Characterization of Acetylxylan Esterase from White-Rot Fungus Irpex lacteus

The carbohydrate esterase family 1 (CE1) in CAZy contains acetylxylan esterases (AXEs) and feruloyl esterases (FAEs). Here we cloned a gene coding for an AXE belonging to CE1 from Irpex lacteus (IlAXE1). IlAXE1 was heterologously expressed in Pichia pastoris, and the recombinant enzyme was purified and characterized. IlAXE1 hydrolyzed p-nitrophenyl acetate, α-naphthyl acetate and 4-methylumbelliferyl acetate, however, it did not show any activity on ethyl ferulate and methyl p-coumarate. We also examined the activity on partially acetylated and feruloylated xylan extracted from corncob by hydrothermal reaction. Similarly, ferulic and p-coumaric acids were not liberated, and acetic acid was only detected in the reaction mixture. The results indicated that IlAXE1 is an acetylxylan esterase actually reacted to acetyl xylan. However, since IlAXE1 was unable to completely release acetic acid esterifying xylopyranosyl residues, it is assumed that acetyl groups exhibiting resistance to deacetylation by IlAXE1 are present in corn cob xylan.     

Principle validation of nuclear fuel material isotopic composition measurement method based on photofission reactions

We have developed a new nondestructive assay technique based on the photonuclear reaction, which is aimed at measuring the isotopic composition of nuclear fuel materials without relying on their self-generated neutron information. This methodology enables measurement of the number of neutrons produced by the photofission reactions at different specific photon energies, and only information on the relative counts and not the absolute values or energies, which are considerably affected by backgrounds, are required for the enrichment induction according to a mathematical process. The present methodology estimated the ²³⁵U enrichment value with 8% accuracy when the cross section uncertainty is 5%.     

Analyses of reactor building by 3D nonlinear FEM models considering basemat uplift for simultaneous horizontal and vertical ground motions

The nonlinear behavior of basemat uplift, which is an important point in seismic designs of nuclear power plants in Japan, has been investigated by arranging joint elements between the reactor building basemat and the soil on a three-dimensional (hereafter referred to as 3D) FEM model of the soil. However, the nonlinearity of the basemat uplift has been investigated separately from the nonlinearity of reactor buildings. These nonlinearities have yet to be taken into account simultaneously in past studies. In this paper, models of the building and the soil using 3D FEM elements with consideration to the nonlinearity of building materials as well as the nonlinearity of the basemat uplift were subjected. The behavior of the building's elements were investigated by carrying out seismic response analyses for horizontal ground motions only, as well as for simultaneous horizontal and vertical ground motions using these models. As a result, it was found that there was little difference in the horizontal response of the building between the horizontal input motions only and the simultaneous horizontal and vertical input motions. The effects of the vertical ground motions on the basemat uplift behavior which is represented by the ground contact ratio were also slight.     

Microstructure of ion-implanted region in TiNi alloy

TiNi alloy samples implanted with various fluences of 3 MeV Cu²⁺ ions were characterized by transmission electron microscope (TEM) and X-ray diffractometer. Cross-sectional TEM images of the samples showed that amorphous region was seen at the fluence of 10¹⁴ ions cm^?2 in case of ion implantation at 300 K of the substrate temperature, but in case of ion implantation at 100 K it did not appear even at 10¹⁵ ions cm^?2. These results were also confirmed by X-ray diffraction profiles of the same samples. Consequently, the extent of microstructure change of TiNi alloy by ion implantation was different depending on the substrate temperature.     

Adaptation to climate change and conservation of biodiversity using green infrastructure

In recent years, we have experienced mega‐flood disasters in Japan due to climate change. In the last century, we have been building disaster prevention infrastructure (artificial levees and dams, referred to as “grey infrastructure”) to protect human lives and assets from floods, but these hard protective measures will not function against mega‐floods. Moreover, in a drastically depopulating society such as that in Japan, farmland abandonment prevails, and it will be more difficult to maintain grey infrastructure with a limited tax income. In this study, we propose the introduction of green infrastructure (GI) as an adaptation strategy for climate change. If we can use abandoned farmlands as GI, they may function to reduce disaster risks and provide habitats for various organisms that are adapted to wetland environments. First, we present a conceptual framework for disaster prevention using a hybrid of GI and conventional grey infrastructure. In this combination, the fundamental GI, composed of forests and wetlands in the catchment (GI‐1) and additional multilevel GIs such as flood control basins that function when floodwater exceeds the planning level (GI‐2) are introduced. We evaluated the flood attenuation function (GI‐1) of the Kushiro Wetland using a hydrological model and developed a methodology for selecting suitable locations of GI‐2, considering flood risk, biodiversity and the distribution of abandoned farmlands, which represent social and economic costs. The results indicated that the Kushiro Wetland acts as a large natural reservoir that attenuates the hydrological peak discharge during floods and suitable locations for introducing GI‐2 are concentrated in floodplain areas developing in the downstream reaches of large rivers. Finally, we discussed the network structure of GI‐1 as a hub and GI‐2 as a dispersal site for conservation of the Red‐crowned Crane, one of the symbolic species of Japan.     

