• Applied Biological Chemistry
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• v.46 no.3
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• pp.251-256
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• 2003

The polyphenol fractions of Korean ginseng were purified using Sephadex LH-20, MCI gel, Bondapak $C_{18}$ TLC, and HPLC from the 60% acetone soluble fraction. Fraction I showed 48.16%, 79.71% and 43.55% inhibition at 150 ppm against lipid oxidation in the presence of copper ion, superoxide and hydrogen peroxidation. Electron donating abilities of fraction II showed 35.17% inhibition at 200 ppm. Fraction III showed 48.49% and 25% inhibition at 150 ppm against lipid oxidation in the presence of ferrous ion and hydroxy radical ion. The phospholipase $A_2$ inhibitory effect of fraction III was 48.9% at the concentration of $60\;{\mu}g/ml$. The cytotoxic effects of fraction II was the highest (73.29% at 0.25 mg/ml) among the tested polyphenol fractions.

• Korean Journal of Food Science and Technology
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• v.30 no.1
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• pp.82-89
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• 1998

This study was performed to elucidate the purification and characterization of pretease from saewoo-jeot, a Korean traditional salt-fermented shrimp product. The protease in saewoo-jeot (Acetes japonicus) were extracted, desalted through electrodialysis and purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-cellulose column chromatography. Purified enzyme had specific activity of 8.4 unit/mg, yield of 14% and purification fold of 9.8. Purified enzyme was confirmed as single band protein by polyacrylamide gel electrophresis and the molecular weight was estimated to be about 24 kDa. The optimal pH and temperature for the enzyme activity were 8.0 and $40^{\circ}C$, respectively. The range of its stability to the pH and temperature were 7.0 to 10.0 and $30^{\circ}C\;to\;60^{\circ}C$, respectively. The activity of enzyme to synthetic substrate showed BAPNA and TAME. The enzyme was activated significantly by manganese ions, while inhibited by STI, TLCK. metals $(K^+,\;Li^+,\;Na^+,\;Ca^{++},\;Co^{++},\;Cu^{++},\;Mg^{++},\;Ba^{++},\;Hg^{++},\;Zn^{++},\;Fe^{+++})$. The Km value of the enzyme was $5.1{\times}10^{-7}\;M$ to hammersten casein. It's suggested that purified protease from saewoo-jeot seemed to be trypsin-like enzyme.

• Food Science of Animal Resources
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• v.24 no.1
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• pp.97-102
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• 2004

The objective of this study was to control Kimchi fermentation using pH sensitive bacteriocin entrapping liposome(bacteriocin-liposome). The liposomes were prepared by the reverse-phase evaporation method from a mixture of DPPC(dipalmitoyl phosphatidylcholine, DPPE(dipalmitoyl phosphatidylethanolamine), DOPC(dioleoyl phosphatidylcholine) and cholesterol in a molar ration of 4:2:1:4. The bacteriocin-liposome was disruptured at pH 4 of buffer and was stable at alkaline pHs(6 and 7). Irrespective of the addition of the bacteriocin-liposomes, the pH of every Kimchi sample decreased to 5 during 5 days storage at 5$^{\circ}C$. Kimchi samples treated with bacteriocin-liposomes maintained pH 4 or higher, while Kimchi samples not treated with bacteriocin-liposomes exhibited pH 3.58 or lower. In general, the pH of Kimchi samples stored at 20$^{\circ}C$ decreased faster, compared to that of Kimchi samples stored at 5$^{\circ}C$. The pH of Kimchi samples treated with the bacteriocin-liposomes was 3.9 during 90 days storage, while that of the samples not treated with the bacteriocin-liposomes was 3.68 and 3.32 during 30 days and 90 days storages, respectively. Lactic acid bacteria in Kimchi treated with the bacteriocin-liposome grew relatively slow at 5$^{\circ}C$. The viable cell number of lactic acid bacteria increased up to 4${\times}$10$\^$7/ cells/ml and then decreased to 8${\times}$10$\^$6/ cells/ml during 90 days storage at 5$^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.5
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• pp.842-849
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• 1997

This study was designed to investigate the antioxidative activity of enzymatic hydrolysates prepared from defatted anchor muscle by various pretenses. In these hydrolysates, the hydrolysates derived from pepsin and Protamex showed the strongest antioxidative activity. Enzymatic hydrolysates also showed the synergistic effects on antioxidative activity of $\alpha-tocopherol$, and almost no change in butylated hydroxytoluene (BHT). Peroxidation of metal ions $(Fe^{3+},\;Cu^{2+})$ was inhibited by enzymatic hydrolysates. Ten fractions (P-1 to P-10) were fractionated from the peptic hydrolysates by Amberlite IR-120 and Bio-gel P-2 column chromatography. The maximum antioxidative activity was observed in the traction P-2 (fraction No. $26\~31$). In amino acid composition of before and after hydrolysis of defatted anchovy muscle and the active fractions (P-2), contents of aspartic arid and glutamic acid were increased, but alanine, cysteine, tyrosine and phenylalanine were decreased.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.309-315
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• 1998

An extracellular levansucrase, which catalyzes the formation of levan from sucrose, from the culture broth of Zymomonas mobilis ZM1 was purified by conventional column purification methods. The final purification yield was 18.3 fold of the crude enzyme from Z. mobilis, with 16.5 % of the enzyme recovered in the preparation step. The molecular weight of the enzyme was estimated to be 91,000 by Superose 12 gel filtration, and 45,000 by SDS-PAGE, indicating that levansucrase is a dimer. The optimum pH for the enzyme activity was around pH 4.0 for sucrose hydrolysis, and was around pH 5.0 for levan formation. The enzyme was inhibited by some metal ions, such as Hg$\^$2+/ and Cu2$\^$2+/, and 50% of inhibition was observed with 5mM EDTA. The enzyme activity was enhanced by the presence of detergent Triton X-100, but inhibited by SDS completely The enzyme catalyzes the liberation of reducing sugars, oligosacccharides and the formation of fructose polymer(levan). The enzyme also catalyzes the transfructosylation reaction of fructose moiety from sucrose to various sugar acceptor molecules, including sugar alcohols.

