• Journal of the Korean Chemical Society
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• v.7 no.1
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• pp.17-24
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• 1963

The fundamental investigations of surface properties of scheelite were made by electrophoretic mobility adsorption and contact angle measurements, and results have been correlated with its floatability obtained by Hallimond tube flotation test. The role of the interfacial electrical condition on the adsorption of collectors on mineral surfaces is discussed with the flotation of scheelite. From electrokinetic measurements made on scheelite, $Ca^{++}$ and $WO_4^{--}$ are identified to act as potential-determining ions, thus controlling the surface properties on this mineral. Therefore, at the fixed pH, the scheelite surface become to be less negatively charged with increasing $Ca^{++}$ concentration and more negatively charged with increasing $WO_4^{--}$ concentration in the pulp. Adsorption of collectors then depends strongly on the concentration of $Ca^{++}$ or $WO_4^{--}$ in the solution; anionic collectors are adsorbed on less negatively charged surfaces and cationic collectors on more negatively charged surfaces, which in turn defines the effective flotation range with respective collectors for this mineral.

• Korean Journal of Pharmacognosy
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• v.22 no.2
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• pp.101-111
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• 1991

Polyphenol oxidase(PPO) was purified from an extract of Puerariae Radix by ammonium sulfate fractionation followed by Sephadex G-150 column chromatography, which resulted in a 56-fold increase in specific activity. The enzyme was optimum of pH 6.5. The optimum temperature of enzymic reaction was about $40^{\circ}$. The enzyme was thermostable with a half-life equal to 32 min at $70^{\circ}$. Km values of the PPO for catechol and pyrogallol from Lineweaver Burk plots were $1.3{\times}10^{-2}M$, $1.16{\times}10^{-2}M$, respectively. The substrate specificity of the Puerariae Radix PPO showed high affinity toward pyrogallol. Reducing reagents such as cysteine, potassium metabisulfite, ascorbic acid, 2-mercaptoethanol completely inhibited the PPO activity at $10^{-2}M$ level. Linewear-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, 4-nitrocatechol, potassium cyanide was competitive with Ki values of $4.3{\times10^{-2}M,\;0.73{\times}10^{-6}M,\;6.9{\times}10^{-6}M,\;6.4{\times}10^{-7}M$, respectively. The browning reaction by PPO was observed to decrease temporarily with the addition of sodium diethyl dithiocarbamate, a well known copper chelating agent. Among the divalent cations, $Cu^{2+}$ ion was strong activator on PPO and $Mn^{2+},\;Co^{2+}$ ions was effect on PPO activity. $Zn^{2+},\;Mg^{2+}$ ions was inhibitor on PPO.

• Journal of the Korean Society of Tobacco Science
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• v.21 no.2
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• pp.160-170
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• 1999

Purification and biochemical experiments on the carboxylesterases-III (CE-III) from the indian meal moth, Plodia interpunctella (Hubner) were carried out to understand their enzymemological characteristics. The CE-III from the fifth instar larvae was purified by means of ammonium sulfate fractionation, gel permeation choromatography and ion exchange choromatography. The optimal temperature for the reaction of the CE-III on the 4 substrates ($\alpha$-Na, $\alpha$-Nb, $\beta$-Na and $\beta$-Nb) was confirmed at 4$0^{\circ}C$. The optimal pH for the reactions on the substrates $\alpha$-Na and $\alpha$-Nb was 7.5. But the optimal pH on the substrate $\beta$-Na and $\beta$-Nb was 8.0. The optimal substrate concentration for the reactions of the CE-III was 3.16 X 10$^{-3}$ M in $\alpha$-Na and $\beta$-Nb. On the substrate $\beta$-Na and $\alpha$-Nb, the optimal substrate concentration was 1.0 X 10$^{-3}$ M for CE-III. The $V_{max}$ and $K_{m}$ values of the carboxylesterases were varied by the substrates as followings: the $V_{max}$ of CE-III was 45.9 for $\alpha$-Na, 52.6 for $\beta$-Na, 36.4 for $\alpha$-Nb, and 83.3 ($\mu$ mol/min/mg protein) for $\beta$-Nb. The $K_{m}$ of CE-III was 1.43 X 10$^{-4}$ M for $\alpha$-Na, 3.57 x 10$^{-5}$ M for $\beta$-Na, 9.17 X 10$^{-5}$ M for $\alpha$-Nb, and 7.14 X 10$^{-5}$ M for $\beta$ -Nb, respectively. The CE-III seemed to have somewhat high thermostability considering that the temperature for effective denaturation on activity was about 5$0^{\circ}C$ ～ 6$0^{\circ}C$.EX>.EX>.

