• Title/Summary/Keyword: Dry cell

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Heterologous Expression of Human Ferritin H-chain and L-chain Genes in Saccharomyces cerevisiae (재조합 효모를 이용한 사람 H-Chain 교 L-Chain Ferritin의 생산)

  • 서향임;전은순;정윤조;김경숙
    • KSBB Journal
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    • v.17 no.2
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    • pp.162-168
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    • 2002
  • Human ferritin H- and L-chain genes(hfH and hfL) were cloned into the yeast shuttle vector YEp352 with various promoters, and the vectors constructed were used to transform Saccharomyces cerevisiae 2805. Three different promoters fused to hfH and hfL were used: galactokinase 1 (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase(GPD) promoter and alcohol dehydrogenase 1(ADH1 ) promoter. SDS-polyacrylamide gel electrophoresis and Western blotting analyses displayed expression of the introduced hfH and hfL. In the production of both ferritin H and L subunits GAL1 promoter was more effective than GPD promoter or ADH1 promoter. Ferritin H and L subunits produced in S. cerevisiae were spontaneously assembled into its holoproteins as proven on native polyacrylamide gels. Both recombinant H and L-chain ferritins were catalytically active in forming iron core. When the cells were cultured in the medium containing 10 mM ferric citrate, the cell-associated concentration of iron was 174.9 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L and 148.8 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L but was 49.4 $\mu\textrm{g}$ Per gram(dry cell weight) in the wild type, indicating that the iron contents of yeast is improved by heterologous expression of human ferritin H-chain or L-chain genes.

PHB Accumulation Stimulated by Ammonium Ions in Potassium-limited Cultures of Methylobacterium organophilum

  • Kim, Seon-Won;Kim, Pil;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.301-304
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    • 1998
  • Methylobacterium organophilum can use nitrogen in the form of ammonium ions ($($NH_4$)_2SO_4\;and\;NH_4Cl) and from nonammonium sources such as glycine, alanine, peptone, and yeast extract. When potassium was limited, significantly more PHB was produced when the ammonium ion was the nitrogen source rather than a nonammonium form. With ammonium, the amount of PHB produced was 0.50-0.53 g PHB/l or $52.0~53.2\%$ of the dry cell weight. If nitrogen was from a nonammonium source, the respective values were 0.04~0.06 g PHB/1 or $8.1~11.3\%$ of dry cell weight. When ammonium sulfate was the sole source of nitrogen under potassium-limited conditions, cell growth and PHB accumulation increased as the pH increased from 6.0 to 7.5. Cell growth and PHB amount at pH 7.5 were 2.50 g dry cell weight/1 and 1.40 g PHB/1, respectively.

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Mammary Cell Turnover under High Temperature during the Dry Period in Dairy Cows

  • Peng, Xiaoqing;Lu, Lin;Li, Yan;Yan, Peishi
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.4
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    • pp.485-492
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    • 2011
  • The influence of high temperature on mammary cell turnover during the dry period is still unclear. The objective of this study was to investigate mammary cell turnover and p53 protein expression in the mammary tissue under high temperature conditions. Mammary gland biopsy samples from 8 dairy cows were obtained at 7, 25, 40, and 53 d during the dry period in summer or spring (n = 4, each season). Cell cycle, cell turnover, and p53 protein expression were analyzed by flow cytometry. During the dry period in summer, the percentage of mammary epithelial cells in the G0/G1 phase was the highest, but those in the S and G2/M phases were lower. However, the proportion of cells in the different stages of the cell cycle was not significantly different among the different biopsy time points, except in the G2/M phase. Under different temperature conditions, the cells were significantly different in their apoptotic rate and proliferation index; moreover, the tendencies of these indicators to change significantly differed. In general, the samples under high temperature conditions showed significantly lower apoptotic rates and proliferation indices. Under high temperature conditions, the apoptotic rate and proliferation index were the lowest (2.17% and 3.26%, respectively) at day 40, and the highest at day 53 (3.67% and 4.61%, respectively). However, under normal temperature conditions, the values of these indicators were the lowest (7.60% and 5.54%, respectively) at day 7, and almost the highest at day 25 (12.85% and 6.47%, respectively). Moreover, p53 protein expression was significantly higher under high temperature conditions than under normal temperature conditions, except at day 25. The level of p53 protein was the lowest (13.10%) under high temperature conditions at day 25, but was the highest (26.07%) under normal temperature conditions. Our findings suggest that high temperature delayed the G2/M phase of the cell cycle and the cell turnover rate, but remarkably increased p53 protein expression. Thus, the results indicate that high temperature extends the recovery period of mammary epithelial cells.

