• Title/Summary/Keyword: Cell lysis

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The Production of Tissue Type Plasminogen Activator from Normal Human Cell tine (정상 인체 세포로부터 조직 플라스미노겐 활성인자의 대량생산)

  • Lee, Hyeon-Yong;Kim, Geum-Soo
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.522-525
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    • 1988
  • A method to produce tissue type Plasminogen Activator (tPA) from normal human fibroblast is developed by cultivating cells in serum free media containing heparin as an inducer. Optimal dose of this inducer was 30$\mu$g/m$\ell$. The composition of serum free medium was also defined to fit to the industrial scale cultivation. 1.42 ug of tPA per 10$^5$ viable cells per ml was produced. 1.1 gram of tPA can be produced every day from this cell line under normal perfusion chemostat operations assuming that same productivity is maintained when the process is sealed up. This method could reduce pro-duction costs and simplify purification processes by using serum free medium. Tissue type PA produced from this cell line has high ability of dissolving clots, based upon fibrin lysis test showing 50mm$^2$ of clearing zones in agarose gel plate. These results were reproducible and in good agreement with results of ELISA assay. tPA from normal human cells will be safer than that from melanoma and recombinant cells in human clinical trials.

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Isolation of Aspergillus fumigatus and Properties of It's Enzyme for Rhodotorula glutinis Cell Wall Lysis (Rhodotorula glutints 세포벽 용해효소를 생산하는 Aspergillus fumigatus의 분리와 그 효소의 특성)

  • 반재구;이준식
    • Korean Journal of Microbiology
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    • v.22 no.4
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    • pp.215-222
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    • 1984
  • A fungus producing cell wall lytic enzyme for Rhodotorula glutinis was isolated from local soil and identified partially as a species of Aspergillus fumigatus group. Thd cell wall lytic enzyme was an inducible exoenzyme and composed of at least lytic polysaccharidase and protease which act cooperatively in the lysis of intact cells. The lytic polysaccharidase was not able to hydrolyze ${\beta}-1,\;3\;and\;{\beta}-1$, 6-glucan which have the same types of bond as found in the cell wall of Ascomycetous yeasts. The lytic polysaccharidase alone was sufficient to hydrolyze the fractionated cell wall (alkali-insoluble residues) of R. glutinis, whereas it showed low activity against intact cells.

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The Pretreatment Effects on Methane Fermentation of Microalgal Biomass (미세조류의 전처리에 따른 메탄발효 특성)

  • Kang, Chang-min
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.849-859
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    • 2000
  • This study was conducted to identify the characteristics of methane fermentation and lysis effects of pre-treated microalgae. Chemical compositions of microalgae showed that the VS(volatile solid) was 86.1% of TS(total solid), and the protein was 63.5% of VS. These values were higher than those of activated sludge. The cell lysis test of raw microalgae biomass was conducted by many physicochemical methods. presenting that the degree of cell lysis was affected by following order: ultrasonic(100min.), alkali(pH 13), ultrasonic(10min), thermal($120^{\circ}C$), thermal($50^{\circ}C$), and acidic(pH 3) treatment. Methane fermentation with many pre-treated samples was performed, showing that the concentration of acetic acid was the highest. followed by propionic acid, butylic acid and valerie acid among all VFA(volatile fattic acid). In methane production. ultrasonic samples were only more effective than untreated one in total gas and methane productivity. but other samples were less effective. Especially. the alkalic sample had an inhibitation effect on methanogens.

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Characterization of yeast cell wall lytic enzyme from Fusarium moniliforme (Fusarium moniliforme이 생산하는 효모세포벽 분해효소의 특성)

  • 장판식;박관화;이계호
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.467-471
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    • 1986
  • Yeast cell wall lytic enzyme was purified from Fusarium moniliforme by ammonium sulfate fractionation and gel column chromatography. The lytic activity was found to consist of three enzyme activities which were resolved on Sephadex G-100. The first peak on chromatogram exhibited proteolytic, lytic and laminarinase activities, and the second had both lytic and laminarinase activities, whereas the third peak was shown to contain lytic activity only. Three enzyme activities showed the synergistic effect and reducing agents accelerated the yeast roil wall lysis. This indicates that lytic, proteolytic and laminarinase activity acted cooperatively in the lysis of intact cells. Tannic acid precipitate of crude enzyme constituted of three enzyme activities had a high lytic activity on viable yeast cell and has proved useful in yeast protoplast formation.

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Comparison of various cell lysis methods for Pichia pastoris

  • Han, Kyung-Ah;Kim, Sun-Yong;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.889-893
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    • 2005
  • Cell lysis method is very important to study intracellular metabolites in microorganisms. In this study, various cell lysis methods were compared to find a good method analysing intracellular metabolites in P. pastoris. P. pastoris was cultivated in YPD medium at 30 $^{\circ}C$, 200 rpm for 24 hours and its morphology as well as the change in strain's shape were observed by microscopy. The UV/vis spectrophotometer was used to measure intracellular protein concentrations.

