記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER  

Vacuum foam drying offers great perspectives to formulate solid-state encapsulated active drugs. Taking into account the specific need of pharmaceutical formulations to keep drug molecules active and dispersed, we show in this paper that vacuum drying of pharmaceutical formulations can be substantially improved by using original rheological approaches. Typically, our formulations totally dried-up in less than 5 min, a delay much shorter than the long hours of the traditional vacuum drying process. Steady-shear rheology was used to evaluate the solution's specific viscosity against the solutions' concentrations, in order to correlate the dilution regime to the foamabiltiy. This rheological approach indicated that in miscible solutions, spontaneous foaming occurred in the semi-dilute entangled regime, near the transition to the concentrated regime; for partially miscible solutions, it occurred in solutions at a concentration near to the percolation concentration. The proposed methodology is versatile, and should provide a simple way to assess the foamability of pharmaceutical formulations.

DOI： 10.1016/j.colsurfa.2021.127174

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.apt.2021.08.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier {BV}  

DOI： 10.1016/j.colsurfa.2021.127137

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.jfoodeng.2020.110325

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Informa UK Limited  

DOI： 10.1080/07373937.2020.1822863

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掲載種別：研究論文（学術雑誌）   出版者・発行元：American Chemical Society (ACS)  

DOI： 10.1021/acs.langmuir.0c00695

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Informa UK Limited  

DOI： 10.1080/07373937.2020.1752711

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.jbiosc.2019.09.008

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Royal Society of Chemistry (RSC)  

<p>A picoliter enzyme reactor using a trypsin immobilized nanochannel realized 25 times faster reaction than the bulk reaction.</p>

DOI： 10.1039/d0an00998a

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記述言語：英語   掲載種別：研究論文（国際会議プロシーディングス）   出版者・発行元：Chemical and Biological Microsystems Society  

The Needs of shotgun proteomics have been increasing in the field of biology and medicine for understanding biological systems. In study, protein sample with pL, which volume was same scale as single cell volume, was successfully digested and separated and detected by integrated device of pL enzyme reactor fL chromatography and UV-POPS detector. This nanofluidic device will greatly contribute to single cell shotgun proteomics.

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.xphs.2019.01.008

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記述言語：英語   掲載種別：研究論文（国際会議プロシーディングス）   出版者・発行元：Chemical and Biological Microsystems Society  

We development a nanofluidic enzyme reactor exceeding the limitation of bulk enzyme reaction. The limit concentration of bulk trypsin solution is approximately 0.5 PM because self-digestion occurs over 0.5 PM solution, which complicates analysis of target proteins by digestion trypsin itself. Using extremely high surface/volume ration of nanospace, trypsin concentration, which indicates number of immobilized trypsin molecules on the nanochannel surface for liquid phase volume, exceed to bulk limit of concentration. As a result, 31 times higher concentration and 33 times faster enzyme reaction compared to bulk was realized. This reactor will be powerful tools which overcomes limitation in bulk and microfluidic method.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier B.V.  

The effect of the properties of a protein on its adsorption to a metal surface in the presence of external electric potential was investigated. Protein adsorption processes at different surface potentials were measured for fifteen types of proteins using an in-situ ellipsometry. The tested proteins were classified into three groups, based on the amount of protein that was adsorbed as a function of the surface potential: In First group of proteins, an increasing trend for the amount adsorbed with a more positive surface potential was found
The amount adsorbed of α-chymotrypsinogen A and ribonuclease A (Second group) were roughly constant and independent of the applied surface electric potentials
In Third group, the amount adsorbed decreased with increasing surface potential. This protein classification was correlated with the isoelectric points of the proteins (First group: ≤9.3
Second group: 9.3–10
Third group: &gt
10). Increasing the pH positively and negatively shifted the surface potentials, allowing ß-lactoglobulin (First group) and lysozyme (Third) to become adsorbed, respectively. The surface potential range for protein adsorption was also markedly shifted depending on the metal substrate type. These findings were interpreted based on the electrostatic interactions among the protein, surface hydroxyl groups, and the applied external electric field.

DOI： 10.1016/j.colsurfb.2018.03.035

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Enzymatic cleaning is a potentially useful method for removing proteinaceous fouling from solid surfaces under mild conditions. Herein, the influence of an external electric field on the enzymatic cleaning of a metal surface fouled with a protein was investigated. The model fouling protein (BSA; lysozyme) was prepared on a stainless steel (St) surface, and the resulting surface subjected to enzymatic cleaning with an electric potential being applied to the St plate. Trypsin, alpha-chymotrypsin, and thermolysin were used as model proteases. The amounts of protein remaining on the plate before and during the cleaning process were measured by means of a reflection absorption technique using Fourier transform infrared spectroscopy. In the case for BSA fouling, the cleaning efficiency of the protease tended to increase at more negative applied potentials. Whereas, there was an optimum applied potential for removing the lysozyme fouling. Atomic force microscopy analyses indicated that applying an adequate range of electric potential enhanced the enzymatic removal of protein fouling inside scratches on the St plate surface. These findings suggest the existence of two modes of electrostatic interactions for the external electric field, one with protease molecules and the other with digested fragments of the fouling protein. (C) 2017 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.colsurfb.2017.07.074

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

Protein-stabilizing characteristics of sixteen proteins during freeze-thawing and freeze-drying were investigated. Five enzymes, each with different instabilities against freezing and dehydration, were employed as the protein to be stabilized. Proteinaceous additives generally resulted in greater enzyme stabilization during freeze-thawing than sugars while the degree of stabilization for basic lysozyme and protamine were inferior to that of neutral and acidic proteins. Freeze-drying-induced inactivation of enzyme was also reduced by the presence of a proteinaceous additive, the extent of which was lower than that for a sugar. In both freeze thawing and freeze drying, the enzymes stabilization by the proteinaceous additive increased with increasing additive concentration. The enhancement of enzyme inactivation caused by pH change was also reduced in the presence of proteinaceous additives. The combined use of a sugar such as sucrose and dextran tended to increase the stabilizing effect of the proteinaceous additive.

DOI： 10.1080/09168451.2016.1274637

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

The technique for homogeneously dispersing hydrophobic drugs in a water-soluble solid matrix (solid dispersion) is a subject that has been extensively investigated in the pharmaceutical industry. Herein, a novel technique for dispersing a solid, without the need to use a surfactant, is reported. A freeze-dried amorphous sugar sample was dissolved in an organic solvent, which contained a soluble model hydrophobic component. The suspension of the sugar and the model hydrophobic component was vacuum foam dried to give a solid powder. Four types of sugars and methanol were used as representative sugars and the organic medium. Four model drugs (indomethacin, ibuprofen, gliclazide, and nifedipine) were employed. Differential scanning calorimetry analyses indicated that the sugar and model drug (100:1) did not undergo segregation during the drying process. The dissolution of the hydrophobic drugs in water from the solid dispersion was then evaluated, and the results indicated that the C-max and AUC(0-60) (min) of the hydrophobic drug in water were increased when the surfactant-free solid dispersion was used. Palatinose and/or alpha-maltose were superior to the other tested carbohydrates in increasing C-max and AUC(0-60 min), for all tested model drugs, and the model drug with a lower water solubility tended to exhibit a greater extent of over-dissolution.

