• Journal of Veterinary Clinics
• /
• v.30 no.3
• /
• pp.218-221
• /
• 2013

A flightless brown hawk owl (Ninox scutulata) weighing 180 g was rescued and referred to the teaching veterinary hospital with humerus open fracture. On physical examination and radiography, open fracture of the left humerus was found. After 3 days, orthopedic surgery was operated with intramedullary pins and wires. The bird died 2 days after surgery with anorexia. On necropsy, multiple green nodules with 2-3 mm in diameter were observed at the surface and inside of the left lung. Numerous conidial heads and spores were seen in the center of foci in the histopathological examination. The mycelia penetrated the surrounding pulmonary parenchyma, showing inflammation and necrosis. The fungus was isolated from the lung and cultured on Sabouraud Dextrose Agar at $30^{\circ}C$ for 7 days. The colony was blue-green color with a powdery surface. The fungus was identified as Aspergillus fumigatus by DNA analysis, including the internal transcribed spacer region, partial ${\beta}$-tubulin, and the calmodulin gene. This case was diagnosed as pulmonary aspergillosis secondary to open fracture of pneumatic bone in a brown hawk owl.

• Korean Journal of Microbiology
• /
• v.46 no.3
• /
• pp.270-277
• /
• 2010

In order to manufacture Bacillus cereus-free fermented soybean products, an antimicrobial agentproducing isolate against B. cereus was obtained from 150 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK 121057 was most closely related to Bacillus licheniformis. The B. licheniformis isolate began to produce the antimicrobial agent after 48 h of incubation. The agent was nonproteinaceous and insensitive to heat, long term storage and protease K. Electron microscopic observation indicated that the agent attacked the membrane of B. cereus, leaving the ghost cell. The isolate inhibited growth of B. subtilis, Lactobacillus brevis and various types of pathogenic strains including Escherichia coli, E. faecalis, Micrococcus luteus, Staphylococcus aureus, Aspergillus flavus, A. ochraceus, and A. parasiticus as well as B. cereus. After coinoculation of B. licheniformis SCK 121057 and B. cereus in the ratio (as the basis of CFU/g sample) of 10 to 1 on the surface of cooked soybeans, cell numbers of B. cereus had been dramatically reduced after 31 days of incubation compared to those of single inoculation of B. cereus.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.6
• /
• pp.1519-1526
• /
• 2010

The behavior of peptide or protein solutes in saline aqueous solution is a fundamental topic in physical chemistry. Addition of ions can strongly alter the thermodynamic and physical properties of peptide molecules in solution. In order to study the effects of added ionic salts on protein conformation and dynamics, we have used the molecular dynamics (MD) simulations to investigate the behavior of Staphylococcus aureus Hfq protein under two different ionic concentrations: 0.1 M NaCl and 1.0 M NaCl in presence and absence of RNA (a hepta-oligoribonucleotide AU5G). Hfq, a global regulator of gene expression is highly conserved and abundant RNA-binding protein. It is already reported that in vivo the increase of ionic strength results in a drastic reduction of Hfq affinity for $Q{\beta}$ RNA and reduces the tendency of aggregation of Escherichia coli host factor hexamers. Our results revealed the crucial role of 0.1 M NaCl Hfq system on the bases with strong hydrogen bonding interactions and by stabilizing the aromatic stacking of Tyr42 residue of the adjacent subunits/monomers with the adenine and uridine nucleobases. An increase in RNA pore diameter and weakened compactness of the Hfq-RNA complex was clearly observed in 1.0 M NaCl Hfq system with bound RNA. Aggregation of monomers in Hfq and the interaction of Hfq with RNA are greatly affected due to the presence of high ionic strength. Higher the ionic concentration, weaker is the aggregation and interaction. Our results were compatible with the experimental data and this is the first theoretical report for the experimental study done in 1980 by Uhlenbeck group for the present system.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.21 no.1
• /
• pp.76-83
• /
• 1992

