• 제목/요약/키워드: Triton X -100

검색결과 321건 처리시간 0.02초

Acinetobacter sp. SY-01로부터 Enantioselective Lipase의 생산 (Production of Enantioselective Lipase from Acinetobacter sp. SY-01)

  • 박대원;박호일;신평균
    • 한국미생물·생명공학회지
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    • 제31권2호
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    • pp.145-150
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    • 2003
  • Acinetobacter sp. SY-01가 생산하는 lipase는 소염진통제로 쓰이는 azolerP 의약품 중간체인 두 enantiomer를 가진 rcemate에 대해 enantioselectivity를 가지는 효소로써 chiral drug를 생산하는데 있어 중요한 역할을 수행한다. Acinetobacter sp. SY-01 lipase 효소의 다량 생산을 위한 여러 가지 최적 조건을 조사하였다. 탄소원으로는 여러 가지 천연 오일을 이용하여 실험한 결과 0.2% olive oil이 가장 좋았으며, 최적 온도와 pH는 30도와 pH 7이였다. EH한 금속이온과 유기용매 안정성에 있어서는 $Fe^{2+}$$Ca^{2+}$ DMSO가 좋은 효과를 나타냇었다. 특히 DMSO는 유기용매가 첨가되지 않았을 때보다 약 2.5배 높은 효소 활성을 나타내었다. 계면활성제 이용에 있어서는 tritom X-100을 이용했을 때 좋은 효과를 나타냈다. Acinetobacter sp. SY-01가 생산하는 lipase 효소의 최적 배지 조성은 0.8% yeast extract, 0.2% olive oil 0.4% triton X-100, 40% DMSO, 0.1 % $NH_4$Cl% 0.4%$K_2HPO_4$ 3.9% $NaH_2PO_4$ 0.03% $CaCl_22H_2O$ 0.01% $FeSO_4$$7H_2O$(pH 7.0)이다.

Fluorescence probe study on the solubilization sites of aniline derivatives in triton X-100 and zephiramine micelles

  • Han, Suk-Kyu;Lee, Yong-Soo
    • Archives of Pharmacal Research
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    • 제9권3호
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    • pp.139-144
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    • 1986
  • The solubilization sites of aniline and its derivatives in micelles were investigated with fluorescence probe technique. The fluorescence probes employed in this study are 12-(9-anthroyl) stearic acid (AS) and 2-p-toluidinylnaphthalene-6-sulfonate (TNS) which are incorporated in the interior of the micelle and attaced to its surface, respectively. As these two probes were effectively quenched by aniline and its surface, respectively. As these two probes were effectively quenched by aniline and its derivatives, the modified Stern-Volmer relationship in micellar system could be applicable to estimate the partition coefficient, $K_{p}$ of the solubilizate between aqueous and micellar phase. Because $K_{p}$ derived by this method reflects the relative proximity of the fluorophore to the quencher, the ratio of $K_{p}$ in the surface area to that in the interior of the micelle is interpreted in terms of the relative location of the solubilizate in micellar aggregate. The results show that the solubilizates are not located in a definite position but distributed in the multiple-sites of the micelle. The solubilization sites of the solubilizates in the icelle are dependent on their structures. As the solubilizate has more numbers of N-substituents of aniline and more numbers, of carbon in the substituent, it tends to incorporate in the interior of the micelle more effectively.

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Reconstitution of Sarcoplasmic Reticulum-$Ca^{2+}$ Release Channels into Phospholipid Vesicles : Investigation of Conditions for Functional Reconstitution

  • Yang, In-Sik;Lee, Hee-Bong
    • BMB Reports
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    • 제28권2호
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    • pp.129-137
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    • 1995
  • The ryanodine-receptor $Ca^{2+}$ release channel protein in the sarcoplasmic reticulum membrane of rabbit skeletal muscle plays an important role in muscle exitation-contraction (E-C) coupling. Various types of detergents were tested, including Chaps, cholate, octylglucoside, Zwittergents, Mega-9, Lubrol PX, and Triton X-100 for solubilization of this protein. Among these, Chaps and Triton X-100 were found to optionally solubilize the channel complex. Optimum conditions for this solubilization were pH 7.4 with a salt concentration of 1 M. The addition of phospholipid in the solubilization step helped in stabilizing the protein. The purification of the receptor was performed using sucrose density gradient centrifugation. Various methods [dilution, freeze-thaw, adsorption (Biobeads), and dialysis] were investigated to incorporate the Chaps-solubilized and purified $Ca^{2+}$ release channel protein into liposomes made from different types of phospholipids. Of these, a combined method consisting of a dialysis, freeze-thaw and sonication steps yielded the best results. Reconstituted vesicles produced by this method with 95% phosphatidylcholine (from soybean extract) had good function.

