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Purification and Characterization of the Lectins from Mushroom Flammulina velutipes (팽이버섯으로부터 Lectin의 정제와 특성)

  • Kim, Hyung-Suk;Son, Seung-Yeol;Hwang, Se-Young;Hong, Bum-Shik
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.304-309
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    • 1999
  • Two Lectins, designated FVL-1 and FVL-2, were isolated and purified from the fruiting bodies of edible mushroom Flammulina veluripes using ammonium sulfate fractionation, ethanol treatment, DEAE-TOYPEARL ion-exchange column chromatography, and TSK-Gel HW-55F column chromatography. Specific activity increased 18 folds for FVL-1 and 7.9 folds for FVL-2 from ethanol treated sample. SDS-PAGE of FVL-1 and FVL-2 gave apparent molecular mass of 10.6 kDa and 37 kDa, respectively. FVL-2 agglutinated all type of human erythrocytes (A, B, AB, and O). However, FVL-1 agglutinated more human erythrocyte type O than type A, B, and AB. The hemagglutination activities of the FVL-1 were effectively inhibited by bovine submaxillary and porcine stomach mucins(BSM and PSM), fetuin, asialofetuin and cations, such as $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Mn^{2+}$ and $Fe^{2+}$. However, FVL-2 was not inhibited by any cations. The hemagglutination activities of the two lectins were not inhibited by the sugar, such as lactose, galactose and sugar derivatives. FVL-1 and FVL-2 were stable at pH $5{\sim}11$ and pH $4{\sim}7$, respectively. FVL-1 was stable below $55^{\circ}C$ and FVL-2 was below $45^{\circ}C$.

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Isolation of Bacillus subtilis GS-2 Producing γ-PGA from Ghungkukjang Bean Paste and Identification of γ-PGA (청국장으로부터 분리한 Poly(γ-glutamic acid)를 생산하는 균주 Bacillus subtilis GS-2의 분리 및 γ-PGA의 확인)

  • Bang, Byung-Ho;Jeong, Eun-Ja;Rhee, Moon-Soo;Kim, Yong-Min;Yi, Dong-Heui
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.1-6
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    • 2011
  • ${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee;Chun, En-Mi;Bae, Myung-Ae;Seu, Young-Bae;Song, Kyung-Sik;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.27-34
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    • 2000
  • The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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Cometabolism of $\omega$-Phenylalkanoic Acids with Butyric Acid for Efficient Production of Aromatic Polyesters in Pseudomonas putida BM01

  • Song, Jae-Jun;Choi, Mun-Hwan;Yoon, Sung-Chul;Huh, Nam-Eung
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.435-442
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    • 2001
  • Poly(3-hydroxy-5-phenylvalerate) [P(3HPV)] was efficiently accumulated from 5-phenylvalerate (5PV) in Pseudomonas putida BM01 in a mineral salts medium containing butyric acid (BA) as the cosubstrate. A nove aromatic copolyester, poly(5 mol% 3-hydroxy-4-phenylbutyrate-co- 95 mol% 3-hydroxy-6-phenylhexanoate) [P(3HPB-co-3HPC)] was also synthesized from 6-phenylhexanoate (6PC) plus Ba. The two aromatic polymers, P(3HPV) and P(3HPB-co-3HPC), were found to be amorphous and showed different glass-transition temperatures at $15^{\circ}C$ and $10^{\circ}C$, respectively. When the bacterium was grown ina medium containing 20 mM 5PV as the sole carbon source for 140 h, 0.4 g/l of dry cells was obtained in a flask cultivation and 20 wt% of P(3HPV) homopolymer was accumulated in the cells. However, when it was grown with a mixture of 2 mM 5PV and 50 mM BA for 40 h, the yield of dry biomass was increased up to 2.5 g/l and the content of P(3HPV) in the dry cells was optimally 56 wt%. This efficient production of P(3HPV) homopolymer from the mixed substrate was feasible because BA only supported cell growth and did not induce any aliphatic PHA accumulation. The metabolites released into the PHA synthesis medium were analyzed using GC or GC/MS. Two $\beta$-oxidation derivatives, 3-phenylpropionic acid and trans-cinnamic acid, were found in the 5V-grown cell medium and these comprised 55-88 mol% of the 5PV consumed. In the 6PC-grown medium containing Ba, seven ${\beta}$-oxidation and related intermediates were found, which included phenylacetic acid, 4-phenylbutyric acid, cis-4-phenyl-2-butenoic acid, trans-4-phenyl-3-butenoic acid, trans-4-phenyl-2-butenoic acid, 3-hydroxy-4-phenylbutyric acid, and 3-hydroxy-6-phenylhexanoic acid. Accordingly, based on the metabolite analysis, PHA synthesis pathways from the two aromatic carbon sources are suggested.

