Directed Evolution of a Cp*RhIII-Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification

Directed evolution of Cp*Rh^III-linked nitrobindin (NB), a biohybrid catalyst, was performed based on an in vitro screening approach. A key aspect of this effort was the establishment of a high-throughput screening (HTS) platform that involves an affinity purification step employing a starch-agarose resin for a maltose binding protein (MBP) tag. The HTS platform enables efficient preparation of the purified MBP-tagged biohybrid catalysts in a 96-well format and eliminates background influence of the host E. coli cells. Three rounds of directed evolution and screening of more than 4000 clones yielded a Cp*Rh^III-linked NB(T98H/L100K/K127E) variant with a 4.9-fold enhanced activity for the cycloaddition of acetophenone oximes with alkynes. It is confirmed that this HTS platform for directed evolution provides an efficient strategy for generating highly active biohybrid catalysts incorporating a synthetic metal cofactor.     

Experimental investigation of structure vulnerabilities to firebrand showers

Attempting to experimentally quantify the vulnerabilities of structures to ignition from firebrand showers has remained elusive. The coupling of two facilities has begun to unravel this difficult problem. The NIST Firebrand Generator (NIST Dragon) is an experimental device that can generate a firebrand shower in a safe and repeatable fashion. Since wind plays a critical role in the spread of WUI fires in the USA and urban fires in Japan, NIST has established collaboration with the Building Research Institute (BRI) in Japan. BRI maintains one of the only full scale wind tunnel facilities in the world designed specifically for fire experimentation; the Fire Research Wind Tunnel Facility (FRWTF). The present investigation is aimed at extensively quantifying firebrand penetration through building vents using full scale tests. A structure was placed inside the FRWTF and firebrand showers were directed at the structure using the NIST Dragon. The structure was fitted with a generic building vent, consisting of only a frame fitted with a metal mesh. Six different mesh sizes openings were used for testing. Behind the mesh, four different materials were placed to ascertain whether the firebrands that were able to penetrate the building mesh assembly could ignite these materials. Reduced scale test methods afford the capability to test new vent technologies and may serve as the basis for new standard testing methodologies. As a result, a new experimental facility developed at NIST is presented and is known as the NIST Dragon's LAIR (Lofting and Ignition Research). The NIST Dragon's LAIR has been developed to simulate a wind driven firebrand attack at reduced scale. The facility consists of a reduced scale Firebrand Generator (Baby Dragon) coupled to a bench scale wind tunnel. Finally, a series of full scale experiments were conducted to visualize the flow of firebrands around obstacles placed downstream of the NIST Dragon. Firebrands were observed to accumulate in front of these obstacles at a stagnation plane, as was observed when the structure was used for firebrand penetration through building vent experiments, due to flow recirculation. The accumulation of firebrands at a stagnation plane presents a severe threat to ignitable materials placed near structures.     

Production of Gentiobiose from Hydrothermally Treated Aureobasidium pullulans β-1,3-1,6-Glucan

We report production of the functional disaccharide gentiobiose β-D-Glcp-(1→6)-D-Glc by a hydrolysis reaction of hydrothermally treated Aureobasidium pullulans β-1,3-1,6-glucan as the substrate and Kitalase as the enzyme. Gentiobiose was produced over the pH range 4−6 and the concentration of gentiobiose produced decreased above pH 7. The maximum value of gentiobiose production was unaffected by the enzyme concentration. The maximum concentration of gentiobiose produced was dependent on the substrate concentration whereas the maximum ratio of gentiobiose to glucose was not. The production of gentiobiose from yeast β-1,3-1,6-glucan was lower than that from A. pullulans β-1,3-1,6-glucan.     

Enantioselective addition Of diketene to aldehydes promoted by chiral schiff base—titanium alkoxide complex. Application to asymmetric synthesis of potential inhibitors of HMG coenzyme reductase

Highly enantioselective addition of diketene to aldehydes was achieved by using novel Schiff base—titanium alkoxide complexes. Up to 92% ee of 5-hydroxy-3-oxoesters was obtained. This procedure provides an efficient method for the asymmetric synthesis of potential inhibitors of HMG coenzyme reductase.     

