• Title/Summary/Keyword: Gel Layer

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Production of Oleamide, a Functional Lipid, by Streptomyces sp. KK90378

  • Kwon, Jeong-Ho;Hwang, Sung-Eun;Han, Jae-Taek;Kim, Chang-Jin;Rho, Jung-Rae;Shin, Jong-Eon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1018-1023
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    • 2001
  • Oleamide (cis-9-octadecenamide) is endogenous primary amide of fatty acid that is produced in small amounts in animal brains. It is known to induce sleep and to lower temperature by destroying the lipid plasma membrane structure of cells, thereby disclosing gap junction channels. To develop a new biological production method for oleamide, a screening program was conducted to isolate a microorganism producing oleamide. Among 1,500 soil microorganisms tested, KK90378 exhibited a potent positive reaction with Dragendoff`s reagent, used to detect the primary amide of oleamide. KK90378 was identified as a Streptomyces species based on cultural and morpohological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extrat. Streptomyces sp. KK90378 produced oleamide 3 days after culture at $28^{\circ}C$, pH 7.2 A series of purification steps, including hexane extraction, silica gel column, and preparative thin layer chromatographies, were performed for the purification of oleamide. A spectrophotometric analysis using $^1H$, $^13C$-NMR, and GC-MS confirmed that the chemical structure of the purified oleamide was identical to that of authentic oleamide.

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An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1001-1010
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    • 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.

Inverse HPLC approach for the evaluation of repulsive interaction between ionic solutes and a membrane polymer

  • Kiso, Yoshiaki;Kamimoto, Yuki;Hosogi, Katsuya;Jung, Yong-Jun
    • Membrane and Water Treatment
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    • v.6 no.2
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    • pp.127-139
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    • 2015
  • Rejection of ionic solutes by reverse osmosis (RO) and nanofiltration (NF) membranes is controlled mainly by electrochemical interaction as well as pore size, but it is very difficult to directly evaluate such electrochemical interaction. In this work, we used an inverse HPLC method to investigate the interaction between ionic solutes and poly (m- phenylenediaminetrimesoyl) (PPT), a polymer similar to the skin layer of polyamide RO and NF membranes. Silica gel particles coated with PPT were used as the stationary phase, and aqueous solutions of the ionic solutes were used as the mobile phase. Chromatographs obtained for the ionic solutes showed features typical of exclusion chromatographs: the ionic solutes were eluted faster than water (mobile phase), and the exclusion intensity of the ionic solute decreased with increasing solute concentration, asymptotically approaching a minimum value. The charge density of PPT was estimated to be ca. 0.007 mol/L. On the basis of minimum exclusion intensity, the exclusion distances between a salt and neutralized PPT was examined, and the following average values were obtained: 0.49 nm for 1:1 salts, 0.57 nm for 2:1 salts, 0.60 nm for 1:2 salts, and 0.66 nm for 2:2 salts. However, $NaAsO_2$ and $H_3BO_3$, which are dissolved at neutral pH in their undissociated forms, were not excluded.

Screening and Characterization of a Novel Cellulase Gene from the Gut Microflora of Hermetia illucens Using Metagenomic Library

  • Lee, Chang-Muk;Lee, Young-Seok;Seo, So-Hyeon;Yoon, Sang-Hong;Kim, Soo-Jin;Hahn, Bum-Soo;Sim, Joon-Soo;Koo, Bon-Sung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1196-1206
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    • 2014
  • A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

Production and Characterization of Lipopeptide Biosurfactant from Bacillus subtilis A8-8

  • Lee Sang-Cheol;Yoo Ju-Soon;Kim Sun-Hee;Chung Soo-Yeol;Hwang Cher-Won;Joo Woo-Hong;Choi Yong-Lark
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.716-723
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    • 2006
  • A biosurfactant-producing bacterial strain was selected from oil-contaminated soil because of its ability to degrade crude oil and tributyrin $(C_{4:0})$. The strain was identified as Bacillus subtilis A8-8 based on its morphological, biochemical, and physiological characteristics. When B. subtilis A8-8 was grown with crude oil as the sole carbon source, the biosurfactant from the strain emulsified crude oil, vegetable oil, and hydrocarbons. Soybean oil was the optimum substrate for the emulsifying activity and emulsion stability of the biosurfactant, both of which were superior to those of several commercially available surfactants. The biosurfactant was purified by a procedure including HCl precipitation, methanol treatment, and silica-gel chromatography. The partially purified biosurfactant was analyzed by TLC (thin-layer chromatography), SDS-PAGE, and HPLC and it reduced the surface tension of water from 72 mN/m to 26 mN/m at a concentration of 30 mg/l. Therefore, the purified lipopeptide biosurfactant has strong properties as an emulsifying agent and acts as an emulsion-stabilizing agent.

