• Title/Summary/Keyword: Bio-column

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Isolation and structural elucidation of the herbicidal active compounds from Ligularia stenocephala M.

  • Lim, Chi-Hwan;Cho, Chong-Woon
    • Korean Journal of Agricultural Science
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    • v.48 no.2
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    • pp.343-351
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    • 2021
  • Screening was conducted using 200 kinds of plant extracts to explore herbicide-activated components of plant origin. We separated and purified active substances and elucidated chemical structures using Ligularia stenocephala M., which has strong activity and has not yet been studied. When the solvent fractions of the leaves of Ligularia stenocephala M. were tested for their herbicidal activity, ethyl acetate and chloroform layer showed an inhibition rate of 95.2% and 94.1%, respectively. In particular, the chloroform layer exerted more than 50% herbicidal activity at 10 ppm. From the chloroform layer with the highest herbicidal activity, we isolated three herbicidal active compounds using stepwise chromatography, specifically silica gel or octadecyl silica (ODS) column chromatography, Sep-pak cartridges, and high performance liquid chromatography (HPLC). Based on the analysis of the active compounds using electron ionization mass spectroscopy (EI-MS), 1H-NMR, and 13C-NMR, we identified the active compounds as euparin, 5,6-dimethoxy-2-isopropenylbenzofuran, and liguhodgsonal. When the herbicidal activity of the identified compounds was tested, euparin showed selective herbicidal activity for lettuce at 10-3 M, and both liguhogsonal and 5,6-dimethoxy-2-isoprophenylbenzofuran exerted selective activity for rice and Echinochloa crus-galli.

Purification and Identification of Apoptosis Modulator Pipernonaline from Piper longum Linn. against Prostate Cancer Cells (필발(Piper longum Linn.)로부터 전립선암 세포사멸물질 pipernonaline의 분리 및 동정)

  • Kim, Kwang-Youn;Kim, Yun-Jin;Lee, Wan;Yu, Sun-Nyoung;Cho, Hyo-Jin;Lee, Sun-Yi;Lee, Han-Seung;Sohn, Jae-Hak;Oh, Hyuncheol;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.19 no.5
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    • pp.671-675
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    • 2009
  • Prostate cancer has been a critical health problem due to an increase of prostate cancer-related deaths worldwide. Also, a frequent treatment option for prostate cancer is androgen ablation, but this treatment has a limited scope, especially for hormone-refractory cancer. There is an urgent need for the identification of alternative therapeutic strategies for prostate cancer. Previously, over one hundred species of dried-plant methanol extracts were tested for inhibitory effects on proliferation. One of them, Piper longum Linn. was selected based on its potent anti-proliferation effect. The dried root of P. longum Linn. was extracted with 100% methanol for 2-3 days and its extract was fractionated using chloroform. The chloroform layer was then subjected to column chromatography on silica gel, reverse phase-18 (RP-18) and Sephadex LH-20, in turn. Finally, the pure compound was obtained and identified as pipernonaline by NMR spectroscopic and physico-chemical analysis. In this study, anti-proliferation and cell cycle arrest effects of pipernonaline on human prostate cancer PC-3 cells were investigated using the MTT and PI staining, respectively. Our findings suggest that pipernonaline represents a dose-dependent growth inhibition pattern on PC-3 cells and, moreover, its growth inhibition is associated with sub-G1 and G0/G1 cell cycle accumulation in PC-3 cells. Also, these results provide an anticancer candidate for human prostate cancer.

Molecular Cloning and Expression of a Gene for Outer Membrane Protein H in Pasteurella multocida (A:3) : Production of Antisera against the OmpH (파스튜렐라 (A:3)외막 단백질 H의 유전자 클론닝$\cdot$발현 및 면역혈청 생산)

  • Kim Younghwan;Hwang Heon;Lee Sukchan;Park Eun-Seok;Yoo Sun-Dong;Lee Jeongmin;Yang Joo-Sung;Kwon MooSik
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.274-280
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    • 2005
  • Pasteurella multocida is known to cause widespread infections in husbandry. To induce homologous and heterologous immunity against the infections, outer membrane proteins (OMPs) in the envelope of P. multocida are thought to be attractive vaccine candidates. Outer membrane protein H is considered as the major component of OMPs. In this study, a gene for OmpH was isolated from pathogenic P. multocida serogroup A. The gene was composed of 1,047 nucleotides coding 348 amino acids with signal peptide of 20 amino acids. The amino acid composition showed about 80 to 98 per cent sequence homologies among other 10 strains of P. multocida serogroup A, reported so far. A recombinant ompH, from which signal peptide was truncated, was generated using pRSET A to name 'pRSET A/OmpH-F2'. The pRSET A/OmpH-F2 was well expressed in E. coli BL21(DE3). The truncated OmpH was purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Its molecular weight was registered to be 40 kDa on SDS-PAGE gel. In order to generate immunesera against the OmpH, 50 ug of the protein was intraperitoneally injected into mice three times. The anti-OmpH immuneserum recognized about $5{\times}10^{-2}$ng quantity of the purified OmpH. It can be used for an effective vaccine production to prevent fowl cholera caused by pathogenic P. multocida (Serogroup A).

Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties (한천분해세균 Cellvibrio mixtus SC-22의 분리 및 효소적 특성)

  • Cha, Jeong-Ah;Kim, Yoo-Jin;Seo, Yung-Bum;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.157-162
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    • 2009
  • An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

Biodegradation of UV Filters in Biological Activated Carbon (BAC) Process : Biodegradation Kinetic (BAC 공정에서의 자외선 차단제 생물분해 특성 : 생물분해 동력학)

  • Seo, Chang-Dong;Son, Hee-Jong;Jung, Jong-Moon;Choi, Jin-Taek;Ryu, Dong-Choon;Jang, Seong-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.11
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    • pp.739-746
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    • 2014
  • In this study, The effects of empty bed contact time (EBCT) and water temperature on the biodegradation of 8 UV filters in biological activated carbon (BAC) process were investigated. Experiments were conducted at two water temperatures (7 and $18^{\circ}C$) and three EBCTs (5, 10 and 15 min). Increasing EBCT and water temperature increased the biodegradation efficiency of UV filters in BAC column. EHMC and BZC were the highest biodegradation efficiency, but BP and 4-MBC were the lowest. The kinetic analysis suggested a first-order reaction model for biodegradation of 8 UV filters at various water temperatures and EBCTs. The first-order biodegradation rate constants ($k_{bio}$) of 8 UV filters ranging from $0.2730{\sim}0.6365min^{-1}$ at $7^{\circ}C$ to $0.4824{\sim}0.8743min^{-1}$ at $18^{\circ}C$. By increasing the water temperature from $7^{\circ}C$ to $18^{\circ}C$, the biodegradation rate constants ($k_{bio}$) were increased 1.5~2.1 times.

Anti-corrugation activity of micosporine-like amino acid mixtures from Chlamydomonas sp. (Chlamydomonas sp. 유래 mycosporine-like amino acid 혼합물의 항주름 활성)

  • Suh, Sung-Suk;Seo, Hyo Hyun;Lee, Jeong Hun;Hwang, Jinik;Park, Mirye;Moh, Sang Hyun;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.8
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    • pp.5393-5399
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    • 2014
  • To examine the effects of a mycosporine-like amino acids (MAAs) mixture from microalgae, Chlamydomonas sp, on the anti-wrinkle activities, the expression levels of genes that are associated with skin aging, including type I procollagen, elastin and involucrin, were analyzed. Asterina 330+palythine (A+P) and shinorine+palythine (S+P) mixtures were purified from Chlamydomonas sp using the following steps: 80% methanolic extraction, column purification, and HPLC analysis. As a result of the MTT assay, A+P and S+P did not induce cellular cytotoxicity with up to 0.1 mg/mL of both MAAs. In addition, the treatment of UV-exposed fibroblasts with A+P (0.05 mg/mL) and S+P (0.01 mg/mL) increased the levels of the PCOLCE mRNAs by 2.7 and 3.6 fold compared to the control group, respectively, The levels of elastin gene expression were elevated 5.59 and 3.1 fold in the A+P and S+P treated (0.01 mg/mL) cells, respectively. In particular, at a concentration of 0.01 mg /mL, the A+P and S+P expression levels of Involucrin mRNAs were increased 3.5 and 2.5 fold in the UV-exposed cells compared to the control, respectively. In conclusion, the MAAs derived from Chlamydomonas sp can be utilized as functional cosmetic materials with anti-wrinkle effects on the skin.

Effects of biodegradable polymer coating urea to nitrogen release in the soil column (생분해성 코팅 요소 종류별 질소 용출 및 온실가스 발생량에 미치는 영향)

  • Jaeyee Choi;JoungDu Shin;HyunJong Cho;Woojin Chung;Sang Beom Lee;Seok In Yun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.32 no.1
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    • pp.49-59
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    • 2024
  • Biodegradable coating urea plays an important role in reducing the non-pollutants in agroecosystems, maximizing the plant nutrient utilization efficiency and slow-releasing nitrogen. Herein, the objective of this study was to investigate the nitrogen-releasing patterns and greenhouse gas emissions on different biodegradable coating urea. The treatments consisted of the control as an application of chemical fertilizers, NBCF as the non-biodegradable coating urea, NB60, and MDS as biodegradable coating urea. As a result of this study, the maximum accumulated total nitrogen (TN) concentration in the NBCF was higher at 33% than one in the NB60 during the precipitation periods. Its leaching period in the NCBF was prolonged for day 10 compared to the NB60. TN and NO3-N releasing patterns in the NBCF and NB60 were fitted well on linear types(R2≥0.991), but their control and MDS were fitted well on Sigmoid curves(R2≥0.994) with high releasing concentration in the MDS compared to the control during leaching periods. For the greenhouse gas emissions, CH4 emissions in the NBCF, NB60, and MDS were increased at 0.38%, 11.36%, and 5.91%, and N2O emissions were also increased at 50.5%, 32.4%, 58.8% as compared to the control, respectively. Therefore, application of biodegradable polymer coating urea might mitigate the non-point pollutants in agro-ecosystem.