Effect of Stage of Growth and Cultivar on Chemical Composition of Whole Maize Plant and Its Morphological Fractions

  • Firdous, R.;Gilani, A.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.3
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    • pp.366-370
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    • 1999
  • Samples of whole plant, leaf and stem of Akbar, Neelum, UM-81 and lZ-31 cultivars of maize fodder harvested up to 14 weeks at different growth stages were drawn and analysed for dry matter contents and various cell wall constituents such as NDF, ADF, hemicellulose, cellulose, lignin, cutin and silica. The dry matter contents of whole maize plant, leaf and stem increased significantly (p<0.01) with advancing plant age. Maximum dry matter was found in the leaf fraction of the plant. The cell wall components continued to increase significantly (p<0.001) in whole maize plant and its morphological fractions as the age advanced. Maximum values for NDF, ADF, cellulose and lignin were observed in stem followed by whole plant and leaf, whereas hemicellulose, cutin and silica contents were higher in leaf fraction of the plant. The cultivars were observed to have some effects on chemical composition of all plant fraction. The results indicated that maturity had a much greater effect on the concentration of all the structural components than did the cultivars. It was concluded that maize fodder should be cut preferably between 8th to 9th week of age (flowering stage) to obtain more nutritious and digestible feed for livestock. Among the maize cultivars, Neelum proved to be the best, due to its higher dry matter contents and lower lignin concentration.

Effects of temperature on the biomass yield and the chemical composition of pseudomonas cells in continuous culture (Pseudomonas sp.의 연속배양에 있어서의 세포의 수율 및 화학적 조성에 미치는 영향)

  • 김창진;이영록
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.163-169
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    • 1983
  • Effects of temperature on the gorwth characteristics and the chemical composition of pseudomonas cells grown under glucose-or methanol-utilizing continuous culture were studied. In a glucose-utilizing continuous culture, optimum dilution rate, agitation, pH, and temperature, for the higher biomass yield were $0.45hr^-$, 7000rpm, pH 7.5, and $30^{\circ}C$, respectively. But in a methanol-utilizing continuous culture, they were $0.125hr^-$, 600rpm, pH 8, and $30^{\circ}C$, respectively. In methanol-utilizing continuous culture, the maximum production rate of the cells was 1.48g, dry wt./1/hr at a dilution rate of $0.45hr^-$, and the cell yield was 0.46g. dry wt./g. glucose. In the methanol-utilizaing continuous culture, the maximum production rate of the cells was 0.33 7g. dry wt./1/hr. at a dilution rate of $0.125hr^-$ and the cell yield was 0.44g dry cell/g. methanol. The contents of protein of the cells increase with the increase ingrowing temperature (from 15 to $30^{\circ}C$), more or less, while the contents of RNA nad carbohydrate of the cells decreased. However, DNA contents of cells growth under the various temperature ranges didn't change. As the temeprature of cultivation rises at a constant dilution rate, the efficiency of RNA in protein synthesis was increased, showing the decreases in the ratio of RNA to protein.

