• Title/Summary/Keyword: thin-layer chromatography

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Purification and Characteriztion of an Antifungal Antibiotic from Bacillus megaterium KL 39, a Biocontrol Agent of Red-Papper Phytophtora Blight Disease. (고추역병균 Phytophthora capsici를 방제하는 길항균주 Bacillus megaterium KL39의 선발과 길항물질)

  • 정희경;김상달
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.235-241
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    • 2003
  • For the biological control of Phytophthora blight of red-pepper caused by Phytophthora capsici, an antibiotic-producing plant growth promoting rhizobacteria (PGPR) Bacillus sp. KL 39 was selected from a local soil of Kyongbuk, Korea. The strain KL 39 was identified as Bacillus megaterium by various cultural, biochemical test and API and Microlog system. B. megaterium KL 39 could produce the highest antifungal antibiotic after 40 h of incubation under the optimal medium which was 0.4% fructose, 0.3% yeast extract, and 5 mM KCl at 30 C with initial pH 8.0. The antifungal antibiotic KL 39 was purified by Diaion HP-20 column, silica gel column, Sephadex LH-20 column, and HPLC. Its RF value was confirmed 0.32 by thin-layer chromatography with Ethanol:Ammonia:Water = 8:1:1. The crude antibiotic KL39 was active against a broad range of plant pathogenic fungi, Rhizoctonia solani, Pyricularia oryzae, Monilinia fructicola, Botrytis cinenea, Alteranria kikuchiana, Fusarium oxysporum and Fusarium solani. The purified antifungal antibiotic KL39 had a powerful biocontrol activity against red-pepper phytophthora blight disease with in vivo pot test as well as the strain B. megaterium KL 39.

Isolation and Characterization of an Agar-hydrolyzing Marine Bacterium, Pseudoalteromonas sp. H9, from the Coastal Seawater of the West Sea, South Korea (서해안 해수로부터 분리한 한천분해 해양미생물 Pseudoalteromonas sp. H9의 동정 및 특성 연구)

  • Chi, Won-Jae;Youn, Young Sang;Kim, Jong-Hee;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.134-141
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    • 2015
  • An agarolytic marine bacterium (H9) was isolated from the coastal seawater of the West Sea, South Korea. The isolate, H9, was gram-negative and rod-shaped with a smooth surface and polar flagellum. Cells grew at 20-30℃, between pH 5.0 and 9.0, and in ASW-YP (Artificial Sea Water-Yeast extract, Peptone) media containing 1-5% (w/v) NaCl. The G+C content was 41.56 mol%. The predominant isoprenoid quinone in strain H9 was ubiquinone-8. The major fatty acids (>10%) were C16:1ω7c (34.3%), C16:0 (23.72%), and C18:1ω7c (13.64%). Based on 16S rRNA gene sequencing, and biochemical and chemotaxonomic characterization, the strain was designated as Pseudoalteromonas sp. H9 (=KCTC23887). In liquid culture supplemented with 0.2% agar, the cell density and agarase activity reached a maximum level of OD = 4.32 (48 h) and OD = 3.87 (24 h), respectively. The optimum pH and temperature for the extracellular crude agarases of H9 were 7.0 and 40℃, respectively. Thin-layer chromatography analysis of the agarase hydrolysis products revealed that the crude agarases hydrolyze agarose into neoagarotetraose and neoagarohexaose. Therefore, the new agar-degrading strain, H9, can be applicable for the production of valuable neoagarooligosaccharides and for the complete degradation of agar in bio-industries.

Studies on the Electrochemical Behaviors, Spectrophotometric Determination of Heavy Lanthanide Ions and Heavy Metal Chelate Complexes with Bidentate Ligands(III) -Synthesis and Characterization of the Tetrakis(5,7-dichloro-8-quinolinato)(2-mercaptopyrimidinato) molybdenum(IV) Complex- (무거운 란탄이온의 전기화학적 거동, 분광학적 정량 및 중금속 이온과 두 자리 리간드 착물에 관한 연구(제 3 보): -테트라키스(5,7-디클로로-8-퀴놀리나토)(2-메르캅토피리미디나토) 몰리브데늄(IV) 착물의 합성 및 특성-)

