• Title/Summary/Keyword: 세포 배양

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Ethanol Production from Tapioca Hydrolysate by Batch and Continuous Cell Retention Cultures (회분 및 연속세포유지 배양에 의한 타피오카당화액으로부터의 에탄올생산)

  • 이용석;이우기
    • KSBB Journal
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    • v.10 no.5
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    • pp.598-603
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    • 1995
  • Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/$\ell$ was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/$\ell$ making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5${\times}$107 cells/ml(about 2.1g/$\ell$). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/$\ell$, about 15g/$\ell$, 81.4g/$\ell$, and 14.7g/$\ell$-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/$\ell$ and the ethanol concentration oscillated around 68g/$\ell$. The volumetric ethanol productivity was about 13.6g/$\ell$-h and the residual reducing sugar concentration about 12g/$\ell$ containing glucose of about 4.5g/$\ell$. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.

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Effect of pH on Organic Acid Production by Anaerobiospirillum succiniciproducens (Anaerobiospirillum succiniciproducens에 의한 유기산 생성에 미치는 pH의 영향)

  • 강귀현;류화원장호남
    • KSBB Journal
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    • v.10 no.5
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    • pp.568-574
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    • 1995
  • To investigate the effect of pH on organic acid production by Anaerobiospirillum succiniciproducens, an anaerobic fermentation was carried out by maintaining the pH of the fermentation broth at 5.8, 6.0, 6.4, 6.8, and 7.2. At various pHs, the concentrations of cell were $1.0∼1.9g/\ell$ which were two to three times higher than those of the other worker's results, and the maximum was obtained at pH 5.8. Substrate consumption was increased by increasing the pH in the range of pH 6.0 to 6.8, while the sugar consumption rate at both pH 5.8 and 7.2 was very slow. The total amount of 2M $Na_2C0_3$ added for adjustment of pH change due to organic acid production was maximum at pH 6.8. Changes of conductivity of the fermentation broth was very simillar to those of 2M $Na_2C0_3$ added at various pHs. Therefore, it is suggested that determination of the amount of organic acid in a broth can be possible by measuring the conductivity. The maximum production yield of lactate based on glucose was 64% for pH 7.2 and 32% for pH 6.8, respectively.

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Supercritical Extraction of Oriental Herb : Anti-aging and Anti-wrinkle Effects (한방원료의 초임계 추출을 이용한 항노화 및 주름개선 효과)

  • Kim, In-Deok;Kwon, Ryun-Hee;Heo, Ye-Young;Jung, Hye-Jin;Kang, Hwan-Yul;Ha, Bae-Jin
    • KSBB Journal
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    • v.23 no.6
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    • pp.529-534
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    • 2008
  • The reactive oxygen species generated by ultraviolet rays causes various types of cutaneous damage, such as lipid peroxidation and denaturation of the extra-cellular matrix. The accumulation of such damage contributes to skin aging, especially the formation of wrinkles. This study was carried out to develop functional cosmatic by using Oriental herb supercritical extracts (OHSE) for prevention of skin. Effects of OHSE on anti-oxidation, collagenase inhibition and collagen synthesis in normal human fibroblast were investigated. OHSE showed antioxidative activity as high as vitamin C, trolox and DL-penicillamine. Also OHSE showed promotive effect on collagen synthesis and inhibitory effect on collagenase activity. From this results, we conclude that OHSE may have the potential to be conveniently used as an additive in cosmetics for prevention and improvement of skin aging.

Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Purification and Characterization of Bacillus subtilis JS-17 Collagenase. (Bacillus subtilis JS-17이 생산하는 Collagenase의 정제 및 특성)

  • Lim Kyoung-Suk;Son Shung-Hui;Kang Ho Young;Jun Hong-Ki
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.657-663
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    • 2005
  • Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

Effect of Three Dimensional Culture of Porcine Endometrial Cells on Their Plasminogen Activity and Pre-implantation Embryo Development after Co-culture (돼지 자궁 세포의 3차원 배양이 Plasminogen 활성과 수정란 발육에 미치는 영향)

  • Lee, Sang-Hee;HwangBo, Yong;Cha, Hye-Jin;Kim, Su-Ji;Kim, Min-Gyeong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.207-219
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    • 2014
  • Three-dimensional (3D) culture system is useful technique for study of in vivo environment and it was used various experiments. This study was investigated to establish of embryo co-culture system and changes of PAs activity in 3D cultured endometrial cells of pigs. In results, growth of stromal cells into gel matrix were detected only with endometrial and myometrial cells. The most rapid growth of stromal cells were confirmed in $2.5{\times}10^5cells/ml$ and gel matrix containing 15% FBS. Expression of urokinase-PA (uPA) after treatment of hCG (0.5, 1.0, 1.5 and 2.0 IU/ml) were higher than without hCG, but, there are not significant difference among the treatment. On the other hand, expression of uPA after treatment of $IL-1{\beta}$ (0.1, 1, 10 and 100 ng/ml) were higher than without $IL-1{\beta}$, but, there are not significant difference. Expression of uPA after treatment of estrogen (0.2, 2, 20 and 200 ng/ml) were not difference, but PA activity was significantly decreased (p<0.05). Blastocyst was producing in PZM-3 medium containing FBS and endometrial cells were grown in PZM-3 medium. When embryos development with cultured endometrial cells, cleavage rates were not significant difference and blastocyst were not produced in co-culture with stromal cells and 3D culture system. 3D culture system had similar activity to in vivo tissue and these features are very useful for study of in vivo physiology. Nevertheless 3D culture system was not proper in embryo co-culture system. Therefore, we suggest that 3D culture system with embryo co-culture need continuous research.

