• Journal of fish pathology
• /
• v.28 no.3
• /
• pp.133-143
• /
• 2015

The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

• Microbiology and Biotechnology Letters
• /
• v.48 no.2
• /
• pp.138-147
• /
• 2020

Various microorganisms play important roles in food fermentation, food spoilage, and agriculture. In this study, the biotype of 54 yeast and bacterial strains having high potential for utilization in food and agriculture, including Candida spp., Lactobacillus spp., and Acetobacter spp., were characterized by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). This characterization using a fast and robust method provides much-needed information on the selected microorganisms and will facilitate effective usage of these strains in various applications. Importantly, the unique protein profile of each microbial species obtained from this study was used to create a database of fingerprints from these species. The database was validated using microbial strains of the same species by comparing the mass spectra with the created database through pattern matching. The created reference database provides crucial information and is useful for further utilization of a large number of valuable microorganisms relevant to food and agriculture.

• Korean Journal of Microbiology
• /
• v.53 no.3
• /
• pp.163-169
• /
• 2017

Strains D4 and D5 were isolated from peach-rotten soil during the peach harvest season. The isolates were identified based on morphological and biochemical characterization, and identification was determined by 16S rRNA gene sequencing. Results showed that D4 has high similarity to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$ at 99.05% and 98.98%, respectively. D5 was also similar to Lactobacillus pentosus ATCC $8041^T$ and Lactobacillus plantarum ATCC $14917^T$ at 98.71% and 98.64%, respectively. In contrast, isolates showed differences in carbohydrate utilization in comparison to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$. In view of this we performed VITEK MS matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, multiplex PCR fingerprinting, and random amplified polymorphic DNA (RAPD)-PCR to further confirm the identification of D4 and D5. The results of these analyses showed that both strains were most similar to Lactobacillus plantarum.

• 한국생물공학회:학술대회논문집
• /
• 2000.04a
• /
• pp.219-222
• /
• 2000

Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

• Biomolecules & Therapeutics
• /
• v.13 no.4
• /
• pp.240-245
• /
• 2005

An antimicrobial molecule was purified from centipede, Scolopendra subspinipes mutilans L. Koch, by reverse phase-HPLC. Its molecular weight was determined to be 1208.5493 by using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Total amino acid composition analysis revealed that it consists of E, G, P, V, L, F, and W. It exhibited a broad antimicrobial spectrum against not only Gram-negative, but also Gram-positive bacteria. Furthermore, it was found to have an antimicrobial activity against vancomycin resistant enterococci (VRE). It may be a useful molecule for a new antibiotic development, especially against drug-resistant bacteria. We suggest that it may playa role in the defense system of this animal. This is the first report of a peptidic antimicrobial substance from centipede.

• Molecular & Cellular Toxicology
• /
• v.4 no.3
• /
• pp.253-259
• /
• 2008

Human follicular fluid (HFF) is the in vivo microenvironment for oocyte maturation and includes a variety of proteins that could be involved in oocyte development and fertilization. We therefore used a proteomic approach to identify new HFF proteins. HFF from mature human follicles was obtained from five women following oocyte collection for in vitro fertilization (IVF). Ethanol-precipitated HFF run on two-dimensional gel electrophoresis (2DE) produced approximately 250 Coomassie brilliant blue-stained spots, 64 of which were identified using matrix-assisted laser desorption/ionization-mass spectrometry (MALDIMS). In this study, several proteins including complement factor H, inter-${\alpha}$ (globulin) inhibitor H4, inter-${\alpha}$-trypsin inhibitor heavy chain H4 precursor, human zinc-${\alpha}$-2-glycoprotein chain B, PRO2619, PRO02044, and complex-forming glycoprotein HC were new proteins that have not been previously reported in HFF using proteomic methods. Additionally, we identified alloalbumin venezia for the first time from trichloroacetic acid (TCA)-precipitated HFF. These HFF proteins could serve as new biomarkers for important human reproductive processes.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2005.10b
• /
• pp.280-281
• /
• 2005

• Proceedings of the PSK Conference
• /
• 2003.04a
• /
• pp.217.3-218
• /
• 2003

A novel lectin has been purified from the fruiting bodies of the mushroom, Fomitella fraxinea, which belongs to bracket fungi by a combination of ion-exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephacryl S-200 HR. The lectin, designated as FFrL, was a homotetrametric protein with a molecular weight of 50 kDa as demonstrated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and MALDI-TOF-MS(matrix assisted UV laser desorption/ionization time-of flight mass spectrometry). (omitted)

• Molecular & Cellular Toxicology
• /
• v.5 no.3
• /
• pp.198-206
• /
• 2009

Microarray and proteomic expression patterns in response to cadmium exposure were analyzed in human lung epithelial cells. Among 35,000 genes analyzed by cDNA microarray, 228 genes were up-regulated and 99 genes were down-regulated, based on a fold change cut-off value of ${\geq}2$. Combining two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-ToF-MS), 25 of 629 protein spots showed fold changes in expression ${\geq}2$ (17 up-regulated, 8 down-regulated). After comparing the cDNA microarray and proteomic analyses, only transglutaminase 2, translation elongation factor 1 alpha 1, and glyceraldehyde-3-phosphate dehydrogenase showed overlapping signals in the cDNA microarray and proteomic analyses, whereas the remaining differentially expressed proteins showed large discrepancies with respect to mRNA expression.

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