect can be neglected.     

Reaction synthesis of spark plasma sintered MoSi2-B4C coatings for oxidation protection of Nb alloy

MoSi₂-B₄C coatings with different B₄C contents were prepared on Nb alloy by spark plasma sintering (SPS) process. Powder mixtures of Mo, Si and B₄C were used as the coating starting materials. Besides MoSi₂ and B₄C phases, small amounts of SiC and MoB are also found in the coatings because of the reactions of Mo, Si and B₄C powders during sintering. Compared with single MoSi₂ coating, the MoSi₂-B₄C coatings show better oxidation resistance at 1450?℃, and dense B₂O₃-SiO₂ oxide scales form after 100?h oxidation. The B₄C or MoB in the MoSi₂-B₄C coatings can serve as the B donor for the formation of B₂O₃. A slight degradation in the microstructure of the MoSi₂-B₄C coatings after oxidation is observed, which can be attributed to the presence of an NbB layer in the inter-diffusion zone of the coatings that retards the inward diffusion of Si from the coating into the substrate alloy. The microstructure development and oxidation behavior of the MoSi₂-B₄C coatings have been discussed.     

Porous Si3N4 ceramics prepared by aqueous gelcasting using low–toxicity DMAA system: Regulatable microstructure and properties by monomer content

In this study, the low–toxicity monomer N, N–dimethylacrylamide (DMAA), serving as both gelling agent and pore–forming agent, was adopted to fabricate porous Si₃N₄ ceramics with a regulatable microstructure and property by aqueous gelcasting. Results indicate that monomer content played an important role in regulating and optimizing the properties of sintered bodies. With increasing monomer content (5.94–30.69?wt%), both slurry viscosity (maximum 0.14?Pa?s at 95.40 s^?1) and green body strength (11.35–49.23?MPa) exhibited monotonic increasing trends, demonstrating superior mechanical properties to those obtained using the neurovirulent acrylamide (AM) gelling system. The increased monomer content not only improved porosity, but also promoted α→β–Si₃N₄ transformation as well as β–Si₃N₄ grain growth through enhancing the connectivity of interlocking pores and accelerating the vapor phase transport during liquid–phase sintering. These variations in phase composition and microstructure derived from the varied monomer content further resulted in monotonic changes in porosity (40.32–51.50%), mean pore size (0.27–0.38?μm), flexural strength (202.77–132.15?MPa), fracture toughness (2.93–2.32?MPa?m^1/2), dielectric constant (3.48–2.78) and loss (3.52–3.09?×?10^?3) at 10?GHz for sintered bodies, displaying an excellent comprehensive properties. This study suggests a promising prospect for DMAA in preparation of high–performance porous Si₃N₄ ceramics by aqueous gelcasting.     

物联网环境下基于情境的语义Web服务选择

当前Web服务海量增加，物联网应用技术快速发展、不断普及，而现有的Web服务选择算法低效、用户匹配度低。针对该问题提出一种物联网环境下基于情境的语义Web服务选择方法。该方法应用QoS参数的无量纲化与语义Web服务动态选择方式，将物联网环境下服务与语义Web服务相结合，并根据用户需求针对QoS选择最优的服务集。实验表明，该方法能有效地提高用户服务动态选择的成功率。     

A Versatile Theranostic Nanoemulsion for Architecture‐Dependent Multimodal Imaging and Dually Augmented Photodynamic Therapy

To design a clinically translatable nanomedicine for photodynamic theranostics, the ingredients should be carefully considered. A high content of nanocarriers may cause extra toxicity in metabolism, and multiple theranostic agents would complicate the preparation process. These issues would be of less concern if the nanocarrier itself has most of the theranostic functions. In this work, a poly(ethylene glycol)‐boron dipyrromethene amphiphile (PEG‐F₅₄‐BODIPY) with 54 fluorine‐19 (¹⁹F) is synthesized and employed to emulsify perfluorohexane (PFH) into a theranostic nanoemulsion (PFH@PEG‐F₅₄‐BODIPY). The as‐prepared PFH@PEG‐F₅₄‐BODIPY can perform architecture‐dependent fluorescence/photoacoustic/¹⁹F magnetic resonance multimodal imaging, providing more information about the in vivo structure evolution of nanomedicine. Importantly, this nanoemulsion significantly enhances the therapeutic effect of BODIPY through both the high oxygen dissolving capability and less self‐quenching of BODIPY molecules. More interestingly, PFH@PEG‐F₅₄‐BODIPY shows high level of tumor accumulation and long tumor retention time, allowing a repeated light irradiation after a single‐dose intravenous injection. The “all‐in‐one” photodynamic theranostic nanoemulsion has simple composition, remarkable theranostic efficacy, and novel treatment pattern, and thus presents an intriguing avenue to developing clinically translatable theranostic agents.     

Four phenolic acids from purple sweet potato and their effects on physicochemical,digestive and structural characteristics of starch

Liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) was employed to investigate free phenols that were released from purple sweet potato (PSP) by alkaline, acid and enzymatic hydrolysis. Four phenolic acids, including ferulic, isoferulic, 4-hydroxybenzoic and caffeic acids, were identified. Based on their effects on the characteristics of purple sweet potato starch (PSPS), the four phenolic acids were studied. Furthermore, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT–IR) and X-ray diffraction (XRD) techniques were employed to explore the microstructures of the complexes of the phenolic acids and PSPS. The obtained results demonstrated that the pasting, thermal, retrogradation, as well as digestive properties of PSPS were all influenced by the phenolic acids which interacted with PSPS through noncovalent hydrogen bonds. The influence of the four phenolic acids on the properties of PSPS was in the descending order: 4-hydroxybenzoic acid > ferulic acid > caffeic acid > isoferulic acid.