DOI： 10.1021/acs.molpharmaceut.6b01048

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DOI： 10.1016/j.colsurfb.2016.07.042

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DOI： 10.1016/j.nbt.2016.06.966

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

The creation of patterned structures with micro/nano-scale lengths on flat surfaces has recently been emerged as an important technique in various fields. In this report, we present a simple method to create nanopatterns of 3-aminopropyltriethoxysilane (APS) on glass substrates by the combination of colloid lithography and vapor deposition. When deposition was conducted at room temperature under natural humidity, an array of nanorings was formed on the substrate by the condensation of APS vapor on the water bridges remaining under the particles. By varying only the deposition temperature, the structure of the ordered arrays could easily be transformed: nanorings were converted to honeycomb and dot like structures by increasing temperature. This transformation proposedly occurred by the condensation and polymerization of APS vapor through the deformed particles of the colloidal monolayer. We also fabricated a patterned polymer brush array and a pore array using the obtained APS nanopattern as a template. (C) 2016 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.colsurfa.2016.01.047

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCI LTD  

A solid dispersion technique to homogeneously disperse hydrophobic ingredients in a water-soluble solid without using surfactant was examined as follows: first, freeze-dried amorphous sugar was dissolved in an organic medium that contained a soluble model hydrophobic component. Second, the mixed solution of sugar and the model hydrophobic component was vacuum dried into a solid (solid dispersion). Methanol and six fat-soluble flavours, including cinnamaldehyde, were used as organic media and model hydrophobic components. The retention of flavours in the solid dispersion during drying and storage under vacuum was evaluated. The amorphised disaccharides dissolved in methanol up to 100 mg/mL, even temporarily (20 s to 10 days) and could be solidified without any evidence of crystallisation and segregation from flavour. The solid dispersion, prepared using a-maltose usually showed 65-95% flavour retention during drying (and storage for cinnamaldehyde), whereas &gt;= 50% of the flavour was lost when the flavour was O/W emulsified with a surfactant and then freeze-dried with sugar. (C) 2015 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.foodchem.2015.11.097

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/btpr.2232

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記述言語：日本語   出版者・発行元：公益社団法人 日本表面科学会  

ELISA，SPRやQCMなどのバイオ分子間相互作用検出系における基板上へのリガンド分子の固定化について，基板親和性ペプチド連結による固定化配向制御ならびにリガンド分子と基板との直接的な相互作用を緩和するクッションタンパク質を用いた機能維持を試みた．そして，クッションタンパク質の様々な分子デザインにより，ペプチド-タンパク質間あるいはタンパク質-タンパク質間相互作用検出を顕著に高感度化できた．

DOI： 10.14886/sssj2008.36.0_37

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DOI： 10.1016/j.apt.2015.10.017

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCI LTD  

An amorphous matrix of a sugar is frequently used as a bulk-forming and stabilizing agent in the food industry but tends to crystallize as the result of water uptake and increase in temperature. Additives and methods used to inhibit the crystallization of amorphous sugar (sucrose) were screened in this study. Freeze-dried amorphous sucrose containing 0.5-5 wt% of additive, including salts, different types of sugars, and polymers, the crystallization temperature (T-cry) and isothermal crystallization characteristics were examined. Certain types of salts markedly increased the Tcry and prolonged the induction period for crystal nucleation. The use of 1 wt% MgCl2 was particularly effective in inhibiting sugar crystallization. The heat treatment of crystalline sucrose under appropriate conditions was also found to result in diminished sucrose crystallization. MALDI-TOF mass spectra of the heat-treated sucrose suggested that sucrose derivatives containing multiple pyranose groups were formed, which would closely relate to the crystallization inhibition. Finally, the protein stabilizing effects of the matrices were evaluated. The results indicated that both the addition of additives and the heat treatment resulted in an improvement of the protein stabilizing effect of amorphous sugar matrix, compared to that of sucrose alone. (C) 2015 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.jfoodeng.2015.01.016

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

The encapsulation of O/W emulsion droplets in a freeze-dried amorphous sugar matrix was investigated, focusing on the impact of the molecular structure of the emulsifying surfactant. O/W emulsions, containing various surfactants, were freeze-dried in the presence of a sugar. Thirty types of surfactants, including eighteen different sugar surfactants and ten types of commercially available sugar ester mixtures, were used. Linoleic acid methyl ester and trehalose were used as the oil phase and sugar. The amounts of oil droplets encapsulated in freeze-dried amorphous sugar matrix were analyzed by Fourier transform Infrared spectroscopy. Sugar surfactants were generally superior to the other classes of surfactants for oil droplet encapsulation during freeze-drying, and there was the optimum alkyl chain length of the sugar surfactant. Sugar esters generally exhibited greater oil encapsulation than sugar ethers. Larger sugar head group appeared to result in better encapsulation in the case of sugar esters, but the opposite tendency was found for sugar ethers. A limited combination of sugar surfactants (15% sucrose mono- and 85% di-stearate) resulted in the maximum oil droplet encapsulation efficiency although these surfactants are individually quite poor in the encapsulation and other tested combinations did not improve the encapsulation efficiency relative to their individual effectiveness. (C) 2015 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.foodres.2015.02.003

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/jps.23988

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC GENERAL MICROBIOLOGY  

A Gram-stain-negative, non-spore-forming, aerobic, oligotrophic bacterium (strain 262-7(T)) was isolated from a crack of white rock collected in the Skallen region of Antarctica. Strain 262-7(T) grew at temperatures between -4 and 30 degrees C, with optimal growth at 25 degrees C. The pH range for growth was between pH 6.0 and 9.0, with optimal growth at approximately pH 7.0. The NaCl concentration range allowing growth was between 0.0 and 1.0%, with an optimum of 0.5%. Strain 262-7(T) showed an unprecedented range of morphological diversity in response to growth conditions. Cells grown in liquid medium were circular or ovoid with smooth surfaces in the lag phase. In the exponential phase, ovoid cells With short projections were observed. Cells in the stationary phase possessed long tentacle-like projections intertwined intricately. By contrast, cells grown on agar plate medium or in liquid media containing organic compounds at low concentration exhibited short- and long-rod-shaped morphology. These projections and morphological variations clearly differ from those of previously described bacteria. Ubiquinone 10 was the major respiratory quinone. The major fatty acids were C-17:1 omega 6c (28.2%), C-16:1 omega 7c (22.6%), C-18:1 omega 7c (12.9%) and C(15:0)2-OH (12.3%). The G+C content of genomic DNA was 68.0 mol%. Carotenoids were detected from the cells. Comparative analyses of 16S rRNA gene sequences indicated that strain 262-7(T) belongs to the family Sphingomonadaceae, and that 262-7(T) should be distinguished from known genera in the family Sphingomonadaceae. According to the phylogenetic position, physiological characteristics and unique morphology variations, strain 262-7(T) should be classified as a representative of a novel genus of the family Sphingomonadaceae. Here, a novel genus and species with the name Polymorphobacter multimanifer gen. nov., sp. nov. is proposed (type strain 262-7(T)=JCM 18140(T)=ATCC BAA-2413(T)). The novel species was named after its morphological diversity and formation of unique projections.