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin (ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. ST producing E. coli KM-7 strain was isolated from the swine and molecular cloning of ST gene of KM-7 strain. Transformant eKT-53 $(ST^+,\;LT^-)$ was selected by infant mouse assay (IMA). The culture supernatant of eKT-53 strain was performed purification by multipled steps. The culture supernatant (crude ST) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Bio-Gel P-6 and preparative polyacrylamide slab gel electrophoresis. About 113-fold purification was achieved with a yield of about 11% of crude ST and the minimum effective dose(MED) of this purified ST was about 2.8ng in IMA. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis.

• The Korea Journal of Herbology
• /
• v.30 no.6
• /
• pp.7-15
• /
• 2015

Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.

• Journal of Life Science
• /
• v.13 no.2
• /
• pp.184-190
• /
• 2003

Myxococcus xanthus is a Gram negative, rod-shaped, soil bacterium that displays a social behaviors, and multicellular development upon nutrient deprivation. The csgA gene encoding a cell surface protein is essential for developmental behaviors including rippling, aggregation, fruiting body formation and sporulation. csgA mutants show normal vegetative growth, but lack all these developmental phenotypes. Expression of the CsgA (C-signal) specific genes are eliminated or dramatically reduced in csgA mutants. In order to identify components of C-signal transduction pathway, second site mutations were introduced into csgA mutants and were identified which can fully or partially restore development of csgA mutants (Rhie, H. G. et. al. 1989. J. Bacteriol. 171, 3268-3276). One of such csgA suppressor mutations, socD500 restores only sporulation to csgA mutants at 15$^{\circ}C$. The socD500 mutaion however eliminates the three basic developmental requirements, starvation, high cell density and a solid surface. Only sporulation, not accompanied with fruiting body formation is induced simply by shifting the temperature of vegetatively growing cells from $32^{\circ}C$ to $15^{\circ}C$. Spores induced by socD500 mutation is not as thick as that of wild-type fruiting body. In socD500 genetic background, two of ten C-signal dependent genes, $\Omega$DK4506 and $\Omega$DK4406 are more highly expressed in growing cells at $15^{\circ}C$. These results indicate that the socD500 mutation may be partly involved in the regulation of expression of two C-signal dependent genes and genes for sporulation in this transduction pathway.

• Molecules and Cells
• /
• v.37 no.11
• /
• pp.833-840
• /
• 2014

Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

• Korean Journal of Microbiology
• /
• v.52 no.2
• /
• pp.148-156
• /
• 2016

This study examined effects of Enterobacter ludwigii SJR3 showing a high 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, on growth of tomato plant and its expression of stress-related genes under drought and salt stress. SJR3 strain was inoculated at $10^6cell/g$ soil to 4-week grown tomato plants, and drought and salt stresses were treated. After additional incubation for 1 week, root length, stem length, fresh weight and dry weight of tomato plants treated with SJR3 increased by 37.8, 37.2, 96.8 and 146.6%, respectively compared to those of uninoculated plants in drought stress environment, and they increased by 19.2, 25.4, 19.5, and 105.8%, respectively in salt stress environment. Proline content in tomato leaves increased significantly under stress conditions as one of a protecting substance against stresses, but proline contents in tomato treated with SJR3 decreased by 62.1 and 54.1%, respectively. Relative expression of genes encoding ACC oxidase, ACO1 and ACO4, ethylene response factor genes ERF1 and ERF4, and some other stress-related genes were examined from tomato leaves. Compared to the non-stressed tomato, expressions of all stress-related genes increased significantly in the stressed tomato, but gene expressions in the inoculated tomato were similar to those of no-stressed control tomato. Therefore, E. ludwigii SJR3 may play an important role in mitigating drought and salt stress in plants, and can increase productivity of crops under various abiotic stresses.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.8
• /
• pp.1339-1348
• /
• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.12
• /
• pp.1636-1643
• /
• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.