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Screening of Anti-Biofilm Compounds from Marine-Derived Fungi and the Effects of Secalonic Acid D on Staphylococcus aureus Biofilm

  • Wang, Jie;Nong, Xu-Hua;Zhang, Xiao-Yong;Xu, Xin-Ya;Amin, Muhammad;Qi, Shu-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1078-1089
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    • 2017
  • Biofilm formation of Staphylococcus aureus is one of its mechanisms of drug resistance. Anti-biofilm screening of 106 compounds from marine-derived fungi displayed that 12 compounds inhibited S. aureus biofilm formation by >50% at the concentration of $100{\mu}g/ml$, and only secalonic acid D (SAD) and B inhibited by >90% at $6.25{\mu}g/ml$ without inhibiting cell growth after 24-h incubation. Meanwhile, it was found that the double bond between C-1 and C-10 of citrinin derivatives and the C-C connection position of two chromone monomers may be important for their anti-biofilm activities. Moreover, SAD slightly facilitated biofilm eradication and influenced its architecture. Furthermore, SAD slowed the cell growth rate in the preceding 18-h incubation and differentially regulated transcriptional expression of several genes, such as agr, isaA, icaA, and icaD, associated with biofilm formation in planktonic and biofilm cells, which may be the reason for the anti-biofilm activity of SAD. Finally, SAD acted synergistically against S. aureus growth and biofilm formation with other antibiotics. These findings indicated that various natural products from marine-derived fungi, such as SAD, could be used as a potential biofilm inhibitor against S. aureus.

A Study on the Carbon Market and Carbon Funds Development. (탄소시장과 탄소펀드 개발에 관한 연구)

  • Son, Woo-Sik;Park, Myong-Sop
    • THE INTERNATIONAL COMMERCE & LAW REVIEW
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    • v.46
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    • pp.265-313
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    • 2010
  • Kyoto Protocol is an international convention on concrete performance program for UNFCCC(United Nations Framework Convention on Climate Change), which regulate and prevent to global warming and officially came into effect on February 16, 2005. Kyoto flexible mechanisms, the agreed environmental system in March 1997 in the Third Conference of Parties in UNFCCC General Assembly, Emission Trading System(ETS), Clean Development Mechanism(CDM) and Joint Implementation(JI), are key policies related to environment. In advanced countries, greenhouse gas emissions should be reduced average 5.2% level compared to 1990 in total emissions during 2008-2012. World leading carbon market finished the trial on the EU ETS I greenhouse gas emissions trading system, EU ETS II is operated regularly after 2008. World Bank leads to make 'Prototype Carbon Fund(PCF)' in April 2004, which is the world first carbon fund and a representative public carbon fund type, World Bank operate various funds including present PCF. Thus, I would like to propose as follows in relation to this study: First, in the validity analysis of carbon funds, it would be needed to analyze the Emission Reduction Cost Efficiency(ERCE) of carbon. The ERCE is a break-even value which brings the Net Present Value(NPV) to zero. NPV approach is used among projects and it enables potential projects to be compared and evaluated the ERCE on the basis of the net present value of net future cash flows. Therefore, according to results of analysis, carbon funds should be developed and invested. Second, it would be necessary to allow of issuing bonds together with carbon funds, carbon finance etc. Third, carbon funds, it would be reasonable to have a relatively enough maturity in project and as a financial derivatives in the international financial markets, it is needed various types of transactions. Fourth, it would be needed to standardize the carbon emissions trading for more efficiently. Fifth, it would be necessary to establish and invest in various kinds of domestic and overseas global carbon funds, including governments, privates, governments and privates sectors. And it is also needed to establish the medium and long term plans for carbon funds. Sixth, it would be needed to foster the advanced trade mechanisms for carbon funds in the most effective ways. Finally, carbon funds should be used in harmony with international societies to reduce global warming as the social responsible investing funds and it should be contribute to sustainable development. In addition, it would seem that carbon funds should be studied on establishing the contributable standard of sustainable development in the future assignment.

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Design of new 1,3-thiazoline derivatives by isosterism and antifungal activity of new 2,4-diimino-1,3-thiazolidines (Isosterism을 이용한 새로운 1,3-thiazoline 유도체의 디자인 및 신규 2,4-diimino-1,3-thiazolidine 유도체의 살균 활성)

  • Hahn, Hoh-Gyu;Nam, Kee-Dal;Lim, Chul-Soo;Mah, He-Duck;Kim, Jin-Cheol;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.7 no.1
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    • pp.51-57
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    • 2003
  • For the purpose of a development of new agrochemical fungicides, new compound 4 in which 1,3-thiazoline scaffold as well as urea moiety in the structure was designed through molecular modification of lead compound, 2-imino-1,3-thiazone based on isosterism. The reaction of N-alkylthiourea 5 and bromoacetonitrile in ethanol gave 2,4-diimino-1,3-thiazolidine hydrobromide 6 regioselectively, which was treated with isocyanates gave the corresponding 8 which is tautomer of 4. Antifungal screening (in vivo) of the synthesized compound 8 against typical plant diseases, which include rice blast, rice sheath blight, cucumber gray mold, tomato late blight, wheat leaf rust, and barley powdery mildew, was carried out. Antifungal activities against rice blast of the compound 8 were weaker than those of 2-phenylimino-1,3-thiazoline 1. Some compounds showed weak antifungal activities against wheat leaf rust.