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Quality Characteristics of Sprout Ginseng Paste added Dry Oyster and Dry Shrimp (굴과 새우를 이용한 새싹삼 페이스트의 품질특성)

  • Jung, Hee-Bum;Seoung, Tae-Jong;Kim, Jeong-Gyun
    • Culinary science and hospitality research
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    • v.23 no.8
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    • pp.206-215
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    • 2017
  • Sprout ginseng paste were prepared with pine nut, dry oyster and dry shrimp to examine the antioxidant properties(total polyphenols, total flavonoids, and electron donating ability) and sensory test(attribute difference and acceptance). Sprout ginseng paste were measured based on color value, pH, viscosity, total bacteria cell numbers for 0 and 20 days at $4^{\circ}C$. The higher total polyphenol and total flavonoid content of sprout ginseng paste added with pine nuts, dry oyster, and dry shrimp were higher antioxidant capacity. DPPH radical scavenging activity was increased from 52.2% (SGP0) to 79.5.0 % with SGP5. The attribute test results reveal that the color intensity, bitter taste, and oily taste were decreased in SGP3 to SGP6. Taste, flavor, and coarseness did not show significant difference in SGP0 to SGP6. Thickness and After taste were increased in SGP4 to SGP6. The preference test results reveal that the appearance, flavor, and texture level did not show significant difference in SGP0 to SGP6. Taste and overall preference were increased in SGP4 to SGP6. L value, pH, decrease while a value and b value show no change in sprout ginseng paste with pine nut, dry oyster and dry shrimp. Total cell number was not detected during storage.

Effect of Aeration-Agitation on Coenzyme Q10 Production Using Rhodobacter sphaeroides

  • Jeong, Soo-Kyoung;Kim, Joong-Kyun
    • Fisheries and Aquatic Sciences
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    • v.11 no.4
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    • pp.224-228
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    • 2008
  • With the aim of increasing the $CoQ_{10}$ production in mass culture, the effect of aeration-agitation on the $CoQ_{10}$ production using Rhodobactor sphaeroides was investigated in a l-L bioreactor. The maximum $CoQ_{10}$ production was 1.69 mg/g of dry cell weight under conditions of 50 Lux, $30^{\circ}C$, 300 rpm, and 5-vvm aeration. The $CoQ_{10}$ production was improved to produce 2.91 mg/g of dry cell weight under reduced conditions of agitation speed (200 rpm) and aeration rate (0.2 vvm). When R. sphaeroides was cultivated under more reduced DO levels during the exponential phase of the cell, the $CoQ_{10}$ production yield of 3.88-mg/g dry cell weight was the maximum obtained. Therefore, poorer conditions of aeration-agitation resulted in higher production of $CoQ_{10}$, and thus DO content was a crucial factor in the production of $CoQ_{10}$. Accordingly, it was necessary to control the DO concentration in order to enhance the $CoQ_{10}$ biosynthesis within a large-scale production.

Study of Methylglyoxal and Phosphorus Stress on Algae (조류의 Methylglyoxal과 인 Stress 연구)

  • 이기태
    • Environmental Analysis Health and Toxicology
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    • v.13 no.3_4
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    • pp.133-142
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    • 1998
  • Effects of phosphorous (P) and methylglyoxal (MG) on the cell number, dry weight, chlorophyll content, photosynthetic and respiratory rate, phosphate uptake and protein content of green algae (Scenedesrnus obliquus) were studied. The algal cell number from the medium treated with 0.5-1.0 mM of MG at 1/2 P or 1/4 P concentration was significantly lower than those of algae treated :with full strength of phosphrous in medium. The inhibitory effect of MG on algal cell division was enhenced at low concentration of phosphorous in medium. At the beginning of logrithmic phase of algal growth, the mean dry weight of algae from the medium without MG-treatment in 1/2 P media was significantly higher than that of algae treated with MG. After logrithmic phase of growth cycle, the mean dry weight of algae from the medium with 1.0 mM of MG-treatment in 1/4 P media was significantly lower than that of algae treated with or without MG. At logrithmic phase of algal growth, there were significant differences in the chlorophyll content among all groups of tested algae with various concentrations of P and MG. At 15 days after inoculation, the mean chlorophyll content per algal cell from the media without MG-treatment in 1/2P was significantly higher than that of other cells from MG-treated media. The adverse effect of MG at concentration of 0.5-1.0mM in 1/2 and 1/4 P media on photosynthetic rate was observed. The mean photosynthetic rate of algal cell without P and MG treatment at 15 days after inoculation was significantly higher than that of MGtreated algae. After logarithmic phase, the algal cell treated with 0.5mM of MG with full strength of phosphorous showed significantly high respiratory rate than that of other cell groups. There were significant differences in mean phosphate uptake rate among all groups of Scenedesmus obliquus at logarithmic phase. At 12 days after inoculation, phosphate uptake rate per each algal cell from the basic media without MG and P treatment was rapidly reduced which shows early introduction to stationary phase.

