• Title/Summary/Keyword: Solvent fraction

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Purification and Characterization of NAD-Dependent n-Butanol Dehydrogenase from Solvent-Tolerant n-Butanol-Degrading Enterobacter sp. VKGH12

  • Veeranagouda, Y.;Benndorf, Dirk;Heipieper, Hermann J.;Karegoudar, T.B.
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.663-669
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    • 2008
  • The solvent-tolerant bacterium Enterobacter sp. VKGH12 is capable of utilizing n-butanol and contains an $NAD^+$-dependent n-butanol dehydrogenase (BDH). The BDH from n-butanol-grown Enterobacter sp. was purified from a cell-free extract (soluble fraction) to near homogeneity using a 3-step procedure. The BDH was purified 15.37-fold with a recovery of only 10.51, and the molecular mass estimated to be 38 kDa. The apparent Michaelis-Menten constant ($K_m$) for the BDH was found to be 4 mM with respect to n-butanol. The BDH also had a broad range of substrate specificity, including primary alcohols, secondary alcohols, and aromatic alcohols, and exhibited an optimal activity at pH 9.0 and $40^{\circ}C$. Among the metal ions studied, $Mg^{2+}$ and $Mn^{2+}$ had no effect, whereas $Cu^{2+},\;Zn^{2+}$, and $Fe^{2+}$ at 1 mM completely inhibited the BDH activity. The BDH activity was not inhibited by PMSF, suggesting that serine is not involved in the catalytic site. The known metal ion chelator EDTA had no effect on the BDH activity. Thus, in addition to its physiological significance, some features of the enzyme, such as its activity at an alkaline pH and broad range of substrate specificity, including primary and secondary alcohols, are attractive for application to the enzymatic conversion of alcohols.

Antioxidant Capacity of Crude Extract and Its Solvent Fractions of Arctic Terrestrial Plant Ranunculus heperporeus (북극식물 Ranunculus hyperboreus의 추출물과 용매분획물의 항산화 활성)

  • Lee, Jung Im;Kim, Hojun;Seo, Hyo-Won;Kong, Chang-Suk;Seo, Youngwan
    • Ocean and Polar Research
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    • v.38 no.3
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    • pp.185-193
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    • 2016
  • In this study, antioxidative potentials of the crude extract and its four solvent fractions from the Arctic terrestrial plant Ranunculus heperporeus were evaluated by using four different activity tests, including the inhibition of intracellular reactive oxygen species (ROS) and lipid peroxidation in Raw 264.7 cells as well as determining the extent of both the scavenging of peroxynitrite ($ONOO^-$) and the oxidative damage of genomic DNA purified from Raw 264.7 cells. Based on a comparative analysis, n-BuOH, and 85% aq.MeOH solvent fractions showed good scavenging effects on the production of intracellcular ROS and inhibited membrane lipid peroxidation and DNA oxidation. In addition, n-BuOH and 85% aq.MeOH fractions exhibited good scavenging effects on both authentic peroxynitrite and one generated from SIN-1. Among the samples tested, the n-BuOH fraction revealed the strongest antioxidant effect.

Detergency and soil Redeposition in a Drycleaning System -The Effect of Surfactant Type and Their Mixture- (드라이클리닝 시스템에서의 세척성과 재오염성 -계면활성제의 종류와 혼합이 미치는 영향-)

  • 김주연;박정희
    • Journal of the Korean Society of Clothing and Textiles
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    • v.23 no.7
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    • pp.1030-1039
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    • 1999
  • The effect of surfactant mixture 9on detergency and soil redeposition in a dry-cleaning system was investigated employing Aerosol OT as an anionic surfactant and Span 80 as a nonionic surfactant. The effect of charge system on soil deposition was also investigated in order to determine the optimum condition at which soil redeposition is minimum,. Soil deposition instead of soil redeposition on cotton, polyester and wool fabrics was measured employing petroleum solvent and perchloroethylene as organic solvents. The results were as follows. 1. Surface tension or interfacial tension was not changed by the addition of any surfactant or surfactant mixtures. In petroleum solvent however interfacial tension between solrent and water decreased when surfactants were added and increased when surfactants were mixed,. 2. The maximum amount of water solubilization increased as the mole fraction of Aerosol OT increased and more water was solubilized in petroleum solvent than in perchloroethylene. 3. The detergency of cotton was greater and the soil deposition rate was lower in Span 80 solution than in Aerosol OT solution. The soil deposition on cotton fabric decreased when water was solubilized in Aersol OT solution 4. The detergency and soil deposition rate of polyester fabric did not change by the surfactant type of the addition of surfactant mixture and soil deposition rate increased bywater solubilization. 5. Soil deposition on wool fabric was very high when Arosol OT was employed in perchloroethylene and the soil deposition did not change greatly by water solubilization.