  • Chang, Choo Hwan;Choi, Won Jong;Park, Keun Su;Son, Pyung Su;Suh, Moo Yul
    • Analytical Science and Technology
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    • v.6 no.5
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    • pp.417-424
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    • 1993
  • Eight-coordinate tetrakis molybdenum(IV) complexes containing 5,7-dichloro-8-hydroxyquinolinol(Hdcq) and 2-mercaptopyrimidine(Hmpd) has been prepared. $Mo(mpd)_4$, $Mo(dcq)(mpd)_3$, $Mo(dcq)_2(mpd)_2$, $Mo(dcq)_3(mpd)$ and $Mo(dcq)_4$ complexes have been isolated by thin-layer chromatography on silicagel plates. These complexes have been charaterized by $^1H-nmr$ spectrum and UV-Vis. spectrum. The chemical shift values of the protons ${\alpha}$ to the nitrogen in the ligands are shifted to down field. The relative intensities of the peaks which are positioned at the same proton of $Mo(dcq)(mpd)_3$ and $Mo(dcq)_3(mpd)$ are observed in 2:1 ratio, in case of $Mo(dcq)_2(mpd)_2$ appears in approximately a 1:1 ratio. The stereochemistry of the complexes in discussed in terms of their nmr spectrum and Orgel's rule. By vertue of the intensities (${\varepsilon}$>10,000~25,000) the low energy($16,600{\sim}19,800cm^{-1}$) bands are observed for the electronic spectra of the complexes are assigned as charge transfer bands.

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In vivo metabolism of carbofuran in resistant and susceptible brown planthoppers, Nilaparvata lugens $St{\aa}l$ (저항성 및 감수성 벼멸구 체내에서의 카보후란 대사)

  • Yoo, Jai-Ki;Ahn, Yong-Joon;Shono, Toshio;Lee, Si-Woo
    • The Korean Journal of Pesticide Science
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    • v.2 no.2
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    • pp.75-82
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    • 1998
  • This study was conducted to find out the biochemical or metabolic resistance mechanism of brown planthopper (BPH) to carbofuran. Differences between resistant ($LD_{50};\;20.3{\mu}g/g$) and susceptible strains($LD_{50};\;0.3{\mu}g/g$) were shown. The amounts of carbofuran metabolite, benzofuranol, and the origin, not developed by Thin Layer Chromatography, were much more in the susceptible strain. But the mother compound, carbofuran, was much more in the resistant strain. The tendencies of metabolism one and three hours after treatment were similar in both strains except for the amounts of metabolites described above. From the study, it is supposed that hydrolytic enzyme, esterase, changes its role from cleaving the esteric bond of carbofuran to making conjugates with carbofuran. This seems to be the main resistance mechanism of BPH to carbofuran. Oxidase and transferase may play little or no role in resistance mechanism. Oxidative and transferring enzymes gave no effects on the metabolism of carbofuran in the resistant strain compared with the susceptible strain.

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Potential Probiotic Properties of Exopolysaccharide Producing Lactic Acid Bacteria Isolated from Fermented Soybean Product (장류유래 Exopolysaccharide 생성 유산균의 잠재적 Probiotic 특성)

  • Ahn, Yu-Jin;Choi, Hye-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.749-755
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    • 2014
  • Exopolysaccharides (EPSs) have been widely used in the food industry as viscofying, stabilizing, and emulsifying agents as well as in the pharmaceutical industry for their immunomodulatory, anti-tumor, and anti-inflammatory effects. A total of 458 lactic acid bacteria (LAB) strains isolated from several kinds of soybean pastes were screened for the production of homo-EPS (HoPS). LAB isolates were primarily screened using thin layer chromatography (TLC) and further screened polymerase chain reaction (PCR) targeting genes involved in HoPS production. Six LAB isolates producing high amounts of HoPS were identified by TLC. Among these isolates, glucansucrase gene was amplified in two strains (JSA57, JSB22), whereas the fructansucrase gene was detected in three strains (JSA57, JSB22, JSB66). After isolating the strains, their morphological characteristics and 16S rDNA sequences were determined. Six species were identified as L. alimentarius HSB15, L. plantarum JSA22, L. pentosus JSA57, L. brevis JSB22, L. alimentarius JSB66, and L. parabrevis JSB89. To evaluate the potential probiotic properties of these LAB, their survival rates against a simulated intestinal environment were determined. After 2 hr of incubation in artificial gastric juice, survival rates of JSA57, JSB90, JSB22, and JSB66 were all greater than 50%. After 2 hr of incubation in bile juice, viable cell count of JSB22 was similar with initial vial cell counts. Growth of the six LAB was screened in arabino-oligosaccharide (AOS)-containing MRS broth. Results showed that growth of the isolates selectively increased after culture in AOS-containing media. Strain JSB22 (6 hr), JSB66 (6 hr), HSB15 (20 hr), and JSA22 (29 hr) showed maximum growth rate. Especially, JSB22 showed the highest growth rate. These results suggest that EPS-producing LAB isolated from Deonjang could be applied as synbiotics.