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Mass Production and Accumulation Characteristics of Polyhydroxyalkanoates by Fed-batch culture of Alcaligenes eutrophus under Phosphate Limitation (인산염 제한하에서 Alcaligenes eutrophus의 유가식 배양에 의한 Polyhydroxyalkanoates의 대량 산과 축적특성)

  • 류희욱;조경숙;장용근
    • KSBB Journal
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    • v.13 no.2
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    • pp.187-194
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    • 1998
  • For mass production of polyhydroxyalkanoates (PHA), high cell density cultures of Alcaligenes eutrophus by fed-batch culture under phosphate-limitation condition has been investigated. PHA accumulation by the regulation by the regulation of initial phosphate concentration could be automatically induced, and high density cell culture above 200 g/L also could be successfully produced. The production of Poly-$\beta$-hydroxybutyrate (PHB) and dry cell weight increased with increasing the initial phosphate concentration. When the initial concentrations of phosphate were in the ranges of 1.5~4.5 g-PO$_4$/L, PHB and dry cell weight obtained were 83~266 g/L and 61~216 g/L, respectively, and PHB productivity was in the ranges of 1.35~3.10 g/L.h. When a mixture of glucose and propionic acid is used as carbon sources, poly(3-hydroxybutyrate-co-poly-3-hydroxyvalerate), P(3HB-co-3HV), could be also successfully produced under phosphate limitation condition. When the mole ratio of propionic acid to glucose in the feeding solution is 0.22, a final dry cell weight of 150 g/L and a P(3-HB-co-3HV) of 90 g/L were produced. Morphological changes and size distribution of PHB granules synthesized in A. eutrophus under phosphate-limitation condition are determined by TEM during the course of fed-batch. Mean granule diameters of PHB produced are in the range of 0.36~0.39 $\mu$m, and mean cell size was elongated from 0.54~0.59 $\mu$m$\times$ 1.3~1.5 $\mu$m to 0.83~0.89 $\mu$m $\times$2.0~2.3 $\mu$m. Phosphate concentration in media did not affect size distribution of PHB granule and cell.

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Studies on the Boron Metabolism of Orchid -Influences on the Cell Wall Structure and its Components- (양란의 붕소 대사에 관한 연구 -특히 세포벽 형성 및 분획 조성에 미치는 영향-)

  • 강영희
    • Journal of Plant Biology
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    • v.22 no.1_2
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    • pp.35-43
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    • 1979
  • The present investigation has been made to study the deficiency symptoms of boron on the formation of cell wall and the development of the individual components of the orchid cell wall. Analytical samples were taken from two sources; one from the individual orchid plants started from an apical meristem culture followed by the generation of the protocorm-like body which was developed into a plant, the other from the plant cultivated in water for 30 days. The amount of boron in the cultrues were controlled and the deficiency symptoms were observed under theelectron microscope, optical microscope with samples taken from the zones of elongation of leaves and compared the dry weight of cell walls and finally the various fractions of the cell wall components. The following results were obtained: (1) The growth of roots and leaves was hampered in the boron deficient plants. (2) In the boron-deficient leaves a severe necrosis and cracks were developed in the tissue of zone of elongation besides the decrease in growth. (3) under the electorn microscope the cell walls of boron-deficient plants showed rough undulated structures unlike the smooth control cell walls. (4) the dry weight of total cells and cell walls of boron deficient plants were higher than the control plants. (5) In the boron deficient plant the amout of pectin and hemicellulose isolated from cell walls were higher and the amount of protein was lower than the controlled plots.

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