• Korean Journal of Food Science and Technology
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• v.26 no.4
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• pp.375-381
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• 1994

A bacterial strain No. KY-126, which produced extracellular cyclodextrin glycosyltransferase(CGTase), was isolated from soil and identified as Bacillus stearothermophilus KY-126. The enzyme was purified by the treatments of ammonium sulfate precipitation, DEAF-Sephadex, Sephadex G-100 column chromatography. The optimal pH and temperature for the enzyme activity were pH 5.5 and $65^{\circ}C$, respectively. And the enzyme was stable at pH values from 6.0 to 11.0 at $55^{\circ}C$ for 30 min and stable up to $60^{\circ}C$ for 30 min.. The enzyme was inhibited by $HgCl_{2}$. The molecular weight of the enzyme was estimated to be 67,000 by using SDS-PAGE. The maximum conversion from starch to cyclodextrin (CD) by CGTase was 43% and obtained at 6 hr reaction and the ratio of ${\alpha}-,\;{\beta}-,\;{\gamma}-$, CD production at this time was 2.9 : 2.1 : 1.0.

• 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.41 no.7
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• pp.522-526
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• 1998

Ferritin from germinated soybean seeds was purified by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, gel filtration chromatography on Sephacryl S-300, and HPLC with Bio-Scale Q2 column. SDS-PAGE analysis showed that the purified ferritin is composed of subunit with an apparent M, 21,000. The molecular mass of the native soybean ferritin estimated by gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be $510{\sim}560\;kDa$. Soybean ferritin contained 833 mol Fe/mol protein, which is 31-fold more iron than pumpkin ferritin and stained positive for iron on non-denaturing gel. Soybean ferritin cross-reacted with anti-soybean rabbit ferritin antiserum.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.132-138
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• 1994

L-Ascorbic acid-producing enzyme in Neurospora crassa was found to exist in mitochondria and the activity of this enzyme was increased by the addition of D-fluconno-${\gamma}$-lactone or L-gulono-${\gamma}$-lactone in the media. L-Ascorbic acid-producin enzyme in N. crassa has been purified with ammonium sulfate precipitation. DEAE Sepharose CL-6B ion exchange chromatography. Sephacryl S-200 gel filtration chromatography and Reactive yellow 3-agarose dye affinity column chromatography. The specific activity of this enzyme was increased to 239.6 fold and the yield was 2.1%. The molecular weight of the native enzyme was 150.000 dalton when it was estimated with Sephacryl S-200 gel filtration chromatography. Its molecular weight appeared as 75.000 dalton by SDS-polyacrylamide gel electrophoresis. which suggested that this enzyme was consisted with two identical subunits. The optimal pH for this enzyme was 9.0 and the $K_m$ value for D-galactono-${\gamma}$-lactone was 0.073 mM.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.8
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• pp.1009-1017
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• 2008

For the effective use of exopeptidase from squid viscera as food processing aids, the viscera of Argentina shortfin squid (Illex argentinus) were fractionated by various methods such as acetone treatment, ammonium sulfate treatment, anion exchange chromatography, and gel filtration. The positive exopeptidase fractions were obtained from the fraction II treated by cold acetone ($30{\sim}40%$, w/w), the fraction V by ammonium sulfate ($60{\sim}70%$ saturation), the fraction II (0.2 M NaCl) by anion exchange chromatography, and the fraction I ($30{\sim}50\;kDa$) by gel filtration. The specific activities of positive fractions from viscera of I llex argentinus against substrates were higher to LeuPNA than to ArgPNA. Total activity and recovery against LeuPNA of positive fraction by gel filtration were 1,867 U and 30.69%, respectively, which were the highest among those of positive fraction. The results suggested that the gel filtration chromatography method was the most efficient method for the fractionation of exopeptidase from viscera of Illex argentinus.