DOI： 10.1099/ijs.0.050005-0

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記述言語：日本語   出版者・発行元：粉体工学会  

DOI： 10.4164/sptj.51.343

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記述言語：英語   出版者・発行元：一般社団法人 日本生物物理学会  

DOI： 10.2142/biophys.54.S209_1

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記述言語：英語   出版者・発行元：一般社団法人 日本生物物理学会  

DOI： 10.2142/biophys.54.S207_6

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

Amorphous matrices, composed of sugars, are markedly plasticized by moisture uptake, which results in physical instability. Our previous studies, in the compression pressure range 443 MPa, indicated that when a matrix is compressed, the amount of sorbed water at given relative humidities (RHs) decreases, whereas the glass transition temperature (Tg) remains constant. Herein, the effect of higher compression pressures than those used previously was explored to investigate the feasibility of using compression to improve the physical stability of amorphous sugar matrix against water uptake and subsequent collapse. Amorphous sugar samples were prepared by freeze-drying and then compressed at 0-665 MPa, followed by rehumidification at given RHs. The physical stability of the amorphous sugar sample was evaluated by measuring Tg and crystallization temperature (Tcry). The amounts of sorbed water, different in the interaction state, were determined using an FTIR technique. It was found that the compression at pressures of 443 MPa decreased the amount of sorbed water, which is a major factor in plasticization and crystallization, and thus markedly increased the Tg and Tcry relative to that for the uncompressed sample. Hence, the compression at several hundreds MPa appears to be feasible for improving the physical stability of amorphous sugar matrix. (c) 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:2187-2197, 2013

DOI： 10.1002/jps.23568

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

The encapsulation of emulsion oil droplets by amorphous sugar matrices, formed by freeze-drying, was investigated, with a focus on the influence of the type of sugar. An oil-in-water emulsion, comprised of linoleic acid methyl ester (LME) and sucrose monolaurate (SML) as an oil phase and surfactant, respectively, were freeze-dried in the presence of different types of sugars. LME-droplet encapsulation during and after freeze-drying were evaluated by FTIR analysis. The loss of LME largely occurred in the early stage of freeze-drying. The size distribution of the encapsulated LME droplets remained unchanged before and after freeze-drying in most cases. The encapsulated fractions of LME droplets could be correlated with the glass transition temperature of the sugars in the fully hydrated state (T-g*), and the existence of an optimum T-g* value for the sugar matrix was predicted. The encapsulation ability of an amorphous sugar matrix was maximized when mono- and polysaccharide were combined so as to give a value for T-g* of approximately -50 degrees C, although, individually, mono- and polysaccharides were quite poor for oil droplet encapsulation. These findings suggest that the structural flexibility of the amorphous sugar matrix is a major determinant in oil droplet encapsulation by an amorphous sugar matrix during freeze-drying. (C) 2012 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.foodres.2012.12.010

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1271/bbb.120583

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1271/bbb.110014

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1271/bbb.100574

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jbiosc.2010.03.010

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN  

We cultivated a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to elucidate the differences of its behaviors by different cultivation methods under the same media, by measuring the growth, secretion of proteases and alpha-amylase, secreted protein level, and gene transcriptional profile by the DNA microarray analysis. The protease activities detected by MSLC and APC were much higher than that by SFC, using both modified Czapek-Dox (mCD) and dextrin-peptone-yeast extract (DPY) media. The alpha-amylase activity was detected in MSLC and APC in a much larger extent than that in SFC when DPY medium was used. On the basis of SDS-PAGE analyses and N-terminal amino acid sequences, 6 proteins were identified in the supernatants of the culture broths using DPY medium, among which oryzin (alkaline protease) and alpha-amylase were detected at a much higher extent for APC and MSLC than those for SFC while only oryzin was detected in mCD medium, in accordance with the activity measurements. A microarray analysis for the fungi cultivated by SFC, APC, and MSLC using mCD medium was carried out to elucidate the differences in the gene transcriptional profile by the cultivation methods. The gene transcriptional profile obtained for the MSLC sample showed a similar tendency to the APC sample while it was quite different from that for the SFC sample. Most of the genes specifically transcribed in the MSLC sample versus those in the SFC sample with a 10-fold up-regulation or higher were unknown or predicted proteins. However, transcription of oryzin gene was only slightly up-regulated in the MSLC sample and that of alpha-amylase gene, slightly down-regulated. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2009.09.004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JOHN WILEY & SONS INC  

An amorphous matrix comprised of sugar molecules is used as excipient and stabilizing agent for labile ingredients in the pharmaceutical industry. The amorphous sugar matrix is often compressed into a tablet form to reduce the volume and improve handling. Herein, the effect of compression on the crystallization behavior of an amorphous sucrose matrix was investigated. Amorphous sucrose samples were prepared by freeze-drying and compressed under different conditions, followed by analyses by differential scanning calorimetry, isothermal crystallization tests, X-ray powder diffractometry, Fourier transform infrared spectroscopy (FTIR), and gas pycnometry. The compressed sample had a lower crystallization temperature and a shorter induction period for isothermal crystallization, indicating that compression facilitates the formation of the critical nucleus of a sucrose crystal. Based on FTIR and molecular dynamics simulation results, the conformational distortion of sucrose molecules due to the compression appears to contribute to the increase in the free energy of the system, which leads to the facilitation of critical nucleus formation. An isothermal crystallization test indicated an increase in the growth rate of sucrose crystals by the compression. This can be attributed to the transformation of the microstructure from porous to nonporous, as the result of compression. (C) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99:1452-1463, 2010

DOI： 10.1002/jps.21890

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2010f.0.1003.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2010f.0.997.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2010f.0.584.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2010f.0.583.0

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記述言語：英語   出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN  

DOI： 10.1016/j.jbiosc.2009.08.436

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

The cathodic electrolysis of H2O2 (H2O2 + e(-) -&gt; OH- + (OH)-O-center dot) on a metal surface in the presence of calcium and phosphate ions results in the formation of calcium phosphate deposits oil the metal surface. In this Study, the deposits formed under various treatment conditions (pHs, concentrations and ratios of calcium/phosphate ions, and so on) were characterized by scanning electron spectroscopy (SEM). and X-ray diffractometry. The exclusive formation of hydroxyapatite, HAP, was observed under comparatively narrow conditions (pH 3-4, [Ca+]/[PO43-] = 25 mM/15 mM), which is clearly different from the reported conditions for the deposition of HAP on titanium substrates. HAP was deposited in the form of a layer. comprised of morphologically amorphous HAP flakes that were less than 20 nm thick. SEM and FTIR analyses of the deposit at different stages of H2O2-electrolysis revealed that a few dozen nanometer-sized spheres of amorphous calcium phosphate were formed in the first step and then fused with each other to form ribbon-like flakes of HAP or broken glass-like brushite, depending on the pH. The pH for HAP formation oil a stainless steel surface was markedly lower than that used for titanium, and the observed process by which amorphous calcium phosphate is converted to HAP was markedly different from that for the electrochemical deposition (electrolysis of water) of HAP on a titanium substrate. (C) 2009 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.colsurfa.2009.09.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

