Thermostable ATP regeneration system using polyphosphate kinase from Thermosynechococcus elongatus BP-1 for D-amino acid dipeptide synthesis

D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 (TmDdl) was a useful biocatalyst for synthesizing D-amino acid dipeptides. TmDdl showed a broad substrate specificity at a high temperature; however, ATP was required for its reaction. One of the methods for an effective ATP supply was the coupling reaction with an ATP regeneration system. However, ATP regeneration systems consisted of enzymes from mesophiles and were difficult to operate at high temperatures. Therefore, an ATP regeneration system that could be used at high temperatures was desired to utilize TmDdl for the effective production of D-amino acid dipeptides. To establish a thermostable ATP regeneration system, polyphosphate kinase from a thermophile, Thermosynechococcus elongatus BP-1 (TePpk), was characterized. TePpk showed thermostability up to 70 degrees C; therefore, it was considered that a thermostable ATP regeneration system could be established using TePpk. In the coupling reaction with purified TmDdl and TePpk at 60 degrees C, the amount of ATP required for D-alanyl-D-alanine synthesis could be reduced to 1% of the theoretical amount required when there was no ATP regeneration. When the coupling reaction was applied to a resting cell reaction, ATP was regenerated from an adenosine scaffold in the cell, and D-alanyl-D-alanine was successfully synthesized in the maximum yield of 80% (mol/mol) without the addition of ATP. Thus, an effective synthesis of D-amino acid dipepitides was achieved using the thermostable ATP regeneration system.     

Energy supply performance and environmental load‐reduction effect of a cogeneration system for an apartment building using distributed heat storage technology

In order to make distributed generation systems for apartment buildings economically viable, it is essential to develop an efficient and low‐cost heat supply system. We are developing a new cogeneration system (Neighboring CoGeneration system: NCG system). The key concept of this system is to install a heat storage unit for the hot water supply, floor heating, and bath heating in each house, and to connect the heat storage units by a single‐loop hot water pipe. The system leads to time leveling of the total heat supply and reduced installation costs. Furthermore, it is expected that the cogeneration can operate according to electrical demand because of the large heat storage capacity of the system. In this study, a dynamic simulation model is developed to evaluate the performance and environmental load‐reduction effect of the NCG system for 50 households. The results show that the NCG system can supply sufficient heat for peak demand in winter and reduce annual CO₂ emissions by 23% on average. © 2013 Wiley Periodicals, Inc. Heat Trans Asian Res, 42(8): 745–757, 2013; Published online in Wiley Online Library ( wileyonlinelibrary.com/journal/htj ). DOI 10.1002/htj.20376     

Studies on fish scale collagen of Pacific saury (Cololabis saira)

We purified and characterized Type I collagen from the scales of the Pacific saury (Cololabis saira) and compared it with collagen from other organisms. Subunit composition of C. saira collagen (2α1 + α2) was similar to that of red sea bream (Pagrus major) and porcine collagen. C. saira collagen did not form a firm gel after neutralization of pH in solution. The temperature of denaturation (24–25 °C) of C. saira collagen was slightly lower than that of P. major collagen (26–27 °C). The contents of proline and hydroxyproline were lower in red sea bream and Pacific saury collagen than in porcine collagen. Circular dichroism spectra and Fourier-transformed infrared spectra showed that heat denaturation caused unfolding of the triple helices in all three collagens.     

Sewage Sludge Dewatering Process Using Liquefied Dimethyl Ether as Solid Fuel

This article describes the dewatering of sewage sludge using liquefied dimethyl ether (DME), which clarified the effect of sludge type and its particle size. Changes in the heating value of the dewatered sludge were also examined. In addition, the change in dewaterability upon reuse of the liquefied DME was investigated experimentally.

Regardless of the nature of the sludge, its dewatering behavior was similar. The amount of dewatered water could be increased by decreasing the diameter of the sludge cake. Complete drying of the sludge cake via DME dewatering resulted in retention of approximately 96% of the heating value (17.7 MJ/kg-sludge) of the original sludge cake dry base (18.4 MJ/kg db). The DME could be recovered after extraction, and reuse experiments showed that liquefied DME dewatering performance was unchanged after five reuses.     

4‐Hydroxy‐3‐methyl‐6‐(1‐methyl‐2‐oxoalkyl)pyran‐2‐one Synthesis by a Type III Polyketide Synthase from Rhodospirillum centenum

The purple photosynthetic bacterium Rhodospirillum centenum has a putative type III polyketide synthase gene (rpsA). Although rpsA was known to be transcribed during the formation of dormant cells, the reaction catalyzed by RpsA was unknown. Thus we examined the RpsA reaction in vitro, using various fatty acyl‐CoAs with even numbers of carbons as starter substrates. RpsA produced tetraketide pyranones as major compounds from one C_10–14 fatty acyl‐CoA unit, one malonyl‐CoA unit and two methylmalonyl‐CoA units. We identified these products as 4‐hydroxy‐3‐methyl‐6‐(1‐methyl‐2‐oxoalkyl)pyran‐2‐ones by NMR analysis. RpsA is the first bacterial type III PKS that prefers to incorporate two molecules of methylmalonyl‐CoA as the extender substrate. In addition, in vitro reactions with ¹³C‐labeled malonyl‐CoA revealed that RpsA produced tetraketide 6‐alkyl‐4‐hydroxy‐1,5‐dimethyl‐2‐oxocyclohexa‐3,5‐diene‐1‐carboxylic acids from C_14–20 fatty acyl‐CoAs. This class of compounds is likely synthesized through aldol condensation induced by methine proton abstraction. No type III polyketide synthase that catalyzes this reaction has been reported so far. These two unusual features of RpsA extend the catalytic functions of the type III polyketide synthase family.     

