• Title/Summary/Keyword: Amberlite XAD

Search Result 81, Processing Time 0.024 seconds

Purification of Heat-Stable Enterotoxin of Enterotoxigenic Escherichia coli eKT-53 (장독성 대장균 eKT-53 균주의 내열성 장독소 정제)

  • Do, Dea-Hong;Kim, Kyo-Chang;Kim, Do-Young
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.21 no.1
    • /
    • pp.76-83
    • /
    • 1992
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin (ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. ST producing E. coli KM-7 strain was isolated from the swine and molecular cloning of ST gene of KM-7 strain. Transformant eKT-53 $(ST^+,\;LT^-)$ was selected by infant mouse assay (IMA). The culture supernatant of eKT-53 strain was performed purification by multipled steps. The culture supernatant (crude ST) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Bio-Gel P-6 and preparative polyacrylamide slab gel electrophoresis. About 113-fold purification was achieved with a yield of about 11% of crude ST and the minimum effective dose(MED) of this purified ST was about 2.8ng in IMA. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis.

  • PDF

Effect of Insulin-like Action and Insulin Sensitizing on 3T3-Ll Adipocytes from Coicis Semen (의이인이 3T3-Ll Adipocytes에서 인슐린성 작용과 인슐린 민감성에 미치는 영향)

  • 김종욱;최용휴;주영승;박선민;이미영;김호경;김홍준;고병섭
    • The Journal of Korean Medicine
    • /
    • v.23 no.1
    • /
    • pp.83-91
    • /
    • 2002
  • Objectives : In this study, water extracts from Coix lachryma-jobi Linne var. mayuen Stapf. were investigated for their effects on insulin-like action and glucose uptake in 3T3-Ll cells. Methods : We examined the effects of insulin-like action on the differentiation of 3T3-Ll fibroblasts. The Coicis Semen was treated with hot water and the extract was freeze-dried. The hot water extract was chromatographed on nonionic polymer resin (Amberlite XAD-4, Sigma) with distilled water (Fr. 1), 20% (Fr. 2), 40% (Fr. 3), 60% (Fr. 4), 80% (Fr. 5), and 100% EtOH (Fr. 6), successively. Results : Total extract of Coicis Semen was fractionated into 0 to 100% MeOH with Amberlite XDA-4 column. Treatment of cells with $10{\;}\mu\textrm{g}/ml$ of total extracts of Coicis Semen significantly increased the differentiation (p<0.05). At $1{\;}\mu\textrm{g}/ml$ level of insulin, the differentiation was accelerated (p<0.01). The effect of extracts plus insulin on the differentiation was greater than that of insulin alone. In 3T3-Ll adipocytes, glucose uptake was higher by 2.7 times with 5 uM of total extract in low dosage of insulin (3 ng/ml) than without total extract. 5 and 50 uM of water and 40% MeOH fractions increased glucose uptake by 3.5 times in 3T3-Ll adipocytes (p<0.00l). Conclusions : Coicis Semen contains compounds which playa role of insulin-like action and insulin sensitizer.

  • PDF

Purification of Total Ginsesides with Macroporous Resins and Their Biological Activities

  • Li, Huayue;Jin, Haizhu;Lee, Dong-Geun;Lee, Jae-Hwa;Lee, Sang-Hyeon;Ha, Bae-Jin;Ha, Jong-Myung
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.20 no.5
    • /
    • pp.1321-1326
    • /
    • 2006
  • Total ginsenosides were purified and their antioxidant, antibacterial and anticancer activities were measured. The crude extracts of ginseng, which were extracted with 75% ethanol by ultrasonification method, were firstly purified on AB-8 macroporous adsorption column to remove water soluble impurities, and decolored on Amberlite IRA 900 Cl anion-exchange column. Then, they were purified on Amberlite XAD16 adsorption column to delete the non-polar impurities. Total ginsenosides contents of the purified extracts were 79.4%, 71.7% and 72.5% in cultured wild ginseng, red ginseng and white ginseng, which were significantly increased than those of crude extracts. All of the three extracts showed concentration-dependant scavenging activities against DPPH radicals, among which white ginseng showed the most powerful activity. Cultured wild ginseng roots showed strongest effect against both B. subtilis PM 125(Gram-positive) and E. coli D31 (Gram-negative) bacteria, while red ginseng and white ginseng only showed the activity against B. subtilis. According to the result of the MTT assay, ail of the three extracts inhibited the growth of U-937 human hohistiocytic lympma cell, which were significantly different (p < 0.05) when compared to the control.

