STD‐NMR‐Based Protein Engineering of the Unique Arylpropionate‐Racemase AMDase G74C

Structure‐guided protein engineering achieved a variant of the unique racemase AMDase G74C, with 40‐fold increased activity in the racemisation of several arylaliphatic carboxylic acids. Substrate binding during catalysis was investigated by saturation‐transfer‐difference NMR (STD‐NMR) spectroscopy. All atoms of the substrate showed interactions with the enzyme. STD‐NMR measurements revealed distinct nuclear Overhauser effects in experiments with and without molecular conversion. The spectroscopic analysis led to the identification of several amino acid residues whose substitutions increased the activity of G74C. Single amino acid exchanges increased the activity moderately; structure‐guided saturation mutagenesis yielded a quadruple mutant with a 40 times higher reaction rate. This study presents STD‐NMR as versatile tool for the analysis of enzyme–substrate interactions in catalytically competent systems and for the guidance of protein engineering.     

Mechanisms of orientational and photoelastic birefringence generation of methacrylates for the design of zero–zero‐birefringence polymers

The intrinsic birefringence Δn⁰ and photoelastic coefficient C of poly(methyl methacrylate), poly(2,2,2‐trifluroethyl methacrylate), poly(phenyl methacrylate), and poly(2,2,3,3,3‐pentafluorophenyl methacrylate) were determined. We categorized these methacrylate polymers into four birefringence‐types, even though their molecular structures differed only by the substituents on the side chains. Based on the results of Δn⁰ and C, novel polymers that exhibit neither orientational nor photoelastic birefringence, i.e., zero–zero‐birefringence polymers, were designed and synthesized by quaternary copolymerization system. Furthermore, we confirmed that the mechanisms of orientational birefringence and photoelastic birefringence generation were different in these methacrylate polymers. The conformation of the repeat unit of the polymers was nearly constant during the generation of orientational birefringence. In contrast, the conformation of the repeat unit of the polymers changed during the generation of photoelastic birefringence in the glassy state. These findings demonstrated the reasonability of evaluating orientational and photoelastic birefringence separately, as well as the adequacy of the classification of polymers into four birefringence‐types. Given these results and the fact that zero–zero‐birefringence polymers could be prepared successfully by four‐birefringence type monomers, we demonstrated the reasonability of the method for designing the zero–zero‐birefringence polymers. POLYM. ENG. SCI., 55:1330–1338, 2015. © 2015 Society of Plastics Engineers     

Effects of Atmospheric Composition on the Molecular Structure of Synthesized Silicon Oxycarbides

The dependence of silicon oxycarbides' chemical composition and molecular structure on their reaction conditions was tested by varying the atmosphere under which pyrolysis was performed. To obtain the silicon oxycarbides, densely cross‐linked silicone resin particles with an averaged diameter of 2 μm were pyrolyzed in various atmospheres of H₂, Ar, and CO₂, in the temperature range 700°C–1100°C. The residual mass of resin after pyrolysis was almost constant at 700°C, although their apparent colors varied distinctly. The sample obtained from the H₂ atmosphere was white, whereas that obtained from the CO₂ atmosphere was dark brown. Fourier‐transform infrared (FT‐IR) spectra of the residues suggested that the Si–O–Si network evolution was accelerated in the CO₂ atmosphere. Beyond 800°C, the chemical compositions of the compounds obtained from a H₂ atmosphere increasingly approached near‐stoichiometric SiO₂–xSiC composition with increasing the pyrolysis temperature. Compounds from a CO₂ atmosphere approached a composition of SiO₂–xC with no free SiC as the pyrolysis temperature increased. In the products from an Ar atmosphere, SiO₂–xSiC–yC compositions were typically obtained. The observed effects of the pyrolysis atmosphere on the resulting chemical compositions were analyzed in terms of thermodynamic calculations. Electron spin resonance (ESR) spectra revealed broad and intense signals from products obtained from either Ar or CO₂. Estimating from the signal intensity, the residual spin concentrations were in the range 10¹⁸–10¹⁹ g^?1. Meanwhile, the spectra from the samples obtained in H₂ showed weak and sharp signals with estimated spin concentrations ranging from 10¹⁶–10¹⁷ g^?1. This signal attenuation may have been due to the hydrogen capping of dangling bond formed during pyrolysis.     

Preparation of porous membranes by selective decomposition of adamantane unit in ABA‐type triblock copolymer

The mechanism of pore formation by selective decomposition of adamantane unit in an ABA‐type triblock copolymer derived from 4,4‐(hexafluoroisopropylidene)diphthalic anhydride‐2,3,5,6‐tetramethyl‐1,4‐phenylenediamine (6FDA‐TeMPD) and poly(2‐methyl‐2‐adamantylmethacrylate) (PMAdMA) was investigated on this basis. This study aims to developing a novel method of material design for high‐precision gas separation membranes and application to electric devices by improving dielectric constant. Scanning electron microscopy images showed that the membrane structure changed considerably after heating; the difference increased with the increase in adamantane content. Interestingly, the internal structure of Block(36 mol%6FDA‐TeMPD/64 mol%PMAdMA) membrane was almost unchanged although its surface structure was changed. These results suggest that the mechanism of formation of porous membrane involves the decomposition of adamantane unit from surface. In addition, more adamantane units inside the membrane were discharged to the surface of the membrane through a path formed by decomposed adamantane units. POLYM. ENG. SCI., 56:1191–1200, 2016. © 2016 Society of Plastics Engineers     

