Reaction mechanism of fluoroacetate dehalogenase from Moraxella sp. B

Fluoroacetate dehalogenase (EC 3.8.1.3) catalyzes the dehalogenation of fluoroacetate and other haloacetates. The amino acid sequence of fluoroacetate dehalogenase from Moraxella sp. B is similar to that of haloalkane dehalogenase (EC 3.8.1.5) from Xanthobacter autotrophicus GJ10 in the regions around Asp-105 and His-272, which correspond to the active site nucleophile Asp-124 and the base catalyst His-289 of the haloalkane dehalogenase, respectively (Krooshof, G. H., Kwant, E. M., Damborsky, J., Koca, J., and Janssen, D. B. (1997) Biochemistry 36, 9571-9580). After multiple turnovers of the fluoroacetate dehalogenase reaction in H218O, the enzyme was digested with trypsin, and the molecular masses of the peptide fragments formed were measured by ion-spray mass spectrometry. Two 18O atoms were shown to be incorporated into the octapeptide, Phe-99-Arg-106. Tandem mass spectrometric analysis of this peptide revealed that Asp-105 was labeled with two 18O atoms. These results indicate that Asp-105 acts as a nucleophile to attack the alpha-carbon of the substrate, leading to the formation of an ester intermediate, which is subsequently hydrolyzed by the nucleophilic attack of a water molecule on the carbonyl carbon atom. A His-272 --> Asn mutant (H272N) showed no activity with either fluoroacetate or chloroacetate. However, ion-spray mass spectrometry revealed that the H272N mutant enzyme was covalently alkylated with the substrate. The reaction of the H272N mutant enzyme with [14C]chloroacetate also showed the incorporation of radioactivity into the enzyme. These results suggest that His-272 probably acts as a base catalyst for the hydrolysis of the covalent ester intermediate.     

Substituent effects on thermal and photo‐alignment behavior of poly(ethylene imine)s carrying azobenzene side groups

A new class of liquid‐crystalline poly(ethylene imine)s (PEIs) having four differently substituted (? CN,? C₄H₉,? OCH₃ and? NO₂) azobenzene side‐chain groups attached through alkyl spacer groups were successfully synthesized using a solution polycondensation reaction. The synthesized polymers were characterized using differential scanning calorimetry, polarized optical microscopy and X‐ray diffraction. The photochemical, thermo‐optical and photo‐orientational behavior of the polymers were investigated in detail. Spin‐coated films of PEIs with azobenzene groups having? C₄H₉,? OCH₃ and? NO₂ substituents showed out‐of‐plane molecular orientation on annealing. Except for the PEI with an azobenzene group having ? NO₂ substituent, all polymers exhibited good photoresponsive properties upon irradiation with UV and visible light. Films of PEIs with azobenzene side groups having? CN,? C₄H₉ and? OCH₃ substituents showed reversible alignment behavior from random state to out‐of‐plane and from out‐of‐plane to random state on annealing and on irradiation with UV and non‐polarized visible light. The reversibility of the molecular orientation of PEIs from random state to out‐of‐plane and from out‐of‐plane to random state greatly depended on the substituent attached to the azobenzene side‐chain group. Copyright © 2010 Society of Chemical Industry     

Expression of Human DNAJ (Heat Shock Protein-40) B3 in Humanized UDP-glucuronosyltransferase 1 Mice

The human DNAJB3 gene encodes a DNAJ (Heat shock protein 40; Hsp40) homolog, subfamily B, member 3 chaperone protein (DNAJB3), which can be down-regulated in disease conditions, as observed in decreased expression of DNAJB3 mRNA in peripheral blood mononuclear cells (PBMC) of obese patients. Recently, humanized UDP-glucuronosyltransferase (UGT) 1 mice (hUGT1 mice) were developed, in which the introduced human UGT1 gene contained a gene encoding human DNAJB3. In the present study, we analyzed the expression of human DNAJB3 mRNA in hUGT1 mice. Among the examined tissues, the testis had the highest expression of human DNAJB3 mRNA, while the lowest expression was observed in the liver. We found that the pattern of tissue-specific expression of mouse Dnajb3 in hUGT1 mice was very similar to that of human DNAJB3. We further demonstrated that the expression of human DNAJB3 in the liver was significantly reduced in high-fat-diet-fed hUGT1 mice compared to the expression level in the control mice, indicating that the expression of human DNAJB3 in hUGT1 mice could be similarly regulated in disease conditions such as obesity. Humanized UGT1 mice might therefore be useful to investigate the physiological role of human DNAJB3 in vivo.     

Extraction of proteins and polymers using reverse micelles and percolation process

Specific interfacial properties, which affect protein extraction in AOT and AOT-lecithin reverse micellar systems (RVMS), have been studied by using their percolation processes. Solubilization of proteins or synthetic polymers into RVMS by the injection method and its effect on the percolation processes could be easily evaluated by the measurement of electrical conductivity. The percolation process is found to be a sensitive and convenient measure of micro-interface of RVMS solubilizing various polymers or proteins, which clearly reflects the polymer(protein)-micellar and micellar-micellar interactions. The stability of RVMS or micellar-micellar interaction was dependent on the kinds, concentration and molecular weight of solubilized polymers. The value of p, defined as the variation of percolation threshold with the concentration of solubilized polymers, can be utilized to evaluate the stability of RVMS solubilizing polymers or micellar-micellar interactions. The values of Β are affected by the hydrophobicity, molecular weight and absolute value of the net charge of the polymers solubilized into the AOT reverse micelles, which were evaluated by using Aqueous Two-Phase Systems (ATPS).     