Volatility of Herbicides Sprayed in Zoysia japonica Turf and Bare Soil (잔디밭과 나지에 산포된 주요 잔디밭용 제초제의 휘산)

  • 김석정;박진희;죽내안지;김길웅;신동현;허영조
    • Asian Journal of Turfgrass Science
    • /
    • v.10 no.3
    • /
    • pp.263-270
    • /
    • 1996
  • This experiment was conducted to investigate the loss of various herbicides by means of vola-tility from the turfgrass field and the hare ground with the different soil moisture contents and temperatures. Different herbicides were applied at the rates of 375 g a.i. /l0a of pendimethalin,250 g a.i. /l0a of napropamide, and 96.4 g a.i. /l0a of dicamba with 200 \ulcorner/10a of spray volume in the turfgrass(Zoysia japonica cut off 5cm) grown in pots(265.8 $cm^2$) and bared soil. The pots were placed in the growth chamber with 10,000 lux of light intensity(12h per day) at 25 and 35˚C for 7days. Amberlite XAD polymeric resin(20/50 mesh) was used as sampling media for herbicide airborne residues. Air flow was maintained at 10 \ulcorner /min by vacuum pump regulated with a factory calibrated flow meter. Herbicide airborne residues were extracted from the XAD resin with 300 ml of 1:1 acetone and hexane. The extracts were concentrated by rotary evaporation at 35˚C and dissolved in 1 ml MeCN for HPLC analysis. The airborne losses of the herbicide applied in the turfgrass and bare soil increased as the temperature and soil moisture contents were increased, regardless of the kinds of herbicide. Higher airborne residues was observed in the turfgrass pots than the bare soil pots. Pendimethalin and dicamba with higher vapor pressure gave rise to the increased loss of airborne herbicides, showing that 6.26 and 6.4% of average airborne loss in pendimethalin and dicamba, respectively, compared to 0.56% in napropamide. The amount of airborne losses in turfgrass was highest at one day after application and then a declined trend was observed as the time was prolonged. Key words. Herbicides, Turfgrass field, Bare ground, Volatility.

  • PDF

Purification and Characterizationn of Biosurfactant from Marine Pseudomonas sp. CHCS-2 (해양으로부터 분리한 Pseudomonas sp. CHCS-2가 생산하는 Biosurfactant의 정제 및 특성에 관한 연구)

  • 류병호;김학주
    • KSBB Journal
    • /
    • v.10 no.5
    • /
    • pp.582-588
    • /
    • 1995
  • A marine microorganism producing biosurfactant was isolated from the oil polluted coast of Chung-Mu in Korea, and was identified as Pseudomonas sp.. It produced the biosurfactanl and its optimum culture conditions for pH and salt concentration were 8.0 and 3.0%, respectively. The productivity of biosurfactant from this strain was affected by the nitrogen source used. For the oil resolvability of the biosurfactant, the residual oil in the culture broth with 2% Kuwait crude oil at each time of 48, 96, and 132hr was investigated by gas chromatography. As result of this experiment, it was verified that the biosurfactant acted on C10-C14, of Kuwait crude oil and so the oil was decomposed. The biosurfactant isolated from the supernatant was purified by adsorption to Amberliter XAD-7 and followed by gel chromatography (Sephadex G-100) and HPLC. The purified biosurfactant showed a high value of emulsifying activity at $40^{\circ}C$ and the emulsifying stability was maintained at the temperature range of $30^{\circ}C$$60^{\circ}C$. The purified biosurfactant reduced the interfacial tension of Kuwait crude oil remarkably and showed improved dispersing ability compared to those of commercial surfactants such as Tween 80, Tween 60 and SDS.

  • PDF

Anti-tumor Activity of the Extract of $Alpinia$ $officinarum$ using Hollow Fiber Assay (Hollow FiberAssay을 이용한 고량강 추출물의 항종양 효과)

  • Lee, Keyong-Ho;Rhee, Ki-Hyeong
    • The Korean Journal of Food And Nutrition
    • /
    • v.24 no.4
    • /
    • pp.496-500
    • /
    • 2011
  • The purpose of this investigation was to evaluate anti-tumor activity and detect what compounds affect its activity form $Alpinia$ $officinarum$ Hance. Two fractions, methanol and ethylacetate, were isolated by Amberlite XAD-2 resin column chromatography from methanol extract of the rhizomes of $Alpinia$ $officinarum$. In hollow fiber assay, the methanol extract and methanol fraction were found to inhibit the tumor growth against colon tumor cell lines such as Colo-320, HCT116 and WiDr. Three diarylheptanoids [5-hydroxy-1,7-diphenyl-3-heptanone, 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone and 3,5-dihydroxy-1,7-diphenylheptane] and two flavonoids [galangin and kaempheride] were isolated and identified from the methanol fraction, which is higher activity than ethylacetate fraction. Among these diarylheptanoids and flavonoids, 3,5-dihydroxy-1,7-diphenylheptane, galangin and kaempheride as active components on anti-tumor activity were mainly posited in methanol fraction.

