• Korean Journal of Agricultural Science
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• v.39 no.2
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• pp.255-261
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• 2012

A bacterium AB-55, isolated from waste mushroom bed of Agaricus bisporus in Sukseong-myeon, Buyeo-gun, Chungcheongnam-do, Korea, was screened onto xylan agar congo-red plate by the xylanolysis method and was used to produce an xylanase in shaker buffle flask cultures containing oat spelt xylans. The phylogenetic analysis using 16S rRNA gene sequence data showed that the strain AB-55 had the highest homology (99.0%) with Bacillus subtilis and it was named as Bacillus subtilis AB-55. A xylanase was purified by ammonium sulfate precipitation (50~80%), gel filtration on sephacryl S-300, and ion exchange chromatography on DEAE sepharose FF. The molecular weight of the xylanase was estimated as 44 kDa by SDS-PAGE. Optimal pH and temperature for the xylanase activity was pH 7 and $50^{\circ}C$, respectively. N-terminal amino acid sequence of the enzyme was identified as Ser-Ala-Val-Lys-His-Gly-Ala-Ile-Val-Phe. The substrate specificity of the enzyme exhibited that it hydrolyzed efficiently oat spelt xylan as well as beechwood xylan, but showed no activity against Avicel and carboxymethyl clellulose (CMC). The enzyme activity was enhanced by $Fe^{2+}$ and $Mn^{2+}$ whereas was entirely inhibited by $Hg^+$.

• Food Science of Animal Resources
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• v.36 no.6
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• pp.769-778
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• 2016

In this study, we improved the eggshell-membrane separation process by separating the shell and membrane with EDTA solution, evaluating effects of three different extraction solutions (acetic acid, EDTA, and phosphate solution), and co-purifying multiple eggshell proteins with two successive ion-exchange chromatography procedures (CM Sepharose Fast Flow and DEAE Sepharose Fast Flow). The recovery and residual rates of eggshell and membrane separated by the modified method with added EDTA solution were 93.88%, 91.15% and 1.01%, 2.87%, respectively. Ovocleidin-116 (OC-116) and ovocalyxin-36 (OCX-36) were obtained by loading 50 mM Na-Hepes, pH 7.5, 2 mM DTT and 350 mM NaCl buffer onto the DEAE-FF column at a flow rate of 1 mL/min, ovocleidin-17 (OC-17) was obtained by loading 100 mM NaCl, 50 mM Tris, pH 8.0 on the CM-FF column at a flow rate of 0.5 mL/min. The purities of OCX-36, OC-17 and OC-116 were 96.82%, 80.15% and 73.22%, and the recovery rates were 55.27%, 53.38% and 36.34%, respectively. Antibacterial activity test suggested that phosphate solution extract exhibited significantly higher activity against the tested bacterial strains than the acetic acid or EDTA extract, probably due to more types of proteins in the extract. These results demonstrate that this separation method is feasible and efficient.

• Journal of Microbiology and Biotechnology
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• v.21 no.3
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• pp.256-262
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• 2011

A 20.9-kDa metalloprotease was isolated from dried fruiting bodies of the wild basidiomycete mushroom Lepista nuda. The N-terminal amino acid sequence of the protease was seen to be ATFVLTAATNTLFTA, thus displaying no similarity with the sequences of previously reported metalloproteases. The protease was purified using a procedure that entailed ion-exchange chromatography on CM-Cellulose, Q-Sepharose, and Mono S, and FPLC-gel filtration on Superdex 75. The protease functioned at an optimum pH of 7.0 and an optimum temperature of $50^{\circ}C$. It was also noted that the protease demonstrated a proteolytic activity of 1,756 U/mg toward casein. The $K_m$ of the purified protease toward casein was 6.36 mg/ml at a pH of 7.0 and with a temperature of $37^{\circ}C$, whereas the $V_{max}$ was 9.11 ${\mu}g\;ml^{-1}\;min^{-1}$. The activity of the protease was adversely affected by EDTA-2Na, suggesting that it is a metalloprotease. PMSF, EGTA, aprotinin, and leupeptin exerted no striking inhibitory effect. The activity of the protease was enhanced by $Fe^{2+}$, but was curtailed by $Cd^{2+}$, $Cu^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ ions. The protease also exhibited inhibitory activity against HIV-1 reverse transcriptase with an $IC_{50}$ value of 4.00 ${\mu}M$. The $IC_{50}$ values toward hepatoma Hep G2 and leukemia L1210 cells in vitro were 4.99 ${\mu}M$ and 3.67 ${\mu}M$, respectively.

• Journal of the Korean Chemical Society
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• v.29 no.6
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• pp.615-622
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• 1985

The chemical species formed by uranium and vanadium and their equilibria have been investigated in the various concentrations of oxalic and acetic acids by the ion exchange chromatography and UV-Vis spectrophotometry. Uranyl and vanadyl ions seem to be form the complex as $UO_2(C_2O_4)_2=$, $UO_2(C_2O_4)_3^{4-}$ and $VO_2(C_2O_4)-2^{3-}$ respectively in the concentration range of 0.005∼0.05M oxalic acid concentration. In the case of acetic acid the equilibria of $UO_2^{2+}+3Ac^-=UO_2(Ac)_3^-$ and $VO_2^++2Ac^-=VO_2(Ac)_2^-$ were existed individually according to the increase of acetic acid concentration.

