• Title/Summary/Keyword: Triton X -100

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

  • 박대원;박호일;신평균
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.145-150
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    • 2003
  • Lipase from Acinetobacter sp. SY-01 plays an important role enzyme that products chiral drug. We investigated optimum condition for mass production of Acinetobacter sp. SY-01 lipase. Addition of among the different oils to medium. olive oil was optimal for enzyme production. When 0.2% olive oil was added as a carbon source, the production of lipase was increased to a maximum. The optimum pH and temperature were pH 7 and $30^{\circ}C$. In the presence of $Fe^{2+}$ and $Ca^{2+}$, the lipase activity was dramatically enhanced by 280% and 160%, respectively. SY-01 lipase was stable in the most of the DMSO among organic solvents. The addition of triton-X 100 increased the SY-01 lipase by 100-fold. The optimum composition of medium for production of the enzyme was 0.8% yeast extract, 0.2% olive oil, 0.4% triton X-100+40% DMSO. 0.1% $NH_4Cl$, 0.4% $K_2HPO_4$ 3.9% $NaH_2PO_4$, 0.03% $CaCl_22H_2O$, 0.01% $FeSO_4$$7H_2O$(pH 7.0).

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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    • v.9 no.3
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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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    • v.28 no.2
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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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    • v.44 no.4
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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.

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

  • 박경숙;이별나
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.503-508
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    • 1997
  • The extraction and separation methods of protein-bound polysaccharides from the mycelium and culture broth of L. edodes were investigated. The use 2% solution of surface active agent, Triton X-100 was effective for extraction of the protein-bound polysaccharide from the mycelium. The extraction of the protein-bound polysaccharides from mycelium with hot water was achieved by 4 hours extraction at 10$0^{\circ}C$. For the separation and partial purification of the protein bound polysaccharides the column chromatography using DEAE-Cellulose, DEAE-Sephadex and Sephadex proved to be effective.

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Effect of Ginseng Saponin, Gypsophila Saponin, and Detergents on Volume Changes and Fragility of Red Blood Cells (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))

  • Lee, Shin-Woong;Lee, Jeung-Soo;Lee, Soo-Kun;Ok, Chang-Kun;Kim, Young-Hie
    • YAKHAK HOEJI
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    • v.33 no.1
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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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Stabilization of Alcohol Oxidase under Electrostimulation; Sugars, Hydrogels and Surfactants Effect (전기자극 하에서 알콜 산화효소의 안정화연구; 당, 히드로젤, 계면활성제 효과)

  • Kim, Beom-Su;Lee, Kang-Min;Biellmann, J.F.;Kim, Kyung-Suk
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.456-460
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    • 2006
  • We investigated the activity and stability of alcohol oxidase from Hansenula sp., Pichia pastoris, and Candida boidinii under the electric stimulation. The activity and stability of alcohol oxidase depended on electric output voltage, electric stimulation time. This inactivation of the enzyme under electric stimulation could be recovered by stabilizing additives such as sugars, surfactants and hydrogels. These alcohol oxidase was more stable in trehalose, Triton X-100, Brji solution and alcohol oxidase from Hansenula is more stable than that from P. pastoris, and C. boidinii. The stabilizing of enzymes against electric stimulation showed a great potential use of enzymes in biotechnology and medical engineering fields.

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

  • 박평규;채희정;김의용
    • KSBB Journal
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    • v.13 no.1
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    • pp.65-70
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    • 1998
  • The physiochemical properties of extracellular lipid produced by an oleaginous yeast, Rhodotorula glutinis K-501 were examined. From the analytical experiments, it was suggested that the extracellular lipid produced is glycolipidic compound. Critical micelle concentration and minimum surface tension of the extracellular lipid in aqueous solution were 89mg/L and 31dyne/cm, respectively. Surface tension was also constant throughout wide range of pH. The emulsifying abilities and dispersing power of the extracellular lipid were much greater than those of commercial surfactants such as Tween 80 and Triton X-100 by factors of 2-3 and 1.3, respectively.

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동전기-생물학적복원기술과 계면활성제를 이용한 phenanthrene 오염토양의 정화

  • 김상준;박지연;이유진;양지원
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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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
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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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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