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.     

Hydrogen and carbon dioxide adsorption with tetra‐n‐butyl ammonium semi‐clathrate hydrates for gas separations

Gas adsorption rates of H₂, CO₂, and H₂‐CO₂ gas mixture (H₂/CO₂ = 3.4) with tetra‐n‐butyl ammonium salt (bromide, chloride, and fluoride) semi‐clathrate hydrate particles were measured at 269 K to assess their properties for gas separation. Equilibrium gas occupancies in the S‐cages of the particles were in order of (high to low) for hexagonal structure‐I, tetragonal structure‐I, and superlattice of cubic structure‐I structures with the maximum fractional occupancy by CO₂ being about 40%. The CO₂ diffusion rate depended on the anion size of the salt, which is attributed to distortion of the S‐cage that is close to the molecular size of CO₂. Simulations of semi‐clathrate hydrate particles with theory showed that H₂/CO₂ selectivities could be as high as 36 (3.0 mol% TBAF) and that selectivities for an ideal membrane (3.3 mol% TBAF) could be >100 (269 K, 0.3–4.5 MPa). Semi‐clathrate hydrates have wide application as separation media for gas mixtures. © 2014 American Institute of Chemical Engineers AIChE J, 61: 992–1003, 2015     

Energy value and digestibility of dietary oil containing mainly 1,3-diacylglycerol are similar to those of triacylglycerol

Diacylglycerol (DAG) is a component of various vegetable oils. Approximately 70% of the DAG in edible oils are in the configuration of 1,3-DAG. We recently showed that long-term ingestion of dietary oil containing mainly 1,3-DAG reduces body fat accumulation in humans as compared to triacylglycerol (TAG) oil with a similar fatty acid composition. As the first step to elucidate the mechanism for this result, we examined the difference in the bioavailabilities of both oils by measuring food energy values and digestibilities in rats. Energy values of the DAG oil and the TAG oil, measured by bomb calorimeter, were 38.9 and 39.6 kJ/g, respectively. Apparent digestibility expressed according to the formula: (absorbed) x (ingested)⁻¹x100=(ingested—excreted in feces)x(ingested)⁻¹x100 for the DAG oil and the TAG oil were 96.3±0.4 and 96.3±0.3% (mean±SEM), respectively. The similarity in the bioavailabilities of both oils supports the hypothesis that the reduced fat accumulation by dietary DAG is caused by the different metabolic fates after the absorption into the gastrointestinal epithelial cells.     

Assessing the kinetic model of hydro-distillation and chemical composition of Aquilaria malaccensis leaves essential oil

This study aimed to model the kinetic of hydro-distillation of Aquilaria malaccensis leaves oil in order to understand and optimize the extraction process. In addition, this study, for the first time, aimed to identify the chemical compositions of the A. malaccensis leave-oil. By assessing both first-order kinetic model and the model of simultaneous washing and diffusion, the result indicated that the model of simultaneous washing and diffusion better describes the hydro-distillation mechanism of the essential oil from A. malaccensis leaves.The optimum time, solid to liquid ratio, and the heating power for extracting the highest amount of essential oil were found to be around 3 h, 1:10(g·ml-1), and 300 W respectively. Yellow essential oil with a strong smell and a yield of 0.05 v/w was extracted by hydro-distillation Clevenger apparatus. Chemical compounds of the essential oil were analyzed using gas chromatography–mass spectroscopy(GC/MS), which resulted in identification of 42 compounds that constitute 93% of essential oil. Among the identified components,Pentadecanal(32.082%), 9-Octadecenal,(Z)(15.894%), and Tetradecanal(6.927%) were the major compounds.Considering the fact that all the identified major components possess pesticidal properties, A. malaccensis leaves can be regarded as a promising natural source for producing pesticides.     

Heterodimerization of Group I Ribozymes Enabling Exon Recombination through Pairs of Cooperative trans‐Splicing Reactions

Group I (GI) self‐splicing ribozymes are attractive tools for biotechnology and synthetic biology. Several trans‐splicing and related reactions based on GI ribozymes have been developed for the purpose of recombining their target mRNA sequences. By combining trans‐splicing systems with rational modular engineering of GI ribozymes it was possible to achieve more complex editing of target RNA sequences. In this study we have developed a cooperative trans‐splicing system through rational modular engineering with use of dimeric GI ribozymes derived from the Tetrahymena group I intron ribozyme. The resulting pairs of ribozymes exhibited catalytic activity depending on their selective dimerization. Rational modular redesign as performed in this study would facilitate the development of sophisticated regulation of double or multiple trans‐splicing reactions in a cooperative manner.     

Human behavior in a staircase during a total evacuation drill in a high‐rise building

Many studies have been conducted on the evacuation behavior on the staircases of buildings, but very little data are available for a situation with many occupants in a crowded high‐rise building. Therefore, this study investigated the evacuation behavior of a large number of evacuees on the staircase of a 25‐story high‐rise building. A total evacuation drill was conducted with 2088 evacuees, and the behavior of 1136 evacuees on the landings of the south staircase was recorded by a video recorder on the ceiling. The relationship between the density and speed of the evacuees on the landings was analyzed from the evacuation data for two situations: without and with merging in the stair flow. The evacuation stair flow in this drill had merging occupants entering from the floors, but no one entered from the lower floors during the latter period of the drill. Therefore, the flow during the latter period was treated as non‐merging flow, for which it was observed that, when the staircase was fully crowded, the density on the landings in the moving situation was different from that in the stopped situation. Moreover, the density on the landings was different from that on the treads. Furthermore, in the merging flow, a merging ratio of approximately 50:50 occurred during the congested evacuation. Copyright © 2016 John Wiley & Sons, Ltd.     

Laboratory evaluation of nanoparticle penetration efficiency in a cylindrical counter flow denuder for non-specific removal of trace gases

This article presents a cylindrical counter-current flow diffusion denuder with high efficiency penetration of nanoparticles, for non-specific removal of trace gases from an air flow. The denuder was designed to exchange gases in the sample flow by diffusion to the purge flow across a cylindrical microporous glass tube. Laboratory test results indicated that removal efficiencies of gases increased with a lower sample flow rate and a higher sample to purge flow rate ratio. Additionally, the pore size of the microporous glass did not affect gas removal efficiency and particle penetration following optimization of sample and purge flow rate conditions. Significantly high particle penetration was obtained for the counter flow denuder technique (94% penetration for 20 nm of polystyrene latex particle [PSL]) that agreed with theoretical estimation attributed to diffusion loss.

Copyright © 2017 American Association for Aerosol Research     

Pilot-Scale Operation of a Concrete Sludge Recycling Plant and Simultaneous Production of Calcium Carbonate

A pilot-scale plant to treat concrete sludge and produce calcium carbonate (CaCO₃) and an environmental purification agent (phosphorus adsorbent derived from concrete sludge, PAdeCS®) was designed, constructed, and operated. Concrete sludge from a concrete pile and pole production plant, boiler gas containing CO₂, and groundwater were used in the plant. The process involved calcium extraction from concrete sludge into water, followed by reaction of the calcium with CO₂ to produce crystalline CaCO₃. The pilot-scale plant was operated for 1 week, and the mass flows, conversion of CO₂ to CaCO₃, and net CO₂ emissions of the process were estimated. High-purity CaCO₃ (>97%) suitable for industrial use was obtained. Based on the power consumption of the process and the amount of CO₂ sequestered into CaCO₃, a net reduction in CO₂ emissions can be achieved using this process. The produced PAdeCS can be used as an inexpensive substitute for calcium series environmental purification agents.