• The Korean Journal of Zoology
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• v.38 no.4
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• pp.490-497
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• 1995

In vitro tissue culture of fat body of Hyphantria cunea in the medium containing [35S]-methionine reveaied that storage protein-i (SP-1) is taken up into fat body of prepupae and 1-day-old pupae. Using Western blotting and ligand binding method, we were able to identify the protein band of the SP-1 receptor protein. For the partial purification, the membrane proteins of fat body cells were solubilixed with 1% Triton X-1OO and applied to anion exchange chromatography. The results revealed the molecular weight of the receptor protein to be about 80 kl)a in SDSPAGE, and the P1 was estimated to be about 6.1. The mobility of the receptor protein in 8D8-PAGE was highly dependent on both temperature during electrophoresls and the condition of samples whether they were in reducing or nonreducing.

• Food Science and Biotechnology
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• v.16 no.1
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• pp.127-132
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• 2007

[ ${\alpha},\;{\alpha}$ ]-Trehalose was efficiently modified by a transgalactosylation reaction of Escherichia coli ${\beta}-galactosidase$ using lactose as a donor to yield two galactosyl trehalose trisaccharides. The reaction products of trehalose by the enzyme were observed by thin layer chromatography (TLC) and high performance anion exchange chromatography (HPAEC) and were purified by BioGel P2 gel permeation chromatography and recycling preparative HPLC. Liquid chromatography-mass spectrometry (LC-MS) and ^{13}C$ nuclear magnetic resonance (NMR) analyses revealed that the structures of the main products were $6^2-{\beta}-D-galactosyl$ trehalose (1) and $4^2-{\beta}-D-galactosyl$ trehalose (2). A reaction of 30%(w/v) trehalose and 15%(w/v) lactose at pH 7.5 and $45^{\circ}C$ resulted in a total yield of approximately 27-30% based on the amount of trehalose used. The galactosyl trehalose products were not hydrolyzed by trehalose. In addition the mixture of transfer products (9:1 ratio of 1 to 2) showed higher thermal stability than glucose, lactose, and maltose, but less than trehalose, against heat treatment over $100^{\circ}C$ at pH 4 and 7. It also exhibited better thermal stability than sucrose at pH 4 alone.

• Preventive Nutrition and Food Science
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• v.18 no.4
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• pp.273-279
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• 2013

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.852-858
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• 2008

An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.

• Korean Journal of Environmental Agriculture
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• v.30 no.4
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• pp.367-376
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• 2011

BACKGROUND: In order to develop effective assessment method for Korean paddy soil microbial community structure, reliable genomic DNA extraction method from paddy soil and quantitative real-time PCR (qRT-PCR) method are needed to establish METHODS AND RESULTS: Out of six conventional soil genomic DNA extraction methods, anion exchange resin purification method was turn to be the most reliable. Various PCR primers for distinguishing five bacterial phylum (${\alpha}$-Proteobacteria, ${\beta}$-Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes), all bacteria, and all fungi were tested. Various qRT-PCR temperature conditions were also tested by repeating experiment. Finally, both genomic DNA extraction and qRT-PCR methods for paddy soil were well established. CONCLUSION: Quantitative real-time PCR (qRT-PCR) method to assess paddy soil microbial community was established.

• Analytical Science and Technology
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• v.17 no.4
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• pp.302-307
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• 2004

Determination of actinides in the environmental sample requires separation of each element. This procedure is tedious and time consuming. And also, the detection limits of some nuclides using alpha spectrometry are rather higher. To overcome the lower detection limit and complicated separation procedure, a simple analytical technique for the determination of actinide isotopes in the environmental samples was developed and applied to IAEA and NIST reference sediment samples. For the separation of actinides from matrix, anion exchange resin and TRU-spec extraction chromatography resin were used and chemical yields were obtained using natural uranium, thorium, $^{242}Pu$ and $^{243}Am$ tracers. For overcoming the higher detection limits of U and Th in alpha spectrometry, neutron activation analysis was applied. Using combined method, the detection limit was increased about 10 times. The activity values of each isotope were consistent with the reference values reported by IAEA and NIST.

