• Journal of Environmental Health Sciences
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• v.27 no.1
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• pp.75-82
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• 2001

Microorganisms can attach firmly to food contact surface material and the resitance of adherent bacteria differ markedly from planktonic cells. Therefore, adherent cells are a potential contamination problem to the food preparation because of their high resistance. to sanitation and heat treatment. This study was carried out in order to investigate growth and decontamination of Listeria monocytogenes attached to stainless steel, glass and plastic. Listeria monocytogenes cells could attach to all types of surface at three temperatures after contact times for 24 hrs. The numbers of adherent cells were greater at higher temperatures, but not increased with incubation time. When recovery of adherent cells was investigated, after 24 grs, the numbers of adherent cells were about 10$^{7}$ , 10$^{10}$ , 11$^{11}$ at 4$^{\circ}C$, $25^{\circ}C$, 3$0^{\circ}C$ repectively. Planctonic cells decreased by 2 log cycles after exposure to the domestic sanitizer. Adherent cells showed high resistance to domestic sanitizers and that was dependent upon surface materials studied, being greatest on plastic followed by stainless steel and glass. Adherent cells were more resistant to heat treatment than planktonic cells. When adherent cells were exposed to the temperature of 5$0^{\circ}C$, 55$^{\circ}C$, 57.5$^{\circ}C$ for 10 min, their populations did not decrease significantly. When the temprature increased to 6$0^{\circ}C$, cells attached to all types of surfaces were completely inactivated for 10 min.

• Bulletin of the Korean Chemical Society
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• v.32 no.6
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• pp.1925-1930
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• 2011

Psychrophilic bacteria have acquired cold-resistance in order to protect themselves against freezing temperatures, which would otherwise be lethal. DnaK/DnaJ/GrpE systems are molecular chaperones which facilitate proper folding of newly synthesized proteins. Efficient folding processes are of great importance especially in a cold environment, such as the Arctic. In order to understand the protection mechanisms of psychrophilic bacteria against cold temperatures, we have explored a genome of KOPRI22215, tentatively identified as Psychromonas arctica, whose genome sequence has not yet been discovered. With an aim of searching for a coding gene of DnaK from KOPRI22215, we have applied a series of polymerase chain reactions (PCR) with homologous primers designed from other Psychromonas species and LA PCR in vitro cloning. 1917 bp complete coding sequence of dnaK from KOPRI22215 was identified including upstream promoter sites. Recombinant plasmids to overexpress PaDnaK along with EcDnaK (DnaK of E. coli) were then constructed in pAED4 vector and the pET-based system to induce PaDnaK expression by IPTG. Characterization assays of expressed PaDnaK were carried out by measuring survival rates upon 4 day incubation at 4 $^{\circ}C$: a refolding assay as molecular chaperone, and ATPase assay for functional activity. Taking account of all the data together, we conclude that PaDnaK was identified, successfully expressed, and found to be more efficient in providing cold-resistance for bacterial cells.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.329-333
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• 2006

Two distinct and stable isolates of Barley mild mosaic virus(BaMMV) designated as Naju82-S(severe) and Naju82-M(mild) were obtained. These two isolates differed in their symptomatology, virus transmission characteristics and cultivar specificity at various temperature. Thus, these isolates were referred to as strains in this study. BaMMV Naju-S strain showed severe mosaic symptoms accompanied by necrosis on the infected leaves. Naju82-S strain is more virulent demonstrated by shorter incubation period and relatively high virus concentration than Naju82-M strain. Five Korean cultivars were tested for their pathogenicity to different strains based on the rate of infection. Results showed that infection rate of cultivars to both strains did not significantly differed from each other. However, under different temperatures, the pathogenicity on the two cultivars such as cultivars Hopumbori and Sessalbori were significantly affected. Hopumbori was moderately resistant to both strains at $10-12^{\circ}C$ and susceptible at $15-18^{\circ}C$. Similarly, Sessalbori was moderately resistant at $10-12^{\circ}C$ to both strains but distinctly differentiated at $15-18^{\circ}C$ wherein it was resistant to mild strain and highly susceptible to severe strain. Other cultivars including Baegdong, Jinyangbori and Neahanssalbori consistently showed susceptible reaction to both strains at varying temperatures tested in this study.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1894-1901
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• 2015

A novel sulfatase gene, ary423 (1,536 bp ORF), encoding a protein of 511 amino acids with a calculated molecular mass of 56 kDa, was identified from Flammeovirga pacifica, which was isolated from deep-sea sediments of west Pacific Ocean. Amino acid sequence analysis revealed that Ary423 possessed a conserved C-X-A-X-R motif, which was recognized as the sulfatase signature. Phylogenetic analysis suggested that Ary423 belonged to arylsulfatases. After heterologous expression in Escherichia coli cells, the recombinant Ary423 was purified with a Ni⁺ affinity column, and was shown to be highly active at a broad range of temperatures from 30° to 70℃, with maximum activity at 40℃. Furthermore, recombinant Ary423 retained more than 70% and 40% of its maximum activity after 12 h of incubation at 50℃ and 60℃, respectively, exhibiting good thermostability at high temperatures. The optimal pH for Ary423 was determined to be 8.0 and the activity of Ary423 could be slightly enhanced by Mg²⁺. The recombinant enzyme could hydrolyze sulfate ester bonds in p-nitrophenyl sulfate (NPS) and Asparagus crude polysaccharides with a specific activity of 64.8 U/mg and 25.4 U/mg, respectively. These favorable properties could make Ary423 attractive for application in the desulfating process of agar production.