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Improved recovery of active GST-fusion proteins from insoluble aggregates: solubilization and purification conditions using PKM2 and HtrA2 as model proteins

  • Park, Dae-Wook;Kim, Sang-Soo;Nam, Min-Kyung;Kim, Goo-Young;Kim, Jung-Ho;Rhim, Hyang-Shuk
    • BMB Reports
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    • 제44권4호
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    • pp.279-284
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    • 2011
  • The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1 : 200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.

표고버섯 배양액으로부터 단백다당류의 추출 및 정제 방법 (Extraction and Separation of Protein-bound Polysaccharide by Lentinus edodes)

  • 박경숙;이별나
    • 한국식품영양학회지
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    • 제10권4호
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    • pp.503-508
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    • 1997
  • L. edodes의 배양액 및 균사체로부터 항암작용이 있는 단백 다당류의 추출 및 정제방법에 관한 실험을 실시하였다. 단백다당류 추출에서 비이온성 계명 활성제인 2% Triton X-100 용액으로 추출할 경우 상온에서 추출하여도 통상의 10$0^{\circ}C$ 열수 추출을 통해 얻을 수 있는 것보다 수율이 높았다. 그러나 계명활성제 용액에 4N NaCl을 첨가하여 추출하면 ethanol 침전과정에서 단백다당류의 침전물을 얻을 수 없었다. 열수 추출에 있어서 추출 온도는 10$0^{\circ}C$가 효과적이고 추출 시간은 4시간 정도면 충분하였다. 열수 추출 결과 ethanol 침전 및 동결건조를 통하여 얻어진 조단백 다당류의 정제 실험에서는 alumina를 충전제로 하는 gel filtration보다 sephadex에 의한 gel filtration 또는 DEAE-cellulose, DEAE-sephadex를 이용한 ion exchange chromatography가 보다 효과적이다.

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인삼(人蔘) Saponin, 은시호(銀柴胡) Saponin 및 계면활성제(界面活性劑)가 적혈구(赤血球)의 용적변화(容積變化) 및 Fragility에 미치는 영향(影響) (Effect of Ginseng Saponin, Gypsophila Saponin, and Detergents on Volume Changes and Fragility of Red Blood Cells)

  • 이신웅;이정수;이수근;옥창건;김영희
    • 약학회지
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    • 제33권1호
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    • pp.15-19
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    • 1989
  • The effects of Gypsophila saponin, sodium dodecylsulfate (SDS) and Triton X-100 on volume changes and fragility of red blood cells were compared to ginseng saponin to elucidate whether there are any difference in their action on membrane lipid. Cell volume was decreased to about 38% in 1M NaCl and increased to about 20% in 1/10M NaCl. Hematocrit value was decreased by Gypsophila saponin, SDS, and Triton X-100 which caused hemolysis in isotonic NaCl solution. These detergents also inhibited increase of cell volume and accelerated hemolysis in hypotonic solution. However, ginseng saponin did not influence to osmotic volume changes and hemolysis of red blood cells. These results suggest that the disruptive effect of plant saponin on membrane barriers induced by removing membrane lipid is different from their source and ginseng saponin has very low affinity to membrane lipid.