• KSBB Journal
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• v.8 no.2
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• pp.164-171
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• 1993

The production of extracellular polysaccharide by Methylomonas mucosa (NRRL B-5696) was investigated. The microorganism uses methanol as the carbon source for their growth and produces exopolysaccharides. The productivity of exopolysaccharides was investigated under various culture modes: batch, fed-batch and continuous culture. In flask culture the growth of cell mass and the production of polysaccharide were inhibited at above 1% (v/v) methanol. At 1%(v/v) methanol maximum specific growth rate was obtained. As C/N ratio (g methanol/g ammonium sulfate) increased, polysaccharide production increased and cells mass decreased. Magnesium ion was also found to be essential for the polysaccharide production. In batch culture the production of polysaccharides was more affected by the specific growth rate than the cell concentration. In fed-batch culture the concentration of polysaccharide was 4 times higher than that of batch culture, but the yield was lower. The productivity of fed-batch with continuous feeding was higher than that of batch or fed-batch with intermittent feeding. This is due to no methanol limitation or inhibition that used to occur in fed-batch culture with intermittent feeding. In continuous culture pure oxygen was supplied to avoid the oxygen limitation. As the dilution rate in- creased up to 0.21 h-1, the yield and productivity increased. The solution viscosity of the produced polysaccharide obtained from above increased exponentially with the concentration of polysaccharide.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.845-852
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• 2001

A microorganism producing fibrinolytic enzyme was isolated from Korean traditional soybean paste and identified as Bacillus sp. KDO-13. The fibrinolytic enzyme was purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-celluose, and gel chromatography on Sephadex G-100 of the culture supernatant of Bacillus sp. KDO-13. The molecular weight of the purified enzyme was estimated to be 44,000 by SDS-PAGE. The optimum pH and temperature for the enzyme activity were pH 8.0 and $50{\circ}C$, respectively. The enzyme activity was relatively stable at pH 7.0-9.0 and temperature below $50{\circ}C$. the activity of the enzyme was inhibited by $AI^{3+}$ and $Hg^{2+}$, but activated by $Co^{2+}$\;and\;Ni^{2+}. In addition, the enzyme activity was potently inhibited by EDTA and 0-phenanthroline. The purified enzyme could completely hydrolyze a fibrin substrate within 6 h in vitro, and had a low $K_m$ value for fibrin hydrolysis. It was concluded that the purified enzyme was a metalloprotease with relatively high specificity for fibrinolysis, and thus, could be applied as an effective thrombolytic agent.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.114-120
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• 1997

Microorganisms producing xylanase were screened for the enzymatic production of xylo-oligo saccharides from xylan. One of the bacteria isolated from compost produced an endoxylanase extracellularly. The bacterium was identified as Bacillus sp. according to its taxonomic characteristics examined. Xylanase production reached upto 5 U/ml after 22 h of culture in LB medium at $30^{\circ}C$. The xylanase was purified by ammonium sulfate precipitation and gel filtration. The molecular weight of the xylanase was estimated to be 20,400 by SDS-PAGE. Optimal temperature and pH for the xylanase activity was $60^{\circ}C$ and 6.5, respectively. The enzyme was stable at temperatures upto $40^{\circ}C$ and pH values from 4 to 10. The xylanase was completely inhibited by the addition of 2 mM mercury ion. Apparent $K_m$ and $V_max$ values for oat spelt xylan were 9.2 mg/ml and 1954 U/mg protein, respectively. For birchwood xylan, the values were 6.3 mg/ml and 1009 U/mg protein. The predominant products of the xylan hydrolysis were xylobiose, xylotriose and xylotetraose, indicating that the enzyme is an endoxylanase. Upto $85{\%}$ of the initially added enzyme (2 U/ml) was bound to 50 mg/ml of the insoluble fraction of oat spelt xylan after incubation at $30^{\circ}C$ for 30 min.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.50-58
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• 1996

Interspecific hybrids between Aspergillus oryzae var oryzae and A. nidulans 514 were obtained by nuclear transfer technique. Several autotrophic mutants isolated from conidiospores of the two strains were mutagenized with ultraviolet and N-methyl-N-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of hybrid formation by nuclear transfer were $3{\times}10^{-5}{\sim}1{\times}10^{-5}$. From observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyptypes are aneuploid. The hybrids showed 1.1~1.4 fold higher xylanase activities than parental strains did. The xylanase of Aspergiilus sp. TAVD514-3 was purified and some of it's enzymatc characteristics were investigated. The enzyme was purified about 85 fold with an overall yield of 17% from the culture medium by ammonium sulfate fractionation, Sephadex G-75 gel permeation chromatography, and CM-sephadex A-50 ion exchange chromatography. The purified enzyme functions optimally at pH 9.0 and 80$^{\circ}C$. The enzymatic activity was increased by the presence of $Mg^{2+}$ and $Mn^2$ ions.