Preconcentration and Determination of Fe(III) from Water and Food Samples by Newly Synthesized Chelating Reagent Impregnated Amberlite XAD-16 Resin

  • Tokahoglu, Serife;Ergun, Hasan;Cukurovah, Alaaddin
    • Bulletin of the Korean Chemical Society
    • /
    • v.31 no.7
    • /
    • pp.1976-1980
    • /
    • 2010
  • A simple and reliable method has been developed to selectively separate and concentrate trace amounts of Fe(III) ions from water and food samples by using flame atomic absorption spectrometry. A new reagent, 5-hydroxy-4-ethyl-5,6-di-pyridin-2-yl-4,5-dihydro-2H-[1,2,4] triazine-3-thione, was synthesized and characterized by using FT-IR spectroscopy and elemental analysis. Effects of pH, concentration and volume of elution solution, sample flow rate, sample volume and interfering ions on the recovery of Fe(III) were investigated. The optimum pH was found to be 5. Eluent for quantitative elution was 10 mL of 2 M HCl. The preconcentration factor of the method, detection limit (3s/b, ${\mu}gL^{-1}$) and relative standard deviation values were found to be 25, 4.59 and 1%, respectively. In order to verify the accuracy of the method, two certified reference materials (TMDA 54.4 lake water and SRM 1568a rice flour) were analyzed. The results obtained were in good agreement with the certified values. The method was successfully applied to the determination of Fe(III) ions in water and food samples.

The Flavonoids from Caragana Chamlagu Leaves (골담초엽의 플라보노이드)

  • 마충운;함인혜;황완균
    • YAKHAK HOEJI
    • /
    • v.43 no.2
    • /
    • pp.143-149
    • /
    • 1999
  • The phytochemical studies of the leaves of Caragana chamlagu were carried out as a sieries of the investigation of medicinal resources. The roots of Caragana chamlagu have been used as neuralgia, arthritis and migraine in the folk medicines of Korea. The methanolic extract of the leaves of Caragana chamlagu was suspended with water and then separated with chloroform. Compound I was isolated from precipitates of these water fraction by recrystalization. The aqueous fraction of MeOH extract was performed to column chromatography on Amberlite XAD-4 and Sephadex LH-20, and three compounds, compound II, compound III, and compound IV were isolated. The structures of the four compounds were elucidated by spectroscopic data of $^1H-NMR$, ^{13}C-NMR$, IR, and FAB-MS. Compound I-IV were tilianine ($acacetin-7-O-{\beta}-D-glycopyranoside$), rutin($quercetin-3-O-{\alpha}-L-rhamnopyranosy(1{\rightarrow}6)-{\beta}-D-glu-copyranoside$), $kaempferol-3-O-{\alpha}-L-rhamnopyranosyl(1{\rightarrow}6)-{\beta}-D-galactopyranoside$, and apigetrin, ($apigenin-7-O-{\beta}-D-glycopyranoside$), respectively.

  • PDF

Escherichia coli에서 발현된 재조합 인간 상피세포 증식인자의 정제 및 특성

  • 박세철;유광현
    • Microbiology and Biotechnology Letters
    • /
    • v.24 no.4
    • /
    • pp.478-484
    • /
    • 1996
  • Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL21 harboring a plasmid pYHB101. The maximum production was 68.7 mg/l when the E. coli strain was cultured at 25$\circ$C for 48 hours in the modified MBL medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD- 7 chromatography, ultrafiltration, and DEAE Sepharose fast flow ion exchange chromatography with an overall yield of 66.6%. The purified rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'- deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.

  • PDF

Isolation of Anti-Herpes Simplex Virus Type 1(HSV-1) Component from Thujae orientalis Semen (백자인(栢子仁)으로부터 항Herpes 바이러스 1형(HSV-1) 물질의 분리)

  • Kang, Eun-Jung;Kang, Bong-Joo;Park, Kap-Joo;Ko, Byoung-Seob;Kim, Ho-Kyoung
    • Korean Journal of Pharmacognosy
    • /
    • v.29 no.4
    • /
    • pp.277-282
    • /
    • 1998
  • In order to search for anti-Herpes simplex virus type 1(HSV-1) agents, we screened 80 specimens of Korean traditional medicine by SRB assay. The methanol extracts of Thujae orientalis Semen (Cupressaceae) showed strong anti-HSV activity among samples tested. From the butanol fraction of Thujae orientalis Semen anti-HSV-1 agent was isolated by chromatographic separation using Amberlite XAD-4 and Sephadex LH-20. The structure was elucidated by spectroscopic methods, and was identified as ${\beta}-sitosterol$ (compound I). Compound I exhibited anti-HSV-1 activity with $EC_{50}$ of 0.6 mg/ml and $CC_{50}$ of 5.99 mg/ml, respectively.

  • PDF