• 미생물학회지
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• 제54권1호
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• pp.9-17
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• 2018

저온 환경에서의 생장에 영향을 주는 지방산 합성 관련 유전자 bkdR, sigL, yplP, des들의 역할을 알아보기 위하여 각각 유전자들이 상실된 Bacillus subtilis CU1065와 JH642 돌연변이들을 제조하였다. 이들 유전자들의 저온 민감성을 확인하기 위해 $37^{\circ}C$와 $15^{\circ}C$에서 세포들의 생장을 측정하였다. $37^{\circ}C$에서 야생형과 결실 돌연변이 균주는 거의 유사한 정도의 생장을 보였으나, $15^{\circ}C$에서 오직bkdR 결실 돌연변이만이 야생형에 비해 매우 느린 생장이 관찰되었으며 sigL, yplP 결실의 경우 야생형에 비해 다소 느리거나 유사한 생장을 보였다. bkdR, sigL, yplP 결실에 대한 이중, 삼중 돌연변이를 만들어 LB agar에서 $20^{\circ}C$로 키워 저온생장을 조사한 결과, bkdR 결실이 포함되지 않은 어떤 이중, 삼중 결실들에서는 저온에 민감한 생장을 보이지 않았다. 온도 민감성 특성을 보다 잘 알아보기 위하여 $37^{\circ}C$에서 $OD_{600}=0.4$까지 키워 $15^{\circ}C$로 온도를 내리는 저온충격 조건에서 생장하는 실험을 진행하였다. 이 실험에서 오직bkdR 결실 돌연변이만이 현저히 낮은 생장을 보였으며 추가적인 des 결실은 저온 민감성을 증가시킨다. bkdR은 branched-chain fatty acid을 합성하는 전구물질인 isoleucine, valine, leucine 아미노산을 생산하는 bkd operon을 활성화한다. bkdR 결실 돌연변이의 저온생장에서 이들 아미노산의 저온생장에 미치는 영향을 조사한 결과 isoleucine은 bkdR 결실에 대한 저온 민감성을 회복시켜주나 valine은 저온 민감성을 회복시켜 주지 못하는 결과를 보였다. isoleucine은 분해되어 anteiso-branched 지방산 합성의 전구물질로 만들어지는 반면에, valine은 iso-branched 지방산 합성의 전구물질로 만들어진다. 따라서 저온생장에서 branched-chain fatty acid 중 anteiso-branched 지방산이 중요한 역할을 하고 있음을 알 수 있었다.

• Journal of Acupuncture Research
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• 제23권5호
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• pp.167-175
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• 2006

Objectives : We studied the effects of Radix Asteris herbal acupuncture solution (RAHAS) on the type I hypersensitivity. Methods : In vivo, we measured compound 48/80 induced active systemic anaphylactic shock, anti-DNP IgE induced passive cutaneous anaphylaxis (PCA) and acetic acid induced microvascular permeability using ICR mice. In vitro, we showed effects on cytotoxicity and ${\beta}$-hexosaminidase release from RBL-2H3 cells. Results : In vivo, RAHAS pretreatments at $BL_{13}$ and optional points inhibited active systemic anaphylactic shock induced by compound 48/80 and microvascular permeability increased by acetic acid. PCA was only inhibited by RAHAS pretreatments at $BL_{13}$. In vitro, RAHAS treatments inhibited ${\beta}$-hexosaminidase release. Conclusion : These results suggest that RAHAS may be beneficial in the prevention of type I hypersensitive inflammatory response.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1217-1226
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• 2004

