• Journal of Microbiology
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• v.41 no.1
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• pp.58-62
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• 2003

An extracellular pretense of Bacillus amyloliquefaciens S94 was purified to apparent homogeneity. The enzyme activity was strongly inhibited by general inhibitor for serine protease, PMSF, suggesting that the enzyme is a serine pretense. The purified enzyme activity was inhibited by leucine peptidase inhibitor, bestatin, suggesting that the enzyme is a leucine endopeptidase. The maximum proteolytic activity against different protein substrates occurred at pH 10, 45$^{\circ}C$ (protein substrate) and pH 8, 45$^{\circ}C$ (synthetic substrate). The purified enzyme was specific in that it readily hydrolyBed substrates with Leu or Lys residues at P$_1$ site. The pretense had characteristics of a cold-adapted protein, which was more active for the hydrolysis of synthetic substrate in the range of 15$^{\circ}C$ to 45$^{\circ}C$, specially at low temperature.

• Food Science and Preservation
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• v.18 no.4
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• pp.612-619
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• 2011

Cronobacter sakazakii has been isolated from a wide range of environmental sources and from several foods of animal and plant origin. The objective of this study was to determine the resistance of C. sakazakii (ATCC 12868, ATCC 29004, and ATCC 29544) in cold, cold-freeze thaw, cold-acid, and cold starvation-freeze thaw stress. The number of C. sakazakii decreased to 1 log CFU/mL at $5^{\circ}C$ (cold storage) for 10 days. When C. sakazakii was cultivated at a low temperature ($13^{\circ}C$), the population of C sakazakii ATCC 12868 and 29004 increased to $10^9$ CFU/mL, and the population of C. sakazakii ATCC 29544 increased to $10^8$ CFU/mL. For C. sakazakii ATCC 12868 and 29004, the cold-adapted cells ($5^{\circ}C$ 24 hr) decreased by 4 log CFU/mL, and the low-temperature-cultivated cells ($13^{\circ}C$) decreased by 0.5 log CFU/mL. In this study, low-temperature cultivation enhanced the freeze-thaw cross-resistance due to the metabolic changes in the cells. Cold stress ($5^{\circ}C$ 48 hr, $13^{\circ}C$ cultivation) enhanced the cold-acid cross-resistance. The cold-starved cells in the sterilized 0.1% peptone water enhanced the freeze-thaw cross-resistance with significant differences (p<0.05). Therefore, the increased tolerance of the cold-adapted or low-temperature-cultivated C. sakazakii cells to freeze-thaw, acid, or starvation suggests that such environments should be considered when processing minimally processed foods or foods with extended shelf life.

• Journal of the Society of Naval Architects of Korea
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• v.50 no.6
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• pp.450-457
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• 2013

For the design guide of a vessel operating in cold region, numerical analysis was carried out to evaluate the weather-tight door which installed the heating cables by using ANSYS 13.0 Transient Thermal. The numerical analysis was performed by considering Advection-Diffusion equation. This study based on the experimental results of 'A study on Anti-Icing Technique for Weather-Tight Door of Ice-Strengthened Vessels'(Jeong, et al., 2011a) in KIOST. For validation of the numerical analysis results, the cold chamber experimental data measured by the heat sensors in certain location of the weather-tight door was used. The external environmental temperature which varies from $5^{\circ}C$ to $-55^{\circ}C$ was considered in numerical analysis. Also three different heating cables which have the heat capacity of 33W/m, 45W/m and 66W/m were adapted for the design parameters to be the most efficient and guidelines for anti-icing design of the weather tight door.

