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The Development of Serum-Free Media for the Betrovirus Production and Using It in Continuous Production (유전자 재조합된 레트로바이러스의 생산을 위한 무혈청 배지의 개발과 레트로바이러스의 연속 생산공정 개발)

  • 안지수;조수형;김병기
    • KSBB Journal
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    • v.16 no.3
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    • pp.274-280
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    • 2001
  • A serum-free media for CRIP/MFG-LacZ retrovirus production was developed and applied to continuous packed-bed culture system. The serum-free media developed by fractional factorial design contains indulin ($10\mug/mL$), transferrin ($5\mug/mL$), BSA (4 mg/mL), EGF (25 ng/mL), and linoleic acid ($10\mug/mL$). Operation of continuous packed-bed reactor using Fibra-cel enabled the packging cell to stably maintain retrovirus titer for about 1 week. The optimal operation conditions for dilution rate and temperature were 0.67(h(sup)-1) and $32^{circ}C$, respectively. Using this media, the retrovirus titer(cfu/mL) in the packed continuous culture was about 50% that of continuous culture with serum containing DMEM media.

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Isolation and Identification of Marine Bacterium Cytophaga sp. AYK301 and Optimal Culture Conditions for the Production of Agarase (해양세균 Cytophaga sp. AYK301의 분리.동정 및 한천분해효소 생산을 위한 최적배양조건)

  • Lee, Won-Gyeong;Kim, Bong-Jo;Ha, Sun-Deuk;Gong, Jae-Yeol
    • KSBB Journal
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    • v.14 no.5
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    • pp.572-577
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    • 1999
  • A marine bacterium with highly effective agar degrading activity was ioslated from the southern sea of Korea (Chonnam, YoChon) and identified as Cytophaga sp. and named as Cytophaga sp. AYK301. This strain produced an extracellular agarase which had a high activity with agar. The optimum culture conditions for the production of agarase have been determined. For the increase of agarase productivity, 0.2% agar, 0.3% beef extract, and 0.05% NH$_4$NO$_3$ were used as carbon, organic and inorganic nitrogen source, respectively. The optimal initial pH, NaCl, culture time and temperature for the agar degrading activity were 7.5, 7.0%, 36 hr and $25^{\circ}C$, respectively. In the optimal conditions, the agarase production was increased up to more than 4.0 folds as compared to that by the basal medium.

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Studies on Cysteinedesulfhydrase produced by Bacteria.(Part 1) Isolation and Identification of Cysteinedesulfhydrase producing Bacterium. (Bacteria가 생산하는 Cysteinedesulfhydrase에 관한 연구(제일보) Cysteinedesulfhydrase 생산균의 분리검색 및 동정)

  • 최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.2 no.1
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    • pp.37-43
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    • 1974
  • 1. In the course of investigation on the metabolism of cysteine by microorganisms, the authors have found that among 70 strains tested Cysteinedesulfhydrase occured most remarkably in the cells of the strain I-3-2 isolated from the soil when it was grown on a medium containing L-Cysteine. The morphological, cultural and physiological properties of this strain were investigated. From the results, the bacterium was identified as a variety of Aerobactor aerogenes. 2. The cultural conditions for the formation of Cysteinedesulfhydrase by the strain I-3-2 were also investigated and the results were as follows: 1). The optium pH of the culture medium was 7.0-7.5. 2). As a carbon source, glycerol was most effective when it was added to the basal medium at 0.1 % concentration. 3). By addition of calcium chloride at 0.2% concentration, the formation of the enzyme remarkably increased. 4). Maximal formation of the enzyme was observed at the end of logarithmic phase of cell growth, thereafter the enzyme activity was diminished rapidly.

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Cultural Conditions for the Improvement in Gibberellic Acid Productivity by a Mutant of Gibberella fujikuroi ATCC 12616-Gibberella fujikuroi G-36 (Gibberella fujikuroi ATCC 12616 으로부터 얻어진 변이주 Gibberella fujikuroi G-36의 Gibberellic Acid 의 배양조건)

