• Title/Summary/Keyword: Thin Layer Chromatography(TLC)

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Isolation of a New Agar Degrading Bacterium, Maribacter sp. SH-1 and Characterization of its Agarase (신규 한천분해세균 Maribacter sp. SH-1의 분리 및 효소 특성조사)

  • Lee, Chang-Eun;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.156-162
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    • 2016
  • In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.

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.

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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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.

A Study on the Lipid Components in Oyster Mushroom (느타리버섯의 지방성분(脂肪成分)에 관한 연구(硏究))

  • Kwon, Yong-Ju;Uhm, Tai-Boong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.2
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    • pp.175-180
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    • 1984
  • Lipids in oyster mushroom (Pleurotus florida) were extracted by the mixture of chloroform-methanol (2: 1, v/v) and fractionated into neutral lipids, glycolipids and phospholipids by silicic acid column chromatography. Components and fatty acid composition of each fraction were deter- mined by thin-layer and gas-liquid chromatographies. Fresh oyster mushroom contained 0.5% total lipid in which the contents if neutral lipids, glycolipids and phospholipids were 33.8%, 19.7% and 45. 6%, respectively, Triglycerides(38.2%), free fatty acids (20%) and free sterol (10%) were the major components among the neutral lipids. Diglycerides, monoglycerides, sterol esters and three unidentified neutral lipids were the minor components. Major components of glycolipids were steryl glycosides(35.9%) and esterified steryl glycosides (23.7%). Digalactosyl diglycerides, mono-galactosyl diglycerides and two unknown components were also present. Of the phospholipids, phosphatidyl cholines and serines (48.2%), and phosphatidyl ethanolamines(44.4%) were the major components. On the other hand, the major fatty acids of neutral lipids we.e linoleic, palmitoleic, oleic and palmitic acid. Linoleic and palmitic acid were the predominant fatty acids of both glycolipids and phospholipids.

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Studies on the Lipid Composition of Bush Clover (Lespedeza bicolor) Seed (싸리종자(Lespedeza bicolor)의 지질성분(脂質成分)에 관한 연구(硏究))

  • Kim, Hyang-Ran;Koh, Moo-Seok;Yang, Hee-Cheon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.3
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    • pp.75-84
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    • 1987
  • Lipids in Bush Clover (Lespedza bicolor) seed were extracted with the mix ture of chloro-form-methanol (2 : 1, v/v) and then fractionated into neutral lipids, glycolipids and phospholipids by silicic acid column chromatography. Components and fatty acid composition of each fraction were determined by thin layer and gas chromatographies. The results were summarized as follows. In Bush Clover seed, the contents of neutral lipids, glycolipids and phospholipids were 71.75%, 23.26% and 4.99% respectively. Triglycerides(61.90%) and free fatty acids(22.04%) were the major components among the neutral lipids. Esterified sterols, free sterols, diglycerides and monoglycerides were the minor components. The major components of glycolipids were monogalactosyl diglycerides(38.19%) the others were esterified steryl glycosides, cerebrosides and digalactosyl diglycerides. The major components of the phospholipids were phosphatidyl cholines(36.46%), phosphatidyl inositols(21.52%) and phosphatidyl ethanolamines(17.29%). The major fatty acid of total lipid, neutral lipids and glycolipids were linoleic acid, linolenic acid, oleic acid and palmitic acid. On the other hand, predominate fatty acid of phospholipids were linoleic acid, palmitic acid, linolenic acid and stearic acid.

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Process Optimization of Ginseng Berry Extract Using Mixed Solvent and its Ginsenoside Analysis (혼합용매를 이용한 진생베리 추출물 최적 공정 개발 및 추출물의 진세노사이드 분석)

