• Korean Journal of Microbiology
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• v.27 no.2
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• pp.117-123
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• 1989

Radioactive N-$\alpha$-p-nitrobenzoxycarbonyl (NBZ)-L-[2,$3-^{3}$H] arginyl diazomethane was used as an affinity label for the vacuolar arginine transporter in Neurospora crassa. Vacuolar matrix proteins were removed by fracturing the membranes with freeze-thaw method in dry ice/ethanol bath. Vacuolar membrane proteins were then wasged with 500mM NaCl to remove ionically bound derivatives and peripheral membrane proteins from vacuolar membranes. After dissolved in 1% Titon X-100, dissolved vacuolar memvrane proteins were separated with molecular sieve column chromatography, anion and cation exchange chromatographies. The arginine transporter was purified giving the purification factor of 1136.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.4
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• pp.816-821
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• 1999

Cholesterol esterase(pCEH, pancreas cholesterol ester hydrolase, E.C.3.1.1.13) which is secreted from pancreas has been known as an important lipase for cholesterol uptake. cholesteryl acyl esters from a diet must be hydrolyzed to free cholesterol and fatty acid by cholesterol esterase before the absorption in small intestine. For the development of inhibitory substances from natural source, we screened many extracts of oriental herbs for the inhibition of cholesterol esterase in vitro. The ethanol extract of Ephedra herba showed strong inhibitory activity. Solvent fractionation and silica gel column chromatography with the extract lead to the purification of the inhibitory principle in Ephedra herba. Crystallized inhibitor was identified as ( ) ephedrine by using UV, FT IR, 1H NMR, 13C NMR and GC/Mass. These results suggest that ( ) ephedrine can be used as a potential lead compound for the development of inhibitor for cholesterol uptake by cholesterol esterase inhibition.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.2
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• pp.227-236
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• 1995

The objective of this research was to characterize fucoidans isolated from Laminaria religiosa and Undaria pinnatifida in Korea to obtain basic data for Production of soluble dietary fiber materials with biological functionality. Fucoidans were successively extracted 3 times at $65\%$ for 1hr with arid solution of pH 2.0, and cetylpyridinium chloride was used for partial purification. The yields of partially purified fucoidans were $2.71\%$ for L. religiosa, $6.65\%$ for sporophylls of U. pinnatifida and $0.40\%$ for blade of U. pinnatifida. The yield from sporophylls of U. pinnatifida was highest among the sample tested, whereas the yield from blade of U. pinnatifida was lowest. It appeared that the fuconidans content in different parts of U. pinnatifida varied. Partially purified fucoidans were separated into 3 fractions by DEAE-Sephadex A-25 ion exchange column and the maior fractions were refractionated with tractional precipitation with ethanol. $60-70\%$ ethanol precipitated fractions of 1. religiosa and sporophylls of U. pinnatifida turned out to be homogeneous by cellulose acetate electrophoresis and gel filteration chromatography. The molar ratios of fucose, galactose, and sulfate in the purified fucoidans(ethanol precipitated fractions) were 1 : 0.31 : 2.43 for L. religiosa and 1 : 0.97 : 1.99 for sporophylls of U. pinnatifida. The averaged molecular weights of the purified fucoidans from L. religiosa and sporophylls of U. pinnatifida were 31,000 and 38,000, respectively.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.25-30
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• 2001

A strain of Actinomycetes producing cathepis B inhibitor was isolated from soil and identified as Streptomyces misakiensis. The product of S. misakiensis inhibited effectively cathepsis B proteinases as well as trypsin and papain. The cathepsin B inhibitor were largely produced with incubation for 4 days. The S. misakiensis was the most growth with incubation for 5 days. The cathepsin B inhibitor was isolated from the extraction of both with ethanol, ethanol and chlorofrom, and following several column chromatography such as sephadex G-15, silica gel 60 and sephadex LH-20 chromatography. The moleculer weight of purfied inhibitor was 138 dalton.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.343-347
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• 2005

The polysaccharides were extracted from fruiting body, mycelia, and cell-free broth of Agaricus blazei Murill. The crude polysaccharides were obtained by the ethanol addtion. They were further purified using ion-exchange chromatography and gel chromatography. Ion-exchange chromatography using DEAE-cellulose column separated neutral and acidic polysaccharides. Neutral polysaccharides were then purified with gel filtration chromatography. For single peak obtained from gel filtration chromatography was molecular weight was measured with Sepharose CL-6B. The same procedure with acidic polysaccharides were performed to get the purified polysaccharides.

• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.523-528
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• 1998

We isolated microorganism secreting antimicrobial substance from tomato and identified as Enterococcus faecium. This substance was completely inactivated by pretense treatment and retained activity after catalase treatment. This result indicated that the antimicrobial activity of this substance was due to proteinaceous substance known as bacteriocin. The bacteriocin inhibited growth of Gram positive bacteria, such as Listeria monocytogenes, Leuconostoc mesenteroides, Lactobacillus plantarum, Streptococcus agalactiae, Streptococcus pyrogenes, and Gram negative bacteria, such as Pseudomonas aeruginosa. Purification of the bacteriocin was achieved by ethanol precipitation, ion exchange chromatography on CM Sepharose CL-6B, and gel filtration on Sephacryl S-100 HR. After these purification steps, the specific activity of the bacteriocin was increased 35.8 fold compared with culture broth. Purified bacteriocin was shown single band on SDS-PAGE and molecular weight was estimated 51 kDa. The residual activity of this bacteriocin was 3.3% at 10$0^{\circ}C$ for 60 min, and this bacteriocin was stable at pH 2~7.