Development of In Vitro Culture System for Male Germline Stem Cells in Porcine (돼지 웅성 생식선 줄기세포의 체외배양기법 개발)

  • Kim, Yong-Hee;Kim, Byung-Gak;Lee, Yong-An;Kim, Bang-Jin;Kim, Ki-Jung;Lee, Myeung-Sik;Im, Gi-Sun;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.171-177
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    • 2009
  • Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from $10{\sim}14$-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and $O_2$ tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, $37^{\circ}C$ temperature, and $O_2$ 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.

Herbicidal Selective Activity of Oxyfluorfen to the Selected Rice Cultivars and Major Paddy Weed Species (벼품종(品種) 및 주요(主要) 논잡초종(雜草種)에 대한 Oxyfluorfen의 선택활성(選擇活性) 연구(硏究))

  • Kim, Y.J.;Guh, J.O.;Pang, S.;Choi, K.J.
    • Korean Journal of Weed Science
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    • v.7 no.2
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    • pp.220-235
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    • 1987
  • The experiment was carried out to find the feasibility of using Oxyfluorfen in the paddy fields by investigating the difference of selective activity of Oxyfluorfen among rice cultivars and major paddy weed species. The dosage of Oxyfluorfen that show selective activity between rice cultivars and weed species ranged from 0.1 to 0.4kg ai/ha. The degree of growth inhibition was in order of whole-plant soaking application > root soaking application > stem bandage application, and in that case $10^{-5}$M Oxyfluorfen was treated after emergence. Especially the growth inhibition of rice cultivars and Cyperus serotinus was low, among others. Photosynthesis was severely inhibited at the Oxyfluorfen level above $10^{-4}$ M in all the tested weeds, but inhibition of respiration was not to be seen. Isolated single cells of two rice cultivars and Cyperus serotinus were tolerant to $10^{-5}$M Oxyfluorfen,but those of Echinochloa crus-galli and Sagittaria pygmaea were susceptible comparatively. The growth inhibition of suspension cultured rice cell induced by the increments of Oxyfluorfen concentration, and the degree of inhibition was higher in C.V. Mushakdanti than in C.V. Aichiasahi.

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Physiological studies on cell division by the technique of synchronous culture of chlorella (II) (클로렐라의 동조배양법에 의한 세포분열의 생리학적 연구 2)

  • 이영녹;심웅섭
    • Korean Journal of Microbiology
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    • v.7 no.1
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    • pp.10-21
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    • 1969
  • The effect of glucose and 2-thiobarbituric acid on the biosynthesis of cell constituents such as protein, carbohydrate, DNA, RNA, phospholipid and PCA-soluble phosphate compounds in Chlorella duing the life cycle was measured, and the changes in the content of these main cellular components of the algal cell were analyzed in connection with the nuclear and cytoplasmic divison. In the normal autotrophic synchronous culture the contents of protein, RNA, and DNA in the cell showed a chracteristic changes according to the progress of cell development, increasing more or less throughout all the life cycle. The synthesis of protein is more prominent in the division period nad that of DNA is more active in the ripening period, while the synthesis of RNA is more rapid in the growing and ripening periods than other developmental stages. The period of division cycle was little affected by glucose in the medium, although the synchrony of the growth and cellular division was disturbed and the n value increased. The cotents of protein, carbohydrate, RNA nad DNA of the cell were increased by the glucose treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment. The synthesis of protein, carbohydrate, DNA, RNA and phospholipid of the cell was also retarded by 2-thiobarbituric acid. In the autotrophic, mixotrophic and 2-thiobarbituric acid-treated cultures, each having different mode cytoplasmic division, a common general schema occurring in the cell during the life cycle may be drawn as follows. The ratio of RNA to protein attains maximum value in the $L_1$-cell stage prior to the nuclear division and thereafter decreases during the periods of ripening and division. The ratio of PCA-soluble phosphate compounds to protein increased from the begining of the culture to $L_4$-cell stage successively and thereafter decreased gradually during the division period, while the ratio of protein to DNA kept almost constant up to the division period and thereafter increased during the division period. Therefore, it is presumed that the increase in the ratio of RNA to protein is to be an inducer of nuclear division and that the cytoplasmic division is induced by the increase in the ratio of protein to DNA.

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Identification and Characterization of an Endophytic Strain of Streptomyces from Rice Roots (Orysa sativa L.) (벼(Orysa sativa L.) 뿌리로부터 분리된 내생 Streptomyces 균주의 동정 및 특성)

  • Kim, Jae-Heon;Lee, Jun-Kwan
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.375-380
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    • 2011
  • We isolated an endophytic actionmycete from root tissues of rice plant collected from paddy field near Dankook University, Cheonan, Korea. Surface sterilized roots were laid on the selective agar plates and incubated. The powdery actinomycete colonies appeared on the root surface after four weeks incubation. We isolated a strain JK-5 among them and could determine its taxonomical position as Streptomyces diastaticus subsp. ardesiacus by using 16S ribosomal DNA sequencing. The chemotaxonomical and morphological studies confirmed the taxonomical position of the strain JK-5. The shape of aerial hyphae was flexible and they contained spore chains with more than 30 smooth spherical spores per chain. Cell walls contained LL-diaminopimelic acid. There was no characteristic sugar in whole-cell hydrolysates. The major fatty acids were anteiso-15:0, anteiso-17:0 and iso-16:0. The specific menaquinones, MK-9 ($H_6$), MK-9 ($H_8$), were detected. The GC content was 72%. Antifungal activities of the strain JK-5 were relatively strong against fungal plant pathogens. The endophytic growth of the strain JK-5 was confirmed by SEM observation of the root and stem of the infected rice plant.