• 제목/요약/키워드: Triton X

검색결과 346건 처리시간 0.026초

Klebsiella pneumoniae 균주의 세포외막에서 분리한 2-furaldehyde dehydrogenase의 특성에 관한 연구 (Characterization of the Outer Membrane-Associated 2-Furaldehyde Dehydrogenase from Klebsiella pneumoniae)

  • 이준우;강사욱;하영칠;한홍의
    • 미생물학회지
    • /
    • 제26권3호
    • /
    • pp.197-206
    • /
    • 1988
  • Klebsiella pneumoniae의 세포외막으로부터 2-furaldehyde를 2-furoic acid로 산화시키는 2-furaldehyde dehydrogenase를 분리하여 그 특성을 조사하였다. 이 효소는 $\beta$-$NAD^{+}$를 특이적으로 요구하였다. 분리과정중의 효소활성도는 2-furaldehyde를 기질로 사용하고 $\beta$-$NAD^{+}$를 조효소로 사용하면서 high performance liquid chromatography에 의해 측정 되었다. 세포외막은 Percoll의 밀도흉배에 의한 초원심분리방법과 $Mg^{2+}$, Triton X-100으로 용해시킨 후, 초원심분리시키는 방법으로 수집되었다. 세포외막단백질은 EDTA와 lysozyme을 처리함으로서 얻어졌고, 효소는 QAE-Sephadex Q-504 S Sephadex G-100-을 사용하면서 column chromatography 방법에 의해 분리되었다. 본 효소는 $85^{\circ}C$, PH9.5, 그리고 1.5% (vol/vol) Triton X-100의 존재하에서 최대활성을 보여주었다. 효소의 분자량은 nondenaturing polyacrylamide gel e electrophoresis의 결과, 88, 000.으로 추정되었고, 2-furaldehyde에 대한 효소의 Km값은 4.72 mM 이였다.

  • PDF

Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer

  • Chung, Hak-Jae;Son, NaRae;Han, Joo-Hee;Park, Chun-Gyu;Kim, Kyung-Woon;Park, Mi-Ryung;Hwang, In-Sul;Park, Jin-Ki;Im, Gi-Sun
    • 한국수정란이식학회지
    • /
    • 제30권3호
    • /
    • pp.225-228
    • /
    • 2015
  • Understanding the behavior of transgenes introduced into oocyte or embryos is essential for evaluating the methodologies for transgenic animal production. To date, many studies have reported the production of transgenic pig embryos with, however, low efficiency in environment of blastocyst production. The aim of present study was to determine the expression and duration of transgene transferred by intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT). Embryos obtained from the ICSI-MGT procedure were analysed for the expression of GFP and then for the transmission of the transgene. Briefly, fresh spermatozoa were bound to exogenous DNA after treatment by Triton X-100 and Lipofectin. When ICSI-MGT was performed using sperm heads with tails removed, the yield of blastocyst (25.3%), treated with Lipofectin (18.8%) and Triton X-100 (19.2%) were observed. Treatments of Lipofectin or Triton X-100 did not further improve the rates of blastocysts. Moreover, the apoptosis rates of embryos were obtained from the control and LIpofectin groups (8.7%, 9.7%, respectively), but were significantly higher in the Triton X-100 group (13.0%). Our results demonstrated that ICSI-MGT caused minimal damage to oocytes that could develop to full term. Moreover, the embryos derived by ICSI-MGT have shown prolonged exogenous DNA expression during preimplantation stage in vivo. However, more efforts will be required to improve the procedures of both sperm treatments cause of high frequency of mosaicisms.

Structural Stability of Bacteriorhodopsin Solubilized by Triton X-100

  • Sasaki, Takanori;Sonoyama, Masashi;Mukai, Yuri;Nakazawa, Chieko;Mitaku, Shigeki
    • Journal of Photoscience
    • /
    • 제9권2호
    • /
    • pp.293-295
    • /
    • 2002
  • The structural stability of bacteriorhodopsin (bR) solubilized by Triton X-100 (TX-100) was studied by measuring the denaturation kinetics in the dark and under illumination, and compared with the structural stability of bR solubilized by octyl-${\beta}$-glucoside (OG). In the dark, bR solubilized by TX- 100 was more stable than bR solubilized by OG. Under illumination, bR solubilized by TX-100 showed light-induced denaturation in the same manner as bR solubilized by OG. These results in the dark well correlated with the experimental results of the visible CD band. Although solubilized bR in the TX-100 concentration range of 2-50 mM showed almost identical positive CD band and did not denature in the dark at 35$^{\circ}$C, the kinetic constant of the photobleaching increased with the increase of TX-100 concentration. These results suggested that photo-intermediates of solubilized bR are destabilized by TX-100 micelles.

