• Title/Summary/Keyword: thin-layer chromatography

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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.

Isolation and Characterization of Inhibition Helicobacter pylori from Culture Media of Fomitopsis pinicola (Fomitopsis pinicola 균사체배양액의 Helicobacter pylori에 대한 저해물질의 분리 및 특성)

  • Lee, Jae-Kyoung;Choi, Seong-Woo;Hwang, Yu-Hyun;Park, Hee-Kuk;Yoo, Jeong-Weon
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.422-427
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    • 2006
  • The culture media from Fomitopsis pinicola were extracted by methanol and examined growth inhibition against Helicobacter pylori. The culture media from 8 days fermentation of F. pinicola showed maximum inhibition activity on H. pylori in 0.25 mg as MIC value. The highest activity against H. pylori by MHCS agar diffusion medium by Fp-P1 in 22.7 mm ID among 3 peaks from methanol fraction of 8 days culture media of Fomitopsis pinicola which purified by ion-exchange chromatography. The Fp-P1 from DEAE-Sephadex A-25 have been analysed by TLC as Fp-T1, Fp-T2 and Fp-T3 by ninhydrin staining. Fp-T3 (Rf value : 0.67) was higher activity against H. pylori in 14.4 mm ID. Fp-T3 was identified as single band by HPLC and TLC. It was assumed to aminosugar by BioLC analysis and TLC staining.

Studies on the Characteristics of Mineral Diluents Affecting the Decomposition of Sumithion in the Dust formulations (Sumithion 분제의 성분 변화에 영향을 미치는 증량제의 특성에 관한 연구)

  • Wuh K. D.;Han S. S.;Keum S. S.;Ahn S. H.;Lee C. N.
    • Korean journal of applied entomology
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    • v.10 no.2
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    • pp.77-83
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    • 1971
  • In order to find out the decomposing factors of Sumithion in mineral diluents, the physical and chemical properties of the diluents such as talcs, bentonites, diatomaceous earth and other clay minerals was studied in relation to the decomposition rate of Sumithion in dust formulations. The total base, moisture adsorption capacity, pH, specific surface, water contents, active $Fe^{++}$, ignition loss and cation exchange capacity were analysed as the properties of mineral diluents, and these properties were correlated with the stability of Sumithion in dust formulations. And in hope of finding out the main factors among those properties of diluents, the decomposed products of Sumithion in dust formulations prepared with standard Sumithion were separated by the methods of column chromatography and investigated by thin layer chromatography. The following results were otbained; 1. The total base, moisture adsorption capacity, specific surface, water contents, active $Fe^{++}$ and cation exchange capacity of mineral diluents were found to be highly effective on the rate of decomposition of Sumithion in dust formulations. 2. Decomposed products of Sumithion in dust formulations were found to be dimethylphosphorothionate, 3-methyl-4-nitro phenol and its derivatives. And one fraction was not dissolved in n-hexane and ethylether, but was soluble in methylalcohol and ethylalcohol. 3. The moat highly correlated properties of diluents with the decomposition rate of Sumithion in dust were found to be the total base and water contents. 4. In regard to the kind of diluents, it was found that the rate of decomposition of Sumithion in dust formulations was higher in order of bentonite, diatomaceous earth, kaolin and talc.

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Propolis from the Stingless Bee Trigona incisa from East Kalimantan, Indonesia, Induces In Vitro Cytotoxicity and Apoptosis in Cancer Cell lines

