• Title/Summary/Keyword: cell production

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Effects of Concentration of Glucose and By-product on the Growth of Bacillus amyloliquefaciens 23350 (B. amyloliquefaciens 세포 성장에 미치는 포도당 및 부산물 농도의 영향에 관한 연구)

  • 차월석;김종균김종수
    • KSBB Journal
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    • v.8 no.5
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    • pp.457-464
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    • 1993
  • In this study, an attempt was made to investigate optimum cell growth and products by Bacillus amyloliquefaciens 23350 in batch culture by varing carbon soures. Maximum dry cell density increased with the increase of initial glucose concentration. Maximum dry cell density was obtained with the highest value of 5.2g/l at 30g/l of initial glucose concentration. By adding acetic acid at 20g/l of initial glucose concentration, the cell growth rate decreased with the increase of initial acetic acid concentration. Among the various carbon sources, maximum $\alpha$-amylase production was obtained with 225unit/ml at 10g/1 of initial glucose concentration. Optimum production of $\alpha$-amylase was obtained with 376unit/ml at 2g/l of initial acetic acid concentration and 20g/l of initial glucose concentration. By 10g/1 of initial glucose concentration, both good maximum specific cell growth rate and maximum $\alpha$-amylase production rate were obtained. In view of the results studied optimum production and specific production rate of $\alpha$-amylase, acetic acid was initially added 2~4g/l with 20g/1 of initial glucose concentration in batch culture.

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Optimization of Induction Conditions for Bacillus-derived Esterase Production by High-cell Density Fermentation of Recombinant Escherichia coli (재조합 대장균의 고농도 배양과 유도조건 최적화를 통한 Bacillus 유래 esterase의 생산)

  • Kang, Seung-Hoon;Min, Byung-Hyuk;Choi, Hong-Yeol;Kim, Dong-Il
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.149-154
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    • 2017
  • To increase the efficiency of esterase production by Bacillus, high cell-density culture of recombinant Escherichia coli through fed batch fermentation was tested. Cells were cultured to $OD_{600}$ of 76 (35.8 g/l DCW) with dissolved oxygen level controlled to least above 30% air saturation by supplying pure oxygen. Cells were cultured to an $OD_{600}$ of 90 (42.4 g/l DCW) with glucose feeding controlled to at least 1 g/l. However, the cells reached stationary phase at the late stage of culture, despite glucose being supplied. Cells were cultured to an $OD_{600}$ of 185 (87.3 g/l DCW) by supplying additional medium with fortified yeast extract. To increase the productivity of the recombinant protein, cell growth and esterase productivity based on induction time were evaluated. Late exponential phase induction for esterase production in fed batch fermentation resulted in maximum optical density $OD_{600}$ of 190 (89 g/l DCW) and maximum esterase activity of 1745 U/l, corresponding to a 5.8-fold enhancement in esterase production, compared to the early exponential phase induction. In this study, we established fermentation methods for achieving maximum production of Bacillus-derived esterase by optimizing IPTG induction time in high-cell density culture by supplying pure oxygen and a nitrogen source.

Effect of Kyejakjimo-tangkami (Guishaozhimu-tangjiawei) on Osteoarthritis (계작지모양가미방(桂芍知母湯加味方)이 골관절염에 미치는 영향)

  • Hong, Seong-Min;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.4
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    • pp.39-57
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    • 2013
  • Objectives The purpose of this study is to prove the effect of Kyejakjimotangkami(KMK) on osteoarthritis. Methods We checked antioxidant activity and measured production of $IL-1{\beta}$, IL-6, TNF-${\alpha}$ in RAW 264.7 cell after treat by KMK. Then we measured hind paw weight of Wister Rat with arthritis induced by MIA after KMK oral administration, checked Prostaglandin E2, IL-$1{\beta}$, IL-6, TNF-${\alpha}$, Osteocalcin, TIMP-1, MMP-9, LTB-4 in serum, ran histopathological test and ${\mu}CT$-arthrography. Results 1. DPPH radical Scavenging was increased depend on concentration of KMK ethanol extract in RAW 264.7 cell. 2. Production of NO was significantly decreased by KMK ethanol extract on concentration of $200{\mu}g/ml$ in RAW 264.7 cell. 3. Production of IL-$1{\beta}$ was significantly decreased by KMK ethanol extract on concentration of $200{\mu}g/ml$. And Production of IL-6, TNF-${\alpha}$ were significantly decreased KMK ethanol extract of every concentration in RAW 264.7 cell. 4. Result of checking hind paw weight when administered KMK ethanol extract to Wister Rat with arthritis induced by MIA was significantly higher than control group and similar to normal group. 5. Production of Prostaglandin E2, IL-$1{\beta}$, Osteocalcin, TIMP-1, MMP-9 and LTB-4 in serum was significantly decreased by KMK ethanol extract after administerd to Wister Rat with arthritis induced by MIA. 6. In Hematoxylin & Eosin staining and Safranin-O staining, we could find inflammation of synovial cell, infiltration of macrophage and granulocyte and degeneration of cartilage and bone were decreased in comparison with control group. 7. When checked cartilage volume to examine degree of cartilage degeneration using ${\mu}CT$-arthrography, volume of cartilage was increased in comparison with control group. Conclusions Comparison of the results for this study showed that KMK ethanol extract have anti-inflammatory effectiveness and can protect cartilage and bone. So we expect that KMK can be used as a effective drugs for osteoarthritis.