  • PDF

Use of Triton X-100 and Sephacryl S-500 HR for the Purification of Cymbidium Mosaic Virus from Orchid Plants

  • Han, Jung-Heon;La, Yong-Joon;Lee, Cheol-Ho
    • The Plant Pathology Journal
    • /
    • 제15권1호
    • /
    • pp.34-37
    • /
    • 1999
  • Cymbidium mosaic virus (CyMV) was purified from CyMV infected orchid plant leaves by Sephacryl S-500 HR column chromatography. Partial purification was done by solubilization with Triton X-100 (alkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG 6,000) followed by ultracentrifugation on 30% sucrose cushion. Based on the spectrophotometric analysis, 33 mg of CyMV could be obtained form 100 g of CyMV-infected orchid plant leaves. The purified CyMV represented one distinct homogeneous band by SDS-PAGE, and electron microscopy revealed that it was highly homogeneous and not fragmented. Bioassay demonstrated that the purified CyMV had a normal infectivity to Chenopodium amaranticolor and orchid plants. Based on these results, the purification method in this work could be served as an improved method for the purification of CyMV and similar viruses with good yield, high purity and native integrity.

  • PDF

Purification and Characterization of Heat-Tolerant Protease Produced by Bacillus polyfermenticus SCD

  • Choi, Gooi Hun;Jo, Mi Na;Kim, Jin-Man;Kim, Cheon-Jei;Kim, Kee-Tae;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • 제23권11호
    • /
    • pp.1554-1559
    • /
    • 2013
  • A protease produced by Bacillus polyfermenticus SCD was purified and characterized as a new detergent material. The protease was purified from supernatant produced by B. polyfermenticus SCD, by ammonium sulfate precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, and finally gel filtration chromatography on Sephadex G-50. The molecular mass of this enzyme was 44 kDa based on SDS-PAGE. The optimum temperature and pH were $50^{\circ}C$ and pH 8.0. The ranges of its stability to the pH and temperature were 7.0 to 9.0 and under $40^{\circ}C$, respectively. The enzyme was highly stable in the presence of the surfactants like Triton X-100 (0.1%), showing a 2-fold increase in its proteolytic activity. However, the enzyme was slightly inhibited by the chelating agent EDTA (1 mM). The enzyme has a maximum activity at $50^{\circ}C$ and the activity can be increased by surfactants such as Triton X-100 and Tween 80.

영가금속에 의한 Endosulfan I과 II의 환원분해에 미치는 계면활성제의 영향 (Effect of surfactants on reductive degradation of Endosurfan I and II by ZVM)

  • 김진영;김영훈;신원식;전영웅;송동익;최상준
    • 한국지하수토양환경학회:학술대회논문집
    • /
    • 한국지하수토양환경학회 2002년도 추계학술발표회
    • /
    • pp.187-190
    • /
    • 2002
  • Reductive dechlorination of endosulfans was studied with zero valent metals (ZVMs) and bimetals in aqueous batch reactors. The effect of surfactants was evaluated. Endosulfan was successfully dechlorinated with zero valent iron. However, a bimetal, palladium coated iron (Pd/Fe) showed a highly enhanced reactivity for both endosulfan I and II indicating palladium act as a dechlorination catalyst on the iron. The effect of surfactants on degradation with ZVM has been very controvertible. Variable concentration of a nonionic surfactant, Triton X-100 and an anionic surfactant, SDS were added into the reactor with ZVM. The reaction rates of endosulfan were increased with both surfactants. In the case of Triton X-100, the reaction rate was increased with the increasing surfactant concentration up to 400 mg/L. Addition of small amount of surfactant under the CMC, the reaction rate was increased. However, the enhancing effect was diminished when a higher concentration of surfactant (1,000 mg/L) was used. Current study implicate that the surfactant adsorbed on the metal surface might increase the surface concentration of endosulfan resulting in the increased reaction rate. However, partitioning of endosulfan into the micelle formed at the high concentration of surfactant diminish the enhancing effect by reducing the contact chance between target compound and the metal surface.

  • PDF

중성 셀룰라제 처리에 의한 데님의 물성 (Effect of Cellulase on Characteristics of Denim)

  • 김지연;송화순;김혜림
    • 한국의류산업학회지
    • /
    • 제11권3호
    • /
    • pp.469-473
    • /
    • 2009
  • Recently, eco-friendly processing has been focused in the textile industry in order to reduce environmental pollutions. Applications of enzyme technology to the textile industry are an example of more environmentally compatible processes. However, there is not enough quantity of referring to denim fabric subjected to enzymatic treatment. In this study, depending on pH, temperature, cellulase concentration, and treatment time, the weight loss of denim fabrics was examined. Characteristics of enzyme-treated fabrics were measured by tearing strength, stiffness, and K/S values. The effect of a non-ionic surfactant (Triton X-100) on characteristics of the enzyme-treated fabrics was evaluated. The cellulase treatment condition on the cotton fabric were optimized to pH 6.0, $50^{\circ}C$, 1%(o.w.f.), and 60minutes. Characteristics of denim fabrics by cellulase treatment in the presence of Triton X-100 did not improve because Triton-X inhibited the activity of enzyme.

