Identification and Characterization of D‐Succinylase,and a Proposed Enzymatic Method for D‐Amino Acid Synthesis

Chiral amino acids are important intermediates for the pharmaceutical industry. We have developed a novel one‐pot enzymatic method for D ‐amino acid synthesis by the dynamic kinetic resolution of N‐succinyl‐dl ‐amino acids using D ‐succinylase (DSA) and N‐succinylamino acid racemase (NSAR, EC 4.2.1.113). The DSA from Cupriavidus sp. P4‐10‐C, which hydrolyzes N‐succinyl‐D ‐amino acids enantioselectively to their corresponding D ‐amino acids, was identified for the first time by screening soil microorganisms. Subsequently, the DSA gene was cloned and overexpressed in Escherichia coli. DSA was shown to comprise two subunits with molecular masses of 26 kDa and 60 kDa. Additionally, the NSAR gene from Geobacillus stearothermphilus NCA1503, which racemizes N‐succinylamino acids, was also cloned and overexpressed in E. coli. The highly purified DSA and NSAR prepared from each recombinant E. coli were characterized and used for D ‐amino acid synthesis. A one‐pot enzymatic method converted 100 mM N‐succinyl‐dl ‐phenylalanine to D ‐phenylalanine in 91.1% conversion with 86.7% ee. This novel enzymatic method may be useful for the industrial production of many D ‐amino acids.

     

Effect of Surface Treatment on the Selective Photocatalytic Oxidation of Benzyl Alcohol into Benzaldehyde by O2 on TiO2 Under Visible Light

The photocatalytic oxidation of benzyl alcohol into benzaldehyde proceeded with high conversion and selectivity on a TiO₂ photocatalyst by O₂ under visible light irradiation. Surface complex formed by the interaction of benzyl alcohol with the Ti sites and/or surface OH groups of TiO₂ play an important role in the absorption of visible light and unique selective photocatalytic reaction.     

Purification, characterization, and steady-state kinetics of a meta-cleavage compound hydrolase from Pseudomonas fluorescens IPO1

2-Hydroxy-6-oxo-7-methylocta-2,4-dienoate (6-isopropyl-HODA) hydrolase (CumD), an enzyme of the cumene biodegradation pathway encoded by the cumD gene of Pseudomonas fluorescens IP01, was purified to homogeneity from an overexpressing Escherichia coli strain. SDS-polyacrylamide gel electrophoresis and gel filtration demonstrated that it is a dimeric enzyme with a subunit molecular mass of 32 kDa. The pH optima for activity and stability were 8.0 and 7.0-9.0, respectively. The enzyme exhibited a biphasic Arrhenius plot of catalysis with two characteristic energies of activation with a break point at 20 degrees C. The enzyme has a K(m) of 7.3 microM and a k(cat) of 21 s(-1) for 6-isopropyl-HODA (150 mM phosphate, pH 7.5, 25 degrees C), and its substrate specificity covers larger C6 substituents compared with another monoalkylbenzene hydrolase, TodR Unlike TodF, CumD could slightly hydrolyze 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (6-phenyl-HODA). A mutant enzyme as to a putative active site residue, S103A, had 10(5)-fold lower activity than that of the wild-type enzyme.     

Application of microalgae hydrolysate as a fermentation medium for microbial production of 2-pyrone 4,6-dicarboxylic acid

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Thermal deformation measurement of the solder joints in electronic packages using electron moiré method

In this paper, electron moiré method is used to measure the thermal deformation of electronic packages. In order to observe the electron moiré fringe, a holographic grid was replicated on the cross section of the BGA type package at 150°C. This grid was fabricated on a glass plate using a moving point holographic system, and was replicated to measured area at high temperature. Under SEM, the holographic grid(specimen grid) and a programmed electron beam scan(master grid) interfere and form electron moiré patterns. The shear strain measurement technique using electron moiré method is described. Using the electron moiré method, the shear strains in the different solder joints were measured and analysed. Some useful results were obtained.     

Classification of particles by centrifugal separator and analysis of the fluid behavior

This study gives fundamental knowledge on the particle classification performance by centrifugal separator.It is found that the cut size of a centrifugal separator decreases as the rotational speed increases and the liquid flow rate decreases. Fitting our experimental results with the theory, they agree with each other at high flow rate. However, the difference between them generates at low flow rate. This is because dead spaces are generated in the centrifugal separator at the low flow rate. Also, the computer simulation of the fluid behavior in the centrifugal separator can find the decrease of the velocity near the wall under the low flow rate, which suggests the possibility of the formation of dead spaces in the separator.     

Catalytic graphitization of an amorphous carbon film under focused electron beam irradiation due to the presence of sputtered nickel metal particles

Electron beam irradiation of Ni particles (diameter: ∼500 nm) in a transmission electron microscope (TEM) formed small secondary particles, which moved randomly on an amorphous carbon film. Catalytic graphitization occurred along the path traced by the moving particles. It is generally well known that heat treatment is needed to produce catalytic graphitization of an amorphous carbon. However, we rapidly produced catalytic graphitization in the presence of sputtered Ni particles by focused electron beam irradiation in the TEM although no external heating was employed.     

The use of a virtual city model for assessing equity in access to views

The development of virtual city models has provided novel possibilities for analyses that require consideration of building heights in urban areas. The study was undertaken to explore these possibilities by using the virtual Kyoto model to examine equity in access to views in the Japanese city. A sample of just over 5000 residences was selected by stratifying for population age and affluence. A series of viewsheds were computed to quantify the visibility of a range of environmental amenities (greenspaces, water bodies, historical buildings, mountains) and disamenities (factories and roads). Evidence of inequity in visual amenity was identified, whereby homes in areas with many old people were much less likely to have views of greenspaces and water bodies, although they were also less likely to see factories and roads and were more likely to view mountains. Homes in more affluent areas had better views of greenspaces, historical buildings, and mountains, and were less likely to see factories and water bodies. We discuss the potential of virtual city models for furthering analyses of the urban environment and raise some caveats regarding their use.     

Particle arrangement and its application

The method by which micrometer-sized particles are arranged, is described. An electrified pattern is drawn on a polished insulating substrate using a SEM with a CAD facility. The substrate is dipped in a suspension of particles. The particles adhere to the substrate and form the pattern corresponding to the electrified pattern by the electrostatic force. The arranged particles are fixed by coating with a fluoride polymer of thin film. The method is applicable to the production of an inflammable gas sensor.