A psychrotrophic bacterial strain, Pseudomonas fluorescens BM07, synthesized unsaturated fatty acids (UFA) from fructose in response to lowering of growth temperature, and incorporated them into both polyhydroxyalkanoic acid (PHA) and membrane lipid. The blocking of PHA synthesis by adding 5 mM 2-bromooctanoic acid to the growth medium, containing 70 mM fructose, was found to be a useful means to profile the composition of membrane lipid by gas chromatography. As the growth temperature changed from 35 to $50^{\circ}C$, the total content of two UFA, 3-hydroxy-cis-5­dodecenoic acid ($C_{12:1}$) and 3-hydroxy-cis-7-tetradecenoic acid ($C_{14:1}$), in PHA increased from 31 to 44 $mol\%$. The growth at lower temperatures also led to an increase in the level of two major UFA, palmitoleic acid (C16:1 cis9) and cis-vaccenic acid (C18:1 cis11), in membrane lipid. A fraction of these membrane-lipid UFA was converted to their corresponding cyclopropane fatty acids (CFA). The CFA conversion was a function of culture time, exhibiting biphasic increase before and after entering the stationary phase. However, pH changes in growth media had no effect on the CFA conversion, which is contrary to the case of E. coli reported. The cells grown at $30^{\circ}C$ responded to a cold shock (lowering the medium temperature down to $10^{\circ}C$) by increasing the level of C16:1 cis9 and C 18: I cis II up to that of $10^{\circ}C$-grown control cells and concomitantly decreasing the relative level of cis-9,10­methylenehexadecanoic acid (the CFA converted from C16:1 cis9) from 14 to 8 $mol\%$, whereas the 10-grown cells exhibited little change in the lipid composition when exposed to a warmer environment of $30^{\circ}C$ for 12 h. Based on this one- way response, we suggest that this psychrotrophic strain responds more efficiently and sensitively to a cold shock than to a hot shock. It is also suggested that BM07 strain is a good producer of two unsaturated 3-hydroxyacids, $C_{12:1}\;and\;C_{141:1}$.

• 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.

• 한국식품저장유통학회지
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• 제19권6호
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• pp.813-817
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• 2012

신선절단 '후지' 사과의 갈변도에 화학물질(ascorbic acid, cystein, sodium chloride, magnesium chloride, calcium chloride, histidine: 1% 용액), 한약재(갈근, 당귀, 산약, 오약, 천궁, 황기: 1% 추출액), 가열(절단과실과 원형과실, $40-60^{\circ}C$ 증류수) 및 코팅(albumin, dextrin, sucrose poly ester, whole soy flour: 0.5-4% 용액) 처리가 미치는 영향을 조사하였다. 각 처리방법별 신선절단 사과의 갈변도는 1% calcium chloride 용액 처리, 황기 추출물 처리, 3% dextrin과 1% whole soy flour 코팅 처리에 의해 가장 낮은 수준을 보였다. 가열처리의 갈변도는 절단과실 처리시에는 $50^{\circ}C$에서, 원형과실 처리시에는 $45^{\circ}C$에서 각각 처리한 시료에서 가장 낮은 수준을 보였다. 이로써 신선절단 사과의 갈변에 대한 한약재 추출물, 가열 및 코팅 처리의 억제 효과와 조건이 확인되었고, 이러한 방법은 화학물질을 부분적으로 대체할 수 있을 것으로 판단된다.

• BMB Reports
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• 제38권5호
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• pp.602-608
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• 2005

As a crucial transcription factor family, heat-shock factors were mainly analyzed and characterized in tomato and Arabidopsis. In this study, we isolated two putative heat shock factors OsHSF6 and OsHSF12 that interact specifically with heat-shock element (HSE) from Oryza sativa L by yeast one-hybrid method. The full-length cDNA of OsHSF6 and OsHSF12 have 1074bp and 920bp open reading frame (ORF), respectively. Analysis of the deduced amino acid sequences revealed that OsHSF6 was a class A heat shock factor (HSF) with all the conserved sequence elements characteristic of heat stress transcription factor, while OsHSF12 was a class B HSF with C-terminal domain (CTD) lacking of AHA motif. Bioinformatic analysis showed that the sequences and structures of two HSFs' DNA binding domain (DBD) had a high similarity with LpHSF24. The results of RT-PCR indicated OsHSF6 gene was expressed immediately after rice plants exposure to heat stress, and the transcription of OsHSF6 gene accumulated primarily in immature seeds, roots and leaves. However, we did not find the transcription of OsHSF12 gene in different organs and growth periods. Our results implied that OsHSF6 might be function as a HSF regulating early expression of stress genes in response to heat shock, and OsHSF12 might be act as a synergistic factor to regulate the expression of down-stream genes.