Electromagnetic Shielding Properties of Woodceramics Made from Wastepaper

Woodceramics are new porous carbon materials, which are made by impregnating woody materials with phenol resin and then thermoformed in a vacuum furnace and these have been shown to have electromagnetic shielding properties. In the recycling of wastepaper, ways of using the wastepaper other than for paper pulp are needed to be developed. In this study, we made Woodceramics from handbill advertisement paper and telephone directory paper, and measured their electromagnetic shielding properties in order to find new uses for wastepaper. The results showed that the Woodceramics made from wastepaper had an electric shielding effectiveness of 30 dB for 100 MHz and 40 to 43 dB for 300 MHz or higher, and had a magnetic shielding effectiveness of 30 dB for 100 MHz and 37 dB for about 400 MHz. An electric equivalent circuit of the pore model in the Woodceramics is introduced. In addition, it is proposed that the excellent electromagnetic shielding effectiveness of the Woodceramics is caused by dielectric loss.     

Sintering Behavior and Surface Microstructure of PbO-Rich PbNi1/3Nb2/3O3–PbTiO3–PbZrO3 Ceramics

The sintering behavior and surface microstructure of PbNi_1/3Nb_2/3O₃–PbTiO₃–PbZrO₃ (PNiNb-PT-PZ) ceramics were investigated. The PNiNb-PT-PZ ceramics with the stoichiometric composition and the addition of excess lead oxide (PbO-rich ceramics) were sintered by liquid-phase sintering in accordance with the solution-reprecipitation mechanism at temperatures below the melting point of PbO. The temperature at which the liquid phase forms fell to near the eutectic point of the PbO–Nb₂O₅ and the PbO–TiO₂ system (868°C) with the addition of 5 mol% PbO. As the calcination temperature influenced the sinterability of the stoichiometric PNiNb-PT-PZ ceramic, unreacted PbO was considered to be the source of the liquid phase in the sintering of the stoichiometric powder. The secondary phase was observed at the surface of PbO-rich ceramics and was suggested to be a liquid phase expelled from inside the ceramic. A sintering scheme of PNiNb-PT-PZ ceramics was proposed, and the high sinterability of PNiNb-PT-PZ ceramics was attributed to the low formation temperature of the liquid phase.     

URINARY 2-HYDROXYFLUORENE AND 1-HYDROXYPYRENE LEVELS IN SMOKERS AND NONSMOKERS IN JAPAN AND THAILAND

2-Hydroxyfluorene (2-OHF) and 1-hydroxypyrene (1-OHP) in urine samples of smokers and nonsmokers were determined separately by using two high-performance liquid chromatography (HPLC) systems with fluorescence detection. Urine samples were collected from the subjects who lived in Japan and Thailand, and were not occupationally exposed to polycyclic aromatic hydrocarbons (PAHs). The mean concentrations of 2-OHF and 1-OHP of Thai smokers (0.75 and 3.03 μmol/mol creatinine) and nonsmokers (0.22 and 0.91 μmol/mol creatinine) were both higher than those of Japanese smokers (0.26 and 0.12 μmol/mol creatinine) and nonsmokers (0.04 and 0.06 μmol/mol creatinine), respectively. The difference between smokers and nonsmokers was more significant for 2-OHF than for 1-OHP, reflecting the higher intake of fluorene in the vapor phase by the smoking. Moreover, the higher urinary levels of both 2-OHF and 1-OHP were observed in Thai nonsmokers than those in Japanese nonsmokers, suggesting the higher background exposure to PAHs of Thai subjects.     

Autogenous shrinkage of high-strength concrete containing silica fume under drying at early ages

The present study investigated experimentally autogenous shrinkage of high-strength concrete containing silica fume under drying at early ages. The influence of drying on hydration of cementitious materials in the high-strength concrete with water-binder ratios of 0.25, 0.35 and 0.45 was evaluated based on bound water content (BWC), which was exposed to drying at the ages of 0.5, 1.0 and 3.0 days, respectively. By establishing the relationship between the BWC and autogenous shrinkage strain under sealed conditions, autogenous shrinkage strain under drying conditions and drying shrinkage strain were separated from total shrinkage strain, and, then, the contribution of autogenous shrinkage in total shrinkage was discussed. The results showed that the percentage of autogenous shrinkage was macroscopically 50-20% based on the present method, while that was 70-30% based on the conventional superposition principle (SP). The latter resulted in overestimating autogenous shrinkage strain under drying conditions.     

Effective Coke Removal in Methane to Benzene (MTB) Reaction on Mo/HZSM-5 Catalyst by H2 and H2O Co-addition to Methane

The catalytic dehydro-aromatization reaction over Mo/HZSM-5 catalyst was drastically stabilized by the co-addition of 5.4% H₂ and 1.8% H₂O to methane feed at 750 °C, 0.3 MPa and methane space velocity of 3000 mL g⁻¹ h⁻¹, suppressing the coke formation effectively, compared with single hydrogen or steam addition.     

The role of aluminum in automotive weight reduction—part III

This series of articles provides, from an international perspective, an analysis of the future of the automotive industry and aluminum’s role in automobile weight reduction. Part I briefly described technological, environmental, and societal issues that are mandating vehicular weight reduction for, among other reasons, purposes of fuel economy. Part II described the status of the weight-reduction technologies that are currently available and projects how they may evolve in the coming years. Part III, the final installment, examines some of the specific strengths and weaknesses of aluminum alloys with regard to their long-term attractiveness to automakers.