• Journal of Microbiology and Biotechnology
• /
• v.16 no.1
• /
• pp.25-31
• /
• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.1
• /
• pp.105-113
• /
• 2019

Although siderophore compounds are mainly biosynthesized as a response to iron deficiency in the environment, they also bind with other metals. A few studies have been conducted on the impact of heavy metals on the siderophore-mediated iron uptake by microbiome. Here, we investigated siderophore production by a variety of rhizosphere fungi under different concentrations of $Zn^{2+}$ ion. These strains were specifically isolated from the rhizosphere of Panax ginseng (Korean ginseng). The siderophore production of isolated fungi was investigated with chrome azurol S (CAS) assay liquid media amended with different concentrations of $Zn^{2+}$ (50 to $250{\mu}g/ml$). The percentage of siderophore units was quantified using the ultra-violet (UV) irradiation method. The results indicated that high concentrations of $Zn^{2+}$ ion increase the production of siderophore in iron-limited cultures. Maximum siderophore production by the fungal strains was detected at $Zn^{2+}$ ion concentration of $150{\mu}g/ml$ except for Mortierella sp., which had the highest siderophore production at $200{\mu}g/ml$. One potent siderophore-producing strain (Penicillium sp. JJHO) was strongly influenced by the presence of $Zn^{2+}$ ions and showed high identity to P. commune (100% using 18S-rRNA sequencing). The purified siderophores of the Penicillium sp. JJHO strain were chemically identified using UV, Fourier-transform infrared spectroscopy (FTIR), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS) spectra.

• 한국정보디스플레이학회:학술대회논문집
• /
• 2008.10a
• /
• pp.1418-1420
• /
• 2008

In this paper, the light emitting efficiency, spectrum, and the lifetime of the phosphorescent devices, whose emission characteristics are strongly dominated not only by the energy transfer but also by the charge carrier trapping induced by the emissive dopant, are explained by differences in the energy levels of the host, dopant, and nearby transport layers. On the basis of our finding on device performance and photocurrent measurement data by time-of-flight (TOF), we investigated the effect of the difference of carrier trapping dopant and properties of the host materials on the efficiency roll-off of phosphorescent organic light emitting diode (OLED), along with a physical interpretation and practical design scheme, such as a multiple host system, for improving the efficiency and lifetime of devices.

• Tribology and Lubricants
• /
• v.15 no.4
• /
• pp.335-342
• /
• 1999

Corrosion of the disk has been an ongoing concern for the manufacturers of hard disk drives. With the advent of magnetoresistive (MR) head, very low levels of corrosion and contamination become more critical since the raised defects and corrosion products on the disk surface-anything that heats the MR sensor due to the contact-can distort the output signal of the head. This phenomenon is called as thermal asperity. In this paper, the effect of corrosion as a form of Co migration on the occurrence of thermal asperity in MR drives was investigated. The corrosion test at high temperature (60$^{\circ}C$) and high relative humidity (80%) was emphasized in this study and the testing results at ambient condition were compared. The corrosion on the disks was characterized as the amount of Co ion migration using an ion chromatography (IC) and a time-of-flight secondary ion mass spectroscopy (TOF-SIMS). It is proved that corrosion on the disk surface after storage testing is closely correlated to the amount of Co ions migration from the magnetic layer to disk surfaces and higher Co migration causes more thermal asperities in the drive. In order to reduce Co migration, several methods such as burnishing process and structure of the carbon overcoat were investigated. It is found that the hydrogenated carbon overcoat shows the least Co migration among different types of overcoat layer. However, the most effective way to reduce Co migration is the application of Cr layer between the overcoat and the magnetic alloy layer.

• Microbiology and Biotechnology Letters
• /
• v.46 no.1
• /
• pp.45-50
• /
• 2018

The chitosanase gene (btbchito) of Bacillus thuringiensis BMB171 was cloned and heterologously expressed in the yeast Pichia pastoris. After purification, about 300 mg of recombinant chitosanase was obtained from the 1-1 culture medium with a specific activity of 240 units/mg. Results determined by the combined use of thin layer chromatography (TLC) and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) showed that the chitooligosaccharides (COSs) obtained by chitosan (N-deacetylated by 70%, 80%, and 90%) hydrolysis by rBTBCHITO were comprised of oligomers, with degrees of polymerization (DP) mainly ranging from trimers to heptamers; high molecular weight chitopentaose, chitohexaose, and chitoheptaose were also produced. Hydrolysis products was also deduced using MS since the COSs (n) are complex oligosaccharides with various acetyl groups from one to two, so the non-acetyl COSs (GlcN)n and COSs with more acetyls (> 2) were not detected. The employment of this method in the production of high molecular weight COSs may be useful for various industrial and biological applications, and the activity of chitosanase has great significance in research and other applications.