We report here on the purification, characterization, molecular cloning, and expression of a new aminoacylase, initially isolated from the supernatant of Streptomyces mobaraensis (Sm-AA). Purified wild-type Sm-AA was found to be a monomeric protein with a molecular mass of 55kDa. The cloned gene of Sm-AA contained an ORF of 1,383 bp, encoding a polypeptide of 460 amino acids. A BLAST search revealed that Sm-AA belongs to the peptidase M20 family, with identities to a hypothetical protein from Streptomyces pristinaespiralis, a putative peptidase from Streptomyces averinitilis, peptidase M20 from Frankia sp., succinyl-diaminopimelate desuccinylase from Hemophilus influenzae, and aminoacylase-1 from porcine kidney at 89, 88, 67, 29, and 25% respectively. The Sm-AA gene was subcloned into an expression vector, pSH19, and was expressed in Streptomyces lividans TK24. The amount of the recombinant Sm-AA expressed in the S. lividans cells was approximately 42-fold higher than that of Sm-AA found in the supernatant of S. mobaraensis. Sm-AA showed high hydrolytic activity towards various N-acetyl-L-amino acids and N-(middle/long)-chain-fatty-acyl-L-amino acids, with a preference for the acyl derivatives of L-Met, L-Ala, L-Cys, etc. with an optimum pH and temperature for reaction of about 7.5 and 50 degrees C (at pH 7.5).

DOI： 10.1271/bbb.90081

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

epsilon-Lysine acylase from Streptomyces mobaraensis (Sm-ELA), which specifically catalyzes hydrolysis of the epsilon-amide bond in various N epsilon-acyl-L-lysines, was cloned and sequenced. The Sm-ELA gene consists of a 1617-bp open reading frame that encodes a 538-amino acid protein with a molecular mass of 55,816 Da. An NCBI protein-protein BLAST search revealed that the enzyme belongs to the YtcJ-like metal-dependent amidohydrolase family, which is further characterized as the metallo-dependent hydrolase superfamily. The Sm-ELA gene was ligated into a pUC702 vector for expression in Streptomyces lividans TK24. Expression of recombinant Sm-ELA in S. lividans was approximately 300-fold higher than that in wild-type S. mobaraensis. The recombinant Sm-ELAs from the cell-free extract and Culture supernatant were purified to homogeneity. The specific activities of the purified Sm-ELAs were 2500-2800 U/mg, which were similar to that obtained for the wild-type Sm-ELA. Using the cell-free extract of the recombinant S. lividans cells, N epsilon-lauroyl-L-lysine was synthesized from 500 mM L-lysine hydrochloride and 50, 100, or 250 mM lauric acid in an aqueous buffer Solution at 37 degrees C. The yields were close to 100% after 6 and 9 1) of reaction for 50 and 100 mM lauric acid, respectively. and 90% after 24 h for 250 mM lauric acid. (C) 2009 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jbiotec.2009.03.008

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記述言語：日本語   出版者・発行元：日本生物工学会  

小麦フスマを原料として用い,市販酵素製剤による糖化反応と酵母によるエタノール発酵を併用した効率的エタノール生産方法の構築のために必要なバイオマスの前処理条件および糖化のための酵素の組み合わせなどの検討を行った.糖化反応条件としては,小麦フスマを140℃で保持時間0hの条件下で水熱処理を行い,反応温度45℃でMeiselaseおよびNovozyme^[○!R]188の2種類の酵素製剤を用いて糖化することが,グルコース生成に対して最も効果的であることがわかった.3.3%(w/v)の水熱処理小麦フスマから理論値の90%に相当するグルコースが生成した.しかし,33%(w/v)の水熱処理小麦フスマを用いた場合には,理論値の72%のグルコースしか生成しなかった.この理由として,反応生成物であるグルコースのβ-グルコシダーゼに対する阻害の影響が考えられた.しかし,酵素糖化を45℃で24時間行った後に,酵母を添加し30℃でエタノール発酵を行うと,糖化反応とグルコース消費が併行して起こるために阻害効果が緩和され,その結果,理論値の88%に相当する約5.2%(w/v)のエタノールが生成した.

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その他リンク： http://dl.ndl.go.jp/info:ndljp/pid/10516714

記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2009f.0.934.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2009f.0.1060.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2009f.0.779.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2009.0.606.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2009.0.608.0

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記述言語：英語  

Proteome analysis plays a key role in the elucidation of the functions and applications for numerous proteins. For proteome analyses, various microplate- and microarray-based techniques have been developed by a number of researchers. Their intent was to immobilize proteins on the surface of a solid substrate in a site-directed manner while retaining structure and native biological function. In this review, we focus on recent advances in immobilization methodology for proteins/enzymes on a surface, including those using the affinity peptides screened by random peptide library systems. We also discuss applications of the affinity peptide-mediated immobilization method in fields related to proteome analysis, particularly our recent work concerning immunoassay and protein-protein interaction analysis. ©2008 Bentham Science Publishers Ltd.

DOI： 10.2174/157016408785909622

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JOHN WILEY & SONS INC  

True density of an amorphous matrix represents the state of molecular packing in the matrix, which is closely related to the physical/chemical properties of the material. Dry gas pycnometry is one possible technique for measuring the true density of an amorphous sugar matrix prepared by freeze-drying. We herein report on the influence of conditions used for pycnometry on the measured density value and propose a protocol for obtaining the true density. The technique is sufficiently accurate to permit values for matrices comprised of different types of sugar to be compared. Using the protocol, the true densities of several amorphous sugar samples containing different types of sugar, freeze-drying conditions (temperature and sugar concentration at the time of freezing of an aqueous sugar solution), pretreatment (compaction and grind) were determined and the results were compared. A model for simulating an amorphous matrix of sugar (trehalose) was constructed using molecular dynamics/mechanics calculations, and the true density of the simulated sugar matrix was found to agree with the value experimentally determined using the proposed protocol. The relationship among the true density, the states of intermolecular interactions, and strain of sugar molecules in the matrix are discussed using the simulated amorphous sugar matrix. (C) 2007 Wiley-Liss, Inc.