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전기자극 하에서 알콜 산화효소의 안정화연구; 당, 히드로젤, 계면활성제 효과 (Stabilization of Alcohol Oxidase under Electrostimulation; Sugars, Hydrogels and Surfactants Effect)

  • 김범수;이강민;;김경숙
    • KSBB Journal
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    • 제21권6호
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    • pp.456-460
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    • 2006
  • 효소는 환경과 조건에 따라 상이한 활성도를 나타내며, 전기 자극에 의하여 비활성화된 알콜 산화효소는 당, 계면활성제, 히드로젤 들에 의하여 안정화되었다. 이들 효소는 trehalose, TritonX-100, Brij에서 보다 안정하였으며, 이들 효소의 안정도는 Hansenula sp., P. pastoris, C. boidinii에서 분리된 효소가 순서적으로 보다 안정하였다. 전기자극하에서 효소의 안정화에 대한 이 연구는 생물공학, 단백질공학, 의료공학 분야에 널리 이용될 수 있으리라 사료된다.

Rhodotorula glutinis K-501에 의해 생산된 세포외지질의 생체계면활성제로서 물리화학적 특성 (Physiochemical Properties of Extracellular Lipid Produced by Rhodotorula glutinis K-501 as a Biosurfactant)

  • 박평규;채희정;김의용
    • KSBB Journal
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    • 제13권1호
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    • pp.65-70
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    • 1998
  • 지질 생산 효모인 Rhodotorula glutinis K -501이 생산하는 세포외지질(extracellular lipid)의 물리화학적 성질을 조사하여 생체 계면활성제로서의 응용 가능성을 검토하였다. 분석실험 결과 생산된 세포외지질은 당지질계 물질임을 알 수 있었다. 세포외지질의 임계미셀농도와 표면장력은 각각 89mg/L와 31dyne/cm로서 계면활성제로서 매우 우수한 표면활성을 가지고 있었으며 넓은 범위의 pH에서 표면장력의 값은 일정하였다. 상용 계면활성제인 Tween 80과 Triton X-100에 비하여 세포외지질은 각각2-3배 높은 유화도와 1.3배 높은 분산력을 가지고 있었다.

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동전기-생물학적복원기술과 계면활성제를 이용한 phenanthrene 오염토양의 정화

  • 김상준;박지연;이유진;양지원
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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    • pp.186-190
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    • 2004
  • The electrokinetic bioremediation employing electrolyte circulation method was carried out for the cleanup of phenanthrene-contaminated kaolinite, and microorganism used in the biodegradation of phenanthrene was Sphingomonas sp. 3Y. The electrolyte circulation method supplied ionic nutrientsand the microorganism into soil, and inhibited the significant pH change of soil by increasing the soil buffering capacity by providing phosphate buffer compounds. When the remediation process was conducted without surfactant, the removal efficiency of phenanthrene, at the initial concentration of 200 ppm, was 69% for only 7 days. Higher microbial population and lower phenanthrene concentration were observed in the anode and middle regions of soil specimen than in the cathode region. The higher density of microorganism was because the microbial movement was in the direction of the anode part due to the negative surface charge. When Triton X-100 and APG of 20 g/1 were used to improve the bioavailability of phenanthrene strongly adsorbed onto soil surface, about 90 and 39% of phenanthrene removal were obtained. Consequently, it was confirmed that the microorganism preferred APC to phenanthrene as carbon source and so the removal efficiency with APG decreased less than that without APG.

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철과 양수성 물질을 이용한 PCE와 크롬 제거에 관한 연구

  • 조현희;천병식;박재우
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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    • pp.68-71
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    • 2004
  • Effects of surfactants and natural organic matter (NOM) on the sorption and reduction of tetrachloroethylene (PCE) and chromate with iron were examined in this study. PCE and chromate reduction by iron depended on the ionic type of the surfactants in this study. The apparent reaction rate constants of PCE with Triton X-100 and hexadecyltrimethyl ammonium (HDTMA) at one half and two times of the critical micelle concentration (CMC) were relatively higher than without surfactants because of the enhanced PCE partitioning and surface concentration. In the presence of sodium dodecyl benzene sulfonate (SDDBS) at 2000 mg/L and NOM at 50 mg/L, the apparent reaction rate constants of PCE increased, but TCE production decreased. The enhanced removal rate of PCE was not due to the dechlorination, and the sorption was dominant iron with SDDBS and NOM. The apparent reaction rate constants of chromate by iron with Triton X-100 and NOM were 1.4-3.1 times lower than without surfactants while that with HDTMA was two times higher than without HDTMA, When the sorbed HDTMA molecules form admicelles, negatively-charged chromate has an affinity for the positively-charged HDTMA head group.

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