• Title/Summary/Keyword: Sephadex LH-20 column chromatography

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Bacillus sp. YJ-63이 생산하는 항곰팡이성 항생물질의 단리

  • Sin, Yeong-Jun;Jeong, Myeong-Ju;Jeong, Yeong-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.509-511
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    • 2000
  • An antifungal antibiotic was produced from Bacillus sp. YJ-63, and antibiotic was purified to a homogeneity by butanol extraction, Silica-gel column chromatography, Sephadex LH-20 gel filtration column chromatography and HPLC. The purified antibiotic showed one spot on Thin Layer Chromatography, which was negative to ninhydrin and positive to iodine. This finding indicated that the antibiotic was a cyclopeptide structure. UV absorption spectrum of the antibiotic in methanol was maximal at 277 nm, which corresponded to the spectrum for all of the Iturin antibiotic. The antibiotic was stable up to $121^{cdot}C$ and in the range of pH $3.O{\sim}l2.0$ with a stable of ${\alpha}-amino$ acid and ${beta}-amino$ acid, and showed high activity for fungi and yeasts,

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Purification and Physical Proerties of Biosurfactant Produced from Rhodotorula muciloginosa (Rhodotorula muciloginosa G-1에서 생산되는 biosurfactant의 정제 및 물리적 성질)

  • 이철수;이병옥;강상모
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.229-235
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    • 1995
  • The surface tension-decreasing biosurfactant was purified from Rhodotorula muciloginosa G-1. The purification procedure was the solvent extraction of culture broth. To ensure complete extraction, the sample was extracted twice with equal volume of ethylacetate. The crude solution was washed with n-hexane to remove unconsumed soybean oil. The crude sample of biosurfactant was applied to Silica gel column chromatography equilibrated with chloroform, and eluted with chloroform : methanol gradient. Serveral solvent system was used to developed the thin layer chromatography (TLC). The purified biosurfactant sample gave one spot (Rf 0.78). It was estimated that biosurfactant was glycolipid about having M.W.1,500 with standard of polyethyleneglycol by Sephadex LH-20 column chromatography.

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Purification and Characterization of Angiotensin I-Converting Enzyme Inhibitor from Porphyra yezoensis (김으로부터 분리한 Angiotensin-I Converting Enzyme 저해제의 정제 및 특성)

  • 최수진;전우진;유광원;신동훈;홍범식;조홍연;양한철
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.719-725
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    • 2000
  • This study focused on the purification and characterization of ACE inhibitor from Porphyra yezoensis. The dried Porphyra yezoensis was ground and hydrolyzed with 2.5 N HCl, followed by neutralization and centrifugation. Then, the subsequential purification of ACE inhibitor was carried out by Amberlite XAD 8, DEAE-Toyopearl 650C, Sephadex LH-20 column chromatography and reverse phase HPLC with C18 column. The purified ACE inhibitor was peptide which consisted of glycine (24.5%), arginine (56.8%) and proline (18.8%). Also, it showed the competitive inhibition pattern to ACE. The apparent molecular mass of purified peptide was 580 dalton, and an IC50 value of ACE inhibitor was 10.6 $\mu\textrm{g}$.

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Stilbenoids of Korean Pine (Pinus koraiensis) Inner Bark

  • Kwon, Dong-Joo;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.5
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    • pp.474-479
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    • 2009
  • Pinus koraiensis inner bark was collected and extracted with 95% ethanol. The extracts were concentrated and then sequentially fractionated using n-hexane, $CH_2Cl_2$, EtOAc, and $H_2O$ to be freeze dried. A portion of EtOAc fraction (6.6 g) was chromatographed on a Sephadex LH-20 column using aqueous methanol to isolate (+)-catechin (1), (-)-epicatechin (2), and trans-pinostilbenoside (3). Resveratrol (4) and trans-pinostilbene (5) were isolated by column chromatography using EtOH-hexane mixture after purification with aqueous methanol. The structures of these stilbenosides and flavans were characterized by spectroscopic tools using NMR and MS.

Production of the Isocyanide Inhibitor of Melanin Biosynthesis by Trichoderma sp. MR-93 (Trichoderma sp. MR-93 균주가 생산하는 Isocyanide 계열의 Melanin 생성 저해물질)

  • Lee, Choong-Hwan;Chun, Hyo-Kon;Chung, Myung-Chul;Lee, Ho-Jae;Bae, Kyung-Sook;Kho, Yung-Hee
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.209-213
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    • 1995
  • During the screening of inhibitors of melanin biosynthesis from microbial secondary metabolites, a fungal strain MR-93 which was capable of producing high level of an inhibitor was selected from plant leaf. Based on taxonomic studies, the fungus could be classified as a strain of Trichoderma sp.. The active compound (MR-93D) was purified from the culture broth by Diaion HP-20 column chromatography, ethylacetate extraction, Sephadex LH-20 column chromatography and HPLC. The inhibitor was identified as 4-hydroxy-8-isocyano-l-oxaspiro[4-4]cyclonon-8-en-2- one by spectroscopic methods of UV, $^{1}$H-NMR, ESIMS and IR. MR-93D showed a strong tyrosinase inhibitory activity with 0.03 $\mu$g/m of IC$_{50}$ value. It also inhibited melanin biosynthesis with 35 mm inhibition zone at 30 $\mu$g/paper disc in Streptomyces bikiniensis, a bacterium used as an indicator organism in this work.

