• Journal of Microbiology and Biotechnology
• /
• v.10 no.1
• /
• pp.27-34
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.32 no.5
• /
• pp.985-990
• /
• 2000

${\zeta}-Carotene$ was subjected to ozonolysis in ice-cold dichloromethane. The ozonolysis products were fractionated with a silica column and the carbonyl fraction was analyzed by ODS-HPLC with a photodiode array detector. ${\zeta}-Carotene$ was solubilized in toluene, and then oxidized by incubating at $37^{\circ}C$, 72 hr under atmospheric oxygen. Carbonyl compound and acidic compound were produced. In comparison with autoxidation and ozonolysis, each compound showed the same retention time and UV-vis spectra were identical to the reference cleavage products prepared by ozonolysis of ${\zeta}-carotene$. Absorption spectrum of acidic compound was similar to that of standard 4,5-didehydrogeranyl geranyl acid which is known to possess biological activity. Thus, eccentric cleavage of ${\zeta}-carotene$ was confirmed to occur in vitro under oxidation condition.

• Korean Journal of Environmental Agriculture
• /
• v.15 no.1
• /
• pp.37-45
• /
• 1996

Abstracts From the previous metabolic study of Pentachlorophenol(PCP), PCP was found to be exclusively transformed into ${\beta}-glucose$ conjugates of PCP in soybean and rice cell suspension cultures. In order to gather structural information of of the glucose conjugate, their aglycons and glycon have been analyzed by GC and GC/MS respectively, after thorough purification by chromatographic techniques. The glucose conjugates were effectively purified through a 1-butanol extraction followed by Silica gel TLC, Sephadex column chromatography and HPLC. Aglycons of the metabolites were identified as PCP, isomeric mixture of tetrachlorophenol, and tetrachlorocatechol and glycon were identified as glucose, suggesting that there are at least three kinds of glucose conjugates with different phenolic moieties. Under controlled conditions, the glucose conjugates were separated into three HPLC peaks which released respective aglycon upon a hydrolytic treatment. These results give valuable information on the structure of the glucose conjugates such that some PCP-driven chlorophenols, in addition to PCP, are also conjugated with glucose.

• Korean Journal of Environmental Agriculture
• /
• v.34 no.2
• /
• pp.139-148
• /
• 2015

BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity and algicidal range of algicide. METHODS AND RESULTS: Among of algicidal bacteria isolated in this study, NH-3 isolate was the strongest algicidal activity against A. catenella. NH-3 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The NH-3 isolate showed over 99% homology with Arthrobacter oxydans, and was designated as Arthrobacter sp. NH-3. The optimal culture conditions were $25^{\circ}C$, initial pH 7.0, and 2.0% (w/v) NaCl concentration. The algicidal activity of Arthrobacter sp. NH-3 was significantly increased to maximum value in the late of logarithmic phase. Arthrobacter sp. NH-3 showed algicidal activity through indirect attack, which excreted active substance into the culture filtrate. When 10% culture filtrate of NH-3 was applied to A. catenella, 100% of algal cells were destroyed within 30 h. In addition, the algicidal activities were increased in dose and time dependent manners. The pure algicide was isolated from the ethyl acetate extract of the culture filtrate of NH-3 by using silica gel column chromatography and high performance liquid chromatography (HPLC). We investigated the algicidal activity of this algicide on the growth of harmful algal bloom (HAB) species, including A. catenella. As a result, it showed algicidal activity against several HAB species at a concentration of $100{\mu}g/mL$ and had a relatively wide host range. CONCLUSION: Taken together, our results suggest that Arthrobacter sp. NH-3 and its algicide could be a candidate for controlling of toxic and harmful algal blooms.

• Applied Biological Chemistry
• /
• v.10
• /
• pp.47-61
• /
• 1968

Sweetpotato infected by Soft-rot, Rhizopus nigricans, has been investigated for its biochemical changes. The results of the present experiment are summarized as follows: 1. From the Soft-rotted tuber, nineteen Ehrlich's positive substances and three fluorescent compounds which had not been found in a healthy tuber were detected with thin layer chromatography. 2, Ipomeamarone and umbelliferone, each of which are predominating substances among the Ehrlich's positive and fluorescent compounds, were isolated by silica gel column chromatography and studied for their identities. 3. Time course biosynthesis of the metabolites was studied and discussed for their possible biosynthetic pathways. 4. Among the metabolites, ipomeamarone and ipomeamarone-like substances have enzyme: inhibitive action against ${\alpha}$-amylase and probably other enzymes; the inhibition type was determined as the uncompetitive one. 5. Ipomeamarone obtained from soft-rotted sweetpotato was proved to have an uncoupling action as 2, 4-dinitrophenol.