Development of Novel Inhibitors for Histone Methyltransferase SET7/9 based on Cyproheptadine

The histone methyltransferase SET7/9 methylates not only histone but also non‐histone proteins as substrates, and therefore, SET7/9 inhibitors are considered candidates for the treatment of diseases. Previously, our group identified cyproheptadine, used clinically as a serotonin receptor antagonist and histamine receptor (H1) antagonist, as a novel scaffold of the SET7/9 inhibitor. In this work, we focused on dibenzosuberene as a substructure of cyproheptadine and synthesized derivatives with various functional groups. Among them, the compound bearing a 2‐hydroxy group showed the most potent activity. On the other hand, a 3‐hydroxy group or another hydrophilic functional group such as acetamide decreased the activity. Structural analysis clarified a rationale for the improved potency only by tightly restricted location and type of the hydrophilic group. In addition, a SET7/9 loop, which was only partially visible in the complex with cyproheptadine, became more clearly visible in the complex with 2‐hydroxycyproheptadine. These results are expected to be helpful for further structure‐based development of SET7/9 inhibitors.     

Microstructure of selectively heated (hot spot) region in Fe3O4 powder compacts by microwave irradiation

An Fe₃O₄ powder compact was irradiated with a 2.45 GHz microwave single-mode applicator at the magnetic field maximum position. Selectively heated regions (hot spot region) having several hundred micrometers to millimeter scale were formed. They exhibited metallic color. The SEM/EDX observations showed no appreciable difference in the compositions between the hot spot regions and the matrix. However, micro-XRD revealed that the hot spot region had a larger fraction of FeO than the matrix did, although the major consisting phase was Fe₃O₄ with a little Fe₂O₃. TEM observations indicated that the observed hot spot regions comprise these oxide phases separated in nano-sized grains, which agrees with our previous report. The larger fraction of FeO phase and flat surface might be related with the metallic color of the hot spot region. Their formation mechanisms and phase constitution were discussed.     

Measurement of self-diffusion coefficients in Li ionic conductors by using the short-lived radiotracer of 8Li

For the effective use of short-lived radioactive beams, soon to be available at the Tokai Radioactive Ion Accelerator Complex, the authors have developed a radiotracer method for diffusion studies in solids. The experimental test was performed by the measurement of the diffusion coefficients of Li in a sample of the compound βLiAl using an α-emitting radiotracer of ⁸Li (T_1/2=0.84 s). It was found that the time-dependent yields of the α particles from the diffusing ⁸Li that was initially implanted in the sample could be used as a measure of the diffusivity of the tracer in a nondestructive way. The method was applied to measure the self-diffusion coefficients of Li in βLiGa, and for investigating how the Li diffusion in the Li ionic conductors is affected by the concentration of atomic defects (i.e., the existence of the atomic vacancies of Li and the defects in Ga sites that are replaced by Li).     

Improvement in mechanical properties of poly(p‐phenylene sulfide) fibers by high‐tension multiannealing method

High‐tension multiannealing (HTMA) was applied to improve the tensile properties of poly(p‐phenylene sulfide) fibers, which was furthermore applied to the fibers produced and improved with the zone‐drawing and zone‐annealing treatments. The HTMA treatment was repeatedly applied to the fibers under the conditions of a 250°C temperature and an applied tension of between 201.0 and 188.0 MPa. As a result, at the 13th treatment the degree of crystallinity increased to 40%. On the other hand, the orientation factor of crystallites increased dramatically to 0.982 during the zone‐drawing treatment, but increased only slightly during the subsequent treatments of zone annealing and HTMA. The finally obtained fiber had a tensile modulus of 10.4 GPa and a tensile strength of 0.73 GPa. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 75: 1569–1576, 2000     

Intelligent tutoring system for electric circuit exercising

An intelligent tutoring system called Circuit Exerciser is described. The system is designed to help university students learn more about electric circuits. It can formulate drill problems, solve them, and infer mistakes in a student's answer. It can also provide helpful comments to the students on how the mistake was made. The system shows the circuit of the presented problem on graphic displays and is student-friendly. The system architecture, pedagogical cycle, and execution of the system are presented     

New type of phenolic resin: Curing reaction of phenol‐novolac based benzoxazine with bisoxazoline or epoxy resin using latent curing agent and the properties of the cured resin

In this study, we aimed to reduce the cure time, and to lower the cure temperature of the benzoxazine compound. Therefore, curing reaction of benzoxazine with bisoxazoline or epoxy resin using the latent curing agent and the properties of the cured resins were investigated. The cure behavior of benzoxazine with bisoxazoline or epoxy resin using the latent curing agent was monitored by differential scanning calorimetry and measurements for storage modulus (G′). The properties of the cured resin were estimated by mechanical properties, electrical insulation, water resistance, heat resistance, and flame resistance. As a result, it was confirmed that by using the latent curing agent, cure time of benzoxazine and bisoxazoline or epoxy resin was reduced, and cure temperature was lowered. And it was found that the curing reaction using phenol‐novolac based benzoxazine (Na) as the benzoxazine compound could proceed more rapidly than that using bisphenol‐A based benzoxazine (Ba) as the benzoxazine compound. However, the cured resins from Ba and bisoxazoline or epoxy resin using the latent curing agent showed good heat resistance, flame resistance, and mechanical properties compared with those from Na and bisoxazoline or epoxy resin using the latent curing agent. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009