• Korean Journal of Microbiology
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• v.21 no.3
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• pp.135-142
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• 1983

The presence of polyphosphate phosphohydrolase (PPPH) and tripolyphosphate phosphohydrolase (TPPH) in Chlorella ellipsoidea were confirmed from the cell-free extract of the algal cells and three forms of PPPH were isolated, purified, and measured Km-Vmax value and inhibitory effect by metal ions, respectively. PPPH was most active at pH7.2, whereas TPPH at pH 7.6. Both enzymes exhibited their maximum activity at $37^{\circ}C$. For the manifestation of catalytic activity, divalent, divalent metal ions are needed, and the best activator for both enzymes was $Co^{++}\;ions\;(10^{-3}M)$. These enzymes were inhibited by $Hg^{++}\;ions\;(10^{-3}M)$ considerably. PPPH from Chlorella ellipsoidea was purified by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-25, and gel filtration on Sephadex G-100, and some properties of the three different fraction with PPPH activity $(PPPH_1,\;PPPH_2,\;and\;PPPH_3)$ were found, i.e, PPPH has multiple form. The Km values of $PPPH_1,\;PPPH_2,\;and\;PPPH_3$ obstained were $6.25{\times}10^{-4}M,\;10^{-4}M-4/M,\;and\;3.33{times}10^{-4}M$ and Vmax were 3.33 mM/min, 3.33 mM/min, and 2.67 mM/min, respectively. It was shown that the types of inhibition of $Hg^{++} on the activities of three forms of PPPH were competitive inhibition.

• Journal of Life Science
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• v.8 no.2
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• pp.145-157
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• 1998

We isolated 100 Vobrio sp. from marine products and sea from July to September, 1997. We attemped on purification of hemolysin produced by Vibrio sp. The growth, hemolysin production patterns by the 100 strains of Vibrio sp. showed identical, in general. V. unlnificus ys-1 produced hemolysis as the higtest titer. The optimal culture conditions for the hemolysin production by the V. vunificus ys-1 are followings; 1. Hemolysin production was optimal dering the late exponetial phage. 2. Maximal growth, hemolysin production were in heart infusion broth. 3. Maximal yields of hemolysin was obtained when the heart infusion broth had an intial pH of 8.0, 3$0^{\circ}C$, 3% NaCL. Hemolysin was purified from culture filtrate of the strain by ammonium sulfate recipitation, ion exchange and hydrophobic interaction chromatography. The results were as follows; 1. Hemogeneity of the purified hemolysin was demonstrated by revealing single band on SDS-PAGE. The molecular weight of purified hemolysin was 45KDa. 2. The absorbance rattern in ultraviolet wsa typical of those seen with most proteinb with 280nm. 3. Purified hemolysin was atable at 5$0^{\circ}C$ but 7$0^{\circ}C$ of the acivity was lost by heating for 30 min at 6$0^{\circ}C$/ Optimal temperature of purified hemolysin was 35$^{\circ}C$. 4. Purified hemolysin was stable at the pH range of 6~9, but in the less the pH5.0. above the pH 9.0, the hemolysin activity was lost completely.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.54-62
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• 1993

Intracellular adenosine deaminase (ADA) from Aspergillus oryzae was purified using ammonium sulfate fractionation, a DEAE-Sephadex A-50 anion exchange chromatography, an ultrafiltration using a PM 10 membrane and two times of Sephadex G-100 gel filtration chromatography. The enzyme was purified 151 fold with a 9% recovery. Purified enzyme gave a single protein band with a molecular weight of 105,000 delton. The enzyme was reasonably stable. The enzyme activity was kept even after 1 hr incubation at 55.deg.C, but decreased significantly at 60.deg.C. The pH optimum was found to be from 6.5 to 7.5. Among tested compounds, the substrate activity was found with adenosine, adenine arainofuranoside, formymcin A, 2'-deoxyadenosine, 3'-deoxyadenosine, 2', 3'-isopropylidene adenosine, 2,6-diaminopurine deoxyriboside, .betha.-nicotinamide adenine dinucleotide (reduced form), 6-chloropurine riboside, 2'-adenine monophosphate (AMP), 3'-AMP and 5'-AMP. The values of Km of adenosine and 2'-deoxyadenosine were calculated to be 500 and .$710\mu$m, respectively. ADA was sensitivite to $Zn^{2+}$, $^Cu{2+}$ and $Fe^{3+}$, p-chloromercuribenzoate and mersalyl acid inactivated the enzyme. The activity of enzyme was not changed when ADA was incubated with dithiothreititol, 2-mercaptoethanol, N-ethylmaleimide, iodoacetic acid and iodoacetamide.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.249-256
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• 1992

Potato juice was found out to have a strong inhibition activity on the growth of Clostridium perf;nngens during work of foodstuffs for the improvement of human intestinal microflora. The anti-bacterial activity of the precipitated protein obtained from the potato juice in 70% ammonium sulfate solution was stable at the range of pH 4 to 10, whereas it was lost by a heat treatment at $60^{\circ}C$ for 10 min. The minimal inhibitory concentration of the precipitated protein on the growth of C1. Pefingens was about 0.2 mg/ml. The potato protein also suppressed the growth of C1. butyrincm and Eubacterium iimosum, while it showed a promoting effect for the growth of Bifdobacterium bifidum, Bif: animalis, Lactobacillus plantarum and Lact. acidophitus. The potato protein was further purified by CM-Sepharose ion exchange column chromatography, Sephadex G-150 gel filtration column chromatography and SDS-polyacrylarnide gel electrophoresis. The purified protein(kCp) was proved to be a glycoprotein by PAS staining and its molecular weight was about 38.7 kd.