• Bulletin of the Korean Chemical Society
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• v.31 no.4
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• pp.887-890
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• 2010

The family of ClpB protein is a molecular chaperone which protects cellular proteins from being aggregated upon exposure to severe environmental stresses in association with DnaK/DanJ/GrpE in the ATP-dependent manner. In a psychrophilic bacterium which survives at a subzero temperature, any functional role of cold-active ClpB protein can be rather crucial. In order to identify a ClpB encoding gene from a cold-adapted bacterium whose genome sequence has not been fully discovered, we have employed a series of PCR technologies, including a gradient PCR with homologous primers, an inverse PCR and a cassette PCR. The full sequence of PaclpB gene was successfully identified and compared with those of other psychrophilic species. We have further cloned the gene in E.coli expression systems and were able to induce PaClpB protein expression by IPTG, which help us understand a molecular mechanism for survival against extremely cold environments.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.2
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• pp.203-211
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• 1995

This study was conducted to know how low temperature response of rice varieties is available for breeding lines adapted to cold weather. Some agronomic characters related to cold tolerance were evaluated for 188 varieties including three varietal types of Japonica, Tongil, and Indica. Cold tolerance of Japonica and Indica type varieties varied in leaf discoloration, heading delay, reduction of culm length, panicle number, spikelet number, panicle exsertion, spikelet fertility, and phenotypic acceptability at maturity, while Tongil type varieties were susceptible in most agronomic characters. In leaf discoloration, most Japonica type varieties were highly tolerant, while Indica type varieties showed various responses from tolerant to susceptibility. Not significant difference among three varietal types was observed on reduction of panicle number and spikelet number due to low temperature.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.128-135
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• 2009

Acinetohacter baumannii BD5 was isolated from waters of Baek-du mountain, and the lipase gene was cloned using a PCR technique. The deduced amino acid sequence of the lipase and lipase chaperone were found to encode proteins of 325 aa and 344 aa with a molecular mass of 35 kDa and 37 kDa, respectively. The lipase gene was cloned and expressed in Escherichia coli BL21(trxB) as an inclusion body, which was subsequently solubilized by urea, and then purified using Ni-affinity chromatography. After being purified, the lipase was refolded by incubation at $4^{\circ}C$ in the presence of a 1:10 molar ratio of lipase:chaperone. The maximal activity of the refolded lipase was observed at a temperature of $35^{\circ}C$ and pH 8.3 when p-NP caprate(C10) was used as a substrate; however, 28% of the activity observed at $35^{\circ}C$ was still remaining at $0^{\circ}C$. The stability of the purified enzyme at low temperatures indicates that it is a cold-adapted enzyme. The refolded lipase was activated by $Ca^{2+},\;Mg^{2+},\;and\;Mn^{2+}$, whereas $Zn^{2+}\;and\;Cu^{2+}$ inhibited it. Additionally, 0.1% Tween 20 increased the lipase activity by 33%, but SDS and Triton X-100 inhibited the lipase activity by 40% and 70%, respectively.

• Horticultural Science & Technology
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• v.29 no.6
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• pp.561-567
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• 2011

Cold tolerance of the native Rhododendron species which are on the verge of extinction in Korean nature were compared with the introduced species and its mechanism were studied physiologically with the investigation of the leaf angle, leaf curling, and photosynthetic activity. The degree of cold tolerance measured with the leaf burning after winter season was higher in the native species, Rhododendron brachycarpum and Rhododendron brachycarpum var. roseum than all the introduced species. 'Nova Zembla', an introduced species, showed high sensitivity to the low temperature. Changes in leaf angle by the low temperature were bigger in 2 native species and 'Parker's Pink' than the other introduced species and small comparatively in 'Nova Zembla' and 'Cunningham's White' cultivar. Leaf curling also occurred strongly in 2 native species by the low temperature. While, it was comparatively little and mild in the other introduced species. Therefore these results suggested that the leaf movement such as leaf angle change and curling adapted to the low temperature is positively related to the cold tolerance of 2 native species. By the way, such relationship is not explainable in the cold-sensitive 'Parker's Pink' cultivar showing comparatively stronger leaf movement. Photosynthetic activity measured before the winter season was high in the cold-tolerant R. brachycarpum and its recovery after winter season was faster in the 2 native species and the introduced 'Cynosure' cultivar than the other introduced species. They were the lowest in the most cold-sensitive 'Nova Zembla'. This phenomena occurred similarly even in the stomatal conductivity, suggesting that the movement of water from the roots to the leaves is better and then the leaf burning after winter season become small in the cold-tolerant species. The recovery of photosynthetic activity and stomatal conductivity was comparatively slower in the cold-sensitive 'Parker's Pink'. From the above results, leaf behavior adapted to the low temperature during the winter season and water movement to the leaves are related collectively to the cold tolerance represented as the leaf burning in the Rhododendron species is suggested.