• Herbal Formula Science
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• v.17 no.2
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• pp.163-174
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• 2009

Reactive oxygen species(ROS) can induce hepatotoxicity and trigger apoptosis in the liver. In this study, we investigated the sulfated polysaccharide A-1 from Opuntia ficus-indica against alcoholic oxidative stress in human liver Hep G2 cell. An antioxidant substance A-1 obtained from the enzymatic extract of Opuntia ficus-indica fruit was purified by DEAE-cellulose ion exchange and sephadex G-100 gel permeation chromatography. The purification yield and molecular weight were 14.3% and 1.8 KDa, respectively. The A-1 predominately contained arabinose, galactose, rhamnose and also sulfate group. The structure of A-1 was investigated by periodate oxidation, FT-IR spectroscopy, $^1H$-NMR spectroscopy. The A-1 mainly composed of alternating unit of ${\rightarrow}4$)-$\alpha$-L- Rapp-2-$SO_3^-$-$\alpha$-L-Galp-($1{\rightarrow}$ and branched linkage of $\beta$-D-Arbp- ($5{\rightarrow}$. The antioxidative activity was measured using the SOD, CAT activity and GSH assay, respectively. The expression of Nrf2 protein was analyzed by western blotting. The viable cell count analyzed by autofluorescence. Oxidative stress induced by ethanol(1 M) were dramatically reduced by A-1 treatment. A-1 also prevented cell death induced by oxidative stress. It also increased expression Nrf2 protein level. We concluded that sulfated polysaccharide A-1 from Opuntia ficus-indica effectively protect Hep G2 liver cell from alcoholic oxidative stress.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.5
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• pp.434-442
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• 1990

Aspergillus sp. CC-29 ws selected for its strong protease activity among various stains of molds found in soil. It was found that the production of alkaline protease reached to maximum when the wheat bran medium containing glucose as carbon source had been cultured for 4 days. Alkaline proteased was purified 36.10 fold from Aspergillus sp. CC-29 The purification procedures included ammonium sulfate fractunation gel filteration on Sepha-dex G-75 G-150 and DEAE-cellulose ion-exchange chromatography, The yield of the purified enzyme was 22.40% The purified enzyme was confirmed as a single band by the polyacryla-mide. When the purified enzyme was applied to SDS-PAGE the molecular weight was estima-ted 24000. The optimum pH for the enzyme activity was 9.0 and the optimum temperature was 4$0^{\circ}C$ The reaction of this enzyme followed typical Michaelis-Menten kinetics with the Km value of 2.10$\times$10-4M with the Vmax of 29.41 $\mu$g/min. The enzyme was reactively stable in alkalic condition and unstable by heat treatment. The activity of alkaline protease was increased by the addition of Ca2+ whereas it was inhibited by Hg2+ Zn2+ at concentration of 1$\times$10-3M.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.624-631
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• 2007

Thermostable protease is very effective to improve the industrial processes in many fields. Two thermostable extracellular proteases from the culture supernatant of the thermophilic fungus Chaetomium thermophilum were purified to homogeneity by tractional ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose, and Phenyl-Sepharose hydrophobic interaction chromatography. By SDS-PAGE, the molecular mass of the two purified enzymes was estimated to be 33 kDa and 63 kDa, respectively. The two proteases were found to be inhibited by PMSF, but not by iodoacetamide and EDTA. The 33 kDa protease (PRO33) exhibited maximal activity at pH 10.0 and the 63kDa protease (PRO63) at pH5.0. The optimum temperature for the two proteases was $65^{\circ}C$. The PRO33 had a $K_m$ value of 6.6mM and a $V_{max}$ value of $10.31{\mu}mol/l/min$, and PRO63 l7.6mM and $9.08{\mu}mol/l/min$, with casein as substrate. They were thermostable at $60^{\circ}C$. The protease activity of PRO33 and PRO63 remained at 67.2% and 17.31%, respectively, after incubation at $70^{\circ}C$ for 1h. The thermal stability of the two enzymes was significantly enhanced by $Ca^{2+}$. The residual activity of PRO33 and PRO63 at $70^{\circ}C$ after 60min was approximately 88.59% and 39.2%, respectively, when kept in the buffer containing $Ca^{2+}$. These properties make them applicable for many biotechnological purposes.