• Title/Summary/Keyword: Triton X -100

Search Result 320, Processing Time 0.026 seconds

Effects of protein concentration and detergent on endotoxin reduction by ultrafiltration

  • Jang, Hyun;Kim, Hyo-Seung;Moon, Seung-Cheol;Lee, Young-Rae;Yu, Kang-Yeoul;Lee, Byeong-Kil;Youn, Hyun-Zo;Jeong, Young-Ju;Kim, Byeong-Soo;Lee, Sung-Ho;Kim, Jong-Suk
    • BMB Reports
    • /
    • v.42 no.7
    • /
    • pp.462-466
    • /
    • 2009
  • Lipopolysaccharide (LPS), found in the outer membrane of Gram negative bacteria, only exerts its toxic effects when in free form. LPS has three major parts, lipid A, the toxic component, along with a core polysaccharide and O-specific polysaccharide. LPS monomers are known to have molecular masses between 10 to 30 kDa. Under physiological conditions, LPS exists in equilibrium between monomer and vesicle forms. LPS removal by 100 kDa ultrafiltration was more efficient (99.6% of LPS removed) with a low concentration of protein (2.0 mg/ml) compared to a high concentration (20.1 mg/ml). In the presence of different detergents (0.5% Tween 20, 1.0% taurodeoxycholate and 1.0% Triton X-100), LPS removal was more efficient at low protein concentrations (2.0 mg/ml) compared to high protein concentrations (20.1 mg/ml).

Purification and Characterization of a Novel Antifungal Protein from Paenibacillus macerans PM1 Antagonistic to Rice Blast Fungus, Pyricularia oryzae

  • Bae, Dong-Won;Kawk, Weon-Sik;Lee, Joon-Taek;Son, Dae-Young;Chun, Sung-Sik;Kim, Hee-Kyu
    • Journal of Microbiology and Biotechnology
    • /
    • v.10 no.6
    • /
    • pp.805-810
    • /
    • 2000
  • An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

  • PDF

Magnetic Orientations of Bull Sperm Separated into Head and Flagellum Treated by DTT or Heparin

  • Suga, D.;Shinjo, A.;Kurnianto, E.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.13 no.2
    • /
    • pp.167-175
    • /
    • 2000
  • This paper describes the magnetic orientation of bull sperm separated into the head and the flagellum treated by DTT or heparin in a 5,400G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4^{\circ}C$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the head area at 1, 3 and 6 days. After measuring the area, each sample was exposed to a 5,400G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the sperms were separated into the head and the flagellum through the DTT treatment. Almost of the separated heads showed that their long axis oriented perpendicularly to the magnetic lines of force, and most of the long axis perpendicularly oriented heads showed that their flat plane oriented perpendicularly in a 5,400G magnetic field. Also, the demembranation of the head tended to increase those perpendicular orientations, while those perpendicular orientations of the head declined with the decondensation of the sperm nuclei. These findings suggest that strong magnetic anisotropy for the perpendicular orientation of the long axis and the flat plane of the head occurs in the sperm nuclei in a 5,400G magnetic field. The separated flagellum showed lower parallel orientation, and the separated and demembranated flagellum showed parallel orientation to the magnetic lines of force in this magnetic field. These findings suggest that weak magnetic anisotropy of the parallel orientation of the flagellum occurs in the inside components in a 5,400G field.

Isolation of an Acinetobacter junii SY-01 Strain Producing an Extracellular Lipase Enantioselectively Hydrolyzing Itraconazole Precursor, and Some Properties of the Lipase

  • Yoon, Moon-Young;Shin, Pyong-Kyun;Han, Ye-Sun;Lee, So-Ha;Park, Jung-Keug;Cheong, Chan-Seong
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.1
    • /
    • pp.97-104
    • /
    • 2004
  • Water-sludge bacteria were screened to find a lipase enantioselectively hydrolyzing itraconazole precursor, which is well known as the starting material of antifungal drug agents. A bacterial strain was isolated and identified as Acinetobacter junii SY-01. After the strain was cultivated, the enzyme was purified 39.4-fold using ultrafiltration and gel filtration through a Sephadex G-100 chromatographic column and the activity yield was 34.9%. The molecular weight of the enzyme was about 40 kDa, as measured by SDS-PAGE, and the optimum pH was 7.0- 9.0 and stable at pH 6.0- 9.0. The optimum temperature was 45- $5^{\circ}C$, and 73% of the enzymes activity remained after incubation at 70% for 1 h. Enzyme activity was enhanced by gall powder, sodium deoxycholate, a cationic detergent Tween 80, and a non-ionic detergent Triton X-100, but was markedly inhibited by metal ions such as $Hg^{2+},Cu^{2+},Ni^{2+}/,Ca^{2+}$, and an anionic-surfactant sodium dodecylsulfate. The $K_{m}$ values for (R)- and (S)-enantiomers of the itraconazole precursor were 0.385 and 21.83 mM, respectively, and the $V_{max} values ($\mu$Mㆍmin^{-1}.)$ were 6.73 and 6.49, respectively. The acetyl group among the different acyl moieties of itraconazole precursor showed the highest enantioselectivity for the hydrolysis by the Acinetobacter junii SY-01 lipase, and the lipase from Acinetobacter junii SY-01 displayed better enantioselectivity than that of commercially available lipases and esterases.

Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
    • /
    • v.41 no.1
    • /
    • pp.51-57
    • /
    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

  • PDF

The Search for Naturally Occurring Herbicidal Compounds - I. Screening Search for Herbicidal Substances in Higher Plants (천연에서부터 제초활성물질의 탐색 - 제1보 식물체에 함유된 제초활성물질의 검색)

  • Ahn, J.W.;Kim, J.S.;Cho, K.Y.
    • Korean Journal of Weed Science
    • /
    • v.9 no.1
    • /
    • pp.69-75
    • /
    • 1989
  • To search germination inhibitors in higher plants, first of all, some experiments containing selection of test weed seeds and effects of solvents and surfactants for bioassay establishment were conducted. Then MeOH-extracts of 45 plants were assayed for germination inhibition activities against Echinochloa crus-galli (L.) Beauv, Digitaria sanguinalis (L.) Scop., Cyperus iria L., Portulaca oleracea L. and Oenothera lamarckiana Ser. seeds. Among them extracts from Rhathiolepis ovata Briat and Picea abies (L.) Karst showed strong inhibitory effect (60-90% inhibition) on the germination of tested weed seeds at 5000 ppm. On the other hand, the extract from Youngia sonchifolia Max stimulated the germination and growth of tested weeds.

  • PDF

Effect of UV-Absorber Treatment on the Mechanical Properties of Cotton Fabrics (자외선 차단 가공에 의한 면직물의 역학적 특성 변화)

  • Kwon, Young-Ah;Kang, Mi-Jung;Cho, Hyun-Hok
    • Journal of the Korean Society of Clothing and Textiles
    • /
    • v.26 no.12
    • /
    • pp.1701-1708
    • /
    • 2002
  • 본 연구의 목적은 자외선 차단 가공 처리 및 자외선 조사처리에 의해 면직물의 역학적 특성 변화를 조사하고 가공제 처리조건에 따른 차이점을 분석하는 것이다. 시료로는 100% 면직물을, 자외선 흡수제로 2,2'-dihydroxy-4,4'-dimethoxy benzophenone을, 첨가제로 Triton X-100, polyethylene glycol 400, MgCl$_2$.6$H_2O$를 사용하였다. 자외선 흡수제 처리는 Atlas Launder-O-meter로 75$^{\circ}C$에서 60분간 흡진법으로 하였다. 미처리 시료 및 처리시료는 모두 xenon 램프에 80시간 동안 노출되었으며, 자외선 조사 전후 시료의 역학적 특성은 KES-F시스템을 사용하여 측정하였다. 본 연구의 결과는 다음과 같이 요약할 수 있다. 자외선 흡수제 처리는 처리 농도에 상관없이 면직물의 선형인장성(LT), 인장레질리언스(RT), 굽힘강성(B), 굽힘이력(2HB), 전단강성(G), 전단이력(2HG5), 표면마찰계수(MIU)등을 증가시켰으며 표면거칠기(SMD)는 감소시켰다. 압축특성은 처리농도의 영향을 받아서 고농도의 자외선 흡수제 처리는 압축특성을 감소시켰으며, 저농도의 처리는 압축특성을 증가시켰다. 자외 선 흡수제 처리는 처리농도에 상관없이 면직물의 fullness/softness를 유의하게 증가시키는 반면 stiffness, crispness및 anti-drape stiffness를 감소시켜서 가공포의 종합태(THV)는 가공 전에 비하여 저하하였다. 자외선 조사는 가공 전 면직물의 경우 B, 2HB, G, 2HG, 2HG5, LC를 감소시켰다. 자외선 조사는 처리농도와 상관없이 가공포의 WT, 굽힘특성 , 전단특성을 감소시켰으며, 저농도의 흡수제 처리포의 경우 SMD를 증가시켰으며, 고농도의 흡수제 처리포의 경우 SMD를 감소시켰다. 자외선 조사는 처리농도와 상관없이 가공포의fullness/softness, stiffness, anti-drape stiffness를 유의하게 감소시켜서 자외선 조사 전 보다 THV가 37% 저하하였고 미 가공포의 THV를 저하율보다는 저하가 낮았다. 자외선 흡수제 처리에 의해 면직물의 태는 가공 전보다 감소하지만, 자외선 흡수제 처리는 자외선 조사에 의한 태 감소율을 낮추는 데 유의 한 효과가 있다.