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The Synergistic Action of the AL-Protease from Arthrobacter luteus on the Lysis of Yeast Cell Walls (Arthrobacter luteus가 생산하는 AL-Protease의 효모세포벽 용해 촉진작용)

  • Oh, Hong-Rock;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.4
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    • pp.401-408
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    • 1985
  • The yeast cell wall lytic action of the alkaline AL-protease, which was found out of the crude Zymolyase that a kind of yeast cell wall lytic $endo-{\beta}-1$, 3-glucanase produced from Arthrobacter luteus, was investigated with the viable cells of S. sake and it's cell wall preparation. AL-protease on the lysis of the viable yeast cells showed very low activities with the alone, but the lytic activities were highly increased with the combination of AL-protease and Zymolyase. On the stepwise treatment of the viable yeast cells with AL-protease and Zymolyase, the cells were lysed highly only by the course having a treatment with Zymolyase after pretreatment with AL-protease. Thus synergistic action of AL-protease was not observed with any some commercial enzymes, known as a type of alkaline and serine protease such as AL-protease, and was also found to be affected greatly by the culture conditions and species of the yeast tested. AL-protease caused the release of some peptide and a lot of sugar from the cell wall preparation, but could not lysed the cell wall more than 66%. Whereas Zymolyase could lysed the cell walls almost completely with alone. On the basis of these results, the synergistic action of AL-protease on the lysis of S. sake cells is hypothesized that at first AL-protease bind to the yeast cell surface layer consisting of mannan and protein, and then changes their conformation to facilitate the penetration of Zymolyase from the outside to the inside framework layer constituted of alkali insoluble ${\beta}-1,\;3-glucan$.

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A POSSIBLE MECHANISM OF POLYACETYLENE: MEMBRANE CYTOTOXICITY

  • Kim, Hyeyoung;Lee, You-Hni;Kim, Shin-Il
    • Toxicological Research
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    • v.4 no.2
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    • pp.95-105
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    • 1988
  • The effects of polyacetylenes on living membrances, rat erythrocyte and murine leukemic L1210 cell as well as artificial lipid bilayer were determined to investigate the cytotoxic mechanism of polyacetylenes against cancer cell lines. As results, panaxydol and panaxynol caused erythrocyte hemolysis dose-dependently while panaxytriol had no lysis. For liposomes composed of phosphatidyl choline (PC) and phosphatidic acid(PA), all three polyacetylenes supressed the osmotic behavior at the same degree.

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Comparative Study of Natural Killer Cell Activity after Red Ginseng Medication on Rat (원산지별 홍삼투여에 따른 백서의 자연살해세포 활성도 변화에 대한 연구)

  • Im, Ju-Hyuk;Cho, In-Young;Min, Kyoung-Yoon;Lee, Hae-Yong;Kim, Seul-Ji;Park, Young-Ju;Lew, Jae-Hwan;Lee, Beom-Joon;Kim, Sung-Wook;Joo, Il-Woo
    • The Journal of Internal Korean Medicine
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    • v.29 no.4
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    • pp.1075-1082
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    • 2008
  • Objectives : The purpose of this experiment is comparing the difference on natural killer cell activity through Korean red ginseng and Chinese red ginseng by $^{51}Cr$ release assay. Methods : Thirty rats were equally divided into a Korean red ginseng group, a Chinese red ginseng group and a control group. Korean and Chinese red ginseng were administrated to the rats at 200mg daily for a weak, while 0.9% normal saline was given to the control. Percent specific lysis (PSL) and lytic units (LU) were calculated from spleen cells by $^{51}Cr$ release assay. Results : Percent specific lysis of the Korean red ginseng group was significantly higher than that of the control in the ratio of 100:1, effector cell:target cell (p<0.05). Percent specific lysis of Korean red ginseng group was also significantly higher than that of the Chinese red ginseng group in the ratio of 25:1, effector cell:target cell (p<0.05). Chinese red ginseng showed no effect on NK cell activity. Conclusions : These findings suggest that Korean red ginseng improves immunologic function and shows superior effects than Chinese red ginseng.

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Effect of Electric fusion Methods on Cell Fusion Rate and Embryo Development by Somatic Cell Nuclear Transfer in Korean Native Cattle(KNC) (한우 체세포를 이용한 핵이식에서 전기융합 방법이 융합율 및 배발달율에 미치는 영향)

  • 김은국;김정욱
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.171-178
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    • 2003
  • This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM+0.05% FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3% BSA or 10% FBS at 37$^{\circ}C$, 90% $N_2$, and 5% $CO_2$in incubator for 6 days. The fusion rates were 51.6%(chamber) and 68.9%(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7%, 11.5%), respectively. The cleavage rates were significantly different between the chamber method(73.2%) and needle method(80.3%), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1%, 61.1% and 18.4%, 26.3% respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7%) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

Molecular Cloning and Expression of a Sodium-Driven Flagellar Motor Component Gene(motX) from Vibrio fluvialis

  • Park, Je-Hyeon;Lee, Jong-Hee;Kim, Young-Sook;Hong, Yong-Ki;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.973-978
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    • 2001
  • The bacterial flagellar motor is a molecular machine that couples proton or sodium influx to force generation, mostly for driving rotation of the helical flagellar filament. In this study, we cloned a gene (motX) encoding a component of the sodium-driven flagellar motor from Vibrio fluvialis. The nucleotide sequence of the motX gene, composed of 633 bp and 211 amino acid residues, was determined. Overexpression of the motX gene in Escherichia coli using a strong promoter induced growth inhibition and cell lysis. The lethal effect of E. coli was suppressed by adding amiloride, as a potent inhibitor for the sodium channel. Electron microscopic observation of the expressed protein indicated that MotX protein induced by isopropyl ${\beta}$-D-thiogalactopyranoside caused the lysis of host cell.

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