• 한국환경과학회지
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• 제14권12호
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• pp.1163-1170
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• 2005

The microbial adsorption characteristics of two different media for biological treatment were studied using attached diverse microbes onto activated carbon and ceramic. The results in the experiments of the characteristics of physical adhesion on two different media with addition of high and low concentrated substrate in the culture were observed that the efficient of adhesion onto F-400 activated carbon was higher over that of ceramic due to the surface area of media. The irradiation treatment by ultrasonication with 400 W power and 3 min retention time on the media without addition substrate conditions and subsequent mixing throughly the culture showed the highest efficiency of cell detachment on the media. Three different microbes, P. ovalis, A calcoaceticus, and B. subtillis were used for the study of the characteristics of microbial adhesion on the media. p ovalis showed the highest adhesion capability while B. subtillis showed the lowest capability adhesion onto media either addition of substrate in the culture. The mixed bacterial culture showed $10\%$ lower removal efficiency of DOC in the low concentrated substrate culture compared to the single pure culture. Whileas, it did not show significant difference between two cultures at high concentrated substrate. It was also observed same population density of microorganism by counting of microbes adhered to microbial media with an ultrasound treatment.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1316-1323
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• 2017

Geosmin and 2-methylisoborneol (2-MIB), responsible for earthy or musty smell, are a major concern for safe drinking water supplies. This study investigated the effects of environmental factors on odorous compound production and cell growth in cyanobacterial strains. Anabaena sp. FACHB-1384, a 2-MIB producer, was sensitive to low temperature (<$20^{\circ}C$). However, geosmin producers, Anabaena sp. Chusori and Anabaena sp. NIER, were sensitive to high light intensity (>$100{\mu}mol/m^2/sec$), but not to low temperature. Geosmin concentrations increased under higher nitrate concentrations, being linearly proportional to cell density. A P-limited chemostat showed that P-stress decreased the geosmin productivity and extracellular geosmin amount per cell in Anabaena sp. NIER. However, only 2-MIB productivity was reduced in Planktothrix sp. FACHB-1374 under P-limitation. The extracellular 2-MIB amount per cell remained constant at all dilution rates. In conclusion, high light intensity and P-stress can contribute to the lower incidence of geosmin, whereas 2-MIB reduction could be attainable at a lower temperature.

• Journal of Applied Biological Chemistry
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• 제43권2호
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• pp.109-111
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• 2000

Effect of the two-stage operation and cell concentration on indirubin production was investigated using bioreactor culture of Polygonum tinctorium. Two-stage culture was operated successfully for 110 days without any adverse effects on continuous indirubin production. Maximum indirubin concentration was found to be at 80 mg/bioreactor. Initial cell concentration significantly affected indirubin production. The indirubin production at 29.2% PCV was improved by 845%, compared to that at 5% PCV. For high-density bioreactor culture of P. tinctorium, a maximum production rate of 10.2 mg indirubin/L day was obtained. Indirubin recovery for bioreactor operation was also examined using XAD-2, XAD-4, XAD-7, and solid silicon. XAD-4 was 1.6-fold more effective than that for solid silicon in indirubin recovery.

• The Korean Journal of Physiology and Pharmacology
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• 제19권1호
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• pp.35-42
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• 2015

In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.213-217
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• 2003

The effects of glucose concentration on the production of PHB by fed-batch culture of Ralstonia eutropha were investigated. In the range of glucose concentration of $2.5\;{\sim}\;40\;g/l$, it was found that the high glucose concentration was not favorable for the PHB formation after the phosphate limitation. It was further confirmed by the specific PHB synthesis rates and yields. The PHB concentration decreased much with the increase of glucose concentration. But if the glucose concentration was very low, e.g. 2.5 g/l, the cell growth and PHB synthesis also could be limited because of inadequate glucose supply. Itwould be better to maintain the glucose concentration at about 9.0 g/l to obtain high DCW, PHB concentration and productivity.

• 한국양식학회지
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• 제11권2호
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• pp.231-240
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• 1998

온도와 염분에 대하여 내성이 강하고 대량배양이 쉬워서 조개류의 인공종묘생산시 먹이로서 많이 사용하고 있는 5종의 Tetraselmis (T. tetrathele, T. suecica, T. subcordiformis, T. sp. (Haeundae), T. sp (China)) 중 우리나라 기후 환경에 대향배양이 가장 적합한 종을 선택하기 위핵서 최적 배양환경 (온도, 염분)을 조사하고 이때의 세포크기와 영양성분을 조사하였다. 5종의 Tetraselmis의 세포크기 및 체적은 T. sp. (Haeundea)가 장경 $17.6{\pm}1.87$${\mu}$m, 체적 727${\mu}$$m^3$로 가장 대형종이었으며 중국산 T. sp.가 장경14.6+- 1.46${\mu}$m, 체적 625${\mu}$$m^3$으로 가장 소형종이였다. Tetraselmis는 광염성, 광온성이나 최적성장을 위한 수온과 염분 구간은 각각 24~$30^{\circ}C.$와 27~30%.였다. 5종의 Tetraselmis에서 고수온에 대한 내성은 중국산 Tetraselmis, 저수온데 대한 내성은 T. tetrathele가 높은 것으로 나타났다. 5종의 Tetraselmis중 배양밀도가 가장 높았던 종은 T. suecica였으며, 이 종은 배양 7일째 $24^{\circ}C.$, 30%.에서 세포수가 $141{\times}10^4$cells/ml (s.g.r., 0.3906)로 성장이 가장 좋았다. 5종의 Tetraselmis의 화학성분중 조단백질 함량은 T. suecica가 44.50%로 가장 높게 나타났고 조지방은 해운대산 T. sp.가 7.13%로 높게 나타났다. 필수아미노산은 해운대산 .T.sp.에서 50.40%로 가장 높게 나타났고, 불포화지방산은 중국산 T.sp.에서 11.7%로 가장 높게 나타났다. 5종의 Tetraselmis의 성장과 영양성분 결과로 보아 우리나라에서 대량배양이 가장 적합한 종은 T. seucica임을 알수 있었다.