  • 오영준
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.152-155
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    • 2000
  • Cultural Conditions for the Improvement in Gibberellic Acid Productivity by a Mutant of Gibberellafujikuroi ATCC 12616-Gibberella fujikuroi G-36 . Dh, Young-Jun. Department of Food and Biotechnolog}'r Dongshm Umversity, Naju 520-714, Korea - A mutant Gibberella jujih/roi G- 36 was selected by metagenesis of G/bberella fitjikuroi ATCC 12616 with mutagens such as N-methy1-N'~nitro~N"nitrosoguanidine and hydroxylamine for improving productivity of gibberellic acid. The mutant strain produced gibberellic acid (70 mg/l) more than that of wilde type. A fermentation medium containing glucose, $NH_4N0_3$, $MgS0_4$, $KH_2P0_4$ and trace elements was deve]oped for the maximal production of a gibberellic acid by the mutanL The Guctuating cultural temperature that was vaded from 300e to 20DC resulted in higher GA yield than that of fixed cu1tura] temperature at $28^{\circ}C$.

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Apoptosis in Human Corneal Epithelial cells induced by Exhausted Medium (Exhausted Medium에 의한 각막상피 세포의 세포고사 유도)

  • Kim, Jae-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.5 no.1
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    • pp.83-87
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    • 2000
  • To investigate exhausted-medium-induced apoptosis in human corneal epithelial(HCE) cells, this study was performed DNA gel electrophoresis, M30 CytoDEATH staining and FAS-FAS ligand ELISA. SV-40 transfected cells were grown to confluency in culture for 7days. The supernatant was harvested and filtered with $0.22{\mu}m$ filter paper. Fresh HCE cells were exposed to the filtered exhausted medium for 1~2 days. Apoptotic cells were prepared for DNA extraction and run the agarose gel for DNA ladder pattern. M30 CytoDEATH was used a tool for easy and reliable determination of very early apoptosis in HCE cells. The control and exhausted medium were assayed for soluble FAS/FAS ligand protein by ELISA. HCE cells exposed to exhausted medium showed a typical DNA ladder pattern. Sporadic M30 CytoDEATH positive cells were detected among HCE cells exposed to exhausted medium. Soluble FAS/FAS ligand levels were not elevated in the exhausted medium compared to the fresh medium control. This study suggests that possible mechanism of exhausted medium induced apoptosis does not include the FAS-FAS ligand system.

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Isolation and Optimal Producing Conditions of Broad Spectrum Antibiotics from Streptomyces sp. Y-88 (광범위 항생물질을 생산하는 Streptomyces sp. Y-88의 분리 및 생산 최적 조건)

  • Bang, Byung-Ho;Jeong, Eun-Ja
    • The Korean Journal of Food And Nutrition
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    • v.22 no.1
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    • pp.103-109
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    • 2009
  • In order to isolate antibiotic producing microorganisms, several actinomycetes were isolated from soil samples. The aerial hyphae of Y-88 strain were gray in color with tree types. Under the microscopic examination, the Y-88 isolate formed a spiral aerial spore mass with a smooth surface and a rectiflexibilis type of spore chain. Y-88 utilized glucose, fructose, arabinose, and sucrose, but not rhamnose, raffinose, mannitol, or inositol. In addition, Y-88 produced melanin on the tyrosine agar and the strain could utilize L-valine, L-phenylalanine, and L-hydroxyproline. Based on these results and the cultural and physiological characteristics described in the Bergey's Manual, the Actinomycetes, Y-88, was identificated as a Streptomyces species. The optimum medium conditions for this antibiotic producing Streptomyces sp. Y-88 was 1.6% soluble starch, 0.6% glucose, 0.6% beef extract, 0.01% $K_2HPO_4$, 0.01% $MgSO_4$ $7H_2O$, and 0.01% $ZnSO_4$ $7H_2O$ at an initial pH of 4.0, and 25$^{\circ}C$.

Screening and Optimal Culture Conditions of Antibiotic-Producing Actinomycetes B-51 for Multidrug Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii에 유효한 방선균 B-51의 탐색 및 이 균주가 생산하는 항생물질 발효 최적 배양 조건)

  • Rhee, Moon-Soo;Kim, Gwan-Pil;Bang, Byung-Ho
    • The Korean Journal of Food And Nutrition
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    • v.23 no.1
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    • pp.63-69
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    • 2010
  • With the increase of the use of antibiotics and invasive procedures, infections caused by multidrug-resistant Acinetobacter baumannii(MRAB) are increasing. We screened the antibiotic producing strain B-51 for antibacterial activity against MRAB from the soils and studied the effects of culture medium on the antibiotic production of B-51. The medium conditions for maximum antibiotic productivity of B-51 was 2% glycerol, 0.5% soybean meal, 0.01% $CaCl_2$, 0.01% $MgSO_4{\cdot}7H_2O$ and 0.01% $KH_2PO_4$ at an initial pH of 6.0, at $30^{\circ}C$ for 76 h.