Characteristic of Nutrient Release by Submerged Plants under Different Water Temperatures in Lake Reservoir (호소저수구역에서 침수식물체의 수온별 영양염류 용출 특성)

  • Lim, Byung-Jin;Lee, Sang-Gyu;Seo, Dong-Cheol;Choi, Ik-Won;Kang, Se-Won;Seo, Young-Jin;Lee, Dong-Jin;Kim, Sang-Don;Heo, Jong-Soo;Cho, Ju-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.657-663
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    • 2012
  • To evaluate effects of water temperatures on nutrient releases of submerged plants in lake reservoir, COD, T-N and T-P releases of submerged plants were investigated for 60 days under different incubation temperatures ($5^{\circ}C$ and $25^{\circ}C$) in columns. The amounts of COD releases by Carex dimorpholepis were $60.4mg\;L^{-1}$ at $5^{\circ}C$ and $78.0mg\;L^{-1}$ at $25^{\circ}C$. In Miscanthus sacchariflorus, the amounts of COD releases were $62.5mg\;L^{-1}$ at $5^{\circ}C$ and $70.5mg\;L^{-1}$ at $25^{\circ}C$. The amounts of T-N releases in Carex dimorpholepis at $5^{\circ}C$ and $25^{\circ}C$ were 45.8 and $60mg\;L^{-1}$, respectively. In Miscanthus sacchariflorus, the amounts of T-N releases were $55.7mg\;L^{-1}$ at $5^{\circ}C$ and $61.0mg\;L^{-1}$ at $25^{\circ}C$. At $5^{\circ}C$, the amounts of T-P releases in Carex dimorpholepis and Miscanthus sacchariflorus were 5.65 and $7.10mg\;L^{-1}$, respectively. At $25^{\circ}C$, the amounts of T-P releases in Carex dimorpholepis and Miscanthus sacchariflorus were 8.70 and $8.18mg\;L^{-1}$, respectively. In the column experiment, the amounts of COD, T-N and T-P releases by submerged plants at $25^{\circ}C$ were generally higher than those at $5^{\circ}C$.

Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

Triglyceride Composition of Some Vegetable Oils 1. Triglyceride Composition of Sesame Oil (식물유의 Triglyceride 조성 1. 참기름의 Triglyceride 조성)

  • Park Yeung-Ho;WADA Shun;KOIZUMI Chiaki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.1-6
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    • 1981
  • In this study, sesame oil was chosen as the experimental sample and analysed for its triglyceride composition by high-performance liquid chromatography(HPLC) in combination with gas liquid chromatography(GLC). The triglycerides were separated from sesame oil by liquid chromatographies on Bio-Beads SX-2 and on Sephadex LH-20, and fractionated into five groups on the basis of their partition numbers by reverse phase HPLC on a column packed with $\mu-Bondapak$ C18 using methanol-chloroform mix-ture as a solvent. Each of these collected fractions gave one to three peaks in the GLC chromatograms according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analysed by GLC. From the results, it was found that the sesame oil consists with twenty one kinds of triglyceri-des, and the major triglycerides of sesame oil are those of $(2\;{\times}\;C18:1,\;C18:2\;;\;17.1\%),\;(C18:1,\;2{\times}C18:2\;;\;17.0\%),$ $(3\;{\times}\;C18:2\;;\;17.0\%),\;(3\;{\times}\;C18:1\;;\;10.9\%),$ $(3\;{\times}\;C18:2\;;\;9.6\%),\;(C16:0,\;C18:1,C18:2\;;\;7.9\%),$ $(C16:0,\;2\;{\times}\;C18:1\;;\;7.4\%),\;(C16:0,\;2\;{\times}\;C18:2\;;\;6.8\%),$ $(C18:0,\;C18:1,\;C18:2\;;\;3.1\%),\;(2\;{\times}\;C18:0,\;C18:2\;;\;1.5\%)$ $(C18:0,\;2\;{\times}\;C18:1\;;\;1.4\%),\;(C16:0,\;C18:0,\;C18:1\;;\;1.3\%),$ $(2\;{\times}\;C16:0,\;C18:1\;;\;1.2\%),\;and\;(C16:0,C18:0,\;C18:2\;;\;1.0\%)$.

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