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Antioxidative and Cytotoxic Effects of Solvent Fractions from Elaeagnus multiflora (뜰보리수 열매의 용매분획별 항산화 및 암세포 증식 억제 효과)

  • Kim, Sung-Ae;Oh, Se-In;Lee, Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.134-142
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    • 2007
  • This study was performed to determine the antioxidative and cytotoxic effects of Elaeagnus multiflora by examining its scavenging effects on the 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical, the inhibition of lipid peroxidation, and its inhibitory effects on cancer cell proliferation in HeLa cells, MCF-7 cells, and SNU-638 cells by MTT assay. Here, dried samples were extracted in ehtanol at room temperature and fractionated into five different solvent types: hexane, dichloromethane, ethylacetate, butanol, and aqueous partition layers. The hexane(62.92${\pm}$2.45%) and dichloromethane(65.25${\pm}$4.74%) fractions of Elaeagnus multiflora's flesh, and the aqueous(94.65${\pm}$0.02%) and ethylacetate(93.83${\pm}$0.02%) fractions of Elaeagnus multiflora's seeds, inhibited DPPH radical production. The DPPH radical scavenging effects of the flesh and seed were different according to solvent fractions. The inhibition of lipid peroxidation by the flesh and seed extracts were 76.11${\pm}$3.66 and 69.57${\pm}$2.27, respectively, for hexane and 67.57${\pm}$2.43 and 62.09${\pm}$0.90, respectively, for the dichloromethane fraction. Among the various partition layers of Elaeagnus multiflora's flesh, hexane and dichloromethane showed the strong cytotoxicities on all the cancer cell lines used in the study. Also all the fractions of Elaeagnus multiflora's seed exhibited significant effects on the inhibition of cancer cell growth(hexane > dichloromethane > ethylacetate > butanol > aqueous partition layers). These results indicate that the haxane and dichloromethane partition layers of Elaeagnus multiflora's flesh and seed extracts have possible antioxidative and anticancer capacities. Although further studies are needed, the present work suggests that Elaeagnus multiflora may be an antioxidative and chemopreventive agent.

Antimicrobial Activities of Green Tea Extract and Fractions on the E. coli O157:H7 (Escherichia coli O157:H7균에 대한 녹차 추출물 및 분획물의 항균효과)

  • Cho Sun-Young;Choi Jae-Ho;Ham Seung-Shi;Oh Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.20 no.1
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    • pp.48-52
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    • 2005
  • This study was investigated to determine the antimicrobial activity of ethanol extract of green tea, and of solvent fractionated ethanol extract on Escherichia coli O157:H7. MIC value of ethanol extracts and solvent fractionated ethanol extract (hexane, ethyl-acetate, chloroform and water) were $2000{\mu}g/ml,\;0,\;500{\mu}g/ml,\;2000{\mu}g/ml,\;1000{\mu}g/ml$, respectively. The antimicrobial activity of ethyl acetate fraction against E. coli O157 :H7 ranged from $250-2000{\mu}g/disk$. The antimicrobial activity was significantly increased as concentration dependent. When E. coli O157:H7 was pH-adjusted TSB containing $1000{\mu}g/ml$ ethyl acetate, E. coli O157:H7 was significantly inhibited at initial pH of 10, whereas the pathogen grew well in the presence of pH 4.5-pH 9. The inhibitory effect of the ethyl acetate fraction on the growth of E. coli O157 :H7 was investigated. Growth of both strain in the tested conditions were rapidly occurred up to 12 h, but no growth was occurred in the presence of $250-1000{\mu}g/mL$ of ethyl acetate fraction for 72 h.