Unique crystal structures and their applications as materials for Li1+x-yM1-x-3yTix+4yO3 (M = Nb or Ta, 0.07 ≤ x ≤ 0.33, 0 ≤ y ≤ 0.175)

In the Li₂O–M₂O₅–TiO₂ (M = Nb or Ta) system, Li_1+x-yM_1-x-3yTi_x+4yO₃ (LMT) forms with a superstructure known as the M-phase, in which the periodical intergrowth layers are inserted in a matrix having a trigonal structure of LiMO₃. We have been investigating this unique material focusing on its crystal structure, photoluminescence, and electrical properties.First, the formation area of the superstructure was compared between Nb- and Ta- systems and the difference was accurately analyzed.Second, to apply this unique material as a host material of phosphor, we synthesized new phosphors with various emission colors. Red phosphor, having a high PL intensity with an internal quantum efficiency of 98%, was successfully synthesized and used as a host material of the solid solution for the Ta system. Its high value was the result of the large centroid-to-cation distance of the Eu³⁺ position in the [(Li, Eu)O₁₂] polyhedral.Further still, toward application of the unique qualities of an electro-ceramic, we successfully fabricated oriented balk ceramics for the Nb system by slip casting in a strong magnetic field of 12 T. As a result, anisotropic electric properties were found along the c-axis, which were caused by the superstructure. We first clarified the mechanism showing that the anisotropic Qf value was due to anisotropic electron conductivity and anisotropic bonding strength within the superstructure.     

Determination of acrylamide in processed foods by LC/MS using column switching

An LC/MS method was developed for the determination of acrylamide (AA) in processed or cooked foods. AA was extracted with a mixture of water and acetone from homogenized food samples after the addition of 13C-labeled acrylamide (AA-1-(13)C) as an internal standard. The extract was concentrated, washed with dichloromethane for defatting, and cleaned up on Bond Elut C18, PSA and ACCUCAT cartridge-columns, and then AA was determined by LC/MS in the selected ion recording (SIR) mode. For the LC/MS analysis, four LC columns were connected in-line and the flow of the mobile phase was switched according to a time-program. Monitoring ions for AA were m/z 72 and 55, and those for AA-1-(13)C were m/z 73 and 56. AA and AA-1-(13)C were determined without interference from the matrices in all samples. The recoveries of AA from potato chips, corn snack, pretzel and roasted tea spiked at the level of 500 ng/g of AA were 99.5-101.0% with standard deviations (SD) in the range from 0.3 to 1.6%. The limits of detection and quantification of the developed method were 9 and 30 ng/g for AA in samples, respectively. The method was applied to the analysis of AA in various processed or cooked food samples purchased from retail markets. High levels of AA were found in potato chips and French-fried potato (467-3,544 ng/g). Fried and sugar-coated dough cakes (karinto) contained 374 and 1,895 ng/g. Corn snacks contained 117-535 ng/g of AA. Roasted foods (such as roasted sesame seed, roasted barley (mugi-cha), roasted tea (hoji-cha), coffee beans and curry powder) contained 116-567 ng/g of AA. Foods made from fish, egg and meat contained lower levels of AA than the plant-based foods. Foods containing much water showed a tendency to have low levels of AA compared with dry foods. The proposed method was applicable to the analysis of AA in variety of processed foods.     

Expression of a gene encoding chitinase (pCA 8 ORF) from Aeromonas sp. no. 10S-24 in Escherichia coli and enzyme characterization

A gene encoding chitinase from Aeromonas sp. no. 10S-24 was expressed using pTrc99A in Escherichia coli JM 105 which yielded a 5-fold higher activity than when pUC19 was used. Three different truncated enzymes (SA-1, SA-2 and SA-3) were obtained after purification. Their isoelectric points were 7.0, 6.9, and 6.7, respectively. The enzymes showed two optimum pHs, 4.0 and 7.0, when incubated with ethylene glycol chitin as the substrate, and were stable over a wide pH range (3.0-9.0). The optimum temperature was 60 degrees C and the enzymes were stable up to 50 degrees C. The chitinases exhibited wide substrate specificities for chitin-related compounds.     

A novel gene delivery system in plants with calcium alginate micro-beads

We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFP. With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 h incubation compared to that with the conventional method using a naked plasmid solution. Transfection was up to 0.22% efficient. These results indicate that bio-beads have a possibility for efficient transformation in plants.