  • Kustiawan, Paula M;Phuwapraisirisan, Preecha;Puthong, Songchan;Palaga, Tanapat;Arung, Enos T;Chanchao, Chanpen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6581-6589
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    • 2015
  • Background: Previously, stingless bee (Trigona spp.) products from East Kalimantan, Indonesia, were successfully screened for in vitro antiproliferative activity against human cancer derived cell lines. It was established that propolis from T. incisa presented the highest in vitro cytotoxicity against the SW620 colon cancer cell line (6% cell survival in $20{\mu}g/mL$). Materials and Methods: Propolis from T. incisa was extracted with methanol and further partitioned with n-hexane, ethyl acetate and methanol. The in vitro cytotoxicity of the extracts was assessed by the MTT assay against human colon (SW620), liver (Hep-G2), gastric (KATO-III), lung (Chago) and breast (BT474) cancer derived cell lines. The active fractions were further enriched by silica gel quick column, absorption and size exclusion chromatography. The purity of each fraction was checked by thin layer chromatography. Cytotoxicity in BT-474 cells induced by cardanol compared to doxorubicin were evaluated by MTT assay, induction of cell cycle arrest and cell death by flow cytometric analysis of propidium iodide and annexin-V stained cells. Results: A cardol isomer was found to be the major compound in one active fraction (F45) of T. incisa propolis, with a cytotoxicity against the SW620 ($IC_{50}$ of $4.51{\pm}0.76{\mu}g/mL$), KATO-III (IC50 of $6.06{\pm}0.39{\mu}g/mL$), Hep-G2 ($IC_{50}$ of $0.71{\pm}0.22{\mu}g/mL$), Chago I ($IC_{50}$ of $0.81{\pm}0.18{\mu}g/mL$) and BT474 (IC50 of $4.28{\pm}0.14{\mu}g/mL$) cell lines. Early apoptosis (programmed cell death) of SW620 cells was induced by the cardol containing F45 fraction at the $IC_{50}$ and $IC_{80}$ concentrations, respectively, within 2-6 h of incubation. In addition, the F45 fraction induced cell cycle arrest at the G1 subphase. Conclusions: Indonesian stingless bee (T. incisa) propolis had moderately potent in vitro anticancer activity on human cancer derived cell lines. Cardol or 5-pentadecyl resorcinol was identified as a major active compound and induced apoptosis in SW620 cells in an early period (${\leq}6h$) and cell cycle arrest at the G1 subphase. Thus, cardol is a potential candidate for cancer chemotherapy.

A Study on Paralytic Shellfish Poison of Sea Mussel, Mytilus edulis -Food Poisoning Accident in Gamchun Bay, Pusan, Korea, 1986- (진주담치의 마비성독에 관한 연구 -1986년 부산 감천만 중독사고를 중심으로-)

  • CHANG Dong-Suck;SHIN Il-Shik;PYEUN Jae-Hyeung;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.4
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    • pp.293-299
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    • 1987
  • At various times and places all over the world men have become ill and some have died after eating shellfish that were intoxicated with paralytic shellfish poison(PSP) caused by Protogonyaulax spp. In late March, 1986, two persons were dead by ingesting wild sea mussels, Mytilur edulis, grown at bottom of an anchored waste ship to be dismantled at Gamchun Bay, Pusan, Korea. The samples were collected from the bottom of the ship during April $1\~April$ 8 of the year to find the cause of the food poisoning accident. The toxicity was estimated by bioassay with ICR male mouse, while the toxins were extracted and characterized. The toxins were extracted with acidified $80\%$ ethanol. The extract was defatted three times with dichloromethane, treated with activated charcoal, and then purified by chromatography on Bio-Gel P-2 and Bio-Rex 70. The toxic fractions obtained were analysed by cellulose acetate membrane electrophoresis, thin layer chromatography and high performance liquid chromatogaphy. The range and the average of PSP-toxicity of the samples were $132\~295\;MU/g$, 203 MU/g respectively. The amount of PSP was $26.4\~58.9{\mu}g/g$ of whole meat in range and $40.6{\mu}g/g$ in average. The toxicity of the digestive gland of the samples was 9 times higher than that of edible meat (except digestive gland) as $439\~979MU/g$, and it was about $70\%$ in total toxin. The compositional analytical results of the paralytic shellfish toxin, Gonyautoxin $1\~4$ were the major part of the PSP and Saxitoxin and neosaxitoxin were detected as the minor component. It was concluded that the food poisoning accident was caused not by Saxitoxins but by Gonyautoxins.