Effect of Alkyl Substituents, Surfactants, and Temperature on the Solubilization of 4-alkylaniline Derivatives by Cationic Surfactants (양이온계면활성제에 의한 4-알킬아닐린 유도체의 가용화에서 알킬치환기, 계면활성제 및 온도의 효과)

  • Lee, Dong-Cheol;Lee, Byung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.2
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    • pp.250-259
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    • 2020
  • The solubilization constant (Ks) was determined by the UV-Vis method to investigate the interaction between organic matter (solubilized substance) and surfactant in solubilization. Solubilization constants and thermodynamic functions, according to the hydrophobic interaction between organic mater (4-alkylanilines with different alkyl substituent length) and cationic surfactants (DTAB, TTAB, and CTAB, having different hydrophobic lengths), were measured and calculated at various temperatures and compared with each other. As a result, the hydrophobic interactions between organic matters and cationic micelles increased with increasing the chain length of solute's substituent as well as surfactant's hydrophobe. However, the hydrophobic effect by the alkyl substituent of organic matter was greater than the hydrophobic effect by the surfactant. In addition, the results of the calculated thermodynamic functions showed that 4-alkylaniline was solubilized at the deep place in the micelle and its solubilization was greatly dependent on both the hydrophobic effects of organic matter and surfactant. At the calculated iso-structural temperature, the difference between the maximum and minimum values was less than 1K within the experimental conditions.

Synthesis and Small Animal Brain PET Study of a Serotonin Receptor Radiotracer, 18F-Mefway (세로토닌 5-HT1A수용체 방사성 추적자 18F-Mefway의 합성과 소동물 뇌 PET 연구)

  • Ahn, Sung-Min;Hong, Tae-Kee;Ryu, Young-Hoon;Choi, Jae-Yong;Kim, Sung-Chul
    • The Journal of the Korea Contents Association
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    • v.9 no.11
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    • pp.262-270
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    • 2009
  • $^{18}F$-mefway has been developed as radioligand for serotonin receptor 5-$HT_{1A}$. The object of this study was to obtain the mefway precursor with the higher yield than previous method and to identify whether $^{18}F$-mefway can bind to 5-$HT_{1A}$ or not. from microPET imaging of small animal brain. Precursor was prepared by a modification of the reported procedure then [$^{18}F$] labeling was performed by adding $^{18}F$ ion at $130^{\circ}C$ in the hot cell for 30min. After purification of reaction mixture using alumina Sep-pak and HPLC, microPET images of small animal brain were determined. The chemical yield of precursor was increased from 9% to 34% using oxalyl chloride and LAH/diethylether. We synthesized a precursor which was successfully labeled with no-carrier-added $^{18}F$-by new synthetic route. This research suggest that $^{18}F$-mefway will be used a radiopharmaceutical for evaluation of central nerve system disorder as imaging a gent for 5-$HT_{1A}$ receptor.

Comparison of Detectable Levels for Screening Residual Antibacterial Agents by Bioassay (잔류 항균물질에 대한 미생물학적 간이검사법의 검출감도 비교)

  • JUNG Sung Hee;KIM Jin Woo;SOHN Sang-Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.3
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    • pp.256-260
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    • 1999
  • Minimun-detectable levels to 28 antibacterial agents used for the prevention and the treatment of fish diseases were determined to establish optimal detective method of bioassay in fish by the EEC 4-plate method, the modified method of EEC 4-plate and the standard method of analysis in food safety regulation. The test organisms used in the methods of bioassay were as follows: Bacillus subtilis BGA (B. subtilis) and Micrococcus luteus ATCC 9341 (M. luteus) in the EEC 4-plate method, B. subtilis, M. luteus and Bacillus cereus var. mycoides ATCC 11778 (B. cereus) in the modified of EEC 4-plate, and B. subtilis, M. luteus, B. cereus and Bacillus stearothermophilis var. calidolactis C-953 (B. stearothermophilis) in the standard method. The standard method showed predominant sensitivity in the detection of penicillins (PCs), and was also highly sensitive to aminoglycosides (AGs). The sensitivity of standard method in the detection of tetracyclines (TCs), marrolides (MLs), nitrofuran derivatives(NFs) and quinolones (QNs) was very low, and against sulfonamides (SAs), however, was extremely low. The modified method of EEC 4-plate showed very high sensitivity to TCs. Both the EEC 4-plate and the modified method of EEC 4-plate showed competitively high sensitivity in the detection of PCs, MLs, NFs, QNs and SAs. All the methods studied in the experiment showed very low sensitivity against chloramphenicol (CMs). Consequently, the modified method of EEC 4-Plate was the best bioassay method with a wide range of sensitivity for the optimal detection of the residual antibacterial agents in fish.

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