• Journal of Pharmacopuncture
• /
• v.20 no.2
• /
• pp.119-126
• /
• 2017

Objectives: Semecarpus anacardium Linn. is a plant well-known for its antimicrobial, antidiabetic and anti-arthritic properties in the Ayurvedic and Siddha system of medicine. This has prompted the screening of this plant for antibacterial activity. The main aims of this study were to isolate compounds from the plant's seeds and to evaluate their antibacterial effects on clinical bacterial test strains. Methods: The n-butanolic concentrate of the seed extract was subjected to thin layer chromatography (TLC) and repeated silica gel column chromatography followed by elution with various solvents. The compound was identified based on observed spectral (IR, $^1H$ NMR, $^{13}C$ NMR and high-resolution mass spectrometry) data. The well diffusion method was employed to evaluate the antibacterial activities of the isolated acyclic isoprenoid compound (final concentration: $5-15{\mu}g/mL$) on four test bacterial strains, namely, Staphylococcus aureus (MTCC 96), Bacillus cereus (MTCC 430), Escherichia coli (MTCC 1689) and Acinetobacter baumannii (MTCC 9829). Results: Extensive spectroscopic studies showed the structure of the isolated compound to be an acyclic isoprenoid ($C_{21}H_{32}O$). Moreover, the isoprenoid showed a remarkable inhibition of bacterial growth at a concentration of $15{\mu}g/mL$ compared to the two other doses tested (5 and $10{\mu}g/mL$) and to tetracycline, a commercially available antibiotic that was used as a reference drug. Conclusion: The isolation of an antimicrobial compound from Semecarpus anacardium seeds validates the use of this plant in the treatment of infections. The isolated compound found to be active in this study could be useful for the development of new antimicrobial drugs.

• The Plant Pathology Journal
• /
• v.15 no.1
• /
• pp.14-20
• /
• 1999

Twenty isolates of Helminthosporium species were obtained from various grass plants and tested for controlling efficacy on the development of plant diseases. An isolate of Helminthosporium sp. TP-4 was chosen and six antibiotic substances were purified from cultures of the fungus by repeated silica gel column chromatography and preparative thin-layer chromatography. They were identified as ophiobolin a, 6-epiophiobolin A, 3-anhydroophiobolin A, 3-anhydro-6-epiophiobolin A, iphiobolin B, and iphiobolin I mainly by mass spectrometry and nuclear magnetic resonance spectrometry. Ophiobolins inhibited the growth of a grampositive bacterium Streptomyces griseus, but were not active against gram-negative bacteria. They also showed an antifungal activity. In in vivo tests, iphiobolin B exhibited potent controlling activities against rice blast, tomato late blight, and wheat leaf rust with control values more than 90% and 70% at concentration of $500\mu\textrm{m}$/ml and 100 ${\mu}{\textrm}{m}$/ml. Ophiobolin A and 6-epiophiobolin A controlled the development of wheat leaf rust more than 80% at concentrations of 100 /ml and $500\mu\textrm{m}$/ml respectively. 3-Anhydro-6-epiophiobolin A was not active against any plant disease. On the other hand, the A-series ophiobolins other than 3-anhydroophiobolin A showed stronger phytotoxic activity in a leaf-wounding assay using 8 plant species than those of 3-anhydroophiobolin A, ophiobolin B, and ophiobolin I. The results indicate that there is little correlation between antifungal activity and phytotoxicity of ophiobolins.

• Journal of the korean society of oceanography
• /
• v.36 no.4
• /
• pp.101-108
• /
• 2001

Polybrominated diphenylether (PBDEs) congeners, 2,4,4'-TrBDE, 2,2',4,4'-TeBDE, 2,2',4,4',5-PeBDE, 2,2',4,4',5,5'-HXBDE, and 2,2',4,4',5,6-HxBDE, were measured in sediments from 52 stations in the southeastern coastal areas of Korea. Sediment samples were analyzed using gas chromatography coupled to mass spectrometer detector (GC/MSD) with positive electron impact (PEI) mode. New analytical methodology for PBDEs by the isotope dilution method was established using a multi-layer silica gel column chromatography. Total PBDEs levels in sediments for Pohang coast ranged from 1,1 to 33.8 ng/g dry weight (mean 5.3 ng/g dry), from 1.6 to 36.4 ng/g dry weight (mean 5.7 ng/g dry) for Ulsan coast, from 0.8 to 20.3 ng/g dry weight (mean 4.9 ng/g dry) for Busan coast, and from 0.8 to 10.3 ng/g dry weight (mean 4.4 ng/g dry) for Jinhae Bay. PBDEs contamination in surface sediments from Korean southeastern coasts was relatively moderate in comparison to that of other marine environment in the world. The predominant PBDE congeners were 2,2',4,4'-TeBDE and 2,2',4,4',5-PeBDE.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.5
• /
• pp.1001-1010
• /
• 2005

Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

• Journal of Ginseng Research
• /
• v.42 no.2
• /
• pp.199-207
• /
• 2018

Background: Ginseng is a well-known traditional Chinese medicine that has been widely used in a range of therapeutic and healthcare applications in East Asian countries. Microbial transformation is regarded as an effective and useful technology in modification of nature products for finding new chemical derivatives with potent bioactivities. In this study, three minor derivatives of ginsenoside compound K were isolated and the inducing effects in the Wingless-type MMTV integration site (Wnt) signaling pathway were also investigated. Methods: New compounds were purified from scale-up fermentation of ginsenoside Rb1 by Paecilomyces bainier sp. 229 through repeated silica gel column chromatography and high pressure liquid chromatography. Their structures were determined based on spectral data and X-ray diffraction. The inductive activities of these compounds on the Wnt signaling pathway were conducted on MC3T3-E1 cells by quantitative real-time polymerase chain reaction analysis. Results: The structures of a known 3-keto derivative and two new dehydrogenated metabolites were elucidated. The crystal structure of the 3-keto derivative was reported for the first time and its conformation was compared with that of ginsenoside compound K. The inductive effects of these compounds on osteogenic differentiation by activating the Wnt/b-catenin signaling pathway were explained for the first time. Conclusion: This study may provide a new insight into the metabolic pathway of ginsenoside by microbial transformation. In addition, the results might provide a reasonable explanation for the activity of ginseng in treating osteoporosis and supply good monomer ginsenoside resources for nutraceutical or pharmaceutical development.