• Microbiology and Biotechnology Letters
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• v.23 no.4
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• pp.446-453
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• 1995

$\alpha $-Arabinofuranosidase was produced by E. coli HB101 haboring the recombinant plasmid pKMG11 which contained the arfI gene of Bacillus stearothermophilus. The maximum production of the enzyme was observed when E. coli HB101 cells were grown at 37$\circ$C for 20 hours in the medium containing 0.5% arabinose, 1.0% tryptone, 0.5% yeast extract, and 1% NaCl. The $\ALPHA $-arabinofuranosidase produced was purified to homogeneity using a combination of 20-50% ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange column chromatography and Sepharose 6B-100 gel filtration. The purified enzyme was most active at 55$\circ$C and pH 6.5. The K$_{m}$ and V$_{max}$ values of the enzyme on $\rho $-nitrophenyl-$\alpha $-arabinofuranoside was determined to be 2.99 mM and 0.43 $\mu $mole/min (319.74 $\mu $mole/min/mg), respectively. The pI value was 4.5. The molecular weight of the native protein was estimated to be 289 kDa. The SDS-polyacrylamide gel clectrophoresis analysis suggested that the functional protein was a trimer of the 108 kDa identical subunits. The N-terminal amino acid sequence of the a-arabinofuranosidase was identified as X-Ser-Thr-Ala-Pro-Arg( \ulcorner )-Ala-Thr-Met-Val-Ile-Asp-X-Ala-Phe.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.38-45
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• 1998

To extract insoluble proteins and to improve funtional properties of abolished proteins, an phytase producing Aspergillus sp. SM-15 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. Phytase production reached to maximum when the wheat bran medium containing 1% mannose, 1% yeast extract, 1% (NH4)2HPO4 and 0.2% calcium chloride was cultured for 4 days. Phytase was purified 17.1 fold and specific activity was 244.32unit/mg by a sequencial process of ammonium sulfate fraction, ion exchange chromatography and gel filtrations Pruified enzyme was confirmed as a single band by the polyacrylamide gel electro-phoresis. The molecular weight of phytase was estimated to be 46,000. The optimum pH and temperature for the phytase activity were 5.5 and 5$0^{\circ}C$. The enzyme is stable in pH 4.5~5.5, 6$0^{\circ}C$. The activity of purified enzyme was inhibited by Hg2+ whereas activited by Pb2+ and Fe2+. The activity of phytase was inhibited by the treatment with iodine. The result indicate the possible involvement of histidine at active site. Km and Vmax of the puridied phytase were 37.037mM/L and 159.87umol/min, respectively.

• The Korean Journal of Ecology
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• v.21 no.1
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• pp.73-80
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• 1998

Peroxidase has been isolated to apparent homogeneity from earthworm, Lumbricus rubellus, using ammonium sulfate fractionation, Sephacryl S-2000 gel filtration, CM-cellulose cation exchange chromatography and native-PAGE elution. Some of its enzymatic characteristics were examined. The optimum pH for gruaiacol oxidation of earthworm peroxidase was determined to be 6.0, and the $K_{m}$ values against guaiacol and $H_2O_2$ were 1.25 mM and 3.4mM, respectively. When various compounds were tested as the possible substrates of the enzyme, o-dianisidine was used as the substrate. However, earthworm peroxidase could not oxidize esculetin and ferulic acid as substrates, suggesting the different characteristics of the enzyme from plant peroxidases. The optimum pH for veratryl alcohol and $H_2O_2$ oxidation was determined to be 2.5 when lignin peroxidation activity was examined. The $K_{m}$ values for veratryl alcohol and $H_2O_2$ were 0.02 mM and 0.13 mM, respectively. Furthermore, the earthworm peroxidase could oxidize phenoxyherbicides such as 2,4-D, 2,4-DP and MCPA as substrates. The optimum pHs for 2,4-D, 2,4-DP and MCPA were determined to be 4.0, 2.0 and 2.0, respectively. The most available substrate was 2,4-DP, followed by MCPA and 2,4-D when their peroxidation activities were compared.

• Carbon letters
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• v.7 no.1
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• pp.1-8
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• 2006

The activated carbon "C" was obtained by carbonization followed by activation with steam at 40% of burn-off. Oxidized carbons C-N, C-P and C-H were obtained by oxidizing the activated carbon C with concentrated nitric acid, ammonium peroxysulfate and hydrogen peroxide, respectively. The textural properties of the carbons were determined from nitrogen adsorption at 77 K. The acidic surface functional groups were determined by pH titration, base neutralization capacity and electrophoretic mobility measurements. The cation exchange capacities of un-oxidized and oxidized carbons were determined by the removal of Cu(II) and Ni(II) from their aqueous solutions. The surface area and the total pore volume decreased but the pore radius increased by the treatment of activated carbon with oxidizing agents. These changes were more pronounced in case of oxidation with $HNO_3$. The surface pH of un-oxidized carbon was basic whereas those of the oxidized derivative were acidic. The removal of Cu(II) and Ni(II) was pH dependent and the maximum removal of the both ions was obtained at pH of 5-6. Cu(II) was more adsorbed, a phenomenon which was ascribed to its particular electronic configuration.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.3
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• pp.279-286
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• 1989

The alkaline protease producing mold isolated from and identified as Aspergillus fumigatus. It was found that the production of alkaline protease reach to maximum was cultured for 3 days at $30^{\circ}C$. The enzyme was purified 86.13 fold and yield of the enzyme purification was 6.4%, The purification procedure include ammonium sulfate treatment, gelfiltration on Sephadex G-25, G-75, G-150 and DEAE-cellulose ion-exchange chromatography. When the purified enzyme was applied sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 63000. This enzyme composed 17 amino acids and main amino acids of this enzyme were glycine and glutamic acid.

• BMB Reports
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• v.29 no.6
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• pp.540-544
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• 1996

Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.