• Proceedings of the Korean Radioactive Waste Society Conference
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• 2005.06a
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• pp.121-128
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• 2005

A study on the selective separation of $^{99}Tc,\;^{94}Nb,\;^{55}Fe,\;^{90}Sr$ and $^{59}Ni(^{63}Ni)$ from various radioactive wastes discharged from the nuclear power plants in Korea is being performed for use in their quantifications which are indispensible for the evaluation of the radionuclide inventory. Separation behaviour of Ce, Ca, Mg, Al, Cr, Ti, Mn and Cu recovered along with Ni during the separation of Re (as a surrogate of $^{99}Tc$), Nb, Fe and Sr by anion exchange and Sr-Spec extraction chromatography was investigated by cation exchange and Ni-Spec extraction chromatography using synthetic radioactive waste dissolved solutions containing matrix elements such as Re, Nb, Fe, Sr, Ni, B, Na, K, Ce, Co, Ca, Mg, Al, Zn, Cr, Pb, Cd, Mo, Mn, Cu, Zr, Ti and U. To purify the Ni fraction recovered and prepare a radionuclide source available for gas proportional counting, an application of the Ni precipitation procedure with dimethylglyoxime in the medium of ammonium citrate and tartaric acid solutions as a masking agent for co-existing metal ions was described in detail.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.574-582
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• 1992

Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

• Journal of Life Science
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• v.18 no.1
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• pp.84-90
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• 2008

Bacterial colonies which were able to oxidize the manganese were isolated from six soil samples in Byungchon area. Among them, one bacterial strain was selected for this study based on its high manganese oxidation activity. This selected bacterial strain was identified as Pseudomonas sp. MN5 through physiological-biochemical test and analysis of its 16s rRNA sequence. This selected bacterial strain was able to utilize fructose and maltose, but they doesn't utilizing various carbohydrates as a sole carbon source. Pseudomonas sp. MN5 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, streptomycin and tetracycline, but a high resistance up to mg/ml unit to heavy metals such as lithium, manganese and barium. Optimal manganese oxidation condition of Pseudomonas sp. MN5 was pH 7.5 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. The manganese oxidizing protein produced by Pseudomonas sp. MN5 was purified by ammonium sulfate precipitation, HiTrap Q FF anion exchange chromatography and G3000sw $_{XL}$ gel filtration chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, three manganese oxidizing protein with estimated molecular weights of 15 kDa, 46.7 kDa and 63.5 kDa were detected. Also, it was estimated that manganese oxidizing protein produced by Pseudomonas sp. MN5 were a kind of porin proteins through internal sequence and N-terminal sequence analysis.

• Applied Biological Chemistry
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• v.34 no.3
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• pp.249-257
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• 1991

To elucidate enzymatic properties of ${\alpha}-galactosidase$ (EC 3, 2, 1, 22) from germinated soybean, changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined. ${\alpha}-Galactosidase$ from germinated soybean was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties was investigated. ${\alpha}-galactosidase$ activity of sobeam was maximized when it was germinated at $25^{\circ}C$ for 120 hour. Raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. Soybean ${\alpha}-galactosidase$ was purified by 6.6 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50, and gel filtration on Sephadex G-150. Its specific activity was 825 Units/mg protein and the yield was 2.5% of the total activity of crude extracts. The purified ${\alpha}-galactosidase$ of soybean was found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. Isoelectric point of soybean ${\alpha}-galactosidase$ was determined analytical isoelectric focusing to be pH 4.8. The soybean ${\alpha}-galactosidase$ was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE. The optimal temperature and pH for the soybeam ${\alpha}-galactosidase$ activity were $40^{\circ}C$ and pH 6.0 and 75% of its activity was lost by heating at $60^{\circ}C$ for 10 min. The enzyme was appeared to have higher affinity to raffinose than to stachyose. The Km value of soybean enzyme was 5.3 mM for ${\rho}-nitrophenyl-{\alpha}-D-galactopyranoside$ and the activation energy on PNPG was calculated to be 13.02 Kcal per mole.

• Korean Journal of Food Science and Technology
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• v.33 no.1
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• pp.117-121
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• 2001

White kidney bean, Phaseolus vulgaris, contains proteinaceous inhibitors of ${\alpha}-amylase$. Two inhibitors have been purified by conventional protein fractionation methods such as ethanol precipitation, ammonium sulfate fractionation, DEAE-Sephadex ion exchange chromatography and Sephadex G-100 gel chromatography. The inhibitors were purified as I-1 and I-2 based on their elution order from the DEAE-Sephadex column. The overall purification ratio were about 15.0 and 14.8 for I-1 and I-2, respectively. The molecular weights of purified ${\alpha}-amylase$ inhibitors were 50,000 and 45,000 determined by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. They contain $17.6{\sim}17%$ of carbohydrate, $70{\sim}80%$ of protein. The carbohydrates were composed of glucose : xylose : mannose : N-acetylglucosamine (5 : 3 : 50 : 42).