• 생명과학회지
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• 제12권5호
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• pp.528-534
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• 2002

우리나라에서 식용으로 뿐만아니라 한방재료로 널리 사용되고 있는 녹두(vigna radiata L. wilczek) 로부터 trypsin inhibitor (Mung bean trypsin inhibitor, MBTI)를 분리정제하여 그 특성을 조사하였다 또한 병태동물모델 즉, septic shock induced guinea pig model을 이용하여 MBTI의 약물학적 효과를 평가하였다. MBTI의 분리 및 정제과정은 Sephadex C-50 chromatography, DEAE-celluloseion exchange chromatography 및 trypsin affinity column 을 차례로 이용하였다. 정제한 MBTI는 전기영동 및 아미노산 서열분석결과 분자량 약 8,000 Da 의 BBI-type (Bowman-birk inhibitor type)임을 알 수 있었으며 이들의 생화학적 특성을 구명하였다. 또한 pseudomonal elastase로 유도된 septic shock guinea pig model에서 MBTI 10 mg/kg를 전처치한 결과 hypotention shock 유발이 억제됨을 알 수 있었다.

• BMB Reports
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• 제31권2호
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• pp.201-208
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• 1998

Amino acid analogs, like other inducers of stress response, induce the synthesis of stress proteins in mammalian cells. In this study, Drosophila Kc cells, in which translation is tightly controlled during stress response, was treated with proline analogs, L-azetidine-2-carboxylic acid (AzC) and 3,4-dehydro-L-proline (dh-P). Kc cells exposed to AzC or dh-P induced the synthesis of several proteins which had the same molecular weights as known heat shock proteins. However, in Kc cells, normal protein synthesis still continued in the presence of amino acids analogs unlike in heat-shocked cells. For the induction of stress response, the incorporation of dh-P into the protein was not essential, but the incorporation of AzC was. The stress protein synthesis was regulated mainly at the transcriptional level by AzC, whereas it was regulated by dh-P at the transcription level and possibly posttranscription level. During recovery, the stress protein synthesis stopped sooner in analog-treated cells than in heat-shocked cells even though the accumulated amount of Hsp70 was much less in proline analogstreated cells. It could be concluded that the proline analogs, AzC and dh-P, induced stress response through a different mechanism from heat shock.

• 한국분말재료학회지
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• 제8권3호
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• pp.174-178
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• 2001

The microstructure and mechanical properties of $Si_3N_4/BN $nanocomposites synthesized by chemical processing were investigated. The nanocomposites containing 15 vol% hexagonal BN (h-BN) were fabricated by hot-pressing $\alpha-Si_3N_4$powders covered with turbostratic BN (t-BN). The t-BN coating on $\alpha-Si_3N_4$particles was prepared by heating $\alpha-Si_3N_4$ particles covered with a mixture of boric acid and urea in hydrogen gas. TEM observations of this nanocomposite revealed that nano-sized h-BN particles were homogeneously dispersed within $Si_3N_4$grains as well as at grain boundaries. The strength and thermal shock resistance were significantly improved in comparison with the $Si_3N_4/BN$ microcomposites.

• 한국동물학회지
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• 제35권2호
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• pp.203-210
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• 1992

Hsp70, a 70 kDa protein, is the maior protein expressed when cells are heat-shocked. A cDNA library from mouse ID13 cells was screened with the human hsp70 gene as a probe, and a positive clone was obtained. The positive clone was subcloned into puc19 and the precise restriction was obtained. The CDNA was sequenced by the Sanger's dideoxv termination method. Single open reading frame that codes for a protein of 70 kDa was found. The DNA sequence of the cloned mouse DNA shows great homology (66-90%) with other mouse hsp70 genes and somewhat less homology (50",) with E. coli hsp70 gene (dnak). With the exception of one amino acid, the protein sequence deduced from the CDNA is identical to the mouse that shock cognate protein 70 (hsc70) that is constitutivelv expressed at normal temperature. The result suggests that the cloned CDNA encodes a hsc70 family rather than a heatinducible family.mily.