Exopolysaccharide Production in Fed-batch and Continuous Culture by Methylomonas mucosa (Methylomonas mnosa에 의한 Exopolysaccharide의 유가식 및 연속 생산)

  • 장호남;권선훈심상준
    • KSBB Journal
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    • v.8 no.2
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    • pp.164-171
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    • 1993
  • The production of extracellular polysaccharide by Methylomonas mucosa (NRRL B-5696) was investigated. The microorganism uses methanol as the carbon source for their growth and produces exopolysaccharides. The productivity of exopolysaccharides was investigated under various culture modes: batch, fed-batch and continuous culture. In flask culture the growth of cell mass and the production of polysaccharide were inhibited at above 1% (v/v) methanol. At 1%(v/v) methanol maximum specific growth rate was obtained. As C/N ratio (g methanol/g ammonium sulfate) increased, polysaccharide production increased and cells mass decreased. Magnesium ion was also found to be essential for the polysaccharide production. In batch culture the production of polysaccharides was more affected by the specific growth rate than the cell concentration. In fed-batch culture the concentration of polysaccharide was 4 times higher than that of batch culture, but the yield was lower. The productivity of fed-batch with continuous feeding was higher than that of batch or fed-batch with intermittent feeding. This is due to no methanol limitation or inhibition that used to occur in fed-batch culture with intermittent feeding. In continuous culture pure oxygen was supplied to avoid the oxygen limitation. As the dilution rate in- creased up to 0.21 h-1, the yield and productivity increased. The solution viscosity of the produced polysaccharide obtained from above increased exponentially with the concentration of polysaccharide.

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Characterization of the Biosurfactant-Producing Bacterium, Pseudoalteromonas sp. HK-3 Isolated from the Crude-Oil Contaminated Areas (원유로 오염된 지역으로부터 분리한 생물계면활성제 생산균주, Pseudoalteromonas sp. HK-3의 특성조사)

  • Cho, Su-Hee;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.346-351
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    • 2010
  • The purpose of this work was to investigate the characteristics of a biosurfactant-producing bacterium isolated from crude-oil contaminated soils. During the incubation of strain HK-3 with 1% crude-oil, bacterial growth pattern, the amount of biosurfactant production, and pH changes were monitored. In order to examine the effect of supplemented carbons on the production of biosurfactant, cultivation of HK-3 cells in BH media with different carbons (e.g. glucose, dextrose, mannitol, citrate, or acetate) revealed that the production of biosurfactant reached the maximal level at the 72 h incubation with mannitol, which the area of clear zone was measured to approximately 7.64 $cm^2$. Identification test using the BIOLOG system, morphology study based on scanning electron microscopy and the 16S rRNA sequence-based phylogenetic analysis assigned strain HK-3 to a Pseudoalteromonas species, designated as Pseudoalteromonas sp. HK-3 which was registered in GenBank as [FJ477041].

Studies on the Isolation and Identification of Xylanase and Mannanase Producing Aspergillus niger (Xylanase와 Mannanase를 생산하는 Aspergillus niger의 분리와 동정에 관한 연구)

  • Kim, Byoung-Suk;Cho, Jin-Kook;Song, Jin-Ook;Lee, Hak-Kyo;Hwang, Seong-Gu
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.427-432
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    • 2009
  • This study was undertaken to screen a high xylanase and mannanase producing microbes. In the first experiment, screening was undertaken against 50 samples of microorganisms having xylanase and mannanase activities from soil and fallen leaves. The screening process has focused on picking out fungi having high xylanase and mannanase activities under the solid-state fermentation. The xylanase and mannanase activities of 6 screened microbes were 0.9~1.6 unit/mL and 0.2~0.4 unit/mL, respectively, under the submerged fermentation condition. However, under the solid-state fermentation, xylanase and mannanase activities were 103.7~220.0 unit/g and 20.1~40.3 unit/g, respectively. Finally one microbe (E-3) was selected and its xylanase and mannanase activities were 197.3 unit/g and 39.9 unit/g, respectively. The morphological and molecular biological classification of E-3 showed 99% homology with the Aspergillus niger.