• Molecules and Cells
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• v.24 no.2
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• pp.268-275
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• 2007

Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

• Proceedings of the PSK Conference
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• 2002.04a
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• pp.166.3-167
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• 2002

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.156.1-156.1
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• 2006

• Korean Journal of Pharmacognosy
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• v.34 no.3 s.134
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• pp.246-249
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• 2003

The major polysaccharide, named EAP, was purified from the aerial parts of Eclipta alba by Sepharose CL-2B ion exchange chromatography and recycling HPLC. The molecular weight of EAP was estimated to be 20911.9 D by MALDI-TOF MS. In addition, the sugar composition was determined to be arabinose (23.6%), mannose (24.8%), galactose (12.3%), and glucose (41.3%), respectively, by GC analysis. The EAP decreased the blood sugar level, which was induced by alloxan in rats, dose dependently.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.763-768
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• 2012

This study has been performed to measure the prevalence and microbial flora on chicken slaughtering as well as the processing process from the months of October to November. Whole-chicken rinsing technique was used in order to analyze the incidence of microorganisms on chicken carcass at the stage before chilling (after evisceration), after chilling and after cutting. The swab technique was used on processing the processed samples, such as working plates and cutting knives. Brine and cooling water from four cooling tubs were taken from each processing processes and were used as samples. Furthermore, the matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for whole cell fingerprinting in combination with a dedicated bioinformatic software tool was used to identify the isolated microorganisms. Of the tested samples and processes, brine ($4.50{\pm}0.64$ Log CFU/mL) and chicken carcass before chilling ($4.15{\pm}0.46$ Log CFU/mL) showed the highest population of microorganisms; the predominant microbial flora of them were Moellerella wisconsensis (54.84%), a member of the Enterobacteriaceae family, and Escherichia coli (60.36%), respectively. However, the predominant microbial flora of cut carcass was changed to Staphylococcus aureus (27.32%), which is a kind of pathogenic microorganism that can cause a food-borne illness. Therefore, the slaughtering and processing procedure of chicken are needed to be controlled more hygienically.

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

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.2919-2923
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• 2013

Esophageal cancer (EC) is one of the most common malignant tumors, and the incidence of esophageal squamous cell carcinoma (ESCC) is highest in China. Early diagnosis and effective monitoring are keys to comprehensive treatment and discovering tumor metastases and recurrence in time. The aim of this study was to confirm serum peptidome pattern utility for diagnosis of ESCC, and assessment of operation success, postoperative chemotherapy results, tumor metastasis and recurrence. Serum samples were collected from 61 patients treated with surgery and chemotherapy and 20 healthy individuals. Spectral data generated with weak cationic-exchanger magnetic beads (WCX-MB) and MALDI-TOF MS by a support vector machine (SVM), were used to construct diagnostic models and system training as potential biomarkers. A pattern consisting of 11 protein peaks, separated ESCC (m/z 650.75), operated (m/z 676.61, 786.1, 786.58), postoperative chemotherapy (m/z 622.77, 650.66, 676.46) and tumor metastasis and recurrence (m/z 622.63, 650.56, 690.77, 676.12) from the healthy individuals with a sensitivity of 100.0% and a specificity of 100.0%. These results suggested that MALDITOF MS combined with MB separation yields significantly higher sensitivity and specificity for the detection of serum protein in patients with EC patients treated with surgery and chemotherapy.

• Food Science and Biotechnology
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• v.14 no.6
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• pp.854-859
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• 2005

Although action modes of equol and genistein have been extensively studied, their precise roles in tumor cells remain elusive. To address possible effects of these compounds on protein expression in mammary tumor cells, proteins modulated in MCF-7 mammary tumor cells when incubated in absence and presence of 10 uM equol or genistein were identified through 2-dimensional gel electrophoresis, MALDI-TOF MS/MS, and NCBInr database search using Mascot software. Most proteins differentially expressed in MCF-7 cells after treatment with 10 uM genistein or equol were identified as being the same. Exposure to both compounds caused decreased cellular expression of RNA-binding protein regulatory subunit and oncogene DJ1 tubulin beta-1 chain, and increased expression of heterogeneous ribonucleoproteins F and L, KH-type splicing regulatory protein, and translation elongation factor EF-Tu precursor. Genistein and equol at dose used in this study showed common action mechanism.

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

• Journal of Life Science
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• v.26 no.3
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• pp.296-301
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• 2016

An antibacterial peptide from skin extract of the catfish Silurus asotus was purified and characterized. The acidified skin extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into flow-through (F.T.), 10% methanol-elute (RM10), 60% methanolelute (RM60), and 100% methanol-elute (RM100) fractions. RM10, RM60, and RM 100 showed antimicrobial activity against Escherichia coli D31. On the other hand, the F.T. fraction did not show antimicrobial activity. Among the various fractions, RM 60 had the highest activity. RM 60 was partially purified on a cation exchange column (CM52) by a stepwise gradient. The ammonium acetate (pH 5.15) 0.02 M – 0.8 M fraction showed antimicrobial activity. Then an antimicrobial peptide was purified using a 0.6M fraction with strong antibacterial activity through a series of five C18 reversed-phase HPLC columns. For the characterization of the purified peptide, the molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of this peptide was about 4182.1 [M+H]⁺. The amino acid sequence of this peptide was partially determined as follows: PALXXKARREAKVKF. These findings suggest that this peptide plays a significant role in the innate defense system of catfish skin.