Mechanism of Magnetite Seam Formation and its Role for FeO Scale Transformation

The phase transformation behavior of a thermally grown oxide scale of FeO on pure-Fe and an Fe–2wt%Au alloy was investigated. Particular attention was paid to formation of a magnetite seam, which is the Fe₃O₄ layer formed at the FeO/alloy interface at an initial stage of the phase transformation, since it has important effects on the overall phase transformation of FeO scale. A thin Au(Fe) layer was found to develop on the Fe–Au alloy at the FeO/alloy interface after 32 min of oxidation at 750 °C in air. This Au(Fe) layer prevented formation of a magnetite seam and accelerated the FeO eutectoid reaction. The Au(Fe) layer acted as a “chemical diffusion barrier” for inward diffusion of Fe from the FeO to the alloy substrate across the FeO/alloy interface and prevented magnetite seam formation.     

An Approach to a Higher‐Power‐Density Power Supply for a 380‐V DC Distribution System

A methodology for realizing a higher‐power‐density DC‐DC converter has been proposed for a power unit installed in a 380‐V DC distribution system. The possibility of the converter design will be strengthened by using the series–parallel connection topology for isolated DC‐DC converters. A converter prototype with a power density of 10 W/cm³ has been fabricated, and the feasibility of the converter design has been confirmed experimentally. This result contributes to the realization of a highly efficient and highly space‐saving 380‐V DC distribution system. © 2013 Wiley Periodicals, Inc. Electr Eng Jpn, 186(3): 51–62, 2014; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.22494     

Suppression of inducible nitric oxide synthase expression and amelioration of lipopolysaccharide-induced liver injury by polyphenolic compounds in Eucalyptus globulus leaf extract

Eucalyptus leaf extract (ELE) is rich in hydrolyzable tannins. We examined the effects of ELE and its constituents on lipopolysaccharide (LPS)-induced liver injury in mice. Mice fed a diet supplemented with 1% ELE were intraperitoneally administered LPS. Six hours later, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were significantly lower in the ELE-supplemented mice than in the controls; LPS-induced hepatic inducible nitric oxide synthase (iNOS) expression was also suppressed. ELE lowered LPS-stimulated iNOS expression in cultured RAW 264.7 macrophages. Furthermore, the aglycones of hydrolyzable tannins, gallic acid (GA) and ellagic acid (EA), inhibited iNOS induction to a greater extent than did ELE (15-fold higher). When mice were fed a 1% GA or EA diet, the increase in the serum ALT and AST activities and hepatic iNOS expression in response to the LPS challenge were significantly attenuated. Thus, hydrolyzable tannins in ELE ameliorate LPS-induced liver injury.     

Fast and Almost Complete Nitridation of Mesoporous Silica MCM-41 with Ammonia in a Plug-Flow Reactor

The title reaction proceeded well to yield silicon (oxy)nitride at 973–1323 K using a plug-flow reactor. The degree of nitridation was studied as a function of temperature and time of nitridation, the sample weight, and the flow rate of ammonia. It was dependent on the reaction temperature and the amount of ammonia supplied per sample weight. The nitridation at 1273 K for 10–25 h yielded the oxynitride with 36–39 wt% nitrogen, which was very close to 40 wt% of Si₃N₄. Characterization with X-ray diffraction, field-emission scanning electron microscopy and transmission electron microscopy measurements, and nitrogen adsorption revealed the conversion of MCM-41 to the corresponding oxynitride without essential loss of the mesoporous structure, the decrements of the lattice constant and the pore diameter by 20–35%, and the increments of the wall thickness by ca. 45%. Solid-state ²⁹Si nuclear magnetic resonance spectra during the nitridation clearly showed fast decrease in SiO₄ species and slow in SiO₃(OH). Various intermediate species, SiO _x N _y (NH₂ or NH) _z , were observed to be formed and finally, ca. 70% SiN₄ species, ca. 20% SiN₃(NH₂ or NH), and ca. 10% SiON₂(NH₂ or NH) were produced, being consistent with the results of the above mentioned elemental analysis.