• Journal of Microbiology
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• 제41권1호
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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.

• 한국식품저장유통학회지
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• 제18권4호
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• pp.612-619
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• 2011

C. sakazakii ATCC 12868, 29004, 29544를 이용하여 저온 저장 중의 변화를 살펴보고 저온과 냉/해동, 저온과 산, 저온에서 starvation한 것과 냉/해동의 교차저항에 대해 알아보았다. C. sakazakii를 $5^{\circ}C$에서 10일간 저장하였을 때 모든 균주들에서 1 log CFU/mL의 사멸을 보였다. C. sakazakii를 $13^{\circ}C$에서 배양한 결과, C. sakazakii ATCC 12868, 29004는 각각 7일째, 5일째에 $10^9$ CFU/mL을 나타냈고 C. sakazakii ATCC 29544는 5일 배양 후부터 30일 동안 $10^9$ CFU/mL에 도달하지 못하고 $10^8$ CFU/mL를 유지하였다. 저온과 냉/해동의 교차저항 결과, C. sakazakii ATCC 12868, 29004에서 대조구와 $5^{\circ}C$에서 2일간 저장한 것은 4 log CFU/mL의 사멸을 보였고 $13^{\circ}C$에서 배양한 것은 일정한 균수를 유지하였다. 저온과 산의 교차저항 실험에서 C. sakazakii ATCC 29544를 제외하고 다른 두 균주에서 대조구에 비해 $5^{\circ}C$ 에서 2일간 저장한 것, $13^{\circ}C$에서 배양한 것은 일정하게 높은 생존율을 보였다. C. sakazakii는 멸균된 $5^{\circ}C$ 0.1% 펩톤수에서 starvation한 것이 $5^{\circ}C$의 TSB배지와 멸균된 증류수에서 starvation한 것보다 냉/해동 환경에서 D값이 가장 높게 나타났다. C. sakazakii는 $5^{\circ}C$에서 균의 생장이 저해되었지만 $13^{\circ}C$에서 C. sakazakii의 생장이 가능하였다. 이 연구 결과는 저온에서 적응하거나 저온에서 배양된 C. sakazakii의 다른 환경에서 교차저항성을 알아보는데 유용한 기초 자료로 사용될 수 있을 것이다.

• 대한조선학회논문집
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• 제50권6호
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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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• 제31권4호
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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.

• 한국작물학회지
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• 제40권2호
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• pp.203-211
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• 1995

벼 품종의 내냉성정도를 판정하는데 기준이 되는 몇가지 형질들에 대하여 품종형에 따른 저온반응의 차이를 구명하고자 오대벼외 187개 품종을 공시하여 실험을 수행한 바 그 결과를 요약하면 다음과 같다. 1. 각 품종형에 따라 대부분의 형질에서 서로 상이하면서 다양한 변이를 나타내었는데, 자포니카형 과 인디카형 에서 는 냉해에 강한 품종이 많은 반면, 통일형 품종은 대부분의 형질에서약한 반응을 나타냈다. 2. 적고현상은 대부분의 자포니카형 품종에서 강한 쪽으로, 통일형 품종은 약한 쪽으로, 그리고 인디카형 품종은 강한 것에서 약한 것까지다앙한 변이분포를 나타냈다. 3. 공시한 품종들간에 증감의 차이가 가장 적게 나타난 형질은 수수와 영화수였으며, 특히 가수는 품종형에 관계없이 188개 품종 중 34개 품종이 냉수구에서 오히려 증가되었다. 4. 출수의 지연, 간장의 단축, 이삭추출도 불량 및 억책율 저하 등에서 통일형 품종은 대부분 그 정도가 심한 반면, 자포니카형 품종과 인디카형 품종에서는 공히 저온반응이 강한 것에서 약한 품종까지 광범위한 변이분포를 나타냈다.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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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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• 제19권2호
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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.

• 원예과학기술지
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• 제29권6호
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• pp.561-567
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• 2011

상록활엽관목으로서 꽃이 화려하고 관상가치가 높으나 멸종위기 수종인 자생만병초와 원예도입종 만병초 품종을 공시하여 내한성의 기작을 잎 운동과 광합성 활성 등의 생리적 반응을 조사하여 구명하고자 하였으며 다음과 같이 요약된다. 월동 후의 엽소 피해 정도로 측정한 내한성 정도는 자생종인 만병초와 홍만병초에서 도입종보다 크게 높았으며, 도입종 중에서도 'Nova Zembla'에서 가장 낮았고, 'Cynosure'와 'Parker's Pink'는 이보다 다소 높았다. 저온에 의한 엽각의 변화는 자생종과 'Parker's Pink' 품종에서 크게 일어났으며, 'Nova Zembla'와 'Cunningham's White'에서의 변화는 적었다. 저온에 의한 잎말림 현상도 자생종에서 크게 일어났고 도입종은 이들보다 작았으며, 도입종 중에서는 'Parker's Pink'가 다소 컸고 나머지 품종들은 작았다. 자생종의 내한성은 저온에 의한 엽각변화와 잎말림현상이 관여하는 것으로 판단되며, 내한성이 약한 'Parker's Pink'에 있어서는 잎운동 활성과의 관련성이 적어 다른 요인이 관여하고 있는 것으로 생각된다. 내한성이 강한 만병초에서 월동 전 광합성 활성이 높았고 월동 후 광합성활성의 회복이 자생종 및 도입종의 'Cynosure'에서 비교적 빨랐으며, 내한성이 가장 약했던 'Nova Zembla'에서 월동 전 광합성 활성과 월동 후 광합성활성의 회복속도가 가장 낮았다. 기공전도도는 광합성활성과 유사한 경향을 보였으며, 내한성이 강한 자생종에서 월동 후 뿌리로부터 잎으로의 수분이동이 원활하여 엽소현상이 적었고, 월동 후 기공전도도가 가장 낮았고 수분이동에 크게 제한을 받은 'Nova Zembla'에서 엽소현상이 크게 나타난 것으로 생각된다. 이상의 결과로부터 만병초 종에 있어서 월동중의 잎 운동과 월동후의 수분이동 및 광합성에 의해 축적되는 수분스트레스 관련 물질들이 종합적으로 관여하여 엽소현상으로서 나타나는 내한성을 결정하는 것으로 생각된다.

• 대한간호학회지
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• 제13권2호
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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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• 제15권4호
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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.