• Title/Summary/Keyword: minimal medium

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Zinc and Selenium Requirements for Glutathione Peroxidase Activity and Cell Survival in Chinese Hamster Ovary Cells Overexpressing Metallothionein

  • Kwun, In-Sook;John R. Arthur;John H. Beattie
    • Preventive Nutrition and Food Science
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    • v.8 no.1
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    • pp.36-39
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    • 2003
  • Many defined cell culture media were formulated over 3() years ago and may be deficient in certain micronutrients whose essentiality has only subsequently been recognised. The objective of this study was to evaluate whether alpha-minimal essential medium (MEM) supplemented with 10% foetal bovine serum contained sufficient selenium for optimal activity of the selenium containing enzymes cytosolic glutathione peroxidase (cGPx) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) in cultured Chinese hamster ovary (CHO) cells. Additionally, the effect of zinc deficiency and metallothionein (MT) overexpression on cGPx and PHGPx activity was studied. The addition of 100 nM of selenous acid to the culture medium increased cGPx expression by 10-fold and PHGPx by about 2-fold in both wild-type CHO-K1 cells and CHO-K1 cells overexpressing mouse MT-1. Zinc deficiency had no significant effect on enzyme activity, but cells overexpressing mouse MT-1 had higher levels of cGPx activity. Zinc deficiency decreased cell survival but overexpression of MT-1 was partially protective, probably because its presence in quantity favoured the uptake, sequestration and cellular retention of any remaining zinc. This study demonstrates that selenium in complete alpha-MEM is insufficient for optimal cGPx and PHGPx activity and may compromise the cellular response to oxidative stress.

Flux Regulation Patterns and Energy Audit of E. coli B/r and K-12

  • Lee, Jin-Won;Goel, Akshay;Ataai, Mohammad-M.;Domach, Michael-M.
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.258-267
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    • 2002
  • A flux determination methodology has been built which enables to develop constrained stoichiometric relationships and metabolic balances. The analysis differs from those developed for anaerobic growth conditions in that cell mass formation is a significant sink for carbon. When combined with experimental measurements, a determined system of equations results yielded tricarboxylic acid (TCA) cycle and glycolytic fluxes. The methodology was implemented to determine the fluxes of E. coli B/r and K12, and it was found that as the growth rate in a glucose minimal medium increased, the cells became increasing glycolytic and the TCA fluxes either leveled off or declined. The pattern identified for the TCA fluxes corresponded to ${\alpha}$-ketoglutarate dehydrogenase's induction-repression pattern, thereby suggesting that the induction-repression of the enzyme could result in significant flux changes. When the minimum flux solution was contrasted to the glycolytic and TCA fluxes determined, two observations were made. First, the minimum flux could provide the cell's biosynthetic ATP requirements. Second, at a high growth rate in a glucose medium, the excess glycolytic flux exceeded that of the TCA cycle, which appeared to more closely match the biosynthetic needs.

Induction of Vitellogenin Synthesis by Androgens in Cultured Hepatocytes of the Eel, Anguilla japonica (간세포 배양을 이용한 뱀장어 Vitellogenin 합성에 대한 웅성호르몬의 영향)

  • 권혁추;박홍양
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.259-269
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    • 1996
  • To establish whether or not androgens is responsible for the induction of vitellogenin(Vg) synthesis and secretion, primary hepatocytes prepared from immature eels were used. The results are follows: 1. Eel hepatocytes were prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in medium for 10 days with minimal cell loss. 2. Estradiol-17$\beta$(E2) alone was insufficient to induce Vg synthesis. The combination of E2 with methyltestosterone(MT) markedly stimulated Vg synthesis. High vg production occurred in MT concentration from 10-6~10-5M in the presence of E2 (10-6M). Testosterone and androsterone were also effective, but progesterone was not effective in inducing Vg synthesis. Neither MT alone nor testosterone and androsterone alone had any effect on Vg synthesis. 3. E2-primed hepatocytes showed Vg synthesis in both media with and without hormones 1 day after culture. In the cultures with the vehicle, MT, or progesterone, the rate of synthesis seemed to decrease with time. But the combination of E2 and MT showed an intense increase in Vg synthesis. Hepatocytes isolated from E2-primed eels also required androgens for continuating of Vg synthesis. 4. These results demonstrate that androgens act together with E2 in synthesis and secretion of eel Vg.

