Development of internal transcribed spacer regions amplification restriction fragment length polymorphism method and its application in monitoring the population of Zygosaccharomyces rouxii M2 in miso fermentation

Recently, the use of the dry yeast of Zygosaccharomyces rouxii M2 for miso (soybean paste) fermentation has been established. A molecular monitoring method was developed and validated in this study to analyze the population of Z. rouxii M2 during the fermentation. The method was based on the restriction patterns of internal transcribed spacer (ITS) regions of the rDNA using HaeIII and HhaI. Among the homologous ITS regions of Z. rouxii strains, Z. rouxii M2 produced diagnostic bands by which it can be differentiated from the other strains used. The specific restriction bands were due to the difference in nucleotide sequence of two different copies of ITS of Z. rouxii M2. Both ITS copies showed 94% sequence similarity but a 13-bp nucleotide substitution and a 19-bp deletion were found in the ITS1 region. Phylogenetic trees were constructed based on ITS and 18S rDNA sequences and it was found that the ITS sequences provide better resolution for the classification of Z. rouxii M2. Since Z. rouxii M2 is a promising strain for use in miso fermentation as a dry starter, the method developed is significant in terms of industrial application in monitoring the growth of Z. rouxii M2 in miso fermentation.     

4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) production in simple media by lactic acid bacterium, Lactococcus lactis subsp. cremoris IFO 3427

4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) was detected in simple media prepared with heated sugar and amino acid solution as a result of the Maillard reaction. However, an increase in the amount of HDMF was observed in the media fermented by Lactococcus lactis subsp. cremoris IFO 3427. 4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) was not detected in all the simple media examined in this study.     

Demineralisation of whey by a combination of nanofiltration and anion‐exchange treatment: a preliminary study

Demineralisation is an important process for the utilisation of whey in the food industry. Sodium (Na⁺) and potassium (K⁺) ions pass through nanofiltration (NF) membranes in ion pairs with chloride (Cl^?) to maintain electroneutrality, but NF processing can only partially remove these ions from whey, because the total Na and K content of whey generally exceeds the Cl content. In this work, a preliminary study of a new demineralisation method was made. A reconstituted sweet whey powder solution was first treated by Cl‐form anion‐exchange resin to increase the molar ratio of Cl to Na and K from ~0.5 to ~1.0. Subsequently, NF removed >90% of Na and K from the anion‐exchange treated whey compared with <60% from the control whey. This new process could be useful for producing 70% demineralised whey with very low levels of the monovalent ions Na⁺, K⁺ and Cl^?.     

Non‐homologous end joining‐mediated functional marker selection for DNA cloning in the yeast Kluyveromyces marxianus

The cloning of DNA fragments into vectors or host genomes has traditionally been performed using Escherichia coli with restriction enzymes and DNA ligase or homologous recombination‐based reactions. We report here a novel DNA cloning method that does not require DNA end processing or homologous recombination, but that ensures highly accurate cloning. The method exploits the efficient non‐homologous end‐joining (NHEJ) activity of the yeast Kluyveromyces marxianus and consists of a novel functional marker selection system. First, to demonstrate the applicability of NHEJ to DNA cloning, a C‐terminal‐truncated non‐functional ura3 selection marker and the truncated region were PCR‐amplified separately, mixed and directly used for the transformation. URA3⁺ transformants appeared on the selection plates, indicating that the two DNA fragments were correctly joined by NHEJ to generate a functional URA3 gene that had inserted into the yeast chromosome. To develop the cloning system, the shortest URA3 C‐terminal encoding sequence that could restore the function of a truncated non‐functional ura3 was determined by deletion analysis, and was included in the primers to amplify target DNAs for cloning. Transformation with PCR‐amplified target DNAs and C‐terminal truncated ura3 produced numerous transformant colonies, in which a functional URA3 gene was generated and was integrated into the chromosome with the target DNAs. Several K. marxianus circular plasmids with different selection markers were also developed for NHEJ‐based cloning and recombinant DNA construction. The one‐step DNA cloning method developed here is a relatively simple and reliable procedure among the DNA cloning systems developed to date. Copyright © 2013 John Wiley & Sons, Ltd.     

Study on the Difference Between the Triple-Point Temperatures of ^{20}Ne and ^{22}Ne Using Sealed Cells

At the National Metrology Institute of Japan (NMIJ), the triple points of \(^{20}\) Ne and \(^{22}\) Ne were realized using modular sealed cells, manufactured by the Istituto Nazionale di Ricerca Metrologica (INRiM) in Italy to measure the difference of the triple-point temperatures of \(^{20}\) Ne and \(^{22}\) Ne. Standard platinum resistance thermometers (SPRTs) were used that were calibrated by NMIJ on the International Temperature Scale of 1990 (ITS-90). In previous reports, sealed cells of \(^{20}\) Ne and \(^{22}\) Ne were mounted one at a time in a cryostat and their triple points were realized in separate cool-downs (the single-cell measurement). In this study, first, the triple point was realized using the single-cell measurement for \(^{20}\) Ne and \(^{22}\) Ne cells. Second, the \(^{20}\) Ne and \(^{22}\) Ne cells were mounted together on the same copper block and their triple points were realized subsequently one after the other in the same cool-down of the cryostat (the double-cell measurement). The melting curves observed by the single-cell and the double-cell measurements were almost identical for each cell. The difference of the triple-point temperatures between the two cells, \(^{22}T -^{20}\!T\) , was estimated, not only using the subrange of SPRTs defined in the ITS-90 from 13.8033 K to 273.16 K (subrange 1) but also that defined from 24.5561 K to 273.16 K (subrange 2). The difference in \((^{22}T-^{20}\!\!T)\) between the subranges 1 and 2 is within 0.06 mK, which is caused by the subrange inconsistency in the ITS-90. The standard uncertainty in \((^{22}T-^{20}\!T)\) due to the subrange inconsistency is estimated to be 0.017 mK. After correction for the effects of impurities and other isotopes in the \(^{20}\) Ne and \(^{22}\) Ne cells, the difference in the triple-point temperatures between pure \(^{20}\) Ne and pure \(^{22}\) Ne is estimated to be 0.146 64 (5) K on subrange 1, which is consistent within the uncertainty with the former studies. When \(^{22}T-^{20}\!T\) for pure \(^{20}\) Ne and pure \(^{22}\) Ne is estimated on subrange 2, \(^{22}T-^{20}\!\!T\) becomes 0.146 60 (5), which agrees very well with the former reports of INRiM evaluating \(^{22}T-^{20}\!T\) on subrange 2.     

