• Title/Summary/Keyword: P-stress

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Construction of hsf1 Knockout-mutant of a Thermotolerant Yeast Strain Saccharomyces cerevisiae KNU5377 (고온내성 연료용 알코올 효모균주 Saccharomyces cerevisiae KNU5377에서 HSF1 유전자의 변이주 구축)

  • Kim Il-Sup;Yun Hae-Sun;Choi Hye-Jin;Sohn Ho-Yong;Yu Choon-Bal;Kim Jong-Guk;Jin Ing-Nyol
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.454-458
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    • 2006
  • HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

Kinetic Modeling of the Enzymatic Hydrolysis of $\alpha$-Cellulose at High Sugar Concentration (순수 섬유소에 대한 고농도 당화공정의 동력학적 모사)

  • 오경근;정용섭홍석인
    • KSBB Journal
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    • v.11 no.2
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    • pp.151-158
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    • 1996
  • For the effective ethanol fermentation, the high concentration of sugar as the substrate of microbial fermentation is required. The most important reason in the inefficient hydrolysis; the easy deactivation of enzyme by temperature or shear stress and the severe inhibition effects of its products. In our work, we comprehended the kinetic characteristics of cellulose and ${\beta}$-glucosidase in the progress of hydrolysis, and observed the potential inhibitory effects of the hydrolyzed products and the deactivation of enzymes. We also tried to present the kinetic model of enzymatic hydrolysis of cellulose, which is applicable to process at the high concentration of sugar. Cellulase and ,${\beta}$-glucosidase exhibit diverse kinetic behaviors. At a level of only 5g/$\ell$ of glucose, the ${\beta}$-glucosidase activity was reduced by more than 70%. This result means that ${\beta}$-glucosldase was the most severely inhibited by glucose. Also at l0g/$\ell$ of cellobiose, the cellulose lost approximately 70% of its activity. ${\beta}$-glucosldase was more sensitive to deactivation than cellulose by about 1.6 times. The comprehensive kinetic model in the range of confidence was obtained and the agreement between the model prediction and the experimental data was reasonably good, testifying to the validity of the model equations used and the associated parameters.

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Analysis of Nutrition and Antioxidants of Yak-Kong Chungkukjang Powder Added Black Foods (블랙푸드가 첨가된 약콩청국장분말의 영양소 및 생리활성물질 분석)

  • Kong, Hyun-Joo;Park, Heyun-Sook;Kim, Tae-Hoon;Shin, Seung-Ryeul;Hong, Ju-Yeon;Yang, Kyung-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1727-1735
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    • 2013
  • This study is carried out to investigate the analysis of nutrition and antioxidants of soybean Chungkukjang powder (SCP), Yak-Kong Chungkukjang powder (YCP) and Yak-Kong Chungkukjang powder added black foods (YCBP) for the management of Alzheimer's disease. The water content of YCBP was higher than that of SCP and YCP. The soluble and crude protein contents of YCP were the highest among three Chungkukjang powders. In SCP, YCP and YCBP, the ratios of unsaturated and saturated fatty acids were 5.17, 5.76, and 5.78, respectively. The mineral content of SCP was higher than that of YCP and YCBP. Antioxidants analysis showed that the content of diadzein was the highest in YCP and genistein, and anthocyanin contents were higher than the others in YCBP. In conclusion, YCP and YCBP had higher contents than SCP in soluble and crude proteins, and unsaturated fatty acids, which are needed for composition and function of the brain tissue. Also, it was found that the contents of diadzein, genistein and anthocyanin with outstanding antioxidative ability were high. Thus, this study suggests that Chungkukjang powder, which is made with Yak-Kong, black sesame, black rice and sea tangle, can be utilized for in vivo experiment for the control of oxidative stress, reported as cause and therapy for Alzheimer's Disease.