Biological Activities of Ethanol Extracts and Fractions of Black Olympia Grape(Vitis Labruscana L.) (거봉 포도종의 에탄올 추출물 및 분획물에 대한 생리활성 효능)

  • 박성진;박부길;이현용;오덕환
    • Food Science and Preservation
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    • v.9 no.3
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    • pp.338-344
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    • 2002
  • This study was conducted to determine biological activities, such as lipid peroxidation inhibition and cytotoxic effect of ethanol extracts of Black Olympia grape seeds and skins, and of organic solvent fractionated ethanol extracts obtained from grape seeds and skins at different temperatures. Among different extraction temperatures, the ethanol extract of grape seed obtained at 30$\^{C}$ had the strongest lipid oxidation inhibition of 60.1%, while the strongest lipid oxidation inhibitory effect of 71.2% was observed in the presence of 20 $\mu\textrm{g}$/㎖ ethylacetate fraction obtained from ethanol extract of grape seeds at 30$\^{C}$. The ethanol extract of grape seeds showed more strong lipid oxidation inhibition than that of skin extracts. Similar results were observed in cytotoxic effects. The ethanol extract of grape seeds at 30$\^{C}$ exhibited more strong cytotoxicity than that of skin extracts on MCF-7, Hep3B, and A549 cell lines. Among organic solvent fractions extracted from the ethanol extracts of gape seeds and skins, the hexane fraction showed the strongest cytotoxic inhibition of 75.15% and 62.50% on MCF-7 and Hep3B cell in the presence of 1.0 $\mu\textrm{g}$/㎖ respectively. On the other hand, the water fraction showed the strongest cytotoxic inhibition of 65.41% on A549 cell in the presence of 1.0 $\mu\textrm{g}$/㎖. Overall, the ethanol extracts and their fractions of Black Olympia grape seeds showed strong lipid oxidation inhibition and cytotoxicity than those of grape skins.

Immunological Activity of Solvent Fractions from $Epimedium$ $koreanum$ Nakai (삼지구엽초 용매별 분획 추출물의 면역관련 활성)

  • Park, Myoung-Su;Kim, Seo-Jin;Wang, Jun;Kim, Gwang-Hee;Oh, Deog-Hwan
    • Food Science and Preservation
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    • v.19 no.1
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    • pp.110-115
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    • 2012
  • $Epimedium$ $koreanum$ Nakai is a wild medicinal plant commonly consumed in South Korea due to its beneficial health effects. In this study, the antimutagenic and immunological activities of $E.$ $koreanum$ Nakai extracts were investigated for their use in food. In the immunomodulating activity, the effects of $E.$ $koreanum$ Nakai on the B cell (Rhamos) and T cell (Molt-4) were investigated. The results showed that the growth and viability of the B and T cells were increased and activated more in the ethylacetate (1.35 and 1.48 times) and water fraction (1.30 and 1.40 times), respectively. In the Ames test, none of the fractions produced a mutagenic effect on $Salmonella$. $typhimurium$ TA98 and TA 100. The ethylacetate fraction showed a strong antimutagenic effect (98%) on and a high butanol fraction (84%) of B(${\alpha}$)P in $S.$ $typhimurium$ TA98 and TA100, respectively. In 4-nitroquinoline-1-oxide (4NQO), all the solvent fractions showed an over 70% antimutagenic effect, except for the chloroform extract. Especially, ethylacetate and butanol showed strong inhibition of the mutagenic effects (80 and 90%) on 4NQO in $S.$ $typhimurium$ TA98 and TA100, respectively. These results provide preliminary data for the development of $E.$ $koreanum$ Nakai as an edible food material.

Physiological activities of water extract and solvent fractions of Grifola frondosa (잎새버섯 물추출물 및 유기용매 분획물의 생리활성)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.192-198
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    • 2015
  • This study was performed in order to analyze the fibrinolytic, thrombin inhibitory, anti-oxidative, acetylcholinesterase inhibitory, and immuno-enhancing activities of the water extract and solvent fractions isolated from Grifola frondosa. Fibrinolytic activity was analyzed using the fibrin plate method, and thrombin inhibitory activity was assayed using the substrate H-D-Phe-piparg- pna. Anti-oxidative activity was estimated using the DPPH assay, and AChE inhibitory activity was measured using the spectrophotometric method. Immuno-enhancing activity was examined using the nitric oxide (NO) production in RAW 264.7 macrophage cells. Cell viability was determined using the MTS assay. Fibrinolytic activities were the highest in water extract (1.55 plasmin units/mL) followed by water fraction (0.85 plasmin units/mL). The thrombin inhibitory activities of the water and ethyl acetate fractions were determined to be 76.43% and 72.59%, respectively. The acetylcholinesterase inhibitory activities of chloroform and hexane fractions exhibited values of 95.14% and 94.74%, respectively. The butanol fraction showed the highest anti-oxidative activity at 94.47%. Anti-proliferating activity against Raw 264.7 cells showed no cytotoxicity. The production of NO in Raw 264.7 cells increased up to 2-fold by adding the water fraction compared to the untreated control. These results suggest that Grifola frondosa may serve as a useful functional food for the enhancement of immune function and the prevention and therapy of cardiovascular diseases.

