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%.     

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.     

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.     

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.     

Potential of predominant activated sludge bacteria as recipients in conjugative plasmid transfer

We investigated the possibility of conjugative plasmid transfer to the predominant bacteria in activated sludge and the factors influencing the transfer frequency in the activated sludge process. We performed conjugative transfers of a self-transmissible, broad-host-range plasmid RP4 from Escherichia coli C600 to activated sludge bacteria by broth mating. Most of the activated sludge bacteria tested could acquire plasmid RP4, although the transfer frequencies varied from 8.8 x 10(-7) to 1.3 x 10(-2) transconjugants per recipient. The transfer frequencies in several strains were similar to, or higher than, that in intraspecific transfer to E. coli HB101. Matings under various environmental conditions showed that factors relevant to physiological activity, such as temperature and nutrient conditions, seemed to affect the transfer frequency. In addition, conjugative transfer was detected even in filtered raw and treated wastewaters. Thus, the predominant activated sludge bacteria seem to have sufficient potential as recipients in conjugative plasmid transfer under the conditions likely to occur in the activated sludge process. Transfer frequency was reduced by agitation in the presence of suspended solid. This may suggest that conjugative plasmid transfer is physically inhibited in aeration tanks.     

Compound-specific factors influencing sorption nonlinearity in natural organic matter

Nonlinear sorption by natural organic matter may have a significant impact on the behavior of organic contaminants in soils and sediments. This study presents a molecular probe approach based on linear solvation energy relationships (LSERs) to identify and quantify the molecular interactions causing concentration-dependent sorption and proposes estimation methods for sorption nonlinearities. Sorption isotherms ranging over concentrations of more than 4 orders of magnitude were determined in batch systems for 23 and 16 chemically diverse probe compounds in a lignite sample and a peat soil, respectively. Each sorbent showed characteristic nonlinear sorption with Freundlich exponents (1/n) being 0.7-1. The LSER-based analysis revealed that the strength of nonspecific interactions did not vary with concentration for both sorbents. In lignite, specific interactions did not affect sorption nonlinearity either, suggesting that compound-independent factors of lignite were responsible for the nonlinear sorption. In the peat soil, by contrast, the specific interactions related to the solute polarizability/dipolarity parameter (S) decreased with increasing concentration. Consequently, compounds of higher S values were more susceptible to nonlinear sorption in the peat soil. Phenol probes have shown that hydrogen bond donating properties of sorbate compounds have a substantial impact on the overall strength of sorption with organic matter, but no significant influence on sorption nonlinearity. Heterocyclic aromatic compounds appear to undergo additional interactions that are not accounted for by the LSER. These additional interactions considerably enhance both sorption capacity and nonlinearity.     

Dietary intake of dioxins in Japan in 2016 with time trends since 1998

Total diet samples collected from seven regions throughout Japan in 2016 were analysed for polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like polychlorinated biphenyls (DL-PCBs), known collectively as dioxins. This led to estimates of the latest dietary intake of these contaminants for the general Japanese population (≥1 year old). The average daily intake of dioxins for a person weighing 50 kg, calculated at non-detected congener concentrations assumed to be equal to zero, was estimated to be 0.54 pg TEQ (toxic equivalents) kg^?1 body weight (bw) day^?1. This value is well below the tolerable daily intake of 4 pg TEQ kg^?1 bw day^?1 for dioxins in Japan. The average intake was highest from fish and shellfish, followed by meat and eggs. The TEQ contribution of the fish and shellfish group to the total dietary TEQs was significant (89%). The DL-PCBs accounted for about 67% of the dioxin intake. The latest dioxin intake level was compared with previous estimates from total diet study results obtained annually since 1998 to determine the time trends in the dietary intake of dioxins in Japan. Overall, the average dioxin intake appeared to be decreasing gradually during the period of study. The previous average intakes of dioxins ranged from 0.58 to 1.9 pg TEQ kg^?1 bw day^?1. The latest average intake was the lowest since 1998 and was about one-third of the average intake in 1998. This decreasing trend in the dietary intake of dioxins was mainly influenced by the decreased dioxin intakes from two food groups, fish and shellfish, and meat and eggs.