A study on the Smoothed Production( II ) (생산평준화에 관한 연구(II))

  • 김학철;송수정;김태호;나승훈;강경식
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.19 no.37
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    • pp.221-231
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    • 1996
  • Applying JIT(Just-In-Time) production system to strength competitiveness power and renovate managent has problems. This study is proposed to solve one of the problems, that mother company has different production system with subcontractor, in order to connect production system of mother company with subcontractor. In the view of the Pull System, production system of mother company, it is possible that the more smoothed production planning is established by developing the algorithm the smoothed production planning preserving the LOTproduction system and comparing the existing research. Also, in the view of subcontractor taking Push System, the possibility of keeping delivery and improving productivity is proved using simulation technique by changing Job shop to GT Cell production system because demand is fluctuating.

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Identification of Putative Embryonic Stem Cells Derived from Embryonic Blastodermal Cells of Fertilized Hen′s Eggs (닭 배반엽세포로부터 유래된 잠정적 배아주세포의 동정)

  • Lee, K.S.;Lee, H.;Kim, K.D.;Park, Seong-Su;Lee, S.H.
    • Korean Journal of Poultry Science
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    • v.27 no.1
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    • pp.73-78
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    • 2000
  • Embryonic stem (ES) cells are pluripotent cell lines, which derived from preimplantation embryo. These cells have been used as a vehicle of foreign DNA for production of transgenic mammals. this experiment was performed to examined the possible use of blastodermal cells derived from hen's egg for germline manipulation. Stage X blsdtodermal cells isolated from fertilized eggs were cultured in DMEM containing 15% fetal calf serum. Blastodermal cells wre co-cultured on the chicken embryonic fibroblast (CEF) or mouse embryonic fibroblast(MEF) cells. to examine the effects of growth factors on stem cell growth, bFGF and LIF were added. There was no significant difference in colony formation of putative ES cells between CEF and MEF as a feederlayer, but the addition of growth factors enhanced the proliferation and inhibited differentiation of blastodermal cells. To characterize the cell colonies as a putative ES cells, putative embryonic cell colonies were stained by periodic acid Schiffs (PAS) reagent. The putative ES cell colonies showed intensive positive reaction similar to the property of undifferentiated PGC upto 20days in vitro, but not in other cell types. this result demonstrates that PAS-positive cell colonies may be used for the study of establishment of chicken ES cell lines for the production of transgenic chicken.

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T cell phenotype and intracellular $IFN-{\gamma}$ production in peritoneal exudate cells and gut intraepithelial lymphocytes during acute Toxoplasma gondii infection in mice

  • Lee, Young-Ha;Shin, Dae-Whan
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.119-129
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    • 2002
  • Although there are many reports on the splenic (systemic) T cell response after Toxoptasma gondii infection, little information is available regarding the local T cell responses of peritoneal exudate cells (PEC) and gut intraepithelial Iymphocytes (IEL) following peroral infection with bradyzoites. Mice were infected with 40 cysts of the 76K strain of T. gondii, and then sacrificed at days 0, 1, 4, 7 and 10 postinfection (PI). The cellular composition and T cell responses of PEC and IEL were analyzed. The total number of PEC and IEL per mouse increased after infection, but the ratio of increase was higher in IEL. Lymphocytes were the major component of both PEC and IEL. The relative percentages of PEC macrophages and neutrophils/eosinophils increased signiflcantly at day 1 and 4 PI, whereas those of IEL did not change significantly. The percentage of PEC NK1.1 and ${\gamma\delta}T$ cells peaked at day 4 PI (p < 0.0001), and CD4 and $CD8{\alpha}T$ cells increased continuously after infection. The percentages of IEL $CD8{\alpha}$ and ${\gamma\delta}T$ cells decreased slightly at first, and then increased. CD4 and NK1.1 T cells of IEL did not change significantly after infection. $IFN-{\gamma}-producing$ PEC NK1.1 T cells increased significantly from day 1 PI, but the other T cell subsets produced $IFN-{\gamma}$ abundantly thereafter. The proportion of IEL $IFN-{\gamma}-producing$ $CD8{\alpha}$ and ${\gamma\delta}T$ cells increased significantly after infection, while IEL NK1.1 T cells had similar $IFN-{\gamma}$ production patterns. Taken together, CD4 T cells were the major phenotype and the important $IFN-{\gamma}$ producing T cell subsets in PEC after oral infection with T. gondii whereas $CD8{\alpha}T$ cells had these roles in IEL. These results suggest that PEC and IEL comprise different cell differentials and T cell responses, and according to infection route these factors may contribute to the different cellular immune responses.

