• Title/Summary/Keyword: co-cultures

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Comparative Production of Green Fluorescent Protein Under Co-expression of Bacterial Hemoglobin in Escherichia coli W3110 Using Different Culture Scales

  • Bassapa Johnvesly;Kang, Dong-Gyun;Park, Suk-Soon;Kim, Ji-Hyun;Cha, Hyung-Joon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.274-277
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    • 2004
  • Production of green fluorescent protein (GFP) as a model foreign protein using different culture scales under co-expression of Vitreoscilla hemoglobin (VHb) in the industrial Escherichia coli strain W3110 (a K12 derivative), was examined. It was found that the VHb co-expressing W3110, exhibited an exceptional and sustained production ability during cell cultures using different scales, while the VHb non-expressing strain showed variable production levels. This high and sustained production ability indicates that the VHb co-expressing E. coli W3110, could be successfully employed for practical large-scale production cultures without the need for serious consideration of scale-up problems.

Production of Salidroside in Rhodiloa sachalinensis A. Bor Callus by the Elicitation and Precursor (참돌꽃 (Rhodiola sachalinensis A. Bor) 캘러스에서 elicitor와 전구체에 의한 Salidroside 생산성의 변화)

  • Lee, Jae-Seung;Kim, Min-Young;Kim, Jae-Heun;Nam, Jong-Hyun;Lee, Hyeon-Young;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.4
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    • pp.268-272
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    • 2008
  • The effect of elicitor and precursor on salidroside production from Rhodiola sachalinensis A.Bor callus cultures was investigated. Callus cultures were treated with yeast extract, soft-ferrite ceramics powder, methyl jasmonate, ascorbic acid, jasmonic acid and $CuCl_2$/$CdCl_2$ as an elicitor. When callus cultures were treated with $0.2g/\ell$ of yeast extract, salidroside production from callus treated with yeast extract is 3.45 times higher than that of the controlled group. Among of them, callus cultures treated with yeast extract produced the highest salidroside. Callus cultures were treated with L-phenylalanine and L-tyrosine as a precursor for 4 days. The result of salidroside content analysis showed that all feeding of precursors not affected salidroside production from callus cultures. In case of L-tyrosine fed into callus cultures, both callus growth and salidroside production decreased at all concentrations.

High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo;Hong, Min-Seon;Shin, Hang-Chel;Lee, Jeewon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.301-305
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    • 2001
  • The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

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Rhus verniciflua Stokes Attenuates Glutamate-induced Neurotoxicity in Primary Cultures of Rat Cortical Cells

  • Jeong, Eun-Ju;Sung, Sang-Hyun;Kim, Jin-Woong;Kim, Seung-Hyun;Kim, Young-Choong
    • Natural Product Sciences
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    • v.14 no.3
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    • pp.156-160
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    • 2008
  • The methanolic extract of Rhus verniciflua Stokes (RVS-T) and its fractions (RVS-H, RVS-C, RVS-E and RVS-B) showed significant neuroprotective activity against glutamate-induced toxicity in primary cultures of rat cortical cells. RVS-B, which showed the most potent neuroprotective activity, was further fractionated to yield RVS-B5. Treatment of cortical cells with the RVS-T, RVS-B and RVS-B5 reduced the cellular ROS level and restored the reduced activities of glutathione reductase and SOD induced by glutamate. Although, the activity of glutathione peroxidase was not virtually changed by glutamate, RVS-B5 increased the glutathione peroxidase activity. In addition, these three tested fractions significantly restored the content of GSH which was decreased by glutamate insult in our cultures. Taken together, it could be postulated that RVS extract, in particular its fraction RVS-B5, protected neuronal cells against glutamate-induced neurotoxicity through acting on the antioxidative defense system.

Ampicillin and co-trimoxazole susceptibility of Shigella isolated in 1978-80 (Shigella $1978{\sim}80$년 분리주의 Ampicillin 및 Co-trimoxazole 감수성)

  • Chong, Yun-Sop;Lee, Sam-Uel Y.;Ryu, Young-Hat
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.33-38
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    • 1980
  • Shigella remains to be an important enteric pathogen in this country for the moment. Moreover, since 1978, most of the isolates have become resistant to ampicillin and co-trimoxazole, which used to be the drugs of choice for shigellosis. Since a disc diffusion technique alone has been used in our routine susceptibility test, the minimum inhibitory concentrations(MIC) of both ampicillin and co-trimoxazole to Shigella have never been known. In order to determine these, 195 isolates were tested by an agar dilution method, all of which were isolated at Yonsei Medical Center during the period of June 1978 to July 1980. The following results were obtained. 1. Sixty cultures(29.7%) were susceptible to ampicillin, being the MIC of 8 ${\mu}g/ml$ or less and 53(27.2%) were susceptible to co-trimoxazole, being the MIC of TMP/SMZ 4/76 ${\mu}g/ml$ or less. S. flexneri type 2 was often resistant to both antimicrobic agents. 2. An increasing rate of resistant isolates was noted, particularly in the year of 1979. 3. Many isolates were resistant to both agents. Somewhat more cultures. were ampicillin susceptible and co-trimoxazole resistant than the other way around. It seems that the determination of species or even serotypes might be of help sometimes to select proper antimicrobic agent to control the infection. A routine antimicrobic susceptibility test of Shigella to both ampicillin and co-trimoxazole would be advisable for a better selection of chemotherapeutic agent.