  • Ha, Yoo-jin;Kim, Mee-Ree;Yoo, Sun-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7794-7800
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    • 2015
  • Ginsenosides in ginseng berry has been known as functional materials showing physiological effect to the human. Specially, ginseng berry contains plenty of ginsenoside Re, but the study of extraction processes were not enough performed. Accordingly, the purpose of this study was to establish the optimized extraction process for obtaining ginsenoside Re from ginseng berry. The extraction process of ginsenosides was performed in 250 mL extraction flask containing 150 solvent and 10 g of dried ginseng berry. The extracted ginsenoside Re, Rg1 and Rd and total crude ginsenosides from ginseng berry were evaluated by TLC according to the treated conditions (the ratio of alcohol to water, extraction temperature, extraction period, and extraction times). Optimized conditions for extraction was 70% to 30% of the ratio of alcohol to water, $80^{\circ}C$ of extraction temperature, 4 h of extraction period, and 2 times of extraction frequency. The amount of total crude ginsenosides of the extract obtained from the optimized process was 88.6 mg/g based on dried ginseng berry. The composition of ginsenosides from the extracted was 5.5% of Rb1, 5.2% of Rc, 14.3% of Rd, 51.5% of Re, 8.1% of Rf, and 15.7% of Rg1. A protopanaxtriol ginsenosides of whole ginsenosides extracted was about 80%.

Physicochemical Properties and Biological Activities of Angelica gigas Fermented by Saccharomyces cerevisiae (유용 효모균주를 이용한 발효참당귀분말 추출물의 이화학적 특성 및 생리활성 효과)

  • Sim, So-Yeon;Park, Woo-Sang;Shin, Hyun-Seung;Ok, Min;Cho, Young-Su;Ahn, Hee-Young
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1136-1143
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    • 2019
  • The purpose of this study was to investigate the biological activities of an aqueous extract of Angelica gigas (Ag) fermented by Saccharomyces cerevisiae (Sc). First, the soluble solids of the F/3 group, in which the Ag was fermented by Sc for 3 days, decreased from $1^{\circ}Bx$ to $0.9^{\circ}Bx$. On the other hand, the pH increased with the number of days of fermentation. The result of a TLC experiment confirmed that it gradually decomposed into a low-molecular weight sugar form upon fermentation. The total phenolic compounds and flavonoid contents were higher in the fermented group than in the non-fermented group. K and Ca contents were increased by fermentation in the following order: F/3, NF, and F/0 groups. Decursin and decursinol angelate contents were highest in the F/3 group. The DPPH (${\alpha}$, ${\alpha}{\prime}$-diphenyl-${\beta}$-picrylhydrazyl) radical scavenging activity of the NF, F/0, and F/3 groups were 41.89%, 39.51%, and 60.26%, respectively. The inhibition activities of tyrosinase and lipoxygenase were stronger in the F/3 group than in the NF group. This experiment showed that the fermentation of Ag Nakai can lead to an increase in its antioxidant ability, physiological activity, whitening and anti-inflammatory effects. Thus, this oriental herbal medicine can be developed into a functional material that can be utilized in the development of cosmetic products in future.

Isolation of an Agarase-producing Persicobacter sp. DH-3 and Characterization of its β-agarase (Agarase를 생산하는 Persicobacter sp. DH-3의 분리 및 β-agarase의 특성)

  • Heo, Da-Hye;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.2
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    • pp.158-163
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    • 2019
  • The purpose of this study was to isolate a new marine agarase-producing bacterium. Agarase can hydrolyze agar and agarose to produce agarooligosaccharides or neoagarooligosaccharides, which possess many physiological functions. Strain DH-3 was isolated from seawater collected from the coast of Yeosu at Jeollanam province, Korea. A 16S rDNA sequence analysis showed this strain to be Persicobacter sp. DH-3. Extracellular agarase was prepared from culture media of Persicobacter sp. DH-3 and used for characterization. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 50, 55, 70, 100, 90, and 50%, respectively. Relative activities at pH 5, 6, 7, and 8 were 75, 100, 90, and 75%, respectively. The enzyme showed maximum activity at $50^{\circ}C$ in a 20 mM Tris-HCl buffer at pH 6. This enzyme could be useful, as agar is in liquid state at $50^{\circ}C$. Agarase activities were maintained at 80% or more for 2 hr at 20, 30, and $40^{\circ}C$. Thin layer chromatography analysis suggested that Persicobacter sp. DH-3 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarohexaose and neoagarotetraose. In addition, zymogram analysis confirmed that Persicobacter sp. DH-3 produces at least three agar-degrading enzymes with molecular weights of 45, 70, and 140 kDa. Therefore, it is expected that agarases from Persicobacter sp. DH-3 could be used to produce functional neoagarooligosaccharides.