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Open Market Strategy of the Business Innovation Platform for SME Informatization based on Cloud Computing (클라우드를 이용한 중소기업정보화 경영혁신플랫폼의 오픈 마켓 전략 연구)

  • Han, Hyun-Soo;Yang, Hee-Dong;Kim, Kiho
    • Journal of Information Technology Services
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    • v.14 no.4
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    • pp.15-30
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    • 2015
  • SMBA (Small and Medium Business Administration) and TIPA (Korea Technology and Information Promotion Agency for SMEs) have operated the Business Innovation Platform for SME Informatization based on cloud computing technology with the cooperation of seven industry cooperatives since 2013. This project will evolve into the open market platform where service providers and users voluntarily participate and transact. This research conducts the literature review about the concept of open market and the empirical analysis through survey for the software providers and the future users regarding the future operation methods. The policy about how the open market strategy for the business innovation platform needs to be designed and implemented are organized as the three differentiated government support strategies. The first is to provide free IT services including specialized core operation support S/W which is developed only for the small or home office group of firms which lack minimal informatization capability and budget. The second is to augment IT platform service through incorporating ERP supplier initiated commercial S/W sales window for those firms having medium level informatization capability. This includes to provide IT support for customization and system integration with existing government subsidized S/W. The third is to provide upgrading services of existing S/W functions to facilitate better system utilization. The results provide useful insight for government role to enhance SME competitiveness using IT.

Utility of solid phase extraction for colorimetric determination of lead in waters, vegetables, biological and soil samples

  • Al-Mallah, Zakia;Amin, Alaa S.
    • Journal of Industrial and Engineering Chemistry
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    • v.67
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    • pp.461-468
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    • 2018
  • A highly sensitive, selective and rapid method for the determination of lead based on the reaction of lead (II) with 5-(4'-chlorophenylazo)-6-hydroxypyrimidine-2,4-dione (CPAHPD) and the solid phase extraction of the Pb(II)-CPAHPD complex with Amberlite XAD-2000 was developed, in the presence of pH 5.6 buffer solution and Triton X-114 medium. CPAHPD reacts with lead to form a violet complex with a molar ratio of 2:1 (CPAHPD to lead). This complex was enriched by the solid phase extraction with Amberlite XAD-2000. An enrichment factor of 500 was obtained by elution of the complex from the resin with a minimal amount of isopentyl alcohol(0.2 mL). In isopentyl alcohol medium,the molar absorptivity of the complex is $1.13{\times}10^6L\;mol^{-1}cm^{-1}$ at 647 nm. Beer's law is obeyed in the range of $5.0-160ng\;mL^{-1}$ in the measured solution. The relative standard deviation for 10 replicate samples of $50ng\;mL^{-1}$ level is 1.26%. The detection and quantification limits reaches 1.5 and $4.7ng\;mL^{-1}$ in the original samples. The presented procedure was successfully applied for determination of lead content in real samples such as vegetables, waters, biological and soil samples with satisfactory results.

Protoplast Fusion of Cellulolytic Aspergillus wentii and Aspergillus niduk (섬유소 분해효소를 생성하는 Aspergillus wentii와 Aspergillus nidulans의 원형질체 융합)

  • 성낙계;이상원;강신권;노종수;정영철
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.460-465
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    • 1990
  • Regeneration of protoplast was effective by preincubating spore suspension containing 30$\mu g$/ml of 2-DG for 4 hours, and CBE medium containing casamino acid, bovine serum albumin, ergosterol and myoinositol was found to be more efficient than any other regeneration medium tested in this experiment. The regeneration frequency was about 30%. Optimal conditions for conidial protoplast fusion were obtained by treatment of protoplasts with 10 mM $CaCl_2$ and 30% polyethylene glycol 4000 (pH 7.5) as fusogenic agent at $37^{\circ}C$ for 10 minutes. The fusion frequency was $8.2\times 10^{-4}$. The higher productivity of enzyme of fusant FWN-56 was achived: 2.3-fold for CMCase, 1.5-fold for avicelase, 1.8-fold for $\beta$-glucosidase and 2.5-fold for xylanase compared to that obtained in two parental strains. The genetic stability of fusant after maintenance on minimal medium for more than 4 weeks was high because segregant rate was below 1%. The conidial DNA content of fusant was 1.4-1.6 times higher than that of the parental strains, The nucleus size of fusants were also higher than that of each parental strains.