• Journal of Korean Academy of Nursing
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• v.13 no.2
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• pp.87-104
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• 1983

It has been well documented that animals exposed to cold show increased activity of thyroid gland. The calorigenic action of thyroid hormone has been demonstrated by a variety of in vivo and in vitro studies. According to Edelman et al., the thyroid thermogenesis is due to activation of energy consuming processes, especially the active sodium transport by the hormone in target tissues. If so, the increase in thyroid activity during cold exposure should induce increased capacity of sodium transport in target tissue and the change in tissue metabolism should be precisely correlated with the change in Na+_K+_ATPase activity of the tissue. This possibility was tested in the present study: in one series, changes in oxygen consumption and Na+_K+_-ATPase activity of liver preparations were measured in rats as a function of thyroid status, in order to establish the effect of thyroid hormone on the tissue respiration and enzyme system in another series, the effect of cold stimulus on the serum thyroid hormone level, hepatic tissue oxygen consumption and Na+_K+_ATPase activity in rats. The results obtained are as follows: 1. The Na+_dependent oxygen consumption of liver slices, the oxygen consumption of liver mitochondria and the Na+_K+_ATPase activity of liver preparations were significantly inhibited in hypothyroidism and activated in hyperthyroidism. Kinetic analysis indicated that the Vmax. of Na+_K+_ATPase was decreased in hypothyroidism and increased in hyperth)'roidism. 2. In cold exposed rats, the serum triiodothyronine (T₃) level increased rapidly during the initial one day of cold exposure, then declined slowly to the control level after two weeks. The serum thyroxine (T₄) level decreased gradually throughout the cold exposure. Accordingly the T₃/T₄ratio increased. The mitochondrial oxygen consumption and the Na+_dependent oxygen consumption of liver slices increased during the first two days and then remained unchanged thereafter The activity of the Na+_K+_ATPase in liver preparations increased during cold exposure with a time course similar to that of oxygen consumption. Kinetic analysis indicated that the Vmax. of Na+_K+_ATPase increased. 3. Once the animal was adapted to cold, induction of hypothyroidism did not significantly alter the hepatic oxygen consumption and Na+_K+_ATPase activity. These results indicate that: 1) thyroid hormone increases capacities of mitochondrial respiration and active sodium transport in target tissues such as liver; 2) the increased T₃level during the initial period of cold exposure facilitates biosynthesis of Na+_K+_ATPase and mitochondrial enzymes for oxidative phosphorylation, leading to enhanced production and utilization of ATP, hence heat production.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.831-837
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• 2005

Low-temperature adaptation and cryoprotection were studied in Leuconostoc mesenteroides SYl, a strain isolated from Kimchi. L. mesenteroides SY1 cells grown in exponential growth phase at $30^{\circ}C$ were exposed to $15^{\circ}C,\;10^{\circ}C$, and $5^{\circ}C$ for 2, 4, and 6 h, respectively, and then frozen at $- 70^{\circ}C$ for 24 h. Survival ratio was measured after the cells were thawed. The freezing-thawing cycles were repeated four times. Preadapted cells survived better than non-adapted control cells, and the highest survival ratio ($96\%$) was observed for cells preadapted for 2 h at $5^{\circ}C$, whereas control cells showed only $22\%$. The 2D gel showed that two proteins (spots A and B) were induced in cells preadapted at lower temperatures. Spots A and B have the same molecular weight (7 kDa), but the pI was 4.6 for spot A and 4.3 for spot B. The first 29 and 15 amino acid sequences from spots A and B were determined, and they were identical, except for one amino acid. A csp gene was cloned, and nucleotide sequencing confirmed that the gene encoded spot A cold shock protein.