• Journal of Applied Biological Chemistry
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• v.59 no.1
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• pp.19-23
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• 2016

A phenol substance was extracted from Pinus koraiensis Siebold et Zucc leaf extracts and its biological efficacy was measured. The highest content of the phenol substance contained in Pinus koraiensis Siebold et Zucc leaves was 13.5 mg/g, which was obtained when it was extracted with 80% ethanol. At a concentration of 200 mg/mL, the phenolic substances extracted with 80% ethanol and water showed antimicrobial activities against Helicobacter pylori, producing clear zones of 10 and 12 mm diameter, respectively. Pinus koraiensis Siebold et Zucc. leaf extracts were separated using a Sephadex LH-20 column and 4 fractions were obtained (fractions A-D). Fractions C and D showed the greatest inhibitory activity against Helicobacter pylori producing 10.1 and 12.3 mm clear zones, respectively. These two fractions were purified using a Sephadex LH-20 and MCI-gel column ($H_2O{\rightarrow}100%$ ethanol). Purified compounds A and B were identified as syringic acid and compound C was identified as p-coumaric acid based on $^1H$-nuclear magnetic resonance (NMR), $^{13}C$-NMR, and fast atom bombardment mass spectrometry spectra. When two or more purified compounds were mixed, a synergistic effect of anti-Helicobacter pylori activity was evident. This result indicates that extracts of Pinus koraiensis Siebold et Zucc leaves could be considered a functional food because of their high antimicrobial properties.

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.293-298
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• 1998

Allithiamine was synthesized when thiamine was mixed with fresh garlic extract made with ethanol at a alkali medium. Allithiamine was isolated and purified using solvents such as ethyl acetate, diethyl ether and benzene. Purified allithiamine was identified by determination of melting point, elemental analyzer and LC/Thermospray/Mass-spectrometer. On the other hand, synthetic conditions of allithiamine from fresh garlic, pH, temperature and ratio of garlic to ethanol were investigated. Synthetic rates of allithiamine under alkali conditions were rapidly increased while those under acidic conditions very slowly increased. The synthetic rates of allithiamine increased as temperature increased, but decreased above $70^{\circ}C$ as reaction time increased. There was no significant difference in synthetic rate of allithiamine when garlic was mixed above 4 times of ethanol. Therefore, optimum condition of pH, temperature and ratio of garlic to ethanol for synthesis of allithiamine were 8, $60^{\circ}C$ and 1 : 4, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.11
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• pp.1640-1646
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• 2010

The optimum condition was investigated for the extraction of glycosaminoglycan (GAG) from sea hare, Aplysia kurodai. The most effective enzyme was Flavourzyme for extraction of glycosaminoglycan. The optimum incubation temperature and time for hydrolysis were $60^{\circ}C$ and 15 hr, respectively. The yield of precipitated polysaccharide depended on Brix and ethanol volume. The most effective concentration of Brix and ethanol were sixty and 5 volume of ethanol, respectively. Most GAG was eluted between 0.5 M and 0.75 M NaCl gradient on DEAE-Sepharose column, and identified by electroconductivity. The contents of hexuronic acid from polysaccharide extract and GAG were 1.0 g/100 g and 6.0 g/100 g, respectively. Hexosamine of polysaccharide and GAG as indicator of GAG component was 5.6 g/100 g and 25.7 g/100 g, respectively. GAG was identified as heparan sulfate compared with bands of other GAG on agarose gel electrophoresis, and its molecular weight was 29.6 kDa on Superdex 200 HR column.

• KSBB Journal
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• v.16 no.1
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• pp.71-75
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• 2001

Acanthoside-D, contained in Acanthopanax chilsanensis is known as a ginseng-like substance. This work was focused to set up analytical and preparative conditions for Acanthoside-D purification. The ethanol extract from the powder of the trunk of Acanthopanax chilsanensis was partitioned with hexane. A $\mu$-Bondapak $C_{18}$ (3.9$\times$300 mm, $10\mu\textrm{m}$) column was used to separate Acanthoside-D from the trunk of Acanthopanax chilsanensis. From the experimental results, the mobile phase used for isolating Acanthoside-D from the extract was water/acetonitrile/methanol=80/14/6 %(v/v). The flow rate of mobile phase was 1.0 mL/min, and UV wavelength was fixed at 210 nm. Finally on a semi-preparative column (3.9$\times$ 300 mm, $15\mu\textrm{m}$, Lichrospher 100RP-18) with the same mobile phase composition, the allowable maximum injection volume increased to 250 $\mu\ell$