• BMB Reports
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• 제38권2호
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• pp.177-181
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• 2005

Effects of common electrophoretic reagents, reducing agents ($\beta$-mercaptoethanol [BME] and DTT), denaturants (SDS and urea), and non-ionic detergent (Triton X-100), on the activity and stability of bovine plasmin (b-pln) and human plasmin (h-pln) were compared. In the presence of 0.1% SDS (w/v), all reagents completely inhibited two plns, whereas SDS (1%) and urea (1 M) denatured plns recovered their activities after removal of SDS by treatment of 2.5% Triton X-100 (v/v). However, reducing agents (0.1 M of BME and DTT) treated plns did not restore their activities. Based on a fibrin zymogram gel, five (from b-pln) and four (from h-pln) active fragments were resolved. Two plns exhibited unusual stability in concentrated SDS and Triton X-100 (final 10%) and urea (final 6 M) solutions. Two bands, heavy chain-2 (HC-2) and cleaved heavy chain-2 (CHC-2), of b-pln were completely inhibited in 0.5% SDS or 3 M urea, whereas no significant difference was found in h-pln. Interestingly, 50 kDa (cleaved heavy chain-1, CHC-1) of b-pln and two fragments, 26 kDa (light chain, LC) and 29 kDa (microplasmin, MP), of h-pln were increased by SDS in a concentration dependent manner. We also found that the inhibition of SDS against both plns was reversible.

• 한국미생물·생명공학회지
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• 제15권5호
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• pp.334-339
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• 1987

Pyruvic acid 생산 미생물로부터 얻어진 pyruvic acid의 tryptophan으로의 효소적 전환을 조사하였다. 발광 미생물인 Beneckea sp.가 pyruvic acid의 생산에 사용되었다. pyruvic acid, indole과 ammonia로 부터 tryptophan을 합성하는 tryptophanase의 효소원으로 Enterobacter aerogenes ATCC10031의 균체가 반응용액에 직접 사용되었다. Tryptophan의 생산량을 증가시키기 위해, 비이온성 계면활성제와 비 수용성 유기용매가 indole의 저장소로 사용되었다. 비이온성 계면활성제의 경우 triton X-100은 매우 효과적이였다. 1.5%의 triton X-100 이 사용되었을 때, 37$^{\circ}C$에서 48시간 동안에 7.7g/$\ell$의 tryptophan이 생산되었다. 비수용성 유기용매의 경우 10%의 benzene이 사용되었을 때, 37$^{\circ}C$에서 48시간 동안에 8.7g/$\ell$의 tryptophan이 생산되었다. 이 tryptophan의 양은 indole과 pyruvic acid를 기준으로 각각 48%와 36%의 전환율에 해당한다.

• 한국토양환경학회지
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• 제1권2호
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• pp.61-72
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• 1996

일련의 회분식 실험 및 실험실 규모의 연속식 실험을 통하여 유기오염물질에 의해 오염된 토양에 in-situ 토양세척기법을 적용하는데 필요한 운전조건을 도출하였다. 실제 토양에 소수성 유기오염물질인 n-dodecane, naphthalene, anthracene을 일정량 오염시킨 후 Tween 80, Triton X-100, Sodium dodecyl sulfate(SDS)가 포함된 용액을 이용하여 세척능력을 비교하였다. 회분식 및 연속식 실험 결과, Tween 80와 Triton X-100가 n-dodecane의 제거에 효과가 우수하였으며, naphthalene과 anthracene의 제거에는 SDS와 Tween 80가 우수하였다. 연속식 실험이 끝난 후, 토양에 잔류하는 세척제의 양은 Tween 80이 제일 적었다. 연속식 실험에서 통과유속을 변화시킨 결과, 7 ml/min에서는 세척에 필요한 접촉시간의 감소로 인하여 세척효과가 3 또는 5ml/min보다 오히려 감소되었다. 본 실험의 적용 범위 내에서는 토양의 초기 pH는 세척이 진행됨에 따라 세척능력에 큰 영향을 미치지 않았다. 세척제의 미생물에 대한 독성실험 결과 Tween 80이 그람 음성균과 양성균의 성장에 가장 영향을 미치지 않는 것으로 판명됐다.

• 한국수정란이식학회지
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• 제30권3호
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• pp.225-228
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• 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.

• Journal of Microbiology and Biotechnology
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• 제23권11호
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• pp.1554-1559
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• 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.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2002년도 추계학술발표회
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• pp.187-190
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• 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.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.233-238
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• 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.

• BMB Reports
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• 제40권1호
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• pp.53-57
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• 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.

• 약학회지
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• 제36권2호
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• pp.129-136
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• 1992

The effects of ginseng saponins, gypsophila saponin, sodium dodecyl sulfate(SDS), and Triton X-100 on membrane $K^+-dependent$ phosphatase activity which is lipid dependent and represents dephosphorylation step of the complete Na+, $K^+-ATPase$ reaction were investigated in this study to elucidate whether the effects of ginseng saponins are due to the detergent action, using sarcolemma enriched preparation isolated from dog ventricle. $Na^+$, $K^+-ATPase$ and $K^+-dependent$ phosphatase activities of cardiac sarcolemma were about $143\;{\mu}mol$ Pi/mg protein/hr and $34\;{\mu}mol$ p-nitrophenol/mg protein/hr, respectively. While ginseng saponins (triol>total>diol) inhibited $K^+-dependent$ phosphatase activity, gypsophila saponin, and low dose of SDS($0.4\;{\mu}g/{\mu}g$ protein), and Triton X-100 ($0.6\;{\mu}g/{\mu}g$ protein) increased the enzyme activity, indicating disruptive effect of detergents on membrane barriers. The activating effect of low doses of Triton X-100 on membrane $K^+-dependent$ phosphatase appeared at concentration decreasing light scattering. However, the inhibitory effect of ginseng saponin appeared before a decrease in light scattering. These results suggest that low concentrations of ginseng saponins inhibit the membrane $K^+-dependent$ phosphatase by interacting directly with enzyme before membrane disruption.

• Journal of Yeungnam Medical Science
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• 제3권1호
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• pp.41-48
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• 1986

사람의 적혈구막으로부터 Band 3를 분리정제하고 이를 liposome 내로 도입시켜 그 결과를 관찰하였다. 적혈구를 약알칼리 저장액으로 용혈시켜 막을 분리한 후, 저이온강도 용액으로 처리하여 Band 4를 추출하였다. Triton X-100 추출액에 p-chloromercuribenzoate를 가하고 sucrose density gradient ultracentrifugation후 fractionation하여 Band 3를 정제하였다. phosphatidyl L-serine과 cholesterol을 1 : 1 molar ratio로 섞고 진공회전 증발기를 사용하여 chloroform을 제거한 후 Triton X-100을 제거한 Band 3용액을 가하고 sonication함으로 liposome(reverse-phase evaporation vesicle)을 만들면서 Band 3를 도입 시켰다 Band 3의 분리정제 및 liposome에 도입되었음은 sodium dodecyl sulfate-polyacrylamide gel 전기영 동 후 coomassie brilliant blue 염색으로 확인할 수 있었다.