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Production, Purification and Antifungal Activity of Antibiotic Substances Produced by Pseudomonas aeruginosa Strain B5

  • Kim, Beom-Seok
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.12-18
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    • 1993
  • Pseudomonas aeruginosa strain B5 with antagonistic activity against Phytophthora capsici and Magnaporthe grisea, was isolated from pepper-growing soil. From the culture of P. aeruginosa strain B5 grown on King's medium B, antibiotic substances were purified using XAD-2 column chromatography. XAD-2 eluates inhibited not only the mycelial growth of P. capsid and M. grisea, but also the development of Phytophthora blight on pepper plants. The crude antibiotic substances were further purified by using silica gel column chromatography, Sephadex LH-20 column chromatography, thin layer chromatography on silica gel plates, and high performance liquid chromatography. Silica gel column chromatogrphy gave good separation of the four antibiotic substances. The pure antibiotics P1, P2, and P3 finally purified by preparative HPLC inhibited the mycelial growth of P. capsici, at concentrations from 7 to 10 $\mu g/ml$. Only P1 and P2 had antifungal activity against M. grisea at 8 $\mu g/ml$. P1 and P3 were highly inhibitory to the mycelial growth of Botryosphaeria dothidea and Botrytis cinerea at relatively low concentrations. However, the three antibiotics had no antifungal activity against Rhizoctonia solani. The chemical structures of these antibiotics are being identified.

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Identification of Streptomyces misakiensis Producing Cathepsin B Inhibitor and the Purification of Inhibitor (Cathepsin B 저해물질을 생산하는 Streptomyces misakinesis의 동정 및 저해물질의 분리)

  • 한길환;김상달
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.25-30
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    • 2001
  • A strain of Actinomycetes producing cathepis B inhibitor was isolated from soil and identified as Streptomyces misakiensis. The product of S. misakiensis inhibited effectively cathepsis B proteinases as well as trypsin and papain. The cathepsin B inhibitor were largely produced with incubation for 4 days. The S. misakiensis was the most growth with incubation for 5 days. The cathepsin B inhibitor was isolated from the extraction of both with ethanol, ethanol and chlorofrom, and following several column chromatography such as sephadex G-15, silica gel 60 and sephadex LH-20 chromatography. The moleculer weight of purfied inhibitor was 138 dalton.

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Anti-inflammatory Effect of Jatrorrhizine from Phellodendron amurense in Lipopolysaccharide-stimulated Raw264.7 Cells (Lipopolysaccharide로 유도된 Raw264.7 cell에서 Phellodendron amurense의 Jatrorrhizine에 의한 염증 억제효과)

  • Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.54 no.2
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    • pp.114-119
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    • 2011
  • n-Butanol extracts from Phellodendron amurense have about 50% inhibitory activity against hyaluronidase. The anti-inflammatory compound was isolated from P. amurense by Sephadex LH-20 and MCI-gel CHP-20 column chromatography with gradient elution. As a the result, its structure was identified as Jatrorrhizine by the interpretation of spectroscopic analyses including $^1H$- and $^{13}C$-NMR. In anti-inflammatory activity, the expression of nitric oxide (NO) was inhibited as above 60% at 100 ${\mu}g/mL$ concentration of extracts and then purified Jatrorrhizine from P. amurense. The inhibitory activities against the expression of inducible NO syntase (iNOS) and cyclooxygenase-2 (COX-2) were 45% and 29%. It seems that the extracts and purified Jatrorrhizine from P. amurense were expected anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated Raw264.7 cells.

Antitumor and Antiinflammatory Constituents from Celtis sinensis

  • Kim Dae Keun;Lim Jong Pil;Kim Jin Wook;Park Hee Wook;Eun Jae Soon
    • Archives of Pharmacal Research
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    • v.28 no.1
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    • pp.39-43
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    • 2005
  • Eight compounds were isolated from the methanolic extract of the twigs of Celtis sinensis through repeated silica gel and Sephadex LH-20 column chromatography. Their chemical structures were elucidated as two triterpenoids, germanicol and epifriedelanol, two amide compounds, trans-N-caffeoyltyramine and cis-N-coumaroyltyramine, two lignan glycoside, pinoresinol glycoside and pinoresinol rutinoside, and two steroids by spectroscopic analysis.

Antioxidative Components from the Aerial Parts of Lactuca scariola L.

  • Kim, Dae-Keun
    • Archives of Pharmacal Research
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    • v.24 no.5
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    • pp.427-430
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    • 2001
  • The antioxidant activity of Lactuca scariola (Compositae) was investigated by measuring the radical scavenging effect on DPPH (1,1-diphenyl-2-picrylhydrazyl) radical. The methanolic extract of the aerial parts of Lactuca scariola showed strong radical scavenging activity. The EtOAc soluble fraction exhibited a stronger activity than the others, and was purified by silica gel and Sephadex LH-20 column chromatography. Quercetin-3-O-$\beta$-D-glucopyranoside, luteolin-7-O-$\beta$-D-glucopyranoside, luteolin, quercetin and kaempferol, together with 11$\beta$,13-dihydrolactucin were isolated from the EtOAc soluble fraction as active ingredients.

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