Physical and Mechanical Properties of Cements for Borehole and Stability Analysis of Cement Sheath (관정 시멘팅 재료의 물리역학물성 및 시멘트층의 안정성 분석)

  • Kim, Kideok;Lee, Hikweon;Kim, Taehee;Kim, Gyo-Won
    • The Journal of Engineering Geology
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    • v.26 no.1
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    • pp.101-115
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    • 2016
  • We carried out laboratory material tests on two cements (KS-1 ordinary Portland and Class G) with changing W/S (Water/Solid) and the content of fly ash in order to evaluate their physical and mechanical properties. The specimens of KS-1 ordinary Portland cement were prepared with varying W/S (Solid=cement) in weight, while those of Class G cement were prepared with changing the content of fly ash in volume but maintaining W/S (Solid=cement+fly ash). The results of the material tests show that as the W/S in KS-1 ordinary Portland cement and the content of fly ash in Class G cement increase, the properties (density, sonic wave velocity, elastic constants, compressive and tensile strengths, thermal conductivity) decrease, but porosity and specific heat increase. In addition, an increase in confining pressure and in the content of fly ash leads to plastic failure behavior of the cements. The laboratory data were then used in a stability analysis of cement sheath for which an analytical solution for computing the stress distribution induced around a cased, cemented well was employed. The analysis was carried out with varying the injection well parameters such as thickness of casing and cement, injection pressure, dip and dip direction of injection well, and depth of injection well. The analysis results show that cement sheath is stable in the cases of relatively lower injection pressures and inclined and horizontal wells. However, in the other cases, it is damaged by mainly tensile failure.

Characteristics of Thermal Variations with the Different Land Covers in an Urban Area (도시 지역에서 토지 피복에 따른 열 변이 특성)

  • Park, Sung-Ae;Kong, Hak-Yang;Kim, Seung-Hyun;Park, Sungmin;Shin, Young-Kyu
    • Ecology and Resilient Infrastructure
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    • v.3 no.1
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    • pp.46-53
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    • 2016
  • This study was conducted to analyze the effect of the different land covers of an urban park (Hyowon park) in downtown Suwon on the urban thermal variations during a hot summer. The effect of the air temperature reduction in the urban park was 4.4%-4.5% for the downtown residence (Maetan-dong). This value was about 0.8% lower than that of the outskirts residence (Sanggwanggyo-dong). The daily mean temperature, daily maximum temperature, summer day and heat wave frequency were measured under the different land covers (cement-block, grass, pine-grass, shading area and mixed forest) showed these values generally decreased under natural land cover types. Daily minimum temperature and tropical night frequency didn't seem to correlate with the land cover types. Means of thermal comfort indices (wet bulb globe temperature, heat index and discomfort index) in the shading area, mixed forest and the pine-grass types were lower than those of cement block and grass types. However the levels of those indices were equal to 'very high' or 'caution' levels in the afternoon (13:00-15:00). In the morning (06:00-08:00), thermal comfort indices of the urban park didn't correlate with land cover types. Therefore, to reduce heat stress and to improve the thermal comfort in urban parks, an increase in the area of natural land cover such as grass, forest and open spaces is required.

Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70 (수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현)

  • Min, Byung Hwa;Hur, Jun Wook;Park, Hyung Jun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.639-647
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    • 2018
  • A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • LEE Mi-Ock;SONG Ki-Hong;LEE Hyun-Kyung;JUNG Ji-Yoon;CHOE Vit-Nary;CHOE Sunghw
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04a
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    • pp.69-75
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    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd/dwf3 were shown to be blocked in $D^4$ reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bri1/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus It is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRI1 could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

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Function of Dual Specificity Kinase, ScKns1, in Adhesive and Filamentous Growth of Saccharomyces cerevisiae (Saccharomyces cerevisiae의 균사형 생장에서 이중 특이성 인산화 효소, ScKns1p의 기능 분석)

  • Park, Yun-Hee;Yang, Ji-Min;Yang, So-Young;Kim, Sang-Mi;Cho, Young-Mi;Park, Hee-Moon
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.110-116
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    • 2011
  • In the previous study with the Saccharomyces cerevisiae S288c strains, no known function of the dual specificity kinase, ScKns1, was reported because its gene deletion did not show any noticeable phenotypic changes. Recent study with fission yeast, however, revealed the involvement of the LAMMER kinase in flocculation, filamentous growth, oxidative stress, and so on. Therefore we made Sckns1-deletion mutants with the ${\Sigma}1278b$-background, with which one can induce filamentous and adhesive growth in contrast to those of the S288c-background. The $Sckns1{\Delta}$ strains of both haploid and diploid showed defect in filamentous growth under conditions for inducing the filamentous growth such as nitrogen starvation and butanol treatment. Both kinds of the deletion mutants also showed decrease in adhesive growth on agar surface. Interestingly enough the defects of the $Sckns1{\Delta}$ strains were suppressed by the over-expression of each gene for the components of the MAPK signaling pathway such as STE11, STE12, and TEC1, respectively, but not by the upstream components, RAS2 and STE20, respectively. Although further investigations are required, these results indicate that the ScKns1 may act in place between the Ste20 and the Ste11 of the S. cerevisiae MAPK cascade.