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Hair Growth Promotion by δ-Opioid Receptor Activation

  • Zheng, Mei;Choi, Nahyun;Balboni, Gianfranco;Xia, Ying;Sung, Jong-Hyuk
    • Biomolecules & Therapeutics
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    • v.29 no.6
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    • pp.643-649
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    • 2021
  • Literature has revealed that the delta opioid receptor (DOR) exhibited diverse pharmacological effects on neuron and skin. In the present study, we have investigated whether the activation of DOR has hair-growth promotion effects. Compared with other opioid receptor, DOR was highly expressed in epidermal component of hair follicle in human and rodents. The expression of DOR was high in the anagen phase, but it was low in the catagen and telogen phases during mouse hair cycle. Topical application of UFP-512, a specific DOR agonist, significantly accelerated the induction of the anagen in C3H mice. Topical application of UFP-512 also increased the hair length in hair organ cultures and promoted the proliferation and the migration of outer root sheath (ORS) cells. Similarly, pharmacological inhibition of DOR by naltrindole significantly inhibited the anagen transition process and decreased hair length in hair organ cultures. Thus, we further examined whether Wnt/β-catenin pathway was related to the effects of DOR on hair growth. We found that Wnt/β-catenin pathway was activated by UFP-512 and siRNA for β-catenin attenuated the UFP-512 induced proliferation and migration of ORS cells. Collectively, result established that DOR was involved in hair cycle regulation, and that DOR agonists such as UFP-512 should be developed for novel hair-loss treatment.

Time Course of $CO_2$ Exchange of Sweetpotato Cultures In Vitro with Different $CO_2$ Concentration under Forced Ventilation (고구마 조직배양묘의 $CO_2$교환량 연속 측정)

  • 오양숙;고재풍수;정순주
    • Proceedings of the Korean Society for Bio-Environment Control Conference
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    • 1997.05a
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    • pp.17-22
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    • 1997
  • 본 연구에서는 고구마 조직배양묘의 $CO_2$ 교환량을 측정하기위해 개방형 광합성측정장치의 원리를 이용해 자체제작한 $CO_2$ 교환량 측정장치를 이용하여 공기 주입구와 배출구의 $CO_2$ 농도를 연속측정하고, 명기와 암기에 있어서의 $CO_2$ 교환량의 일변화를 분석했다. 내용적 1.48$\ell$의 배양기에 32개의 식물체를 이식했을 때 식물체당 일일 $CO_2$ 교환량은 19일째에 포화에 달했다. 식물체 생장에 있어서는 자연환기구보다 강제환기구의 생장이 촉진되었고, 강제환기구에서는 $CO_2$ 시용구가 무시용구보다 생장이 촉진되었다. 본 연구의 결과는 고구마 조직배양묘를 강제환기와 $CO_2$ 시용등 배양환경의 개선함으로써 순환과정을 거치지 않고 포장정식이 가능한 배양묘나 삽수의 생산이 가능함을 시사한다고 사료된다.

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Characteristics of Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Production by Ralstonia eutropha NCIMB 11599 and ATCC 17699

  • Song, Jae-Yong;Kim, Beom-Soo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.603-606
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    • 2005
  • Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

Protective Effects of the Water Extracts of Hovenia dulcis Thunb Against Ethanol-Induced Toxicity in Primary Cultured Rat Hepatocytes (랫드 일차 배양 간세포에서 에탄올의 독성에 대한 헛개나무 물추출물의 보호효과)

  • Kim, Jong-Ho;Seo, Young-Min;Kim, Ju-Hyun;Hyun, Sun-Hee;Lee, Sang-Kyu;Kim, Chun-Hwa;Kang, Mi-Jeong;Jeon, Tae-Won;Yoon, Soo-Hong;Jeong, Tae-Cheon
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.56-61
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    • 2008
  • The hepatoprotective effects of the water extracts of Hovenia dulcis Thunb (HD) were investigated in vitro. Following the induction of hepatotoxicity by ethanol in primary cultures of rat hepatocytes, the protective effects of four different water extracts of HD were determined through serial dose-response and time-dependent studies. The individual extracts used in these studies were prepared from fruits, seeds, leaves and tubes. Treatment of hepatocyte cultures with the water extracts of HD provided a significant protection from the increased lactate dehydrogenase activity induced by ethanol. Particularly, the fruits extract was the most effective against ethanol-induced hepatotoxicity in the primary cultures of rat hepatocytes. The results demonstrated that the extracts might have the protective effect against ethanol-induced toxicity in hepatocyte cultures.

High-yield Production of Functional Human Lactoferrin in Transgenic Cell Cultures of Siberian Ginseng(Acanthopanax senticosus)

  • Jo, Seung-Hyun;Kwon, Suk-Yoon;Park, Doo-Sang;Yang, Kyoung-Sil;Kim, Jae-Whune;Lee, Ki-Teak;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.442-448
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    • 2006
  • Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.