Correlations between the Fatty Acid Composition of Serum Phospholipids and Blood Pressure

  • Lee, Yang-Cha-Kim;Shim, Yoo-Jin;Chung, Eun-Jung;Kim, Soo-Yeon;Yangsoo Jang;Lee, Jong-Ho
    • Nutritional Sciences
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    • v.5 no.2
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    • pp.68-74
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    • 2002
  • Studies of the relationship between the composition of serum fatty acids and blood pressure are complex and controversial. Fatty acids, important constituents of biological membranes, could potentially affect vasoreactivities including blood pressure. In this study the compositions of fatty acids in serum phospholipids were compared between three types of hypertensive subjects (men, pre-menopausal women, and post-menopausal women) and their respective nrmotensive controls. Serum lipids were extracted and phospholipids were separated by thin layer chromatography. The percentage of palmitic acid (16 : 0) in serum phospholipids was significantly higher and the percentage of stearic acid (18 : 0) was significantly lower in all three hypertensive groups, compared with their corresponding control groups. Only in the group of post-menopausal women, palmitic acid was closely associated wish increases in both systolic (SBP) and diastolic blood pressure (DBP), while stearic acid was associated with decreases in both SBP and DBP. The polyunsaturated fatty acids in serum phospholipids behaved differently from saturated fatty acids. The ratios of products / precursor fatty acids, such as $\sumLCPUFA\omega6/18 : 2\omega$6, 20 : 4$\omega$6/18 : 2$\omega$6, ∑LCPUFA$\omega$3/18 : 3$\omega$3 and 22 : 6$\omega$3/20 : 5$\omega$3, were all clearly associated with both SBP and DBP in hypertensive, post-menopausal women. Desaturation and elongation in fatty acid metabolism could affect the bioavailability of eicosanoid precursors. Changes in the constituent fatty acids of phospholipids and eicosanoid precursors may also influence fluidity, ionic transport, hormone receptors and enzyme activities in biological membranes. In conclusion, both systolic and diastolic blood pressure in post-menopausal women was positively associated with the level of palmitic acid, and negatively associated with the level of stearic acid, in serum phospholipids. The relationships between serum phospholipid-$\omega$6 and $\omega$3 series fatty acids and blood pressure in women, especially in post-menopausal women, require further investigation by taking into consideration hormonal status and eicosanoid metabolism. Funker study is needed to determine the value of dietary manipulation of fatty acid constituents of serum phospholipids, relating to hypertension in women.

Characterization of Nitrile-hydrolyzing Enzymes Produced from Rhodococcus erythropolis (니트릴 분해효소 생산균인 Rhodococcus erythropolis의 발굴 및 효소 특성 연구)

  • Park Hyo-Jung;Park Ha-Joo;Uhm Ki-Nam;Kim Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.204-210
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    • 2006
  • Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.

Effects of Steroid Hormones on $In$ $Vitro$ GVBD and Oocyte Steroidogenesis in Blacktip Grouper, $Epinephelus$ $fasciatus$

  • Hwang, In-Joon;Kim, Seol-Ki;Choi, Sang-Jun;Lee, Chi-Hoon;Lee, Young-Don;Kim, Hyung-Bae;Baek, Hea-Ja
    • Development and Reproduction
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    • v.16 no.1
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    • pp.39-45
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    • 2012
  • To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, $Epinephelus$ $fasciatus$, we incubated vitellogenic oocytes (0.41 and 0.50 mm in average diameter) in the presence of exogenous steroid precursor ($[^3H]17{\alpha}$-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-$17{\beta}$, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 mm were more abundant than those of 0.41 mm. Also, we investigated the $in$ $vitro$ effects of human chorionic gonadotropin (HCG; 5, 50 and 500 $IU/m{\ell}$), $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) and $17{\alpha},20{\beta}$-trihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}21P$; 5, 50 and 500 $ng/m{\ell}$, respectively) on oocyte maturation. In the oocytes of 0.41 mm, treatment with 50 IU HCG stimulated GVBD ($55.30{\pm}1.20%$) compared with controls ($32.41{\pm}3.13%$, $p$<0.05). In the oocytes of 0.50 mm, treatment of $17{\alpha}20{\beta}P$ (50 and 500 $ng/m{\ell}$) stimulated GVBD ($50.13{\pm}2.52$ and $51.77{\pm}5.91%$, respectively) compared with controls ($36.81{\pm}2.89%$, $p$<0.05). Treatment with 500 IU HCG also stimulated GVBD ($49.59{\pm}5.15%$) compared with controls ($p$<0.05). Taken together, these results suggested that both HCG and $17{\alpha}20{\beta}P$ were effective on in vitro oocyte maturation and $17{\alpha}20{\beta}P$ may act as a maturation inducing hormone in blacktip grouper.