Extracts from Pinus densiflora Siebold et Zuccarini Sawdust Inhibit the Mycelial Growth of Lentinus edodes (Berk.) Sing., Edible Mushroom

  • Hong, Seong-Cheol;Choi, Myung-Suk;Lim, Bu-Kug;Lee, Jong-Yoon;Rinker, Danny L.;Yang, Jae-Kyung
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.349-354
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    • 2004
  • The extracts of Pinus densiflora sawdust by n hexane, ethyl acetate and methanol solvent were investigated to identify their mycelial growth inhibition against Lentinus edodes. The yields of n hexane soluble fraction, ethyl acetate-soluble fraction, and methanol soluble fraction from P. densiflora sawdust were obtained 1.36%, 2.21% and 4.03% using organic solvent, respectively. The mycelial growth inhibition of L. edodes was the greatest for n hexane extract, ranging from 36.5% to 47.6% at concentrations of 125 ppm to 1,000 ppm, with the values for all concentrations significantly different from one another. After direct extraction of P. densiflora sawdust using n hexane, ethyl acetate and methanol, each extract was separated into three fractions by silica gel column chromatography and then the fractions were isolated on the values of $R_f$ by thin layer chromatography. The mycelial growth inhibition against L. edodes was recognized in the fractions II (33.5%) and III (37.6%) of n hexane extract, the fraction II (21.4%) of ethyl acetate extract and the fraction II (26.4%) of methanol extract. The fractions III of n-hexane extract showed the highest growth inhibition among the nine fractions of the organic solvent extract.

Gas Permeation Characteristics of Silica Membrane Prepared by Ultrasonic Spray Pyrolysis (초음파 분무 열분해법에 의해 합성한 실리카 막의 기체 투과 특성)

  • Lee Kew-Ho;Youn Min-Young;Park Sang-Jin;Lee Dong-Wook;Sea Bongkuk
    • Membrane Journal
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    • v.15 no.2
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    • pp.105-113
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    • 2005
  • Silica membranes were prepared on a porous metal sheet by ultrasonic spray pyrolysis method for gas separation at high temperatures. In order to improve the permselectivity, silica was deposited in the sol-gel derived $silica/\gamma-alumina$ intermediate layer by pyrolysis of tetraethyl orthosilicate (TEOS) at 873 K. The pyrolysis with forced cross flow through the porous wall of the support was very effective in plugging mesopores, Knudsen diffusion regime, that were left unplugged in the membranes. At permeation temperature of 523 K, the silica/alumina composite membrane showed $H_2/N_2$ and water/methanol selectivity as high as 17 and 16, respectively, by molecular sieve effect.

Micro Emulsion Synthesis of LaCoO3 Nanoparticles and their Electrochemical Catalytic Activity

  • Islam, Mobinul;Jeong, Min-Gi;Ghani, Faizan;Jung, Hun-Gi
    • Journal of Electrochemical Science and Technology
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    • v.6 no.4
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    • pp.121-130
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    • 2015
  • The micro emulsion method has been successfully used for preparing perovskite LaCoO3 with uniform, fine-shaped nanoparticles showing high activity as electro catalysts in oxygen reduction reactions (ORRs). They are, therefore, promising candidates for the air-cathode in metal-air rechargeable batteries. Since the activity of a catalyst is highly dependent on its specific surface area, nanoparticles of the perovskite catalyst are desirable for catalyzing both oxygen reduction and evolution reactions. Herein, LaCoO3 powder was also prepared by sol-gel method for comparison, with a broad particle distribution and high agglomeration. The electro catalytic properties of LaCoO3 and LaCoO3-carbon Super P mixture layers toward the ORR were studied comparatively using the rotating disk electrode technique in 0.1 M KOH electrolyte to elucidate the effect of carbon Super P. Koutecky-Levich theory was applied to acquire the overall electron transfer number (n) during the ORR, calculated to be ~3.74 for the LaCoO3-Super P mixture, quite close to the theoretical value (4.0), and ~2.7 for carbon-free LaCoO3. A synergistic effect toward the ORR is observed when carbon is present in the LaCoO3 layer. Carbon is assumed to be more than an additive, enhancing the electronic conductivity of the oxide catalyst. It is suggested that ORRs, catalyzed by the LaCoO3-Super P mixture, are dominated by a 2+2-electron transfer pathway to form the final, hydroxyl ion product.