• Archives of Plastic Surgery
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• 제37권5호
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• pp.509-518
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• 2010

Purpose: Major drawbacks of conventional bone marrow stromal cells (BSCs) transplantation method are mainly caused by direct transplanted cell to host cell interactions. We hypothesized that separation of the transplanted cells by a microporous membrane might inhibit most of the potential adverse effects and induce superior effect. The purpose of the study is to determine the optimal condition of the microporous membrane. Methods: First, BSCs were placed in polyethylene terephthalate (PET) transwell inserts with 3, 8, or $12{\mu}m$ pore size, and cultured in 24 well culture plates. After 5 days, bottoms of the plates were observed for presence of attached BSCs in monolayer and cell numbers were evaluated. Second, BSCs were placed PET, polycarbonate (PCT), and mixed cellulose esters (MCE) transwell inserts with 3 and $8{\mu}m$ pore size, and cultured in 24 well culture plates. After 3 days, the supernatants of the media left in culture plate were analyzed for collagen, vascular endothelial growth factor (VEGF), platelet derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF). Third, BSCs were placed in 15% and 70% of the PET membrane with $3{\mu}m$ pore size. All the experimental conditions and methods were same as the second study. Results: The optimal pore sizes to prevent BSC leakage were $3{\mu}m$ and $8{\mu}m$. The amounts of type I collagen and three growth factors tested did not show significant differences among PET, PCT, and MCE groups. However, the collagen, VEGF, and bFGF levels were much higher in the high (70%) density group than in the low (15%) density group. Conclusion: This study revealed that the optimal pore size of membrane to prevent direct BSC to recipient cell contact is in between $3{\mu}m$ and $8{\mu}m$. Membrane materials and pore sizes do not influence the collagen and growth factor passage through the membrane. The most striking factor for collagen and growth factor transport is pore density of the membrane.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.225-230
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• 2004

A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.301-304
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• 2002

참당귀 (Angelica gigas Nakai) 현탁세포의 고농도 배양을 위하여 perfusion 배양을 한 결과 세포의 생장이 증진됨을 확인하였다 . 배양 5 일째부터 sucrose를 4배 농축한 배지로 2 mL 씩 교환해준 결과, 세포의 생장이 계속 유지되었고 최대 세포농도도 23.7 g/L 로 대조구에 비하여 1.7 배 증가하였다. 초기 당농도를 50 g/L 로 하여 배지 교환을 한 결과, 최대 세포농도도 23.8 g/L 로 30 g/L 당농도 대조구에 비하여 1.6 배 증가하였고, 배지교환을 하지 않은 50 g/L 대조구에 비하여 1.1 배 증가하였다 . 따라서 초기 당농도를 높이고 연속적으로 배지교환을 해준다면, 고농도의 세포를 얻을 수 있으며 결과적으로 참당귀 세포가 생산하는 다당류의 생산량을 증진시킬 수 있을 것이다.

• 생명과학회지
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• 제11권2호
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• pp.173-180
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• 2001

This study was conducted to estimate the cultural characteristics, the production of antibiotic, and the selection of optimal media for mass culture of Bacillus licheniformis N1 isolate which was previously reported as an antagonistic bacterium to Botrytis cinerea. We investigated initial pH, temperatures and shaking speed for good cultural conditions and antibiotics production by N1 isolate. According to the results, the optimal conditions of initial pH, temperatures, and shaking speed were determined to be pH 5.0~5.5, 30~35$^{\circ}C$ and 250 rpm, respectively. Also, the optimal conditions for the antagonism by N1 isolate highly appeared in the initial pH as 5.0, and the mycelial growth inhibition was high when the substances used such as glucose or corn starch as carbon sources, and biji(soybean curd residue) flour as a nitrogen source. Furthermore, inhibitory area was significantly expanded, when 3% or 5% of corn starch was added into 5% of Biji flour as nitrogen source, were respectivley selected for mass culture of N1 isolate. Among them, 5% Biji flour medium showed higher cell density more than 10 times that in NB medium after 48 hour incubation. Therefore, the optimal medium was determined as 5% biji flour added 3~5% of corn starch for high density of cells.