DOI： 10.1002/jps.21202

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JOHN WILEY & SONS INC  

The impact of a polymer additive (polyvinylpyrrolidone, PVP) on hydrogen bonding in amorphous sugar matrices as well as on the glass transition temperature, T-g, were examined by temperature scanning Fourier transform infrared spectroscopy (TS-FTIR). An amorphous sugar matrix containing PVP was prepared by air-drying an aqueous solution of a sugar-PVP mixture. The hydrogen bonds in the sugar-PVP mixture (sugar-PVP and sugar-sugar hydrogen bonds) were analyzed from the IR peak positions corresponding to the stretching vibration of C=O groups of PVP and O-H groups of the sugar and the temperature dependence of the peak position of the O-H stretching vibration band. The addition of PVP to amorphous mono and disaccharides significantly lowered the extent of hydrogen bond formation while interactions between sugars and the PVP tended to prevent the disruption of hydrogen bonds due to increasing temperature, the magnitude of which was larger for larger oligomers. The T-g value for the amorphous sugar was increased by the addition of PVP in many cases. As the size of sugar molecule became larger, the relative magnitude of the increased T-g by PVP to the difference between the T-g values for sugar alone and PVP alone became larger and then reached a certain level; it was slight in the case of glucose. Collectively, these results demonstrate that the magnitude of the impact of PVP on an amorphous sugar matrix strongly vary and are dependent on the types of sugar. (C) 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:519-528, 2008.

DOI： 10.1002/jps.21075

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008.0.418.0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS INC  

In an amorphous mixture of sugar and polyvinylpyrrolidone (PVP), PVP carbonyl groups form hydrogen bonds with sugar hydroxyl groups, thereby improving the physical stability of the amorphous matrix against a glass-to-rubber transition. Herein, Fourier self-deconvolved IR bands due to the C=O stretching vibration of PVP in sugar-PVP mixtures were analyzed. The C=O groups in sugar-PVP mixtures generally had four vibrational states, corresponding with free and hydrogen-bonded C=O in three different modes. Changes in these vibrational states induced by increasing the temperature were compared among various sugar-PVP mixtures. Formation and thermal disruption characteristics of different modes of sugar-PVP hydrogen bondings are discussed.

DOI： 10.1080/00387010802370959

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008.0.339.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008.0.345.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008.0.378.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008f.0.175.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2008f.0.955.0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

A Ser/Thr phosphatase gene cloned from Aspergillus oryzae, aoppt, revealed that the tetratricopeptide repeat (TPR) and catalytic domains of the full-length AoPPT are located at the N- and C-terminal regions, respectively, similar to those of human Ser/Thr phosphatase 5 (PP5) and yeast Ppt1. Four different regions of AoPPT, namely, a full-length polypeptide, the catalytic domain, the catalytic domain plus C-terminal 15 amino-acid residues and the TPR domain were expressed in Escherichia coli and their roles in dephosphorylation activity were examined, using p-nitrophenyl phosphate as the substrate. The full-length AoPPT showed the highest dephosphorylation activity while the catalytic domain had the lowest activity. The activity of the catalytic domain was not inhibited by the presence of the TPR domain and arachidonic acid did not increase the activity of the full-length enzyme. These findings suggest that the integrity of the entire enzyme would be necessary for its full activity to be expressed. (c) 2007 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.ijbiomac.2007.03.005

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

Penicillin V acylase from Streptomyces mobaraensis (Sm-PVA) showed high acyl-transfer activity in reactions using methyl esters of carboxylic acid (acyl donor) and amino compounds (nucleophile), to produce the corresponding amides. Moreover, Sm-PVA had broad substrate specificity, as indicated by the fact that it catalyzed the efficient synthesis of fl-lactam antibiotics, capsaicin derivatives, and N-fatty-acyl-amino acid/N-fatty-acyl-peptide derivatives.

DOI： 10.1271/bbb.70052

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

We report on the molecular cloning and characterization of penicillin V acylase (PVA) from an actinomycete, Streptomyces mobaraensis (Sm-PVA), which was originally isolated as an acylase that efficiently hydrolyzes the amide bond of various N-fatty-acyl-L-amino acids and N-fatty-acyl-peptides as well as capsaicin (8-methyl-N-vanillyl-6-nonenamide). In addition, the purified Sm-PVA hydrolyzed penicillin V with the highest activity (k(cat)) among the PVAs so far reported, penicillin G, and 2-nitro-5-phenoxyacetamide benzoic acid. The BLAST search revealed that the Sm-PVA precursor is composed of a polypeptide that is characteristic of enzymes belonging to the beta-lactam acylase family with four distinct segments; a signal sequence (43 amino acids), an a subunit (173 amino acids), a linker peptide (28 amino acids), and a beta subunit (570 amino acids). The mature, active Sm-PVA is a heterodimeric protein with alpha and beta subunits, in contrast to PVAs isolated from Bacillus sphaericus and B. subtilis, which have a homotetrameric structure. The amino acid sequence of Sm-PVA showed identities to PVA from S. lavendulae, N-acylhomoserine lactone-degrading acylase from Streptomyces sp., cyclic lipopeptide acylase from Streptomyces sp., and aculeacin A acylase from Actinoplanes utahensis with 68, 67, 67, and 41% identities, respectively. (c) 2007 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jbiotec.2006.12.017

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

A sandwich ELISA method using peptide tags showing a specific affinity to a hydrophilic polystyrene surface (PS-tags), PS 19 composed of RAFIASRRIKRP and KPS19R10 of KRAFIASRRIRRP and a hydrophilic polystyrene (phi-PS) plate was used to analyze protein-protein interactions. An Escherichia coli cysteine synthase complex, in which serine acetyltransferase (SAT) interacts with O-acetylserine sulfhydrylase-A (OASS) was used as a model system. When the interaction was detected by the conventional sandwich ELISA method using a hydrophobic polystyrene (pho-PS) plate, for the exclusive use of ELISA, the signal intensity was barely detectable due to conformational change of the ligand protein, OASS in the adsorbed state. On the contrary, when OASS, genetically fused with PS19 (OASS-PS19) or chemically conjugated with KPS19R10 (OASS-KPS19R10), was immobilized on the phi-PS plate, a high signal intensity was detected. Furthermore, by applying the two-step sandwich ELISA, in which OASS-PS19 or OASS-KPS19R10 formed a complex with SAT in the blocking solution before immobilization on the phi-PS plate, the signal intensity was further increased with a much shorter operational time, because SAT in the blocking solution formed a complex with OASS-PS19 or OASS-KPS19R10 without any steric hindrance. (c) 2006 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jbiotec.2006.09.018

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Glutathione S-transferase genetically fused with an affinity peptide tag, PS 19 (RAFIASRRIKRP) having a specific affinity for a hydrophilic polystyrene (PS) surface, was preferentially immobilized on a hydrophilic PS (phi-PS) plate without suffering from interference by coexisting protein molecules. Furthermore, rabbit IgG chemically conjugated with a peptide, KPS 19R10, in which (10)Lys in PS 19 was replaced with Arg and one Lys residue was added at the N-terminus as a coupling site for glutaraldehyde, showed a higher immobilization affinity to the phi-PS plate than that conjugated with the PS 19 peptide. On the basis of these findings, the use of a phi-PS plate and peptide tag-linked ligand proteins permitted a one-step or two-step enzyme-linked immumosorbent assay (ELISA) to be achieved, resulting in a substantial reduction in operational time compared with the conventional ELISA method using a hydrophobic PS (pho-PS) plate, while maintaining a high sensitivity. Furthermore, the sensitivity was increased to a greater extent compared to the conventional ELISA meihod when the one-step ELISA was applied to the detection of bovine insulin in a sandwich mode, due to the reduced number of washing and incubation steps. The method proposed here would be a versatile method for use in various ELISA techniques such as sandwich and competitive ELISAs using an antigen, an antibody and streptavidin that are genetically fused or chemically conjugated with the PS-specific affinity peptide as the ligand protein. (c) 2006 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jbiotec.2006.07.011