• Biomedical Science Letters
• /
• v.21 no.4
• /
• pp.233-240
• /
• 2015

Some virulence proteins of Helicobacter pylori, such as vacuolating cytotoxic protein A (VacA) and cytotoxin-associated gene protein A (CagA) have been reported to be causative agents of various gastric diseases including chronic gastritis, gastric ulcer or gastric adenocarcinoma. The expression level of these virulence proteins can be regulated when H. pylori is exposed to the antibacterial agent, cyanidin 3-O-glucoside (C3G) as previously reported. In this study, we analyzed the quantitative change of various virulence proteins including CagA and VacA by C3G treatment. We used 2-dimensional electrophoresis (2-DE) to analyze the quantitative change of representative ten proteome components of H. pylori 60190 ($VacA^+/CagA^+$; standard strain of Eastern type). After 2-DE analysis, spot intensities were analyzed using ImageMaster$^{TM}$ 2-DE Platinum software then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). Next, we selected major virulence proteins of H. pylori among quantitatively meaningful ten spots and confirmed the 2-DE results by Western blot analysis. These results suggest that cyanidin 3-O-glucoside can modulate a variety of H. pylori pathogenic determinants.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.49 no.4
• /
• pp.342-348
• /
• 2004

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the $60\%$ isopropanol extract of soybean(Glycine max [L.] Merr.) seed revealed two abundant proteins with molecular masses of 19 and 10 kDa. Amino acid analysis revealed that the isopropanol-extractable protein fraction was rich in cysteine. Two-dimensional gel electro-phoretic analysis indicated that the 19kDa and 10kDa proteins had pI of 4.2 and 4.0 respectively. Peptide mass fingerprints of trypsin digests of the two proteins obtained using matrix-assisted, laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy revealed the 19kDa protein was Kunitz trypsin inhibitor and the 10kDa protein was Bowman-Birk proteinase inhibitor. When resolved under non-denaturing conditions, the isopropanol-extracted proteins inhibited trypsin and chymotrypsin activity. Results presented in this study demonstrate that isopropanol extraction of soybean seed could be used as a simple and rapid method to obtain a protein fraction enriched in Kunitz trypsin and Bowman-Birk proteinase inhibitors. Since proteinase inhibitors are rich in sulfur amino acids and are putative anticarcinogens, this rapid and inexpensive isolation procedure could facilitate efforts in nutrition and cancer research.

• Proceedings of the Korean Institute of IIIuminating and Electrical Installation Engineers Conference
• /
• 2006.05a
• /
• pp.298-301
• /
• 2006

This paper describes the electron energy distribution function characteristics in $SF_6-Ar$ gas calculated for range of E/N values from $50\sim700[Td]$ by the Monte Carlo simulation(MCS) and Boltzmann equation(BE) method using a set of electron collision cross sections determined by the authors and the values of electron swarm parameters are obtained by time of flight(TOF) method. In this dissertation the results of the combined experimental and theoretical studies designed to understand and predict the spatial growth and transport coefficients for electrons in $SF_6$ and $SF_6-Ar$ mixtures have described. The ionization and attachment coefficients in pure $SF_6$ and $SF_6-Ar$ mixtures have been calculated over the range of 10$SF_6$ molecule and for Ar atom proposed by other authors. The results obtained in this work will provide valuable information on the fundamental behaviors of electrons in weakly ionized gases and the role of electron attachment in the choice of better gases and unitary gas dielectrics or electro negative components in dielectric gas mixtures.

• BMB Reports
• /
• v.53 no.7
• /
• pp.349-356
• /
• 2020

Mass spectrometry (MS) is an ideal tool for analyzing multiple types of (bio)molecular information simultaneously in complex biological systems. In addition, MS provides structural information on targets, and can easily discriminate between true analytes and background. Therefore, imaging mass spectrometry (IMS) enables not only visualization of tissues to give positional information on targets but also allows for molecular analysis of targets by affording the molecular weights. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS is particularly effective and is generally used for IMS. However, the requirement for an organic matrix raises several limitations that get in the way of accurate and reliable images and hampers imaging of small molecules such as drugs and their metabolites. To overcome these problems, various organic matrix-free LDI IMS systems have been developed, mostly utilizing nanostructured surfaces and inorganic nanoparticles as an alternative to the organic matrix. This minireview highlights and focuses on the progress in organic matrix-free LDI IMS and briefly discusses the use of other IMS techniques such as desorption electrospray ionization, laser ablation electrospray ionization, and secondary ion mass spectrometry.

• Food Science and Biotechnology
• /
• v.18 no.2
• /
• pp.519-525
• /
• 2009

A bacterial strain (Strain N-08) capable of extracellularly producing high level of non-reducing oligosaccharide (NR-OS) isolated from soil. The strain was identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Arthrobacter crystallopoietes. The high production of NR-OS was observed in the basal culture medium containing maltose as a sole carbon source. The NR-OS in culture supernatant was purified by glucoamylase treatment and Dowex-1 (OH.) ion exchange chromatography and its structure was characterized. This oligosaccharide consisted of only glucose. Methylation analysis indicated that this fraction was composed mainly of non-reducing terminal glucopyranoside. Matrixassisted laser-induced/ionization time-of-flight (MALDI-TOF) and electrospray ionization-mass spectrometry (ESI-MS)/MS analyses suggested that this oligosaccharide comprised non-reducing disaccharide unit with 1,1-glucosidic linkage. When this disaccharide was analyzed by $^1H$-NMR and $^{13}C$-NMR, it gave the same signals with $\alpha$-D-glucopyranosyl-(1,1)-$\alpha$-Dglucopyranoside. These results indicated that the NR-OS produced by A. crystallopoietes N-08 was ${\alpha}1$,${\alpha}1$-trehalose. This is the first report of the trehalose which can be produced directly from maltose by A. crystallopoietes N-08.