• The Korean Journal of Ecology
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• v.18 no.3
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• pp.323-332
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• 1995

Effects of wintering and temperature on nitrogen fixation activity of nodules of Melilotus suaveolens Ledeb. grown in the field and growth chamber conditions were investigated. The biennial plants transfered to the growth chamber from winter field recovered the activity in 3 weeks of incubation and attained the maximum rate of $153{\mu}mol\;C_2H_4{\cdot}g$ fr wt $nodule^{-1}{\cdot}h^{-1}$ in 5 weeks. When root nodules which adapted to different temperatures, were pretreated with 10, 20 and $30^{\circ}C$ for 1 hour, and then transfered to $30^{\circ}C$, nitrogen fixation activity was promoted in the nodules exposed to lower field temperature ($12^{\circ}C$) with 1$0^{\circ}C$ pretreatment. M. suaveolens maintained nitrogen fixation activity in the wide range of temperatures, and was more tolerant to lower temperature than those of other woody leguminous plants, Diurnal changes of nodule activity showed increase with sunrise and decrease with sunset during spring and autumn, but the activity was inhibited during July and August because of high temperature with stron irradiation. Nitrogen fixation activity of annual plant appeared in mid-April, and showed two peaks (104 and 43 mol $C_2H_4{\cdot}g$ fr wt $nodule^{-1}{\cdot}h^{-1}$) in July and September, and then disappeared after October. Nitrogen fixation activity of biennial plant reappeared in mid-March after wintering and attained two peaks (102 and 82 ${\mu}mol\;C_2H_4{\cdot}g$ fr wt $nodule^{-1}{\cdot}h^{-1}$) in April and June of flowering period, and then disappeared after July due to plant withering by severe drought.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.123-131
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• 2008

Vitrification methods are convenient for cryopreserving plant specimens, as the specimens are plunged directly into liquid nitrogen (LN) from ambient temperatures. However, tissues and species with poor survival are still not uncommon. The development of vitrification solutions with high survival that cover a range of materials is important. We attempted to develop new vitrification solutions using bromegrass cells and found that VSL, comprising 20% (w/v) glycerol, 30% (w/v) ethylene glycol, 5% (w/v) sucrose, 10% (w/v) DMSO and 10 mM $CaCl_2$, gave the highest survival following cryopreservation, as determined by fluorescein diacetate staining. However, the cryopreserved cells showed little regrowth, for unknown reasons. To check its applicability, VSL was used to cryopreserve gentian axillary buds and the performance was compared with those of conventional vitrification solutions. Excised gentian stem segments with axillary buds (shoot apices) were two-step precultured with sucrose to induce osmotic tolerance prior to cryopreservation. Gentian axillary buds cryopreserved using VSL following the appropriate preculturing approach exhibited 78% survival (determined by the regrowth capacity), which was comparable to PVS2 and PVS1 and far better than PVS3. VSL had a wider optimal incubation time (20-45 min) than PVS2 and was more suitable for cryopreserving gentian buds. The optimal duration of the first step of the preculture was 7-11 days, and preculturing with sucrose and glucose gave a much higher survival than fructose and maltose. VSL was able to vitrify during cooling to LN temperatures, as glass transition and devitrification points were detected in the warming profiles from differential scanning calorimetry. VSL and its derivative, VSL+, seem to have the potential to be good alternatives to PVS2 for the cryopreservation of some materials, as exemplified by gentian buds.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.1
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• pp.117-126
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• 1984

Aflatoxins are toxic and carcinogenic secondary metabolites which are produced by trains of A. flavus and A. parasiticus during their growth on foods and feedstuffs. Aflatoxins are a group of closely related heterocyclic compounds of which $B_1$, $B_2$, and $G_2$ are the major members. Aflatoxins are synthesized via a polyketide pathway in which the general steps are acetate, an-thraquinones, xanthone and aflatoxins. Aflatoxin formation is favored by high moisture or high $a_w$(0.95${\sim}$0.99). The limiting $a_w$ for aflatoxin production on agricultural commodities is 0.83. Optimum temperature for aflatoxin production by the molds is $25{\sim}30^{\circ}C$ and the incubation time for the maximum production of the toxin is 7${\sim}$15 days. The limiting temperatures for aflatoxin production are ${\leq}7.5^{\circ}C\;and\;\geq40^{\circ}C$. Cycling temperatures may or may not stimulate aflatoxin production depending on the amplitude of cycling, substrate and strains of molds. Aflatoxin pro-ducing molds are aerobic organisms and thus have a requirement for oxygen. A decreasing $O_2$ concentration and/or increasing concentrations of $CO_2$ or $N_2$ depress the mold growth and aflatoxin formation. A. flavus grows competitively or associatively in the presence of other microorganisms and occasionally loses the competition with other microorganisms. Some lactic acid bacteria have been shown to reduce growth and aflatoxin production by A. parasiticus. Carbon source is the most important nutritional factors affecting aflatoxin formation by the molds. Sucrose, fructose and glucose are the most favorable carbon sources. Food substrates of plant derived products which have high carbohydrate content such as agricultural commodities and their products are most vulnerable to contamination by aflatoxins.