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Inhibitory Effect of Green Tea Extract on Collagenase Activity and Growth of Fish Pathogenic Bacteria (녹차 추출물을 이용한 어병세균의 collagen 분해효소 및 생육 억제)

  • Park, Sun-Mee;Park, Soo-Il;Huh, Min-Do;Hong, Yong-Ki
    • Journal of fish pathology
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    • v.12 no.2
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    • pp.83-88
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    • 1999
  • Green tea extract has been suggested to possess properties of collagenase inhibition and antimicrobial effect to fish pathogenic bacteria. Most fish pathogenic bacteria showed collagenase activities of 0.08-0.7 unit/ml. Among them, Edwardsiella tarda produced large amount of collagenase in the ST medium at $25^{\circ}C$ after 16 hrs. Biosynthesis of the enzyme from E. tarda was decreased to 1/3 by addition of 0.08-0.8 mg/ml of tea extract in the culture medium. The collagenase activity was inhibited almost 100% in the reaction mixture by 0.2 mg/ml of the extract. Then, the minimal inhibition concentration against E. tarda growth appeared as 8 mg/ml of the tea extract in the culture medium.

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Saccharomyces cerevisiae Strain Improvement Using Selection, Mutation, and Adaptation for the Resistance to Lignocellulose-Derived Fermentation Inhibitor for Ethanol Production

  • Jang, Youri;Lim, Younghoon;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.667-674
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    • 2014
  • Twenty-five Saccharomyces cerevisiae strains were screened for the highest sugar tolerance, ethanol-tolerance, ethanol production, and inhibitor resistance, and S. cerevisiae KL5 was selected as the best strain. Inhibitor cocktail (100%) was composed of 75 mM formic acid, 75 mM acetic acid, 30 mM furfural, 30 mM hydroxymethyl furfural (HMF), and 2.7 mM vanillin. The cells of strain KL5 were treated with ${\gamma}$-irradiation, and among the survivals, KL5-G2 with improved inhibitor resistance and the highest ethanol yield in the presence of inhibitor cocktail was selected. The KL5-G2 strain was adapted to inhibitor cocktail by sequential transfer of cultures to a minimal YNB medium containing increasing concentrations of inhibitor cocktail. After 10 times of adaptation, most of the isolated colonies could grow in YNB with 80% inhibitor cocktail, whereas the parental KL5 strain could not grow at all. Among the various adapted strains, the best strain (KL5-G2-A9) producing the highest ethanol yield in the presence of inhibitor cocktail was selected. In a complex YP medium containing 60% inhibitor cocktail and 5% glucose, the theoretical yield and productivity (at 48 h) of KL5-G2-A9 were 81.3% and 0.304 g/l/h, respectively, whereas those of KL5 were 20.8% and 0.072 g/l/h, respectively. KL5-G2-A9 reduced the concentrations of HMF, furfural, and vanillin in the medium in much faster rates than KL5.

Biodegradation of JP-8 by Rhodococcus fascians Isolated from Petroleum Contaminated Soil (유류 오염 토양에서 분리된 Rhodococcus fascians를 이용한 JP-8의 분해 특성)

  • Nam, Bo Hyun;Park, Bong Je;Yun, Hyun Shik
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.819-823
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    • 2008
  • Microorganisms isolated from petroleum contaminated site were tested for their ability to grow on JP-8 by culturing them on the culture medium that contains JP-8 as a carbon source. The microorganism which grew on JP-8 containing minimal salt medium was separated and identified as Rhodococcus fascians. Changes in JP-8 biodegradation of R. fascians that was isolated from petroleum contaminated site was investigated with various inoculums sizes, JP-8 concentrations, medium pHs, and culture temperatures. The amount of JP-8 was analyzed by TPH using Gas Chromatography.