Effects of Solvent-extracted Fractions from Salicornia herbacea on Anti-oxidative Activity and Lipopolysaccharide-induced NO Production in Murine Macrophage RAW264.7 Cells

  • Lee, Whi-Min;Sung, Hye-Jin;Song, Jae-Chan;Cho, Jae-Youl;Park, Hwa-Jin;Kim, Suk;Rhee, Man-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.161-168
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    • 2007
  • Salicornia herbacea L. (Chenopodiaceae: S. herbacea) is a salt marsh plant, which has long been prescribed in traditional medicines for the treatment of intestinal ailments, nephropathy, and hepatitis in Oriental countries. In order to elucidate the mechanisms of this herb, we conducted an anti-oxidative activity, the inhibition of nitric oxide (NO) production, and the suppression of the pro-inflammatory cytokine genes, with the solvent-extracts of S. herbacea. We found that both ethyl acetate and n-butanol tractions showed potent anti-oxidative effects in comparison to other fractions using xanthine oxidase assay with $IC_{50}$ values of $66.0{\pm}0.5\;{\mu}g/ml$ and $82.5{\pm}3.8\;{\mu}g/ml$, respectively. In addition, both ethyl acetate and n-butanol fractions showed more electron donating activity (EDA) than other tractions, according to DPPH (2, 2-Diphenyl-lpicrylhydrazyl radical) assay. The EDA of ethyl acetate fraction ($IC_{50}$ values of $117.5{\pm}3.8\;{\mu}g/ml$) is more significant than that of n-butanol fraction ($IC_{50}$ values of $375.0{\pm}12.5\;{\mu}g/ml$). Among potential anti-oxidative tractions, ethyl acetate traction dose-dependently suppressed lipopolysaccharide (LPS, $0.1\;{\mu}g/ml$)-induced nitric oxide (NO) production in RAW264.7 cell, while n-butanol did not. As expected, ethyl acetate fraction suppressed the expression of inducible NO synthase (iNOS) in RAW264.7 cell stimulated by $0.1\;{\mu}g/ml$ of LPS. Moreover, the ethyl acetate traction suppressed the expression of interleukin-1 $(IL)-1{\beta}$ and granulocyte/macrophage colony-stimulating factor (GM-CSF) mRNA in LPS-stimulated RAW264.7 cells. Therefore, these results suggest that S. herbacea may have anti-oxidative and anti-inflammatory activities by modulating radical-induced toxicity and various pro-inflammatory responses.

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Anti-inflammatory and anti-cancer activities of sterol rich fraction of cultured marine microalga Nannochloropsis oculata

  • Sanjeewa, Kalu Kapuge Asanka;Fernando, Ilekuttige Priyan Shanura;Samarakoon, Kalpa W.;Lakmal, Hetti Handi Chaminda;Kim, Eun-A;Kwon, O-Nam;Dilshara, Matharage Gayani;Lee, Joon-Baek;Jeon, You-Jin
    • ALGAE
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    • v.31 no.3
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    • pp.277-287
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    • 2016
  • Five fractions separated from Nannochloropsis oculata using solvent-solvent partition chromatography of 80% methanolic extract of N. oculata (NOM) followed by the open silica column chromatography of its hexane fraction (NOMH) for the anti-inflammatory on RAW 264.7 cells and anti-cancer activities on HL-60, A-549, HEP-3B, HCT-116, and SW-480 cancer cells. All the five fractions showed potential anti-inflammatory activities against lipopolysaccharide-stimulated RAW 264.7 macrophages cells with IC50 values less than 6.25 μg mL−1. Moreover, 90% n-hexane column elution of NOMH (NOMH90) down-regulated lipopolysaccharide-stimulated protein levels of inducible nitric oxide synthase and cyclooxygenase-2. Furthermore, NOMH90 showed marked cytotoxic effect on the HL-60 cells with IC50 value of 23.58 ± 0.09 μg mL−1. In addition, Hoechst 33342 cell permeable dye used to visualize the apoptosis nucleus and cell cycle analysis measured Sub-G1 DNA contents to confirm reduction of the cell viability in NOMH90 treated cells due to induction of apoptosis in HL60. These results are quite related to the phytosterol contents of the NOMH fractions and the results suggest N. oculata extracts might be useful as potential sources of natural anti-inflammatory and anti-cancer compounds. In conclusion, the sterol content in N. oculata might provide a promising role in future medicines in anti-inflammatory and anti-cancer.