• Korean journal of food and cookery science
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• v.11 no.2
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• pp.153-157
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• 1995

The effect of low concentrations of ethanol(3∼7%, v/v) in culture broth as an antibacteriaB agent against Vibrio parahaemolyticus was tested at -20, 5, 35, 45 and 5%. Increasing concentrations of ethanol progressively inhibited initial growth of t: parahaemelyticus at 35$^{\circ}C$. Growth occured at 5% ethanol, but only after a prolonged lag period. At 7% ethanol, the number of viable cells of V parahae-molyticus declined during incubation. Culture broth containiilg 3∼7% ethanol was inoculated with 106∼107'cells/uu of V Parahaemolyticus and incubated at low temperatures(5$^{\circ}C$, -20$^{\circ}C$) and high tem-peratures(45$^{\circ}C$, 50$^{\circ}C$). In the presence of 5 or 7ft of ethanol, the viability in the cells incubated at high temperatures decreased rapidly. Rate of death increased with increasing concentration of etha-nol.

• Korean Journal of Plant Resources
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• v.27 no.3
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• pp.263-270
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• 2014

Loranthus tanakae (Franch. & Sav.) is an endangered species of mistletoe, distributed in Korean peninsula. The objective of our research is to determine the effect of storage duration and conditions [air flow (AF) and air tight (AT)] at different temperatures for survivability and germination of mistletoe seeds, and also to monitor the effect of seed scarification on germination in vitro. The result revealed that the seeds stored in natural conditions (no stratification) showed highest survival rate of 100% and retained up to 93.3% even after two months of storage in natural conditions and showed higher germination percentage (90%) compare to after ripened seeds. However, the seed stored at $0^{\circ}C$ decreased the germination percentage (ranged from 63 to 73%). Therefore, it can be confirmed that mistletoe does not need after ripened treatment to promote germination. Our research also showed that the storage of L. tanaka seeds in freezing temperatures of $-20^{\circ}C$ and in room temperature for long time either in AT or AF conditions caused the loss of survival and germination rate. On the other hand, the chemical scarification (0.01N HCl incubation for 12 hrs. at $38^{\circ}C$) method was proven more effective to enhance germination percentage of L. tanakae. Regarding the temperature regime, $22^{\circ}C$ showed early germination of mistletoe seeds in vitro.

• Parasites, Hosts and Diseases
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• v.36 no.1
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• pp.15-22
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• 1998

An in uitro culture technique was established for harvesting Strongwloides venezuelensis free-living infective larvae using a nutrient broth medium as a substitute for rat-feces in polyvinyl culture bags ($10{\;}{\times}{\;}12{\;}cm$). The egg hatch rate V) in sterile saline at different incubation temperatures (X) was expressed as the quadratic function, Y = $-0.192X^2$ + 8.673x - 19.550 (r = 0.901). The highest (100%) egghatch rate was observed at $25^{\circ}C$. A significant difference (p<0.05) in development rate W) of free-living infective larvae was observed between different concentrations of nutrient broth (X) which was highest (20.6%1 in 0.12% nutrient broth concentrations, incubated at $20^{\circ}C$ for 5 days [Y = $-864.032X^2$ + 245.995X- 0.560 (r = 0.875)]. Yields (Y) of infective larvae were observed relatively high when the culture medium was incubated at higher temperatures (X) which peaked at $25^{\circ}C$ (20.0%) than at lower temperatures. $15^{\circ}C$M (10.9%) and $20^{\circ}C$ (18.1%) [Y = $-0.189X^2$ + 8.387x- 72.795 (r = 0.981)]. The period W) required for the development of infective larvae decreased with higher incubation temperatures (X) [Y = $0.035X^2$ - 2.025X + 32.375 (r = 0.995)] The highest yield (19.2%) of infective larvae was obtained from culture bag inoculated with 15.000 eggs than with below and over 15,000 eggs in 0.12% nutrient broth and incubated at $25^{\circ}C$ for 4 days. The newly adapted culture method (from egg to third-stage larva) may be useful as a bio-bar/bioassay system for screening new chemical products, anthelmintics and pesticides, as well as for parasito immunological studies with Strongwloides species.