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Atomospheric Concentration and Mutagenicity of Nitroarenes of Suspended Particulates in Seoul (대기부유분진중 nitroarenes 오염도와 그 돌연변이원성에 관한 조사연구 - 서울시 일부지역을 중심으로 -)

  • Jo, Seong-Joon;Choi, Yong-Wook;Lim, Young-Wook;Shin, Dong-Chun;Chung, Yong
    • Environmental Analysis Health and Toxicology
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    • v.5 no.3_4
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    • pp.19-28
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    • 1990
  • Nitroarenes are ubiquitous environmental pollutants displaying potent mutagenicity in bacteria and carcinogenicity in mammal. In this study, the concentration of nitroarenes in coarse and fine particles and mutagenicity of POC$\_$N/ fraction was investigated in suspended particulates at the Shinchon and Bulkwang area of Seoul. The suspended particulates were collected bimonthly by a high volume cascade impactor air sampler from July 1987 to May 1988. Extractable organic matter was obtained by ultrasonic extraction on diethly ether/cyclohexane (8/2, v/v). Neutral fraction was obtained by liquid-liquid extraction. Polar neutral organic compounds (POC$\_$N/) was fractionated by thin-layer chromatography. Finally, the concentrations of nitroarenes in POC$\_$N/ fraction were measured and determined by capillary gas chromatography. Direct and indirect mutagenicity of POC$\_$N/ fraction were measured using Salmonella typhimurium TA 98. The result were as follows: 1) Major nitroarenes at the Shinchon area was 1-nitropyrene and at the Bulkwang area it was 2,7-dinitro-9-fluorenone during the year. 2) Average concentration of total nitroarenes measured was 67.26 ng/m$^3$in fine particles which was 1,3 folds higher that in coarse particle (52.30 ng/m$^3$). 3) Annual pattern of nitroarenes concentrations revealed that concentration during heating season (Feb., Jan., Mar.) was 2.2 folds higher than that in non heating season (May, Jul., Sep.). Concentration of each season has 157.68 ng/m$^3$and 80.39 ng/m$^3$. 4) The mutagenic activity of POC$\_$N/ fraction from fine particles was higher compared to that of coarse particles and was increased when metabolically activated, with 59 mixture. Mutagenicities, Metabolically activated, were significantly different between Shinchon and Bulkwang area, 322.8 rev/250 $\mu\textrm{g}$/plate and 286.8 rev/250 $\mu\textrm{g}$/plate, respectively. 5) Annual pattern of mutagenicity of POC$\_$N/ fraction revealed that mutagenicity during the heating season was 1.7 folds higher at Shinchon area and 1.2 folds higher at Bulkwang than during the non heating season. The variable contents and levels of nitroarenes in suspended particulates may affect human health significantly. Further studies such as risk assessment should be conducted on the basis of these kind of studies.

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Lipid Composition and Protein Pattern of Prunus Tomentosa Thunberg Seed (앵두(Prunus Tomentosa Thunberg)씨의 지방질 조성 및 단백질 패턴)

  • Yoon, Hyung-Sik;Park, Jin-Sang
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.248-252
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    • 1985
  • Prunus tomentosa Thunberg seed was investigated to evaluate its possibility for use as food resources of fats and proteins. The seed contained 40.38% of crude fat and 26.59% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.49% of neutral lipids, whereas compound lipids were only 4.51%. Among the neutral lipid components by thin-layer chromatography, triglycerides were 89.86%, sterols, monoglycerides, sterol esters, free fatty acids and diglycerides were 4.14%, 2.98%, 1.77%, 1.07%, and 0.18%, respectively. Oleic acid (65.06-66.05%) and linoleic acid (26.56-28.40%) were the main fatty acids in the total lipid, neutral lipid and triglyceride fractions. In the glycolipid and phospholipid fractions, predominant fatty acids were oleic acid (40.55-51.46%), linoleic acid (20.26-30.89%) and palmitic acid (17.64-21.43%). The extractability of salt soluble protein of seed was 60%, and recovery rate of main protein fraction separated by Sephadex G-200 was about 46.5%. The electrophoretic analysis showed 7 bands in seed protein.