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007.0.368.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007.0.126.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007.0.417.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007.0.427.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007f.0.990.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007f.0.702.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007f.0.842.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2007f.0.841.0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN  

We identified 22 genes from Aspergillus oryzae that are preferentially expressed in membrane-surface liquid culture (MSLC), among which Ser/Thr protein kinase (aopk1) and phosphatase (aoppt) genes were cloned. We also revealed that aopk1 encodes a protein with an N-terminal sequence 150 amino acid residues longer than that predicted from the registered sequence in GenBank.

DOI： 10.1263/jbb.102.470

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

Temperature scanning Fourier transform infrared, TS-FTIR, spectroscopy of various amorphous sugar matrixes was conducted to investigate the relationship between the glass transition temperature, T-g, of an amorphous sugar matrix and the nature of the hydrogen bonds in the matrix. An amorphous sugar matrix was prepared by air-drying an aqueous solution of sugar, and the degree of formation of hydrogen bonds in the matrix was evaluated at different temperatures using the peak positions of the IR band corresponding to the O-H stretching vibration at around 3400 cm(-1). The T-g value increased with increasing peak position of the O-H stretching vibration at T-g and were correlated reasonably well with the magnitude of the peak shift by the temperature increase (from 25 degrees C) to the T-g value. This demonstrates that the amorphous sugar matrix, in which the segments are fixed by fewer hydrogen bonds, has a higher thermal resistance. The glycosidic linkage largely contributes to the restriction of the segments, pyranose ring, rather than a hydrogen bond. As the degree of polymerization of pyranose rings increases, the degree of hydrogen bond formation needed to hold the matrix in a fixed position decreases. However, the magnitude of the restriction of pyranose rings by a glycosidic linkage changes depending on the type: the restrictions imposed by alpha-1,1 and -1,6 glycosidic linkages are the tightest and most flexible of all of the types of glycosidic linkages, respectively.

DOI： 10.1021/jp057527o

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BLACKWELL PUBLISHING  

We have carried out a genetic analysis on pyruvate kinase (Pyk(Tk)) and phosphoenolpyruvate synthase (Pps(tk)) in the hyperthermophilic archaeon, Thermococcus kodakarensis. In principle, both enzymes can catalyse the final step of the modified Embden-Meyerhof (EM) pathway found in Thermococcales, the conversion of phosphoenolpyruvate (PEP) to pyruvate, with the former utilizing ADP, while the latter is dependent on AMP and phosphate. Enzyme activities and transcript levels of both Pyk(Tk) and Pps(Tk) increased in T. kodakarensis under glycolytic conditions when compared with cells grown on pyruvate or amino acids. Using KW128, a tryptophan auxotrophic mutant with a trpE gene disruption, as a host strain, we obtained mutant strains with single gene disruptions in either the pyk(Tk) (Delta pyk strain) or pps(Tk) (Delta pps strain) gene. Specific growth rates and cell yields were examined in various media and compared with the host KW128 strain. The results indicated that both enzymes participated in pyruvate metabolism, but were not essential. In the presence of maltooligosaccharides, the Delta pyk strain displayed a 15% decrease in growth rate compared with the host strain, indicating that Pyk(Tk) does participate in glycolysis. However an even more dramatic effect was observed in the Delta pps strain in that the strain could not grow at all on maltooligosaccharides. The results clearly indicate that, in contrast to the conventional EM pathway dependent on pyruvate kinase, PEP synthase is the essential enzyme for the glycolytic conversion of PEP to pyruvate in T. kodakarensis. The physiological roles of the two enzymes under various growth conditions are discussed.

DOI： 10.1111/j.1365-2958.2006.05287.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

O-Acetylserine sulfhydrylase-B (OASS-B, EC 2.5.1.47) is one of the two isozymes produced by Escherichia coli that catalyze the synthesis Of L-Cysteine from O-acetyl- L-serine and sulfide. The cysM gene encoding OASS-B was cloned and the enzyme was overexpressed in E coli using pUC19 with a lacUV5 promoter. The enzyme was purified to homogeneity, as evidenced by SDS-PAGE. Approximately 300 mg of purified OASS-B was obtained from 1600 mL of culture broth with a purification yield of 60% or higher. The purified OASS-13 was characterized and its properties compared with OASS-A. OASS-13 did not form a complex with E. coli serine acetyltransferase (SAT, EC 2.3.1.30) and showed a wide range of substrate specificity in nonproteinaceous amino acid synthesis. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.pep.2006.01.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

Cysteine synthase from Escherichia coli is a bienzyme complex comprised of serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase A. The site of interaction of a SAT molecule was investigated by gel chromatography and surface plasmon technique using various mutant-type SATs, to better understand the mechanism involved in complex formation. The C-terminus of SAT, Ile 273, along with Glu 268 and Asp 271, was found to be essential for complex formation. The effects of O-acetyl-L-serine and sulfide on the affinity for the complex formation were also studied using a surface plasmon technique. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2006.01.054

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

Dodecapeptides that exhibit a high affinity specific to a polystyrene surface (PS-tags) were screened using an Escherichia coli random peptide display library system, and the compounds were used as a peptide tag for the site-specific immobilization of proteins. The various PS-tags obtained after 10 rounds of biopanning selection were mainly composed of basic and aliphatic amino acid residues, most of which were arranged in close proximity to one another. Mutant-type glutathione S-transferases (GSTs) fused with the selected PS-tags. PS19 (RAFIASRRIKRP) and PS23 (AGLRLKKAAIHR) at their C-terminus, GST-PS19 and GST-PS23, when adsorbed on the PS latex beads had a higher affinity than the wild-type GST, and the specific remaining activity of the immobilized mutant-type GSTs was approximately 10 times higher than that of the wild-type GST. The signal intensity detected for GST-PS19 and GST-PS23 adsorbed on hydrophilic and hydrophobic PS surfaces using an anti-peptide antibody specific for the N-terminus peptide of GST was much higher than that for the wild-type GST. These findings indicate that the mutant-type GSTs fused with the selected peptide tags, PS19 and PS23, could be site-specifically immobilized on the surface of polystyrene with their N-terminal regions directed toward the solution. Thus, the selected peptide tags would be useful for protein immobilization in the construction of enzyme-linked immunosorbent assay (ELISA) systems and protein-based biochips.