A comparative evaluation of $CO_2$ and erbium-doped yttrium aluminium garnet laser therapy in the management of dentin hypersensitivity and assessment of mineral content

  • Belal, Mahmoud Helmy;Yassin, Abdulaziz
    • Journal of Periodontal and Implant Science
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    • v.44 no.5
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    • pp.227-234
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    • 2014
  • Purpose: Dentin hypersensitivity is a potential threat to oral health. Laser irradiation may provide reliable and reproducible treatment but remains controversial. The present study aimed to evaluate the effects of $CO_2$ or erbium-doped yttrium aluminium garnet (Er:YAG) laser therapy, and to assess mineral content. Methods: Eighteen human single-rooted teeth affected with advanced periodontitis were obtained. Buccal and lingual surfaces were planed to form 36 specimens. Ethylenediaminetetraacetic acid gel (24%) was applied to remove the smear layer and simulate hypersensitive teeth. The experimental groups were: group 1, control (no irradiation); group 2, $CO_2$ laser (repetitive pulsed mode, 2 W, $2.7J/cm^2$); and group 3, Er:YAG laser (slight contact mode, 40 mJ/pulse and 10 Hz). To evaluate dentinal tubule occlusion, six specimens per group (2-mm thickness) were prepared and observed using scanning electron microscopy (SEM) for calculation of the occlusion percentage. To evaluate the mineral content, six specimens per group (0.6-mm thickness) were used, and then the levels of Ca, K, Mg, Na, and P were measured by inductively coupled plasma-atomic emission spectrometry. In addition, the surface temperature of the specimens during laser irradiation was analyzed by a thermograph. Results: The SEM photomicrographs indicated melted areas around exposed dentinal tubules and a significantly greater percentage of tubular occlusion in the $CO_2$ and Er:YAG laser groups than the control, and in the Er:YAG group than the $CO_2$ laser group. In addition, no significant differences were noted among the experimental groups for the mineral elements analyzed. The $CO_2$ laser group showed an evident thermal effect compared to the Er:YAG group. Conclusions: $CO_2$ and Er:YAG laser are effective in treating dentin hypersensitivity and reducing its symptoms. However, the Er:YAG laser has a more significant effect; thus, it may constitute a useful conditioning item. Furthermore, neither $CO_2$ nor Er:YAG lasers affected the compositional structure of the mineral content.

Alkaloids from the Tuber of Aconitum chiisanense (지리바꽃 괴경의 알카로이드)

  • Lee, Moo-Taek;Sung, Hwan-Kil;Whang, Wan-Kyunn;Kim, Il-Hyuk
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.161-173
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    • 1997
  • Tuber of Aconitum chiisanense(Ranunculaceae) a specific medicinal plant in Korea, which is known to have the activity to recover reduced metabolism of feeble patients and has been used to symptoms such as pain, paralysis, atonia and coldness of extremities, etc. were studied. The powdered tubers of the plant were extracted with 10% EtOH 3 times and the combined extract was dissolved in 1N HCl solution and washed with ethyl acetate. The aqueous layer was basified with solid $Na_2CO_3$ and extracted with $CHCl_3$ to obtain an alkaloidal fraction. The alkaloidal fraction was subjected to column chromatography using silica gel, alumina and Sephdex LH 20, etc. From the alkaloidal fraction, five diterpene alkaloids, mesaconitine, aconitine, hypaconitine, 8-O-ethyl 14-benzoylmesaconine and talatizamine, were isolated and identified on the basis of their physico-chemical properties and spectroscopic evidences($^1H$-, $^{13}C$-NMR, EI-MS, IR, 2D-NMR) respectively. Especially the Compound IV, 8-O-ethyl 14-benzoylmesaconine, was assumed to be an artifact resulting from mesaconitine during extraction procedures. The contents of mesaconitine, aconitine and hypaconitine in the mother tuber of this plant were 0.300%, 0.024%, and 0.068%. And that of the attached tuber(new one) of this plant were 0.336%, 0.034% and 0.240% respectively.

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