Isolation of Growth Inhibition Substance on Food borne Microorganisms from Hypericum ascyron L. and Application to Food Preservation (물레나물(Hypericum ascyron L.)의 식중독 미생물 증식 억제 물질의 분리 및 식품적용)

  • Han, Ji-Sook;Lee, Ji-Young;Baek, Nam-In;Back, Il-Woung;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.274-282
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    • 2002
  • The ethanol extract and n-hexane fraction from Hypericum ascyron L. showed strong growth inhibition at 25 ppm on 5 strains of Listeria monocytogenes for 72 hr at $32^{\circ}C$. The purified substance, H2-5-2 fraction, was isolated by silica gel column and preparative thin layer chromatography from n-hexane fraction of Hypericum ascyron L. The H2-5-2 fraction showed a strong bacteriostatic activity on 5 strains of L. monocytogenes at 10 ppm in tryptic soy broth, and the viable cell was reduced 1 log cycle compared to initial cell number. The n-hexane fraction of Hypericum ascyron L. showed strong growth inhibition at 25 ppm on Bacillus cereus and Staphylococcus aureus, and at 50 ppm on Vibrio parahaemolyticus for 72 hr. The purified antimicrobial substance, the H2-5-2 fraction, was assumed as high unsaturated sterol by $^1H-NMR$ and $^{13}C-NMR$. On application test using minced Alaska pollack and ground beef, the n-hexane fraction of Hypericum ascyron L. at the level of 250 ppm was applied at $32^{\circ}C$ and $5^{\circ}C$. At $32^{\circ}C$ storage condition, the antimicrobial substances did not reduced L. monocytogenes ATCC 19113, meanwhile at $5^{\circ}C$ storage condition, L. monocytogenes ATCC 19113 was reduced in viable number.

$^{99m}Tc$ Labeling and Biodistribution of PRODD as a Cerebral Blood Flow Imaging Radiopharmaceutical (뇌혈류 영상용 방사성의약품 PRODD의 $^{99m}Tc$ 표지 및 생체내분포)

  • Chung, Soo-Wook;Jeong, Jae-Min;Lee, Dong-Soo;Cho, Jung-Hyuck;Oh, Seung-Joon;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.3
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    • pp.328-331
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    • 1995
  • $^{99m}Tc$ labeled PnAO(propylene amine oxime) derivatives have been widely studied as brain perfusion agents. We synthesized and characterized a PnAO derivative, (RR/SS/ meso)-4,8-diaza-3,9-dimethylundecane-2, 10-dione bisoxime(PRODD). Proton-NMR spectroscopy and thin layer chromatography(silica gel) were performed for its characterization. PRODD was labeled with $^{99m}Tc$ using stannous chloride as reducing agent. The labeling efficiency was determined to be about 85%. Brain uptakes of $4.16{\pm}0.42$ %ID/g and $3.24{\pm}0.13$ %ID/g were found after 10min and 30min after intravenous injection. Brain-to-blood ratios were 1.17 and 0.75 at 10 and 30 minutes, which were lower than 1.3 and 1.9 of the ratios with commercial ${\pm}$-HMPAO. Autoradiographs of rat brain sections showed the gray matter to white matter ratios of $1.13{\pm}0.10$ at 30 min after intravenous injection, which was lower than $1.94{\pm}0.19$ of commercial $^{99m}Tc$-HMPAO. With the above findings, we concluded that the lipophilic $^{99m}Tc$-PRODD complex was able to cross the blood-brain barrier, however the complex showed lower uptake than $^{99m}Tc$-HMPAO in mouse or rat brains. We could suggest possibility that PRODD could be used as another $^{99m}Tc$ chelator.

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