• BMB Reports
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• v.30 no.2
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• pp.150-156
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• 1997

Tyrosinase was purified from Solanum melongena by ammonium sulfate precipitation, Sephadex G-150 and DEAE-Sephacel column chromatography. The molecular weight of the purified tyrosinase was approximately 88,600 daltons with 805 amino acid residues. The amino acid composition showed the characteristic high contents of glycine, glutamic acid and serine residues. The enzyme had high substrate specificity towards (+)-catechin. The $K_m$, value for L-DOPA was 20.8 mM. L-ascorbic acid, ${\beta}-mercapto-ethanol$, sodium diethyldithiocabamate, KCN and $NaN_3$ had strong inhibitory effects on enzyme activity. Sodium diethyldithiocabamate was a competitive inhibitor of the enzyme with a $K_i$ value of $5.2{\times}10^{-2}\;mM$. The optimum pH of the enzyme was 9.0 and the optimum temperature was $65^{\circ}C$ with L-DOPA as a substrate. In addition, the activity was enhanced by addition of $Ca^{+2}$ or $Cu^{+2}$, but decreased in the presence of $Fe^{2+},Fe^{3+}$ and $Zn^{2+}$ ions.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.109-114
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• 1997

The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.506-512
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• 1995

A cell aggregation factor produced by Aspergillus sp. LAM 94-142 was purified and partially characterized. The factor was purified about 15 folds from culture broth by IRA 420 and IRC 120 treatment, 1% NaCl added acetone precipitation, and Sepharose 4B column chromatography with overall yield of 48%. It was heteropolysaccharide consisted of mannose, arabinose, and glucose with a molar ratio, 31:17:2, and its molecular weight was estimated to be about 900,000 daltons by Sepharodse 4B gel filtration method. The optimum pH and temperature was 8 and 40$\circ$C, respectively. The factor was stable in pH range of 3-9 and at 100$\circ$C for 90 min. The cell aggregation activity of the factor was inhibited by the addition of Hg$^{2+}$, Fe$^{2+}$, Cu$^{2+}$, and some polypeptides such as milk casein or hemoglobin. The factor aggregated Bacillus subtilis, B. macerans, B. turingiensis, E. coli, Peudomonas aeruginosa, P. fluorescens, P. malophilia, and weakly aggregated Staphylococcus sp., Sarcina lutea, P. putida and Cryptococcus neoformnans, but it didn't aggregate various strains of Candida sp. and Saccharomyces sp.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.573-579
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• 1995

A Bacillus strain capable of producing an extracellular malto-oligosaccharides forming $\alpha $-amylase was isolated from soil and designated as Bacillus sp. SUH4-2. The enzyme was purified by ammonium sulfate fractionation, DEAE-Toyopearl and Mono-Q HR 5/5 column chromatographies using a FPLC system. The specific activity of the enzyme was increased by 16.1-fold and the yield was 13.5%. The optimum temperature for the activity of $\alpha $-amylase was 60-65$\circ$C and more than 50% of initial activity was retained after the enzyme was incubated at 60$\circ$C for 40 min. The enzyme was stable over a broad pH range of 5.0-8.0 and the optimum pH was 5.0-6.0. The molecular weight of the enzyme was determined to be about 63.6 kD and isoelectric point was around 5.8. The enzyme activity was strongly inhibited by Mn$^{2+}$, Ni$^{2+}$, and Cu$^{2+}$ ; slightly by Ca$^{2+}$. The purified enzyme produced starch hydrolyzates containing mainly maltose and maltotriose from soluble starch. The starch hydrolyzates were composed of 11% glucose, 59% maltose, 25% maltotriose and 5% maltotetraose.

• YAKHAK HOEJI
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• v.13 no.1
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• pp.1-15
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• 1969

A new method of qualitative and quantitative determination of barbiturates in the pharmaceuticals by means of $\alpha$-picoline-copper (II) was studied. Barbiturates in the pharmaceuticals were dissolved in the mixed solvent of 33% $\alpha$-picoline-Carbontetrachloride to yield Complex Compounds of barbiturates-copper (II)-$\alpha$-picoline. Complex Compounds of barbiturates show uniformly maximum absorption at the wavelength of 540m.mu. and wre to be identified at the concentration of 1 X 10$^{-4}$ Mole, and also was to be quantitatively determined at the concentration of 1 X 10$^{-3}$ Mole. By this method barbiturates in the pharmaceuticals could be determined in the presence of various compounds such as sulpyrine, isopropylantipyrine, antipyrine, phenacetin and etc. But Barbiturates could be also determined by this method after seperation with aminopyrine, acetaminophen, acetylsalicylic acid and etc. by column chromatography. And barbiturates and acetylsalicylic acid could be also determined by simultaneous equation while their complex compounds show uniformly each maximum absorption at the Wavelength of 540 m${\mu}$ and 620 m${\mu}$. I.R. spectra of these complex compounds show identification of Barbiturates derivatives. The composition ratio of these complex compounds were : barbiturates : Cu : ${\alpha}$-picoline=2:1:2.