Effects of Impurity Iron Content on Characteristics of Sintered Reaction‐Bonded Silicon Nitride

The effects of impurity iron content on characteristics of sintered reaction‐bonded silicon nitrides were examined by adding iron powder to a high purity raw Si powder. Powder compacts of the raw Si powder doped with 2 mol% Y₂O₃ and 5 mol% MgSiN₂ as sintering additives and Fe as impurity (0 mass%, 0.1 mass%, 1.0 mass% and 5.0 mass%) were nitrided at 1400°C for 8 h under a N₂ pressure of 0.1 MPa, followed by post‐sintering at 1900°C for 6 h under a N₂ pressure of 0.9 MPa. All the SRBSN (Sintered Reaction‐Bonded Silicon Nitride) specimens had about the same 4‐point bending strength of 730–770 MPa. The fracture toughness of the specimens was gradually decreased with increasing Fe additive amount due to the inhibition of development of rodlike β‐Si₃N₄ grains by SiFe_x particles formed during nitridation process. The thermal conductivity was also decreased with an increase in Fe amount. It seems that the increasing oxygen in grain‐boundary phase caused by the oxidation of Fe during milling resulted in the increase in lattice oxygen of β‐Si₃N₄ grains, which caused phonon scattering and thereby decreased thermal conductivity of β‐Si₃N₄. There was little change in the dielectric breakdown strength of the specimens: 24, 22, 22, and 21 kV/mm for the specimens without Fe, and with 0.1 mass%, 1.0 mass% and 5.0 mass% Fe, respectively. The surface resistivity of the specimens with 0 mass%, 0.1 mass% and 1.0 mass% Fe was in the range of 10¹³ Ω, but the specimen with 5 mass% Fe was about one order lower than the others.     

Synthesis of Well‐Defined Diphenylvinyl(cyclopropyl)phosphine‐Palladium Complexes for the Suzuki–Miyaura Reaction and Buchwald–Hartwig Amination

The well‐defined diphenylvinylphosphine‐palladium complex 1 and the diphenylcyclopropylphosphine‐palladium complex 2 were successfully synthesized. The crystal structures of these complexes were obtained by X‐ray crystallographic analysis. Both complexes were air‐ and moisture‐stable, and could be prepared on a gram scale. These palladium complexes catalyzed the Suzuki–Miyaura reaction of aryl bromides [turnover numbers (TON) up to 196,000] and aryl chlorides (TON up to 50,000). Furthermore, complex 2 catalyzed the Buchwald–Hartwig amination of aryl chlorides and aromatic/aliphatic amines with a low catalyst loading. These complexes showed different reactivities for the coupling of 2‐chloropyridine, and the origin of this difference is discussed.     

Niche Differentiation of Phenol-Degrading Microorganisms in UASB Granular Sludge as Revealed by Fluorescence in situ Hybridization

A microbial community structure of granules harvested from an anaerobic sludge blanket reactor treating phenolic wastewater was investigated using fluorescence ...     

Acetate and ethanol production from H2 and CO2 by Moorella sp. using a repeated batch culture

The growth inhibition of Moorella sp. HUC22-1 by undissociated acetic acid was analyzed using a non-competitive inhibition model coupled with a pH inhibition model. In the cells grown on H2 and CO2, the inhibition constant, K(p) of the undissociated acetic acid was 6.2 mM (164 mM as the total acetate at pH 6.2, pKa = 4.795, 55 degrees C), which was 1.5-fold higher than that obtained in cells grown on fructose. When a pH-controlled batch culture was performed using a fermentor at pH 6.2 with H2 and CO2, a maximum of 0.92 g/l of dry cell weight and 339 mM of acetate were produced after 220 h, which were 4.4- and 6.8-fold higher than those produced in the pH-uncontrolled batch culture, respectively. In order to reduce acetate inhibition in the culture medium, a repeated batch culture with cell recycling was performed at a constant pH with H2 and CO2. At a pH of 6.2, the total acetate production reached 840 mmol/l-reactor with 4.7 mmol/l-reactor of total ethanol production after 420 h. When the culture pH was maintained at 5.8, which was the optimum for ethanol production, the total ethanol production reached 15.4 mmol/l-reactor after 430 h, although the total acetate production was decreased to 675 mmol/l-reactor.