카드뮴 내성 효모의 세포내 카드뮴 축적 기작 (Mechanism of Cadmium Accumulation into the Cell of Cadmium-Ion Tolerant Yeast)

  • 유대식;송형익;정기택
    • 한국미생물·생명공학회지
    • /
    • 제18권3호
    • /
    • pp.233-238
    • /
    • 1990
  • The mechanism of intracellular accumulation of cadmium in a cadmium-ion tolerant yeast, Hansenula ammala B-7, which is an extreme cadmium tolerant strain and has the ability to take up a large amount of cadmium was investigated. The amounts of cadmium taken up by the scalded yeast cells were 2 to 3 times more than the value of the living cells. The living Hansenula anomala B-7 cells adsorbed 74% of cadmium taken up onto the other layer of the cells and 26% of it accumulated inside the cells. But the scalded cells adsorbed 98.3% of cadmium taken up and accumulated 1.7% of it inside the cells. A cadmium uptake and its accumulation were accelerated up to 162.3% and 275.4% by Triton X-100 in the living cells, respectively. Whereas in the scalded cell cadmium uptake was not affected by Triton X-100. Furthermore the cadmium uptake and its accumulation were strongly inhibited by metabolic inhibitors like 2,4-dinitrophenol, sodium azide and potassium cyanide in the living cells, but in the scalded cells cadmium uptake was not affected by metabolic inhibitors. These results suggested that the intracellular accumulation of cadmium by the cadmium-tolerant Hansenula anomala B-7 cells was apparently dependent of biological activity, and also gave evidence of the existance of energy-dependent system.

  • PDF

Highly Selective and Simple Zero and First Order Derivative Spectrophotometric Determination of Palladium by Using α-Benzilmonoxime in Triton X-100 Micellar Solution

  • Eskandari, Habibollah;Karkaragh, Gholamhassan Imanzadeh
    • Bulletin of the Korean Chemical Society
    • /
    • 제24권12호
    • /
    • pp.1731-1736
    • /
    • 2003
  • The reaction of ${\alpha}$-Benzilmonoxime with palladium(II) produces a green complex in triton X-100 micellar media. Palladium has been determined using zero and first derivative spectrophotometric methods. The absorbances of Pd(II)- ${\alpha}$--benzilmonoxime complex at 441.8 and 677.0 nm in 0.10 M perchloric acid solution were monitored and linear working ranges of 0.3-12.0 and 0.7-20 ${\mu}$g mL$^{-1}$ with detection limits of 0.07 and 0.10 ${\mu}$g $mL^-1$ were obtained, respectively. Also, recoveries in the range of 92.8 to 100.1% and relative standard deviations in the range of 0.4 to 7.1% were obtained. First derivative spectrophotometry has also been applied for palladium determination under the optimum condition. The linear dynamic range of 0.2-24.0 ${\mu}$g $mL^{-1}$ palladium with relative standard deviations of 0.6-6.9% and recoveries in the range of 94.9-102.5% has been obtained by first derivative spectrophotometry. The method shows high selectivity because of the high concentration of acid used, which prevents formation of complexes of ${\alpha}$--benzilmonoxime with the other cations. The palladium complex formed was stable at least one day. The method was successfully applied to the determination of palladium in some synthetic palladium alloys and palladium-charcoal powder.

HQNO-sensitive NADH:Quinone Oxidoreductase of Bacillus cereus KCTC 3674

  • Kang, Ji-Won;Kim, Young-Jae
    • BMB Reports
    • /
    • 제40권1호
    • /
    • pp.53-57
    • /
    • 2007
  • The enzymatic properties of NADH:quinone oxidoreductase were examined in Triton X-100 extracts of Bacillus cereus membranes by using the artificial electron acceptors ubiquinone-1 and menadione. Membranes were prepared from B. cereus KCTC 3674 grown aerobically on a complex medium and oxidized with NADH exclusively, whereas deamino-NADH was determined to be poorly oxidized. The NADH oxidase activity was lost completely by solubilization of the membranes with Triton X-100. However, by using the artificial electron acceptors ubiquinone-1 and menadione, NADH oxidation could be observed. The activities of NADH:ubiquinone-1 and NADH:menadione oxidoreductase were enhanced approximately 8-fold and 4-fold, respectively, from the Triton X-100 extracted membranes. The maximum activity of FAD-dependent NADH:ubiquinone-1 oxidoreductase was obtained at about pH 6.0 in the presence of 0.1M NaCl, while the maximum activity of FAD-dependent NADH:menadione oxidoreductase was obtained at about pH 8.0 in the presence of 0.1M NaCl. The activities of the NADH:ubiquinone-1 and NADH:menadione oxidoreductase were very resistant to such respiratory chain inhibitors as rotenone, capsaicin, and $AgNO_3$, whereas these activities were sensitive to 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO). Based on these results, we suggest that the aerobic respiratory chain-linked NADH oxidase system of B. cereus KCTC 3674 possesses an HQNO-sensitive NADH:quinone oxidoreductase that lacks an energy coupling site containing FAD as a cofactor.