DOI： 10.1021/bp050331l

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

A novel enzyme that catalyzes efficient hydrolysis of capsaicin (8-methyl-N-vanillyl-6-nonenamide) was isolated from the culture broth of Streptomyces mobaraensis. The enzyme consisted of two dissimilar subunits with molecular masses of 61 and 19 kDa. The enzyme was activated and stabilized in the presence of Co2+. It showed a pH optimum of about 8 and was stable at temperatures of up to 55 degrees C for 1 h at pH 7.8. The specific activity of the enzyme for the hydrolysis of capsaicin was 10(2)-10(4) times higher than those for the enzymes reported to date. In an aqueous/n-hexane biphasic system, capsaicin analogues such as octanoyl, decanoyl, and lauroyl vanillylamides were synthesized from the corresponding fatty acids and vanillylamine at yields of 50% or greater. In addition, the enzyme catalyzed the deacylation of N-lauroyl-L-amino acids and N-lauroyl-L-dipeptides and the efficient synthesis of N alpha-lauroyl-L-lysine, N epsilon-lauroyl-L-lysine, and various N-lauroyl-peptides in aqueous solution in both the absence and the presence of glycerol.

DOI： 10.1021/jf052102k

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2006f.0.413.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2006f.0.996.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2006f.0.995.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2006f.0.949.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2006f.0.1002.0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

A novel aminoacylase was purified to homogeneity from culture broth of Streptomyces mobaraensis, as evidenced by SDS-polyacrylamide gel electrophoresis (PAGE). The enzyme was a monomer with an approximate molecular mass of 100 kDa. The purified enzyme was inhibited by the presence of 1,10-phenanthroline and activated by the addition of Co2+. It was stable at temperatures of up to 60 degrees C for 1 h at pH 7.2. It showed broad substrate specificity to N-acetylated L-amino acids. It catalyzed the hydrolysis of the amide bonds of various N-acetylated L-amino acids, except for N epsilon-acetyl-L-lysine and N-acetyl-L-proline. Hydrolysis of N-acetyl-L-methionine and N-acetyl-L-histidine followed Michaelis-Menten kinetics with K-m values of 1.3 +/- 0.1 mm and 2.7 +/- 0.1 mm respectively. The enzyme also catalyzed the deacetylation of 7-aminocephalosporanic acid (7-ACA) and cephalosporin C. Moreover, feruloyl-amino acids and L-lysine derivatives of ferulic acid derivatives were synthesized in an aqueous buffer using the enzyme.

DOI： 10.1271/bbb.69.1914

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s11746-005-1121-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

The hydrogen (H-2) production potential of the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1 was evaluated at 85 degreesC. In batch cultivation using a complex medium supplemented with elemental sulfur (S-0), evolution of H2S and CO2 was observed in the gas phase. When So was omitted and pyruvate or starch was added in the medium, the cells produced H-2 at high levels instead of H2S. As the level of H-2 appeared to correlate with the specific growth rate, analysis in continuous cultures was performed to develop a continuous H-2 production system. In a steady-state condition at a dilution rate of 0.2 h(-1), a continuous H-2 production rate (per gram dry weight, gdw) of 24.9 and 14.0 mmol gdw(-1) h(-1) was observed in media supplemented with pyruvate and starch, respectively. In both cultivations, a high accumulation of acetate and alanine was found as metabolites. When the dilution rates were elevated in the medium with pyruvate, steady-state growth was observed up to 0.8 h(-1), and a maximum H-2 production rate of 59.6 mmol gdw(-1) h(-1) was obtained. Based on the experimental results along with data of the entire genome sequence, the metabolic pathway of the strain relating to starch and pyruvate degradation is discussed. (C) 2004 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jbiotec.2004.11.002

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.294.0

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記述言語：英語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.271.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.293.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.242.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.270.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005.0.292.0

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記述言語：日本語   出版者・発行元：公益社団法人 化学工学会  

DOI： 10.11491/scej.2005f.0.311.0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC MICROBIOLOGY  

Fructose-1,6-bisphosphatase (FBPase) is one of the key enzymes in gluconeogenesis. Although FBPase activity has been detected in several hyperthermophiles, no orthologs corresponding, to the classical FBPases from bacteria and eukaryotes have been identified in their genomes. An inositol monophosphatase (IMPase) from Methanococcus jannaschii which displayed both FBPase and IMPase activities and a structurally novel FBPase (Fbp(Tk)) from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 have been proposed as the "missing" FBPase. For this study, using T. kodakaraensis, we took a genetic approach to elucidate which candidate is the major gluconeogenic enzyme in vivo. The IMPase/FBPase ortholog in T. kodakaraensis, Imp(Tk), was confirmed to possess high FBPase activity along with IMPase activity, as in the case of other orthologs. We therefore constructed Deltafbp and Deltaimp strains by applying a gene disruption system recently developed for T. kodakaraensis and investigated their phenotypes. The Deltafbp strain could not grow under gluconeogenic conditions while glycolytic growth was unimpaired, and the disruption resulted in the complete abolishment of intracellular FBPase activity. Evidently, fbp(Tk) is an indispensable gene for gluconeolgenesis and is responsible for almost all intracellular FBPase activity. In contrast, the endogenous imp(Tk) gene could not complement the defect of the fbp deletion, and its disruption did not lead to any detectable phenotypic changes under the conditions examined. These facts indicated that imp(Tk) is irrelevant to gluconeogenesis, despite the high FBPase activity of its protein product, probably due to insufficient transcription. Our results provide strong evidence that the true FBPase for gluconeogenesis in T. kodakaraensis is the FbpTk ortholog, not the IMPase/FBPase ortholog.

DOI： 10.1128/JB.186.17.5799-5807.2004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC MICROBIOLOGY  

Numerous bacteria and mammalian cells harbor two enzymes, phosphopentomutase (PPM) and 2-deoxyribose 5-phosphate aldolase (DERA), involved in the interconversion between nucleosides and central carbon metabolism. In this study, we have examined the presence of this metabolic link in the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1. A search of the genome sequence of this strain revealed the presence of a closely related orthologue (TK2104) of bacterial DERA genes while no orthologue related to previously characterized PPM genes could be detected. Expression, purification, and characterization of the TK2104 protein product revealed that this gene actually encoded a DERA, catalyzing the reaction through a class I aldolase mechanism. As PPM activity was detected in T. kodakaraensis cells, we partially purified the protein to examine its N-terminal amino acid sequence. The sequence corresponded to a gene (TK1777) similar to phosphomannomutases within COG1109 but not COG1015, which includes all previously identified PPMs. Heterologous gene expression of TK1777 and characterization of the purified recombinant protein clearly revealed that the gene indeed encoded a PPM. Both enzyme activities could be observed in T. kodakaraensis cells under glycolytic and gluconeogenic growth conditions, whereas the addition of ribose, 2-deoxyribose, and 2'-deoxynucleosides in the medium did not lead to a significant induction of these activities. Our results clearly indicate the presence of a metabolic link between pentoses and central carbon metabolism in T. kodakaraensis, providing an alternative route for pentose biosynthesis through the functions of DERA and a structurally novel PPM.