Changes in the Myocardial Antioxidant Enzyme System by Post-Ischemic Reperfusion During Corontory Artery Bypass Operations (관상동맥우회술시 심근허혈후 재관류에 의한 활성산소 방어효소계의 변화)

  • 김응중;김기봉
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.850-860
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    • 1996
  • Oxygen free radicals and their metabolites have been implicated as possible causes of reperrusion injury In animal models. Their role in the clinical setting is still controversial. The aim of this study was to evaluate the degree of tissue damage, oxidative stress. and changes in the antioxidant enzyme system in patients undergoing cor nary artery bypass graft operations(CABG) with myocardial protection by cold blood cardioplegia. In patients undergoing CABG(n:10). the levels of lactate dehydrogenate(LDH), creatine phosphokinase MB fraction(CK-MB), and malondialdehyde(M DA) were measured In the coronary sinus effluent before aortic cross clamping and 20 minutes after reperfusion. At the same time, the myocardial tissue activities of superoxide dismutase(SOD). catalase(CAT), glutathione peroxiddse(GSHPX), glutathione reductase (GSSGRd), and glucose 6-phosphate dehydrogenate(GfPDH ) were determined in the right atrial auricle excised before aortic cross clamping and in the left atrial auricle excised 20 minutes after reperfuslon. The levels of increased significantly after reperrusion(p< U.05). There were no significant changes in CAT and CfPDH levels. Western blot analysis was performed to study the induction of antioxidant enzyme and demonstrated increased amount of Cu,Zn-SOD.

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Effects of Vitamin E and Selenium on the Antioxidative Defense System in Streptozotocin-induced Diabetic Rats (Vitamin E와 Selenium이 Streptozotocin 유발 당뇨쥐의 항산화계에 미치는 영향)

  • 이순재
    • Journal of Nutrition and Health
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    • v.29 no.1
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    • pp.22-31
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    • 1996
  • The purpose of this study was to investigate the effect of vitamin E and selenium on the antioxidative defense mechanism in the liver of streptozotocin(STZ)-induced diabetic rats. Sprague-Dawley male rats(120$\pm$10gm) were randomly assigned to one control and five STZ-diabetic groups. Diabetic groups were classified to STZ-0E (vitamin E free diet), STZ-40E(40mg vitamin E/kg of diet), STZ-400E(400mg vitamin E/kg of diet), STZ-S(0.5ppm Se/kg of diet) and STZ-400ES(400mg vitamin E and 0.5ppm Se/kg of diet) according to the level of vitamin E and selenium supplementation. Diabetes was experimentally induced by intravenous adminstration of 55mg/kg of STZ in citrate buffer(pH 4.3) after 4-weeks feedng of six experimental diets. Animals were sacrificed at the 4th day of diabetic states. Activities of the serum glutamic oxaloacetate transaminase(GOT) and the glutaminc pyruvate transaminase(GPT) in STZ-0E, STZ-40E and STZ-S rats were higher than those of control. Liver xanthine oxidase activities were similar to serum GOT and GPT. Liver superoxide dismutase(SOD) activities were higher in STZ-0E and STZ-40E groups by 33%, 22%, respectively than that of control. Glutathione S-transferase(GST) activities of liver were similar to GSH-Px activities. The contents of vitamin E in liver tissue were significantly lower STZ-0E, STZ-40E and STZ-S groups by 50%, 36%, 45% than that of control. Reduced glutathione(GSH) contents of liver were lower STZ-0E, STZ-40E, STZ-400E, STZ-S and STZ-400ES groups by 57%, 51%, 19%, 18%, 12% than that of control. Lipid peroxide values (LPO) in liver were higher 5.6, 2.3 and 2.3 times in STZ-0E, STZ-40E and STZ-S group than that of control. The present results indicate that STZ-induced diabetic rats are more sensitive to oxidative stress, leading to the acceleration of lipid peroxidation process, which can be more accelerated by feeding the low level of dietary vitamin E. In the coincident supplementation of high dietary vitamin E and selenium antioxidative enzymes activities and physiolosical antioxidants were increased more than those of the separate supplementation of vitamin E or selenium. Therefore, dietary vitamin E and selenium reduced peroxidative damage of tissue, promoting antioxidative defense mechanism against lipid peroxidation by diabetes.

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