Production of Xylooligosaccharides by Yeast Cell Surface-Displayed Endoxylanase (효모 세포 표면 발현된 Endoxylanase를 이용한 Xylooligosaccharides의 생산)

  • Kim, Hyun-Jin;Lee, Jae-Hyung;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.307-313
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    • 2008
  • The yeast surface expression system, pCTXYN (6.8 kb), of Bacillus endoxylanase gene (xynB, 642 bp) was constructed and introduced into Saccharomyces cerevisiae EBY100 cell. The transformed yeast cell showing the highest endoxylanase activity was selected through the active staining of colonies grown on YPDG medium containing xylan. With the yeast transformant, EBY100/pCTXYN, grown on galactose containing medium, it was found that the endoxylanase was successfully displayed on the yeast cell surface and the xylooligosaccharides were efficiently produced from xylan. The most of endoxylanase activity was detected in the cell fraction and reached about 1.9 unit/mL after 48 h cultivation. The optimized conditions for xylooligosaccharides production from xylan were determined as follows: substrate and its concentration, oat spelt xylan 6%; concentration of yeast whole-cell, 5 unit/mL; temperature, $50^{\circ}C$, and reaction time $2{\sim}4\;h$. When the oat spelts xylan and corncob xylan were hydrolyzed by treatment with cell surface-displayed endoxylanase, xylotriose was formed as a main product.

Growth conditions of symbiosis for production of single cell protein (단세포단백질 생산을 위한 혼합배양의 생육조건)

  • Lee, Hae-Kyoung;Chung, Yung-Gun;Kwon, Oh-Jin
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.343-348
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    • 1996
  • Experiments were carried out to find possibility of economic production of SCP in mixed culture by Cellulomonas sp. KL-6 and E. coli LI-10. The best cell growth was obtained at the ratio of 1 : 1(v/v) in mixed culture. When these strains were mixed culture, cell growth was increased to about 63%, compared with those of single culture of strain KL-6. It was found that the majority of the population during growth in mixed culture consisted of strain KL-6. $CaCO_3$ added to the medium as the ratio of 0.1% was enhanced medium pH. Cell growth increased in that circumstances. These strains produced much amounts of cellobiose, but glucose was not detected in filter paper medium. When these organisms were cultured under the optimal medium for 4 days, cell mass was produced $1.0\;g/{\ell}$. The results showed the increase of cell mass up to 53% than those produced in CMC medium.

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Effect of Levamisole on Immunomodulation of Eels (Anguilla japonica) In Vitro (In vitro에서 Levamisole이 양식뱀장어의 면역조절작용에 미치는 영향)

  • Choi, Min-Soon;Park, Kwan-Ha;Joung, Kyung-Min;Shim, Hyun-Bin;Yun, Sung-Ho
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.16-23
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    • 1999
  • The immunomodulatory effects of levamisole (LMS) were evaluated in leucocytes of eels in vitro. Proliferation of lymhocytes treated with T-cell mitogen (Con A or PHA) was markedly inhibited by LMS in a dose dependent manner. B cell mitogen (LPS), in contrast, slightly increased the proliferaion. On the other hand, production of MIF and MAF when treated with Con A was increased in a dose-dependent way. NK cell activities were somewhat increased when LMS was pretreated and this augmentation was due to an increase in binding capacity of effector-target cell, but not due to the target cell lytic activity of effector cells. Phagocytic activity, superoxide anion formation, hydrogen peroxide formation and lysozyme activity of leucocytes were enhanced by LMS in a dose related-manner. These results suggest that LMS might modulate the immmune responses by activation of cytokine production and by augmentation of leukocyte activity but not by increment of immunocompetent cell numbers.

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