Differentiation and Distrbution of the Choline Acetyltransferase-immunoreactive Nerve Cells in the Magnocellular Preoptic Nucleus of the Rat Forebrains during the Postnatal Development

  • Chung, Young-Wha;Choi, Yoon-Jin
    • Animal cells and systems
    • /
    • v.1 no.3
    • /
    • pp.483-489
    • /
    • 1997
  • This study was performed to investigate the differentiation and distribution of choline acetyltransferase (ChAT}-immunoreactive cells in the magnacellular preoptic nucleus (MCPO) of the postnatal and adult rat forebrains, utilizing techniques of immunocytochemistry. According to the cell shape and the ratio of long axis versus short axis of cell soma, the ChATimmunoreactive nerve cells in the MCPO were classified into six types: 1) round, 2) oval, 3) elongated, 4) fusiform, 5) triangular, and 6) polygonal types. Frequency distributions of the oval and round nerve cells on the postnatal day (PND) 0 were observed to be high. But in the adult, frequency distributions of the same cells were shown to decrease. Compared to those of the postnatal rats, frequency distributions of elongated, fusiform, triangular, and polygonal nerve cells in the adult were increased. The total mean volumes of ChAT-immunoreactive cell somata in the MCPO of PND 0 rat were the lowest, while those in the PND 17 rat were shown to be the highest and decreased in the adult. The soma volumes of the immunoreactive cells at the PND 17 were evenly distributed, but those in the other developmental stages (e.g. PND 7 and adult) appeared to exhibit unimodal distributions. On the electron micrography, the free ribosomes, polysomes, and rough endoplasmic reticula (RER) of the nerve cells in the MCPO of PND 21 rat forebrains were immunoreactive to ChAT in the tissues untreated with triton X-100. According to the observations in the present study, it is considered that the ChAT-immunoreactive nerve cells in the MCPO of the rat forebrains are differentiated throughout the following processes during the postnatal development: 1) increase in cell soma volumes, 2) development of neurites, 3) increase in the frequency of differentiated cell types, and 4) decrease in cell soma volumes. The ribosomes, polysomes, and RER are considered to be closely related to the intracellular localization and biosynthesis of the ChAT but not Golgi complex.

  • PDF

Photocatalytic activity of rutile TiO2 powders coupled with anatase TiO2 nanoparticles using surfactant (계면활성제를 이용하여 anatase TiO2 나노 입자와 결합된 rutile TiO2 분말의 광촉매 특성)

  • Byun, Jong Min;Park, Chun Woong;Kim, Young In;Kim, Young Do
    • Journal of Powder Materials
    • /
    • v.25 no.3
    • /
    • pp.257-262
    • /
    • 2018
  • The coupling of two semiconducting materials is an efficient method to improve photocatalytic activity via the suppression of recombination of electron-hole pairs. In particular, the coupling between two different phases of $TiO_2$, i.e., anatase and rutile, is particularly attractive for photocatalytic activity improvement of rutile $TiO_2$ because these coupled $TiO_2$ powders can retain the benefits of $TiO_2$, one of the best photocatalysts. In this study, anatase $TiO_2$ nanoparticles are synthesized and coupled on the surface of rutile $TiO_2$ powders using a microemulsion method and heat treatment. Triton X-100, as a surfactant, is used to suppress the aggregation of anatase $TiO_2$ nanoparticles and disperse anatase $TiO_2$ nanoparticles uniformly on the surface of rutile $TiO_2$ powders. Rutile $TiO_2$ powders coupled with anatase $TiO_2$ nanoparticles are successfully prepared. Additionally, we compare the photocatalytic activity of these rutile-anatase coupled $TiO_2$ powders under ultraviolet (UV) light and demonstrate that the reason for the improvement of photocatalytic activity is microstructural.

Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus (통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성)

  • KIM, In-Won;KANG, Ok-Hwa;KONG, Ryong;KWON, Dong-Yeul
    • The Korea Journal of Herbology
    • /
    • v.30 no.5
    • /
    • pp.15-21
    • /
    • 2015
  • Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.