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Studies on antimicrobial agent produced by lactobacillus acidophilus (Lactobacillus acidophilus가 생산한 항균물질(抗菌物質)에 관한 연구(硏究))

  • Kim, Dong-shin
    • Korean Journal of Veterinary Research
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    • v.24 no.2
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    • pp.149-162
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    • 1984
  • The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

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Quantitative Fractionation of Total Lipids and Their Fatty Acid Composition in Korean Yam Tubers. (한국산 마지질의 분획 정량과 지방산조성)

  • Kim, Yong-Sun;Kim, Sang-Soon;Kim, Chul-Jai;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.652-657
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    • 1995
  • To investigate the lipid components of Korean yams(Dioscorea batatas and Dioscorea aimadoimo), silicic acid column, thin layer and gas chromatographies were applied. Total crude lipid contents in D. batatas and D. aimadoimo were 1.70% and 1.31%, and total purified lipid contents were 1.35% and 0.99% on a dry weight basis. Lipids were fractionated and quantitated by silicic acid column chromatography. The contents of neutral lipids, glycolipids and phospholipids were 43.33, 39.60 and 17.07% in D. batatas and 43.94, 34.74 and 21.32% in D. aimadoimo. Neutral lipids were the most abundant, in which triglycerides and sterol ester were found as the major components. Glycolipids were mainly composed of monogalactosyl and trigalactosyl diglycerides. Phosphatidyl choline and phosphatidyl ethanolamine were the major components of phospholipid. Fatty acids of the total lipids were mainly linoleic and palmitic acids. The fatty acid compositions of neutral lipids, glycolipids and phospholipids were the same tendency as that of total lipids. However, the content of linolenic acid in glycolipids was higher than in neutral lipids and phospholipids when fractionated.

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Study of Tannin Reducing Effect of Aronia by Yeast Isolated from Jeotgal (젓갈에서 분리된 효모를 이용한 아로니아의 탄닌 성분 저감화 효과에 관한 연구)

  • Shin, Hyo-Ju;Byun, Ock-Hee;Kim, Yu-Jin;Bang, Bo-Yeon;Park, Jung Min;Jeong, Yong-Seob;Bai, Dong-Hoon
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.247-252
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    • 2015
  • Aronia (Black chokeberry, Aronia melanocarpa) belonging to the Rosaceae family, is native to eastern North America. Aronia contain high levels of flavonoids, mostly anthocyanins and proanthocyanidins, which are known as condensed tannins. The dominant proanthocyanidins in aronia are (-)-epicatechin and (+)-catechin. The concentration of proanthocyanidins in aronia is higher than in other berries, however due to the astringent taste it is not desirable for consumption. Therefore, the purpose of this study is to evaluate the effect of aronia on the reduction in tannins by yeast isolated from regional Jeotgal. We isolated strains of yeast with high ${\beta}$-glucosidase activity from Jeotgal, with the MTY2 strains exhibiting a reduction in final tannin concentration according to thin layer chromatography (TLC) analysis. MTY2 was confirmed as Kazachstania servazzii using an 18S rDNA sequence and named as K. servazzii MTY2. K. servazzii MTY2 showed most significant growth when K. servazzii MTY2 was cultured in a solution of 10% (w/v) glucose, 3% (w/v) tryptone and 0.1% (w/v) sodium chloride. According to the high performance liquid chromatography (HPLC) analysis, the (+) - catechin peak is present, but (-) - epicatechin peak was reduced at culture condition added with 10% glucose in medium.