Enzyme-catalyzed poly(3-hydroxybutyrate) synthesis from acetate with CoA recycling and NADPH regeneration in Vitro

We established a novel enzyme-catalyzed poly(3-hydroxybutyrate) [P(3HB)] synthesis system capable of recycling CoA on the basis of the P(3HB) biosynthetic pathway in Ralstonia eutropha. The system includes purified beta-ketothiolase (PhaA), NADPH-dependent acetoacetyl-CoA reductase (PhaB), PHA synthase (PhaC), acetyl-CoA synthetase (Acs) and glucose dehydrogenase (GDH). In this system, acetyl-CoA was synthesized from acetate and CoA by Acs and ATP, and then two molecules of acetyl-CoA were condensed by PhaA to synthesize acetoacetyl-CoA, which was converted to (R)-3-hydroxybutyryl-CoA (3HBCoA) by PhaB and NADPH. The 3HBCoA was polymerized by PhaC and converted to P(3HB). In this system, the CoA molecules that were released during the condensation and polymerization reactions catalyzed by PhaA and PhaC, respectively, were reused successfully for the synthesis of acetyl-CoA. In addition, NADPH, which was consumed in the reduction of acetoacetyl-CoA, was regenerated by the action of GDH. In this system, the yield of P(3HB) synthesized from acetate as the substrate was 5.6 mg in a 5-ml reaction mixture, and the weight-average molecular weight and polydispersity were 6.64 x 10(6) and 1.36, respectively. Furthermore, CoA was reused at least 26 times, and NADPH was also regenerated at least 26 times during 24 h of reaction.     

Formation of BCC TiFe hydride under high hydrogen pressure

We investigated the hydrogenation of a binary TiFe alloy at 5 GPa and 600 °C by in situ synchrotron radiation X-ray diffraction measurements. After formation of a solid solution of hydrogen in TiFe, an order–disorder phase transition in the metal lattice of TiFe occurred, which yielded a BCC TiFe hydride. The unit cell volume of the BCC hydride increased by 21.0% after the hydrogenation reaction. The volume expansion was larger than that of a γ-hydride TiFeH_1.9 prepared by hydrogenation near ambient conditions.     

Hysteresis phenomena on the crystal lattice of Ti0.8Zr0.2Mn1.5 in the hydrogenation and dehydrogenation process

The crystal structure changes in the hydrogenation and dehydrogenation process have been investigated by the in-situ synchrotron X-ray diffraction measurement to elucidate the mechanism of hysteresis phenomena in equilibrium hydrogen pressure. The hysteresis phenomena were observed on the crystal lattice. The lattice constants and volume of α-phase (hydrogen solid solution) and β-phase (hydride) in the dehydrogenation process with lower equilibrium pressure were smaller than those in the hydrogenation process with higher equilibrium pressure. This phenomenon is opposite to the general relationship between the lattice volume and the equilibrium pressure. The hysteresis phenomenon was also observed in the diffraction peak widths. The peak widths of β-phase in the dehydrogenation process were wider than those in the hydrogenation process. The hysteresis phenomena may be related to lattice defects and the hydrogen content in crystalline regions.     

Excess heat evolution from nanocomposite samples under exposure to hydrogen isotope gases

Anomalous heat effect by interaction of hydrogen isotope gas and metal nanocomposites supported by zirconia or by silica has been examined. Observed absorption and heat evolution at RT were not too large to be explained by some chemical processes. At elevated temperatures of 200–300 °C, most samples with binary metal nanocomposites produced excess power of 3–24 W lasting for up to several weeks. The excess power was observed not only in the D-Pd·Ni system but also in the HPd·Ni system and HCu·Ni system, while single-element nanoparticle samples produced no excess power. The Pd/Ni ratio is one of the keys to increase the excess power. The maximum phase-averaged excess heat energy exceeded 270 keV/D, and the integrated excess heat energy reached 100 MJ/mol-M or 90 MJ/mol-H. It is impossible to attribute the excess heat energy to any chemical reaction; it is possibly due to radiation-free nuclear process.     

Development research on composite adsorbents applied in adsorption heat pump

A novel adsorbent design technique base on the concept of Kelvin equation was proposed to develop hydrophilic adsorbent applicable to water vapor adsorption heat pump (AHP) for high performance. In the process, the composite adsorbent was prepared after silica gel was synthesized in the pores of activated carbons by impregnating activated carbons in sodium silicate solution. Two kinds of activated carbons were tested to produce composite adsorbent and to investigate the performance by measuring the adsorption isotherms of water vapor and pore structure characteristics. All adsorption isotherms of the silica impregnated activated carbons prepared shifted to a lower region of water vapor pressure compared to those of the raw activated carbons. The volume-based amount of adsorption in the AHP operation range (φ = 0.1–0.4) for the adsorbents prepared at sodium silicate solution concentration of 10 wt.% and impregnating time of 48 h are 5.88 and 2.62 times that of the raw activated carbons (AC1 and AC2), respectively. Based on the Kelvin equation, it is clarified that the contact angle and the volume of pore radius greater than 1.2 nm decrease with the increase of sodium silicate solution concentration for the novel composite adsorbents, which contributes the isothermals shift to lower relative pressure range.     

Construction of a reporter yeast strain to detect estrogen receptor signaling through aryl hydrocarbon receptor activation

The activation mechanism of estrogen receptor (ER) signaling by association with the aryl hydrocarbon receptor (AhR) was elucidated recently (Ohtake, et al., Nature 2003, 423, 545). In the present study, we established a reporter yeast strain to evaluate this ER signaling by association with the activated AhR. This yeast strain expresses human ER and AhR, and has a reporter plasmid with estrogen response elements. With this yeast strain we assayed ER activation by various AhR ligands, i.e., 2,3,7,8-tetrachlorodibenzo-p-dioxin, benzo[a]pyrene, 3-methylcholanthrene, beta-naphthoflavone, and indirubin. All these ligands induced ER activation dose-dependently and their EC50 values were 60, 180, 130, 26, and 0.5 nM, respectively. Then, we measured the activity in water collected at 5 localities in the Ishizu River system in Japan. The activities of water samples ranged from 4.8 pmol/L (1.3 ng/L) to 52 pmol/L (14 ng/ L) (17beta-estradiol (E2) equivalent). These values were higher than those measured with the yeast for ER activation through direct ligand binding to ER. The direct ER ligand binding activities of the water samples ranged from 2.5 to 5.3 pmol/L (E2 equivalent). We also measured AhR activation of the water samples using a reporter yeast for AhR ligand activity. The activities ranged from 102 to 472 pmol/L (beta-naphthoflavone equivalent). These results indicate that the water samples contain substances that bind to AhR, and these substances contribute to ER signaling through AhR activation in the yeast reporter strain. This yeast reporter strain should be a useful tool to evaluate direct and indirect ER activation by environmental samples.     

Anode-supported micro tubular SOFCs for advanced ceramic reactor system

In this study, micro tubular SOFCs under 1 mm diameter have been fabricated and investigated at 450–550 °C operating temperature with H₂ fuel. The performance of the 0.8 mm diameter tubular SOFC was 110–350 mW cm⁻² at 450–550 °C operating temperatures. To maximize the performance of the cell as well as to optimize the geometry of tubular cells, a current collecting method used in the experiment was examined. A model was proposed to estimate the loss of performance for single cell due to the current collecting method as functions of anode tube length and thickness. The results showed that the losses of performance were calculated to be 0.8, 2.0, and 4.6% at 450, 500, and 550 °C operating temperatures, respectively, for the 0.8 mm diameter tubular SOFC with the length of 1.2 cm.