DOI： 10.1128/JB.186.13.4185-4191.2004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN  

The fructose-1,6-bisphosphate (FBP) aldolase gene from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 was cloned. The gene encoding FBP aldolase (Tk-fba) was expressed in Escherichia coli and the purified recombinant protein was characterized at high temperature. Tk-Fba is a homodecamer with a subunit molecular mass of 31,283 Da. The amino acid sequence, decameric conformation, formation of a Schiff-base intermediate, and stimulation (286%) of FBP cleavage activity by citrate suggested that Tk-Fba belonged to Class IA, a subtype of the classical Class I aldolases. The specific activity for the FBP cleavage reaction was 18.9 U/mg, which was much higher than those of other Class IA type FBP aldolases. Tk-Fba was extremely thermostable since the optimum temperature seemed to be above 100degreesC. The optimum pH for Tk-Fba was determined to be 5.0 in the absence of citrate, while it shifted to around 7.0 in the presence of citrate. Tk-Fba accepted FBP and fructose-1-phosphate as substrates and K-m values were determined to be 0.063 mM and 4.37 mM, respectively. In addition to citrate, phosphoenolpyruvate and pyrophosphate were also found to be potent activators of Tk-Fba, enhancing activities up to 346% and 201%, respectively. Erythrose-4-phosphate acted as an inhibitor and caused a decrease in the activity to 49%. Tk-Fba also catalyzed the condensation reaction with a similar activity level (14.9 U/mg) to that for FBP cleavage. However, none of the above compounds seemed to have a significant effect on the condensation reaction by Tk-Fba. These results suggest a regulatory function of Tk-Fba toward the catabolic direction of sugar metabolism in T. kodakaraensis KOD1.

DOI： 10.1263/jbb.94.237

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

Fructose-1,6-bisphosphatase (FBPase) is one of the key enzymes of the gluconeogenic pathway. Although enzyme activity had been detected in Archaea, the corresponding gene had not been identified until a presumable inositol monophosphatase gene from Methanococcus jannaschii was found to encode a protein with both inositol monophosphatase and FBPase activities. Here we display that a gene from the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1, which does not correspond to the inositol monophosphatase gene from M. jannaschii, displays high FBPase activity. The FBPase from strain KOD1 was partially purified, its N-terminal amino acid sequence was determined, and the gene (Tk-fbp) was cloned. Tk-fbp encoded a protein of 375 amino acid residues with a molecular mass of 41,658 Da. The recombinant Tk-Fbp was purified and characterized. Tk-Fbp catalyzed the conversion of fructose 1,6-bisphosphate to fructose 6-phosphate following Michaelis-Menten kinetics with a K-m value of 100 mum toward fructose 1,6-bisphosphate, and a k(cat) value of 17 s(-1) subunit(-1) at 95degreesC. Unlike the inositol monophosphatase from M. jannaschii, Tk-Fbp displayed strict substrate specificity for fructose 1,6-bisphosphate. Activity was enhanced by Mg2+ and dithioerythritol, and was slightly inhibited by fructose 2,6-bisphosphate. AMP did not inhibit the enzyme activity. We examined whether expression of Tk-fbp was regulated at the transcription level. High levels of Tk-fbp transcripts were detected in cells grown on pyruvate or amino acids, whereas no transcription was detected when starch was present in the medium. Orthologue genes corresponding to Tk-fbp with high similarity are present in all the complete genome sequences of thermophilic Archaea, including M. jannaschii, Pyrococcus furiosus, Sulfolobus solfataricus, and Archaeoglobus fulgidus, but are yet to be assigned any function. Taking into account the high FBPase activity of the protein, the strict substrate specificity, and its sugar-repressed gene expression, we propose that Tk-Fbp may represent the bona fide FBPase in Archaea.

DOI： 10.1074/jbc.M202868200

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

A corona-discharge reactor was applied to remove benzene and p-dichlorobenzene from nitrogen and a nitrogen-oxygen mixture. Although benzene was not effectively removed from nitrogen by electron attachment, the removal efficiency was improved greatly by mixing oxygen. On the other hand, the high removal efficiencies of p-dichlorobenzene were obtained in nitrogen or a nitrogen-oxygen mixture. The removal mechanism was studied based on the contribution of the ozone reaction and the analysis of the deposit on the anode of the reactor. As a result, the ozone reaction does not contribute to the removals. An FT-IR measurement and thermogravimetry suggest that benzene or p-dichlorobenzene is decomposed by dissociative electron attachment and deposits as polycyclic aromatic compounds of a high boiling point on the anode surface.

DOI： 10.1021/ie980071v

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2016045901

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：WILEY-BLACKWELL  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=305474

記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2014247513

記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2013125671

記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2013125672

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   掲載種別：研究発表ペーパー・要旨（全国大会，その他学術会議）  

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記述言語：英語   出版者・発行元：Faculty of Engineering, Okayama University  

We used modified Czapek-Dox (mCD) or dextrin-peptone-yeast extract (DPY) media to cultivate a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to identify the differences in cultivation behaviors and gene transcriptional profiles. The fungi cultivated by APC or MSLC secreted a greater number of different proteins/enzymes in larger quantities compared with fungi cultivated by SFC, particularly when DPY medium was used. In particular, the amounts of protease secreted by fungi cultivated via MSLC or APC were much greater compared with SFC. When mCD medium was used, α-amylase activity was barely detectable in all cultures while the activity was detected in MSLC and APC in a quantity that was several times higher than that in SFC using DPY medium. SDS-PAGE analysis and N-terminal amino acid sequences confirmed 6 proteins in the culture supernatants when DPY medium was used. Among these proteins oryzin (an alkaline protease) and α-amylase were detected at much higher levels in APC and MSLC compared with SFC, which was consistent withthe measured activity of the secreted enzymes. However, when mCD medium was used, only oryzin was detected in significant amounts in MSLC and APC. Microarray analyses of the fungi cultivated by SFC, APC or MSLC using either mCD or DPY media indicated that the gene transcriptional profile of the MSLC sample was similar to that of the APC sample but different from that of the SFC sample. When mCD medium was used, most of the genes that were up-regulated 10-folds or greater in the MSLC sample relative to the SFC sample were unknown or predicted proteins. Transcription of the oryzin gene was only slightly up-regulated in the MSLC sample while transcription of the α-amylase gene was slightly down-regulated. On the other hand, when DPY medium was used, many known genes including the oryzin gene were up-regulated in the MSLC sample versus the SFC sample.

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：英語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=304106

記述言語：英語   出版者・発行元：公益社団法人日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2009066377

記述言語：日本語   出版者・発行元：すかいら-くフ-ドサイエンス研究所  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：日本語   出版者・発行元：日本生物工学会  

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その他リンク： http://search.jamas.or.jp/link/ui/2008290979

記述言語：日本語   出版者・発行元：日本生物工学会  

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記述言語：英語   出版者・発行元：IEEE  

DOI： 10.1109/MHS.2005.1589975

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その他リンク： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=301726

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