• Korean Journal of Microbiology
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• v.30 no.2
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• pp.71-77
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• 1992

A 50 KD metalloprotease of Serratia marcesrens ATCC 21074 was purified by ammoniumsulfate precipitation. DEAE-cellulose ion exchange chromatography, and sephadex ti-100gel filtration. Optimal pH and temperature of enzyme were pH 8.0 and 37"C, respectively.This enzyme was stable in the ranges of 10-37$^{\circ}$C and pH 5.0--11.0. Thermal denaturationwas investigated by differential scanning calorimetry. Onset temperature of denaturationand endothermic peak temperature were 376$^{\circ}$C and 43.2"C. re:,pectively. The denaturationenthalpy was -8.4mJimg. The purified metalloprotease was ri.sistant to autodigestion for24 hr at 30$^{\circ}$C. Metalloprotease in culture supernatant was also resistant to autodigestionin this conditions. Heat-denatured enzyme. however. was rapidly digested by the nativeenzyme. The metalloprotease was stable to proteolytic digestion by mammalian proteasessuch as trypsin. a-chymotrypsin, and elastase. But the enzyme was easily digested bybacterial protease. thermolysin.bacterial protease. thermolysin.

• Korean Journal of Food Science and Technology
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• v.11 no.2
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• pp.77-80
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• 1979

Effects of interfering substances on the determination of L-ascorbic acid (AsA) in green tea and it's extracts by 2,4-dinitrophenylhydrazine (DNP) method was studied. and the removal of these interfering substances was also investigated. Under the condition prescribed for DNP method, AsA content of green tea are effected by some sugar, reductones, dicarbonyl compounds, organic acids, amino acids and others. All interfering substances except amino acids were eliminated by the chloroform extraction after adding o-phenylendiamine to sample solution. and remaining amino acids were eliminated almost completely by the treatment with ion exchange resin$(Amberlite{\;}IR-120H^{+})$. After removing the interfering substances by the above mentioned procedure, total AsA in green tea was determined by DNP method. The values obtained by this method were in good agreement with those by thin layer chromatography (TLC) method. and the method was more rapid and simple than TLC method.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.348-356
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• 1989

An extracellular peroxidase found in culture broth of Pleurotus ostreatus was induced by syringic acid. This enzyme was fractionated by DEAE Sephadex A-50 ion exchange chromatography and gel filtration chromatogrphy on Sephadex G-150. The enzyme is a glycoprotein containing 35.7% carbohydrate. The results of SDS-linear polyacrylamide gradient gel electrophoresis and gel filtration indicate that the enzyme is a dimer consisted of identical subunits (Mr=72,400). The absorption spectrum of the enzyme indicates the presence of one mole of iron protoporphyrin IX per one mole of subunit. Isoelectric point of the enzyme is 4.26 and $K_m$ values for $H_2O_2$ is $7.2{\mu}M$. The enzyme showed its optimal activity at pH 3.5-4.0 and at $40^{\circ}C$. The Km values of this enzyme for ferulic acid and sinapic acid are 2.4 and 12.3 times higher than those of horseradish peroxidase, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.2
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• pp.187-194
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• 1992

Rhizopus oryzae CJ-2114 was selected for its strong polygalacturonase activity among various strains of mold found in soil. It was found that the production of polygalacturonase reached to maximum when the wheat bran medium containing 1% albumin, 1% sorbitol and 0.2% (NH$_4$)$_2$C$_2$O$_4$was cultured for 96 hrs at 3$0^{\circ}C$. Polygalacturonase was purified 11.13 fold from Rhizopus oryzae CJ-2114. The purification procedures include ammonium sulfate treatment, gel filtration on Sephadex G-75, G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 40.3% .Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-poly-acrylamide gel electrophoresis, the molecular weight was estimated to be 47,000. The amino acid composition indicated relatively high contents of glutamic acid and glyrine.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.26-32
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• 2002

For commercial production of Korean fermented anchovy sauce through rapid fermentation, a bacterial strain which showed the high protease activity was isolated from a commercially fermented anchovy sauce. The isolate was Bacillus amyloliquefaciens, and named as B. amyloliquefaciens HTP-8. The incubation temperature, initial pH, and cultivation time for optimal production of protease by B. amyloliquefaciens HTP-8 were $30^{\circ}C$, 7.0, and 3 days, respectively. In jar fermenter, B. amyloliquefaciens HTP-8 showed higher protease activity when grown at pH 7.0. The protease was partially purified by 80% ammonium sulfate precipitation and CM-Sephadex C-50 ion exchange chromatography. The partially purified enzyme had specific activity of 103.3 units/mg, yield of 0.4%, and purification fold of 43.0. The optimal pH and temperature for the protease activity were 10.0 and $50^{\circ}C$, respectively. The protease was relatively stable at the pH range of 7.0~12.0 and at the temperatures below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{＋}$ /, $Ba^{2＋}$ and selectively inhibited by PMSF, suggesting that it is a serine protease.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.196-201
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• 2006

Vitellogenin, which is found in the serum of female and male fishes exposed to environmental endocrine disrupter or estrogen hormone, is used as a biomarker for environmental contamination with an endocrine disrupter. In order to produce antibody against vitellogenin, a synthetic peptide for partial vitellogenin was injected into rabbits. In addition, by using ion exchange chromatography on DE-52, vitellogenin was purified from the serum of carp induced with $17{\beta}$-estradiol. Polyclonal antibody against purified vitellogenin reacted well with vitellogenin in the serum of carp induced with $17{\beta}$-estradiol and the serum of female carp, whereas polyclonal antibody against the vitellogenin peptide did not react with proteins in those samples. This may indicate that vitellogenin proteins, covalently modified largely, could not be detected by Western blotting with the polyclonal antibody against the synthetic vitellogenin peptide.