• Journal of the Korean Institute of Gas
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• v.21 no.1
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• pp.1-5
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• 2017

In the event of toxic gas accidents, soil deposition is a main factor which has an effect on extent of the damage. In this study, it presents the influence of soil deposition properties according to the change of soil depth and the organic matter contents in soil. In this experimentation, the soil deposition device developed in Air Force Research Laboratory in USA is recreated. The tested samples of mixing soil have each value of the organic matter contents. After a variety of synthetic soil were exposed to constant Cl2 concentration, the chlorinity is measured using an anion exchange chromatography(ICS-1100) to quantify the mount of deposition. As the results, the increase of soil depth causes an decreased soil deposition and the increase of exposure time causes an increased soil deposition in surface. Also, the increase of soil deposition mainly depended on the organic matter contents in surface.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.5
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• pp.1321-1326
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• 2006

Total ginsenosides were purified and their antioxidant, antibacterial and anticancer activities were measured. The crude extracts of ginseng, which were extracted with 75% ethanol by ultrasonification method, were firstly purified on AB-8 macroporous adsorption column to remove water soluble impurities, and decolored on Amberlite IRA 900 Cl anion-exchange column. Then, they were purified on Amberlite XAD16 adsorption column to delete the non-polar impurities. Total ginsenosides contents of the purified extracts were 79.4%, 71.7% and 72.5% in cultured wild ginseng, red ginseng and white ginseng, which were significantly increased than those of crude extracts. All of the three extracts showed concentration-dependant scavenging activities against DPPH radicals, among which white ginseng showed the most powerful activity. Cultured wild ginseng roots showed strongest effect against both B. subtilis PM 125(Gram-positive) and E. coli D31 (Gram-negative) bacteria, while red ginseng and white ginseng only showed the activity against B. subtilis. According to the result of the MTT assay, ail of the three extracts inhibited the growth of U-937 human hohistiocytic lympma cell, which were significantly different (p < 0.05) when compared to the control.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.787-793
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• 2008

A major polyphenolic compound extracted from omija (Schisandra chinensis) fruit was structurally identified, and its composition of major nutrients was investigated as well in this study. A dominating high performance liquid chromatography (HPLC) peak of water-extracted anthocyanin represented 94.1% of total absorbable compounds at 520 nm, which was further identified with HPLC-mass spectrometry (MS). As a result, mass-to-charge ratio (m/z) of the predominant anthocyanin was determined to be 727, and it was identical to molecular mass of cyanidin-3-xylosylrutinoside (Cya-3-O-xylrut). This is the first report that colorant of omija is predominantly composed of Cya-3-O-xylrut. Omija fruit contained exclusively 3 types of monosaccharide such as glucosc (0.68 g), galactose (0.01 g), and fructose (0.52 g) per 100 g of fruits. Several organic acids, citric (3.29 g), malic (1.4 g), acetic (0.4 g), and succinic acids (0.36 g) per 100 g of fruits, were detected by high performance anion exchange chromatography (HPAEC) analysis. During the compositional analysis of tree amino acid by HPLC, it was noticed that omija fruit contained substantial amount (0.01 g/100 g of fruits) of $\gamma$-amino butyric acid (GABA).

• Journal of the Korean Magnetic Resonance Society
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• v.17 no.2
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• pp.67-75
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• 2013

ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic $F_oc$-ring ($TF_oc$-ring) for NMR measurements. The expression of $TF_oF_1$ is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of $TF_oF_1$. $TF_oc$-rings were purified from $TF_oF_1$ according to a reported method. We modified purification procedures to improve purity and yield of $TF_oc$. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the $TF_oc$-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the $TF_oc$-ring structures in these membranes with solid-state NMR.