Growth and characterization of indium phosphide nanowires on transparent conductive ZnO:Al films

The epitaxial growth of indium phosphide nanowires (InP NWs) on transparent conductive aluminum-doped zinc oxide (ZnO:Al) thin films is proposed and demonstrated. ZnO:Al thin films were prepared on quartz substrates by radio frequency magnetron sputtering, then InP NWs were grown on them by plasma enhanced metal organic chemical vapor deposition with gold catalyst. Microstructure and optical properties of InP nanowires on ZnO:Al thin films were investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectric spectroscopy (XPS), photoluminescence and Raman spectroscopy at room temperature. SEM shows that randomly oriented and intersecting InP nanowires were grown to form a network on ZnO:Al thin films. Both wurtzite (WZ) and zincblende (ZB) structures coexist in the random orientation InP NWs on ZnO:Al thin film had been proved by XRD analysis. XPS result indicates Zn diffusion exists in the InP NWs on ZnO:Al. The photoluminescence spectra of InP nanowires with Zn diffusion present an emission at 915 nm. Zn diffusion also bring effect on Raman spectra of InP NWs, leading to more Raman-shift and larger relative intensity ratio of TO/LO.     

甜菜红色素的色谱行为及其波谱表征

目的分离与分析甜菜红色素成分.方法通过HPLC法探讨甜菜红色素在不同洗脱液(乙腈-甲酸、乙腈-乙酸)浓度、不同柱温下0DS柱上的色谱行为;通过LC-ESI-MS和UV光谱鉴定甜菜红色素成分.结果推导出甜菜主要红色素成分间分辨率与酸浓度、乙腈浓度、柱温间的相互关系方程为YRe=6.3931-0.3868Xc-0.2155Xac-0.0384Xt(乙酸)和YRe=5.0501-0.0249Xc-0.1444Xac-0.0236Xt(甲酸);获得了甜菜红色素主要单组分制备物,其主成分为甜菜苷和异甜菜苷,此外,还鉴定出17-脱羧-新甜菜苷等组分.结论色谱分离关系方程和主成分表征对甜菜红色素具有质控意义.     

Helically π‐Stacked Conjugated Polymers Bearing Photoresponsive and Chiral Moieties in Side Chains: Reversible Photoisomerization‐Enforced Switching Between Emission and Quenching of Circularly Polarized Fluorescence

Novel multifunctional conjugated polymers, [poly(p‐phenylene)s and poly(bithienylene‐phenylene)s with (R)‐ and (S)‐configurations], which have fluorescence, chirality, and photoresponsive properties, have been designed and synthesized. The polymers are composed of π‐conjugated main chains, where poly(p‐phenylene) and poly(bithienylene‐phenylene) are fluorescence moieties, and the side chains of the photochromic dithienylethene moiety are linked with chiral alkyl groups. The polymer films exhibit right‐ or left‐handed circularly polarized fluorescence (CPF) and also show reversible quenching and emitting behaviors as a result of photochemical isomerization of the dithienylethene moiety upon irradiation with ultraviolet and visible light. This is the first report realizing the reversible switching of CPF using chirality and photoresponsive properties.     

Thermal conductance calculations of silicon nanowires: comparison with diamond nanowires

We present phonon thermal conductance calculations for silicon nanowires (SiNWs) with diameters ranging from 1 to 5 nm with and without vacancy defects by the non-equilibrium Green’s function technique using the interatomic Tersoff-Brenner potentials. For the comparison, we also present phonon thermal conductance calculations for diamond nanowires. For two types of vacancy defects in the SiNW, a ‘center defect’ and a ‘surface defect’, we found that a center-defect reduces thermal conductance much more than a surface defect. We also found that the thermal conductance changes its character from the usual behavior, in proportion to the square of diameter (the cross-sectional area) for over 100 and 300 K, to the unusual one, not dependent on its diameter at all at low temperature. The crossover is attributed to the quantization of thermal conductance.     

Smoothing of PV system output by tuning MPPT control

A PV system's power output is not stable and fluctuates depending on weather conditions. Using a battery is one feasible measure to stabilize a PV system's power output, but it requires additional costs and results in additional waste of used batteries. In this paper, we propose a new measure for smoothing the short‐term change in a PV system's power output, which is performed by tuning the characteristics of Maximum Power Point Tracking (MPPT) control without additional equipment. In our proposed measure, when the insolation increases rapidly, the operation point of MPPT control changes to a new point where the maximum power is not generated with the current insolation, so that the rate of increase in the PV system's power output is limited to a certain value. In order to evaluate the effect of the proposed measure on the operation of electric power systems, we evaluated the required capacity of generators for load‐frequency control (LFC). It was revealed that the additional LFC capacity is not required even in the case where penetration of PV systems reaches 5% of the utility system's total capacity if the proposed measure is applied to all PV systems. © 2005 Wiley Periodicals, Inc. Electr Eng Jpn, 152(2): 10–17, 2005; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20106